Waste Management 27 (2007) 117–129 www.elsevier.

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Technical paper

Water hyacinths as a resource in agriculture and energy production: A literature review
Carina C. Gunnarsson
a

a,*

, Cecilia Mattsson Petersen

b

Department of Biometry and Engineering, Swedish University of Agricultural Sciences, P.O. Box 7032, SE-750 07 Uppsala, Sweden b Department of Mathematics, Science and Environment, Dalarna University College, SE-781 88 Borlange, Sweden ¨ Accepted 6 December 2005 Available online 31 March 2006

Abstract Water hyacinths are becoming a problem in lakes, ponds and waterways in many parts of the world. This paper contains a literature study of different ways to use water hyacinths, mainly in agricultural or alternative energy systems. The literature review indicated that water hyacinths can be rich in nitrogen, up to 3.2% of DM and have a C/N ratio around 15. The water hyacinth can be used as a substrate for compost or biogas production. The sludge from the biogas process contains almost all of the nutrients of the substrate and can be used as a fertiliser. The use of water hyacinth compost on different crops has resulted in improved yields. The high protein content makes the water hyacinth possible to use as fodder for cows, goats, sheep and chickens. Water hyacinth, due to its abundant growth and high concentrations of nutrients, has a great potential as fertiliser for the nutrient deficient soils of Africa and as feed for livestock. Applying the water hyacinths directly without any other processing than sun drying, seems to be the best alternative in small-scale use due to the relatively small losses of nutrients and workload required. To meet the ever-growing energy demand, biogas production could be one option but it requires investments and technological skills that would impose great problems in developing countries where the water hyacinth is often found. Composting as an alternative treatment has the advantage of a product that is easy to work into the soil compared with dried water hyacinths, because of the decomposed structure. Harvesting and transport of water hyacinths can be conducted manually on a small scale and does not require a new harvesting technique to be introduced. Transporting of fresh water hyacinths means, if used as fertiliser in amounts large enough to enhance or effect crop growth, an unreasonably large labour requirement. Based on the labour need and the limited access to technology, using dried water hyacinths, as green manure is a feasible alternative in many developing countries. Ó 2006 Elsevier Ltd. All rights reserved.

1. Introduction The water hyacinth (Eichhornia crassipes) is a free floating aquatic weed originated in the Amazon in South America (Bolenz et al., 1990) where it was kept under control by natural predators (Lee, 1979). The plant has, through introduction by man, spread throughout the whole tropical zone (Aweke, 1993). Due to its fast growth and the robustness of its seeds, the water hyacinth has since then caused major problems in the whole area, e.g., a reduction of fish. Other effects of the fast growth are physical interference
*

Corresponding author. Tel.: +46 18 672578. E-mail address: Carina.Gunnarsson@bt.slu.se (C.C. Gunnarsson).

with fishing, obstruction of shipping routes and losses of water in irrigation systems due to higher evaporation and interference with hydroelectric schemes and increased sedimentation by trapping silt particles. It also restricts the possibilities of fishing from the shore with baskets or lines (Aweke, 1993) and can cause hygienic problems (Moursi, 1976a; Becker et al., 1987; Abdelhafiz, 1989, from Abdelhamid and Gabr, 1991). Attempts to control the weed have caused high costs and labour requirements, leading to nothing but temporary removal of the water hyacinths. Since the most favourable conditions for the growth of the water hyacinth often are found in developing countries, very limited resources have been put into curbing them. Fighting the water hyacinth

0956-053X/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.wasman.2005.12.011

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C.C. Gunnarsson, C.M. Petersen / Waste Management 27 (2007) 117–129

generates neither food nor income, and the weeds are therefore left to cover the lakes. Fast growth is a feature valued in crops grown by man. The water hyacinth would, therefore, have a great potential if seen as raw material for industries or if incorporated into agricultural practice. This paper contains a literature review on utilisation of water hyacinths, mainly in an agricultural or alternative energy system. 2. The water hyacinth 2.1. Growth and harvest Water hyacinths regenerate prolifically from fragments of stems and the seed can remain viable for more than six years. These ways of regeneration make it very difficult to control the weed (Lee, 1979). The number of plants can more than double in seven days in conditions of high temperature and humidity (Lareo and Bressmi, 1982, from Tag El-Din, 1992) and up to 140 ton of DM/ha and yr are produced (Abdelhamid and Gabr, 1991). The plant normally forms cohesive floating mats and can cover large areas of the water surface. The spreading of the water hyacinth is also thought to be enhanced by winds (Gay, 1960, from Aweke, 1993). The plant flourishes in nutrient-rich waters and on shallow shores with mud rich in nutrients. To estimate how much of a certain water hyacinth product (soil amendment, fertiliser, gas, fodder, etc.) can be produced, it is necessary to make approximations of how much biomass can be harvested. Thomas and Eden (1990) estimate the possible harvesting of water hyacinths to 320 ton of DM/ha and yr. The figure above is based on conditions in Bangladesh, and nothing is said about how much water hyacinth was present before the harvest started. In order to not over-estimate the yield, we assume that no more than the yearly production of biomass can be harvested, i.e., 140 ton of DM/ha and yr (Abdelhamid and Gabr, 1991). The water hyacinth mats are driven by the wind. The harvest possibilities will, therefore, depend on the local conditions and winds. 2.2. Chemical analysis The water hyacinth has an excellent ability to take up nutrients and other chemicals from its environment, and the chemical composition of the water hyacinths depends strongly on its environment (Musil and Breen, 1977, from Poddar et al., 1991). Poddar et al. (1991) reported a nitrogen content of 1.78% (db) in water hyacinths growing in marshy land where the nitrogen level in the water was only 2.40 mg/L. Abdelhamid and Gabr (1991) state the nutrient content is lower in the stem and root compared with the nutrient content in the leaves. Several studies on the chemical composition of the water hyacinths have been reported. Abdelhamid and Gabr (1991) and Bolenz et al. (1990) made studies to evaluate the water hyacinth’s nutritional value for ruminants.

Chanakya et al. (1993) and Patel et al. (1993a) analysed water hyacinths as a substrate for anaerobic digestion. The analysis by Poddar et al. (1991) compared the nutrient content of water hyacinths grown in different habitats. The figures in Table 1 are for water hyacinth grown in a pond. Polprasert et al. (1980) evaluated water hyacinth as a substrate for compost. In the analysis of water hyacinths from Lake Victoria, Kenya (Gunnarsson and Mattsson, 1997) the plants were not washed; hence the analysis also included sand etc. found in the roots. In Table 1 the chemical composition of the water hyacinths from different sources is summarised. 2.3. Health aspects The increased growth rate of the water hyacinths has led to worsened health conditions for the people living in the affected areas. The floating water hyacinth mats can serve as a breeding ground for vector organisms carrying malaria, bilharziosis and river blindness (Moursi, 1976a; Becker et al., 1987; Abdelhafiz, 1989, from Abdelhamid and Gabr, 1991). At some places precautions against water snakes, hippos and crocodiles need to be taken. The water hyacinths consume so much oxygen when decaying that it leads to less oxygen remaining in these waters. The decreased oxygen content in the water leads to less oxygen in the fish. This, combined with fewer algae and other food sources for the fish, cause the meat of the fish to go bad faster than before. Decreased possibility to store fish leads to lower income and food security (Sunday Standard, Kenya, 12/1-1997). This means that decreasing the amount of water hyacinths could hopefully improve the health situation. When working in water hyacinth infested areas, one of the problems is the risk of catching waterborne diseases, in the case of Lake Victoria especially Schistosomiasis. The problem is that the snails that serve as a host for the bilharzia parasites are very likely to be found in water hyacinths. If drying the water hyacinths is to be a successful way of eliminating the risk of catching bilharzia depends on if the water hyacinth can be harvested without snails. 2.4. Harvesting Harvesting the water hyacinths means a mechanical control of growth. Presently, the water hyacinths are only harvested to control their propagation where chemical or biological (e.g., introduction of water hyacinth eating insects) methods are prohibited or unsuccessful. This is, according to Petrell and Bagnall (1991), because mechanical harvesting is too expensive and time-consuming. The advantages of mechanical control of water hyacinths are, according to Verbandt (1990):  The removal of superfluous nutrients.  The immediate result without damage to the ecosystem.

C.C. Gunnarsson, C.M. Petersen / Waste Management 27 (2007) 117–129 Table 1 Chemical analysis of water hyacinths according to different sources Parameter (% on DM basis) Abdelhamid and Gabr (1991)a Bolenz et al. (1990) Chanakya et al. (1993) Patel et al. (1993a) Poddar et al. (1991)b Polprasert et al. (1980)

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Gunnarsson and Mattsson (1997) Fresh Driedc

Dry matter (% on wb) Organic matter (VS) Crude protein Ether extract Crude fibre Nitrogen free extract Ash C/N ratio Neutral detergent fibre Acid detergent fibre Hemicellulose Cellulose Lignin Water soluble Phosphorus Carbon Nitrogen Magnesium Calcium Potassium Metabolizable energy for ruminants (MJ/kg)
a b c

9.5 74.3 20 3.47 18.9 31.9 25.7 – 62.3 29.0 33.4 19.5 9.27 – 0.53 – 0.17 0.58 – 6.35

6.2 – – – – – 15 – – – 22 31 7 – – – – – – –

9.4 83.65 – – – – – – – – 33.97 18 26.36 21.68 – – – – – –

– – 11.9 – – – 20.2 – – – 43.4 17.8 7.8 – – – – – – –

– 83.61 16.25 1.61 16.34 49.41 16.39 – 56.14 37.72 18.42 25.61 9.93 – 0.53 2.76 – 2.29 2.44 –

– – – – – – – 15.8 – – – – – – 0.5 2.9 – – – –

35.6 23.5

52.07 25.1

0.26 27.6 1.18

0.32 18.54 0.74

4.53

2.27

Abdelhamid and Gabr (1991) made the chemical analysis on water hyacinths collected from a canal and a ditch at Mansoura, Egypt. The cell wall composition was fractioned according to Van Soest (1963) and Van Soest and Wine (1967) from Poddar et al. (1991). Dried 13 days with full natural aeration. 

Waterbodies can be used more widely (e.g., for irrigation of agriculture areas and drinking water supply).  Mechanical methods are possible in open flowing as well as in closed water systems. Another advantage of harvest is that it makes it possible to use the water hyacinths in, for example, agricultural practice. A weed screen cleaner with continuously moving rakes is being developed in Belgium. The plan is to construct a back-raked screen that is mobile by being mounted on a converted military amphibian (Verbandt, 1990). Another possibility might be to use agricultural harvesting machinery such as conveyor belts. In the Philippines they are reported to use barges with some kind of hand powered winch when harvesting open-water water hyacinths (Tho´ ren, 1997). When the hyacinths are close to the shoreline, one person can, in an easy and sustainable tempo, harvest approximately 200 kg of fresh water hyacinths per hour (Gunnarsson and Mattsson, 1997). If the water hyacinths are not growing directly on the shoreline, they must be transported there. Petrell and Bagnall (1991) conducted a study to determine drag properties of water hyacinths. The maximum towing velocity, before the mat became unstable and the leading edge started to roll forward under the water, was found to be 0.40 m/s. The maximal size of the mats in this experiment was 2.44 by 1.22 m.

2.5. Transport When using water hyacinths in agricultural practise, transport of the fresh or treated plant is necessary. The need and design of the transport vary depending on the treatment. The design also depends on the scale of the operation. In anaerobic digestion, the water hyacinths will have to be transported to the biogas digester that also requires a low dry matter concentration. The sludge from the process, also having a high water content, must be transported to the field for spreading. In alternatives with compost or green manure, it is advantageous if the treatments are taking place close to where the water hyacinths are harvested. That means less transport of the water hyacinths while they still have a high water content, and thus less transport of water that might need to be added to the compost. The compost product could be transported to the field in baskets carried on the head (Eklund, 1996). The dried water hyacinths used as green manure could be carried in nets or pieces of cloth. In Table 2 is a summary of the transport requirements for providing 1 kg of plant available nitrogen. 3. Treatment 3.1. Carbonisation The main product from this three-stage process (gasification, pyrolysis and carbonisation) is charcoal, as a

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Table 2 Transport requirements for providing 1 kg plant available N through water hyacinth products (Gunnarsson and Mattsson, 1997) Parameter Fresh water hyacinth to harvest Dry matter to harvest Dry matter after process Dry matter of product Amount to spread wh = water hyacinth. Anaerobic digestion 3676 349 243 10 2378 Compost 8351 795 459 70 649 Green manure 2973 284 270 85 324 Unit kg wh (wb) kg wh (DM) kg wh (DM) % kg wh product (wb)

by-product gas is obtained that can be used for internal processes (Thomas and Eden, 1990). There are two problems in using the water hyacinth for making charcoal. First, there is a need to reduce the water content, and second the ash content (40% according to Thomas and Eden, 1990) of air-dried water hyacinth is too high to get a good fuel as an end product. The high investments and technological level necessary also make carbonisation an unfavourable alternative in developing countries (Thomas and Eden, 1990). 3.2. Hydrolysis and fermentation Hydrolysis together with fermentation will give a liquid fuel, for example, ethanol. The process is well suited for material with a high moisture content, which would make aquatic weeds a good substrate. Hydrolysis and fermentation also require yeast fermentable sugars that are available only to a very low extent in water hyacinth and other aquatic weeds. Some kind of pre-treatment is, therefore, needed to make the sugar more easily available for chemical hydrolysis. The pre-treatment requires a relatively high temperature, strong acids and pressurised reactors. Enzymatic hydrolysis is an option, but difficult because of the high lignin content. Slesser and Lewis (1979, from Thomas and Eden, 1990) reported a negative energy balance of such a process. Thomas and Eden conclude that hydrolysis of water hyacinths to produce fuel is, because of the negative energy balance, only feasible in situations where there is a high need for ethanol as a liquid fuel. 3.3. Fodder and silage Water hyacinth as roughage is an interesting alternative to reduce the shortage of animal feed. Tag El-Din (1992) conducted a study to see the effect of substituting bean straw with water hyacinth hay when feeding sheep. Norton (1982, from Tag El-Din, 1992) considered 9% crude protein, on a DM basis, to be the minimum in the fodder for ruminants. Abdelhamid and Gabr (1991) give a crude protein content of 20% on DM basis showing that water

hyacinth in that aspect is a good roughage for ruminants. Tag El-Din (1992) found that using water hyacinth hay as a sole roughage greatly reduced the average daily weight gain. They concluded that for growing sheep, up to 30% of the roughage (bean straw) can be substituted with hay made from dried water hyacinths, without a loss in growth rate. According to Bolenz et al. (1990), the stalk tissues of the water hyacinths contain intercellular spaces filled with air, which soak up water while the animals are digesting. That leads to excessive water consumption and the animals feel replete, although having little material of nutritional value in their rumens. Bolenz et al. (1990) also found, when examining the water hyacinth tissue in microscope, sharp needles formed of calcium oxalate. Bolenz et al. (1990) assumed that these needles could damage the digestive tract of animals fed with water hyacinths, if they were not dissolved by digestive acid. To avoid these problems, Bolenz et al. (1990) suggested the following preparation of the water hyacinths used for feeding animals: Chop the tissues to eliminate the air included and to negate its ability to absorb water. After chopping, the solid material should be separated from the soluble components in the juice by pressing and centrifuging. The solid phase could be washed with acid to eliminate the acid-soluble calcium-oxalate and then processed to a ruminant fodder. The juice could be concentrated, dried and used as a protein enriched fodder component (Bolenz et al., 1990). It is possible to produce silage from water hyacinths, but the water hyacinths need to be chopped into fine pieces to remove the air in the tissues; otherwise these could lead to growth of aerobic moulds during the fermentation. Also sorbic acid can be added to suppress the moulds. For successful fermentation the pH needs to be lowered (below 4), which is achieved by adding sugar. The water hyacinths were calculated to contain 0.52% fermentable sugar in relation to fresh weight so an addition of 0.4% sugar was enough to reach the desired pH value (Bolenz et al., 1990). 3.4. Drying Drying is a pre-treatment step done to get the right moisture content for the subsequent treatment, but also to decrease the weight and thereby make transport easier. It can also be a treatment in itself. Drying the water hyacinths makes it possible to store them for later use and make them easier to decompose once put in the soil. This might be achieved by sun drying, i.e., simply spreading the water hyacinths directly on the ground. Experiences in Egypt of sun-drying of water hyacinths reported no observed mould or contamination during the drying period (Tag El-Din, 1992). Polprasert et al. (1980) report that sundrying water hyacinths for a few days reduce the weight by about half. Apori (1994) found the when plantain and cassava peels were sun dried, the material had to be dried for four days to attain a dry matter content of 87.0% (limit for storability) or above. There were no significant losses of

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nutrients, the maximum loss in crude protein was 0.3% (Apori, 1994). In Kenya water hyacinth roots dried with full natural aeration (hung over a fence) reached a dry matter content of 87% after 13 days. The leaves of the same plants did not reach more than 66% DM. It was also found that the nutrient level in the water hyacinths decreased during the drying process. It should be pointed out that these trials were carried out during the rainy season, and it is likely that the whole plants would reach a dry matter content sufficient for storage and the nutrient losses can be decreased if dried during the dry season (Gunnarsson and Mattsson, 1997). 3.4.1. Incineration Sun drying and direct burning is used on a small scale in certain parts of the world. Fresh water hyacinth has a moisture content of about 90% (Abdelhamid and Gabr, 1991). Even when the moisture level is reduced to 10% the energy density is not more than 1.3 GJ/m3 (Thomas and Eden, 1990). This may be compared with 9.8 GJ/m3 in firewood and does not make the water hyacinth very attractive for direct incineration. The major part of the fresh water hyacinth is water, and any processing or transport is therefore worthwhile only if the water content can be reduced to 10–15% with ‘‘relative ease’’ (Thomas and Eden, 1990). 3.4.2. Briquetting Thomas and Eden (1990) mention briquetting as a possible treatment. The briquettes are made by sun-drying the water hyacinth for a few days, disintegrating, screening and chopping the dried water hyacinths to pieces about 6 mm long. The shredded water hyacinth can then be compressed into briquettes or pellets. The material resulting after briquetting water hyacinth has an energy density of 8.3 GJ/m3, which is comparable to charcoal that has 9.6 GJ/m3. This process requires initial investments for machinery and a rather large area for the drying, which might be expensive. 3.5. Anaerobic digestion Anaerobic digestion is the biological process by which organic matter is degraded in the absence of oxygen and biogas is produced. The three-step process results in a gas that can be used directly for cooking, heating or production of electricity and a nutrient-rich slurry. Biogas is a form of energy that has very useful by-products and positive impacts on public health and pollution. This, together with the growing shortage of firewood and rising cost of fossil fuels, has made anaerobic digestion increasingly interesting. These advantages of the process might make it well suited for use in developing countries. According to Gunnersson and Stuckey (1986) ‘‘Plants such as water hyacinths. . .can be degraded easily, and give quite a high gas yield. In these cases, digestion of these weeds can solve the problems caused by excess growth in canals and

provide energy as well.’’ Day et al. (1990) see biogas as a reliable energy source that can improve the environment both on a large and on a small scale, e.g., deforestation and smoke reduction in kitchens. However, technical requirements might limit the possibility to use anaerobic digestion as a treatment for water hyacinth in rural areas. Lack of water and cow manure as a substrate has been mentioned as other limitations. When water hyacinths are intended as the main substrate, large amounts of animal manure are not needed and, due to their growth place, water is generally available. 3.5.1. Pre-treatment Chopping the water hyacinths increases the specific surface of the substrate and thereby enhances the access of microbes to the plant material, which is important for a well-working biogas process (Haug, 1993). Moorhead and Nordstedt (1993) conducted experiments with different particle sizes, nitrogen contents and inoculum volumes, in a mesophilic process (35 °C). The total biogas and methane production was largest for water hyacinth when the plant material was chopped into 6.04 mm pieces (compared with 1.6 and 12.7 mm). Water hyacinths have a high content of hemicellulose and cellulose, but the existing hemicellulose has a rather strong association with the lignin in the plant, which makes it unavailable for the microorganisms (Patel et al., 1993a). To optimise biogas production, the plants must undergo some kind of pre-treatment. Patel et al. (1993a) used thermochemical pre-treatment to solve these problems and thereby increased the gas production. Patel et al. (1993b) found that the addition of metal ions: Fe3+, Zn2+, Ni2+, Co2+, and Cu2+, will enhance gas production and increase the methane content in the produced gas and also result in better process stability. The water hyacinths used in these experiments were, however, taken from a pond that did not receive municipal or industrial effluent. Water hyacinth from polluted water might already contain sufficient amounts of heavy metals. Geeta et al. (1990) reported increased biogas production when nickel was added to water hyacinth or a mixture of water hyacinth and cow dung. El-Shinnawi et al. (1989) produced biogas from water hyacinth mixed with cow dung, and found the cow dung to provide enough microorganisms to serve as inoculum. The conclusion from these reports is that in developing countries it is probably better to not use costly pre-treatment and instead use a longer residence time. 3.5.2. Digestion product 3.5.2.1. Sludge. Gunnersson and Stuckey (1986) write that the sludge from the anaerobic process is rich in nutrients and organic matter and provides a good way to recycle these nutrients. A wet biogas process has a dry matter content of 2–10% (Thomas and Eden, 1990; El-Shinnawi et al., 1989; Madamwar et al., 1991). Essentially all of the nutrients contained in biomass used for anaerobic

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methane generation remain in the digester sludge (Hons et al., 1993; Stout, 1983) as long as it is not de-watered and stored airtight. Nutrient concentration will increase slightly during digestion because of the loss of volatile solids, associated with methane generation. The high concentration of nutrients gives the sludge a high potential as fertiliser (Hons et al., 1993). Due to the anaerobic conditions, most of the nitrogen in the sludge will be found in organic form, followed by ammonium (NHþ ) (20–50% or 4 more) and a very small part as nitrate (NOÀ ) (Hons 3 et al., 1993). Anaerobic sludge is easily de-watered and there have been thoughts on marketing the dried sludge as a fertiliser. This practice is only feasible for very large feedlots (Stout, 1983). Using the sludge as a fertiliser without de-watering will mean more effort in transport of the material. Much of the nitrogen in anaerobic sludge is in ammonium (NHþ ) form and therefore is less likely to leach from 4 the soil than nitrite (NOÀ ) and nitrate (NOÀ ). The decision 2 3 whether ammonium (anaerobic sludge) or nitrite and nitrate (aerobic sludge or chemical fertilisers) are to be preferred will be based on the soil type (Stout, 1983). From China an increase in agricultural productivity by 30% over farmyard manure is reported when using anaerobic sludge as a fertiliser (van Buren, 1979, from Gunnersson and Stuckey, 1986). This is probably due to the nitrogen in the sludge being more accessible than, in for example, farmyard manure (Gunnersson and Stuckey, 1986). There is a risk of contaminating nearby watercourses (Stark and Clapp, 1980) and decreasing seed germination by applying too much sludge (Hons et al., 1993). Ammonia volatilisation can lead to significant losses if the material is spread on the surface of the soil or stored in containers that are not airtight. Some authors report only negligible volatilisation from sludge. How much is lost will depend on the characteristics of the sludge, method of application, and soil properties (Hons et al., 1993). The risk of ammonia volatilisation increases with high ammonium concentration and pH (Moorhead and Nordstedt, 1993). If the sludge from a biogas process is to be applied as a soil fertiliser in areas with high temperatures, it must be worked into the soil or in some other way covered to minimise the nutrient losses due to ammonia volatilisation. The nitrogen
Table 3 Summary of reported gas yields with water hyacinth as a main substrate Source

losses might otherwise be as high as 70–80% (Thyselius, 1997). Parker and Sommers (1983) report that 15% of the organic nitrogen remaining in the sludge will be available to the plants during the first growing season. They also state that the risk of nitrogen immobilisation in the soil is highest when the sludge has a C/N ratio above 20. 3.5.2.2. Biogas. Chanakya et al. (1993) found that water hyacinth has a high content of fermentable matter and therefore a high potential for biogas production, but the high lignin content can reduce the actual production. The low bulk density could result in large voids with poor compaction and low feed rates (Chanakya et al., 1993) as a result. El-Shinnawi et al. (1989) conducted trials with anaerobic digestion of agricultural waste. Rice straw, maize stalks, cotton stalks and water hyacinths mixed with cow dung were digested in different containers. The mixture of water hyacinth and cow dung was found to produce more biogas per kilogram VS added than maize and cotton stalks, but the total biogas production per kilogram DM added was lower for the water hyacinths. The low values for total gas production was probably mostly due to the high lignin content and low percentage of volatile solids in the water hyacinths (El-Shinnawi et al., 1989). Table 3 is a summary of reported gas yields where water hyacinth made up all or the major part of the substrate. This shows that water hyacinths can compete well with any kind of animal manure as a substrate for biogas production. The gas produced during the process consists mainly of methane, carbon dioxide and ammonia, but small amounts of hydrogen sulphide may occur. The proportion of methane in the produced gas is usually up to 60%, but depends on the substrate (Gunnersson and Stuckey, 1986). Temperature, pH and pressure may alter the gas composition slightly. The specific gravity decreases when the methane content increases (Constant et al., 1989). 3.6. Composting Another possible treatment method for water hyacinths is aerobic decomposition, i.e., composting. Due to the lack of infrastructure and capital, small-scale composting is the

Biogas production (l/g DM) (l/g VS) 0.348 0.292 0.19 0.4 0.24 0.20–0.28 0.293 0.286 0.4

Residence time (days)

Methane (%)

Substrate

Chanakya et al. (1993) Chanakya et al. (1993) Chynoweth et al. (1983, from Moorhead and Nordstedt, 1993) ˚ Ellegard et al. (1983) Hanisak (1980, from Moorhead and Nordstedt, 1993) Moorhead and Nordstedt (1993) Patel et al. (1993a) Patel et al. (1993b) Madamwar et al. (1991)

0.291 0.245 – – – – 0.190 0.143 0.19

300 300 – – – 15–60 8 8 8

60 60 – – – 63–67 62–66 – 65

W (fresh) W (dry) W W W W W W WC

W = water hyacinth as only substrate. WC = a mix of water hyacinth and cattle dung was digested.

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main interest in developing countries. Composting can be defined as the biological decomposition and stabilisation of organic substrates, under conditions that allow development of thermophilic temperatures as a result of biologically produced heat. The final product should be stable, free of pathogens and plant seeds, and beneficial when applied to land (Haug, 1993). To decrease evaporation and losses of nitrogen, as ammonia, the compost can be covered with a layer of, for example, straw, grass or plastic. The advantage of using straw is that the microorganisms use straw as a source of energy and then catch the escaping ammonia in order to fill their nitrogen demand (Claesson and Steineck, 1991). Covering the compost with straw is also advantageous since it decreases the losses of nutrient ´ from rainwater leaching through the compost pile (Ulen, 1991). 3.6.1. Pre-treatment Enhancement of the access of the microbes to the plant material by chopping the fresh water hyacinths is important for a well-functioning compost (Dalzell et al., 1979). To enhance bacterial decomposition, Polprasert et al. (1980) mention that the water hyacinths were shredded into about 5-cm long pieces before they were composted. In a study by Elserafy et al. (1980), the fresh water hyacinths were also chopped before being composted. Composting the water hyacinth without size reducing them might, on the other hand, decrease the need for additional structure-supporting material and decrease the labour requirement. 3.6.2. Amendments For a well-working compost process, conditioning of the feed substrate is sometimes needed. Amendments can be added to prevent lack of energy, nutrients or other chemical substances (Haug, 1993) or to establish a suitable micro-fauna and increase degradation of, for example, cellulose and lignin (Adhikary et al., 1992). In a study of composting water hyacinths conducted in Egypt by Elserafy et al. (1980), lignin and cellulose were reported to remain undegraded after 185 days. The water hyacinths were spread alternating with a microbial activator consisting of ammonium sulphate, superphosphate and lime, in order to keep the process slightly alkaline (Elserafy et al., 1980). Haug (1993) mentions that a cellulose-rich substrate may lack the nutrients necessary to sustain rapid microbial growth rates. Adhikary et al. (1992) investigated the microflora associated with different plant wastes, e.g., water hyacinth, during composting. It was shown that a mixture of fungi, actinomycetes and bacteria added to the compost increased both the cellulose and lignin degradation compared with the untreated control (Adhikary et al., 1992). Since water hyacinths have a relatively high lignin content, preparation of the compost with microorganisms is of interest.

3.6.3. C/N ratio The optimal C/N ratio for the microbes is 15 to 30 according to Haug (1993), who claims that a decreased ratio is no problem for the composting process but leads to losses of excess nitrogen via ammonia volatilisation. Others claim the optimal range for bacterial decomposition to be a C/N ratio of 20–40 (Achraya, 1950, from Polprasert et al., 1980). A balanced nutrient availability for the microorganisms is important for a high decomposition rate (Polprasert et al., 1980). The same authors also noticed a slower composting rate for the piles prepared with water hyacinths compared with a control consisting of cow dung and leaves. The reason, they concluded, might be that leaves consist mainly of hemicellulose and cellulose, compounds reported to be more biodegradable than lignin, the major component of water hyacinths (Karim, 1968, from Polprasert et al., 1980). The water hyacinths have a C/N ratio of about 16 according to Abdelhamid and Gabr (1991), whereas Dalzell et al. (1979) report a C/N ratio of 20. This means that water hyacinths need an addition of cellulose material, such as leaves (C/N 60.8, Polprasert et al., 1980) to keep the ammonia losses low for the microbial decomposition. 3.6.4. Moisture content Elserafy et al. (1980) report an optimal moisture content of about 60% for the composting process, and Dalzell et al. (1979) give an optimal range of moisture content of 50– 60%. Elserafy et al. (1980) claim that the moisture content of the fresh water hyacinth is too high and hence little additional water is needed during the composting. Also the evaporation rate is high in hot climates, i.e., where the water hyacinths grow. Therefore the high moisture content is probably not a big problem when composting water hyacinths. To avoid too low of a moisture content in piles when composting in tropical areas with hot climates, the correct location of the compost operation is important. The preparation of 1 ton of compost product may require up to 2700 L of water. Composting can be carried out either in a pit or in a pile. Dalzell et al. (1979) suggest that compost be produced in pits during dry seasons and, to avoid water logging, in piles during rainy seasons. The compost should be protected from the wind to decrease moisture losses (Dalzell et al., 1979; Njoroge, 1994). Placing the compost pile out of direct sunlight can also reduce the water requirements according to Dalzell et al. (1979). 3.6.5. Pathogen reduction As a measure of bacterial destruction, Polprasert et al. (1980) mention an initial coliform concentration between 5 · 103 and 180 · 103 MPN (most probable number)/g compost mixture and a die-off of 70–90% after 10 week when composting water hyacinths together with nightsoil. As comparison, the US Environmental Protection Agency requires that the density of faecal coliforms must be less than 1000 MPN/g dry matter for sludge-based compost

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intended for public distribution and marketing (Haug, 1993). Such low levels of pathogens were not reached in the trial by Polprasert et al. (1980) where the highest reported temperature was 60 °C. 3.6.6. Use of compost as a soil amendment As the composting process proceeds, the readily degradable organics in the substrate are oxidised and gradually turned into increasingly less degradable humic material (Haug, 1993). If the compost product is not mature enough it has been observed to contain metabolites that are toxic to plants. If the compost applied to soils still has a high C/N ratio, i.e., decomposes rapidly, it may rob the soil of nitrogen needed by the plants. If, on the other hand, the C/N ratio of the organic matter is low, the excess nitrogen released as ammonia may become phytotoxic to plants (Haug, 1993). The major nutrients important for the fertilising qualities of the compost are nitrogen (N), phosphorus (P) and potassium (K). The oxidation of nitrogen in the compost process ultimately yields nitrate, which is not normally lost from the compost pile (Polprasert et al., 1980). Since phosphorus and potassium are physico-chemically less mobile than nitrogen, these compounds remain in the compost unless lost through leaching. They concluded that composting of the water hyacinths in developing countries is a feasible method because of its ability to retain most of the nitrogen, phosphorus and potassium in the compost and attain a satisfactory degree of composting within a relatively short period of time, i.e., 30 days (Polprasert et al., 1980). In a longer perspective, adding compost regularly enhances the quality of the soil by improving the soil structure. Advantages with compost application are (Eklund, 1996):  Increased water-holding capacity of the soil,  Improved soil structure by binding sand particles, making soils less prone to erosion,  Adding nutrients to the soil and thus giving higher crop yields, and  Contributing to re-circulation of organic material. In Africa, many resource-poor farmers cannot afford to purchase fertilisers. They seldom use organic waste products for compost (Abdel-Sabour and Abo El-Seoud, 1996). In a study in India (Sharma and Mittra, 1990), partially decomposed water hyacinth compost was applied to a sandy clay loam in a pot experiment with rice. The grain yield increased with increasing rate of application up to 15 ton/ha. The decomposition of water hyacinths resulted in active mineralisation of nutrients (Kumada, 1977, from Sharma and Mittra, 1990), which enhanced tillering and grain yield. Application of all organic materials increased the organic carbon and available nitrogen, phosphorus and potassium contents of the soil after harvest of the first crop. The higher grain yield of the second crop was associated with increasing soil fertility, owing mainly to increases

in organic carbon and available nitrogen concentrations. It was concluded that yield may be enhanced by incorporating water hyacinth compost up to 10 days before transplanting the rice to the field (Sharma and Mittra, 1990). Water hyacinth compost also had positive effects on sesame growth in Egypt (Abdel-Sabour and Abo El-Seoud, 1996). Primary analysis indicated that compost addition increased the levels of extractable nitrogen, phosphorus and potassium in the top 25 cm of the soil (Abdel-Sabour and Abo El-Seoud, 1996). Composted water hyacinths were also tried as an organic source of nitrogen on fibre yield of white jute. The water hyacinth compost was found to contain 0.55% nitrogen (on DM) and 45% moisture. Application of 40 kg N/ha, through water hyacinth compost alone, increased the fibre yield significantly compared with the control (Thakuria et al., 1991). 3.7. Green manure One option is to use the water hyacinths as green manure. Green manuring involves spreading plant material (with a high nitrogen content) on the fields and sometimes also working it into the soil (van der Werff et al., 1995). Wivstad (1997) reports that the most important features of a green manure are a large dry matter production and a high ability to fix nitrogen. The chemical analyses found in literature indicate a high nutrient content of the water hyacinth, 20% crude protein (Abdelhamid and Gabr, 1991), but values as low as 7.26% have been reported (Elserafy et al., 1980). They also have a very high dry matter production (140 ton of DM/ha and yr, Abdelhamid and Gabr, 1991). This should make them suited for use as green manure. Drying the water hyacinths before spreading them on the fields ought to be done primarily to minimise the risk of bilharzia (Thors, 1997) and secondly to decrease the labour required for transportation. Drying might lead to losses in biomass and thereby nutrients. For many plants their leaves have a higher nutrient content than the rest of the plant. This is also true for the water hyacinths. The nitrogen content of the leaves and stem is 3.7% (DM) and 2.7% (DM), respectively (Abdelhamid and Gabr, 1991). The leaves and other fragile parts will usually make up the largest part of the mechanical losses (Grant, 1990). Leaf-losses may, therefore, lead to a decrease in the average nutrient content calculated on the total dry matter. According to Gupta et al. (1996), leaves do not make up more than 25% of the total biomass of water hyacinths. 3.7.1. Application of water hyacinth To minimise the losses of nitrogen through volatilisation once the plant material is spread in the field, it should be covered by soil (Dalzell et al., 1979). One of the easiest ways to do this would be to, while ploughing, put the water hyacinths in the plough furrow and allow the next furrow slice to cover them. Gunnarsson and Mattsson (1997)

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found that partially dried water hyacinths used as green manure continued to grow in soil, which is why it would be desirable to somehow disintegrate the plants before usage. These methods require a certain level of mechanisation. If all of the agricultural work is done by hand, the water hyacinth could be hoed into the soil while preparing it for sowing. A method must be developed so that the working hours spent on the fields do not drastically increase, but exactly how this could be done depends on the level of mechanisation. Nitrogen mineralization is the transformation from the organic state into the inorganic forms of ammonium or nitrate, thereby making it available to the plants. No figures on mineralization of dried water hyacinths were found in literature, but the nitrogen and lignin content of water hyacinths are similar to that of subterranean clover described in an incubation experiment to examine carbon and nitrogen mineralization from green manure legumes decomposing in the soil (Marstorp and Kirchmann, 1991). Also the nitrogen uptake from decomposing legume material by subsequent crops was determined. During 115 days of incubation, approximately 30–35% of total nitrogen in subterranean clover was mineralised. The net nitrogen mineralization correlated well with the C/N ratio of the legumes; the species with high C/N ratio had lower mineralization. Calculations of nitrogen mineralised from the same green manure legumes under field conditions indicated that the potentially mineralisable amount of nitrogen (N0) decreased with plant age (Kirchmann and Marstorp, 1991). 4. Discussion To establish a well-operating chain for use of the water hyacinth, all the steps involved in the process – harvest, transport, pre-processing and processing – must be considered. A participatory approach is important for adjusting the systems to be introduced to the needs and possibilities (technological and economic) of the people. If the system is to be successful, it has to be accepted in the area and people must benefit from it. In Table 4 an evaluation of different methods for utilisation of water hyacinths in agriculture is presented. The evaluation is based on nitrogen losses and how much water hyacinth product must be spread to provide the available nitrogen needed. A method for harvesting the plant material must be developed to get the weeds out of the water. In many areas, the supply of water hyacinth is almost unlimited. A good estimation of the amount of water hyacinths available for harvest is 140 ton of dry matter per hectare, which is equivalent to the annual production per hectare (Abdelhamid and Gabr, 1991). The nitrogen content of the water hyacinths is between 1.2% and 3.2% on a dry matter basis. Harvest of the water hyacinths on a large scale can be done with specially designed machines. If the water hyacinths have to be transported to the place of harvest, Petrell and Bagnall (1991) found the maximum towing velocity to

Table 4 Nitrogen losses and amounts needed to provide 1 kg plant available nitrogen Parameter Mechanical N-losses Process losses Losses during spreading Total N loss DM loss due to processing N mineralised at spreading Anaerobic digestion 10 0 15 24 30 14 Compost 0 39.6 0.6 40 44 4 Green manure 15.8 0.3 0 16 13 0 Unit % of initial N % of initial N % of initial N % of initial N % of initial DM % of initial N as NOÀ or 3 NHþ 4 % of remaining org, N % of initial N kg wh (DM) kg wh (wb) kg wh product (DM) % of wet weight kg wh product (wb)

N mineralised first season Plant available nitrogen Amount to harvest Fresh water hyacinth to harvest Amount dry matter to spread Dry matter in product Amount to spread

15

10

30

23 135 1432 95

9 338 3568 192

25 124 1305 108

10 957

70 273

85 127

Calculations are based on a dry matter content of 9.5% and a nitrogen content of 3.2% (db) in the fresh water hyacinths (Gunnarsson and Mattsson, 1997). wh = water hyacinth.

be 0.40 m/s. Towing of the water hyacinths might, however, not be necessary as they often are transported directly to the shore by winds. On a smaller scale, harvesting by hand is preferable since it is a simple and inexpensive method. Harvesting of the water hyacinths would improve the health situation for people living in water hyacinth infested areas. Direct negative effects of water hyacinths are the increased occurrence of diseases like malaria and bilharzia. The quality of the fish caught in areas with lots of water hyacinths has also deteriorated (Sunday Standard, Kenya, 12/1-1997). The harvesting process in itself might impose health problems. Harvesting will lead to contact with the water and thereby the risk of being infected by bilharzia. The total time the plants are kept dry, or at least out of the lake, must exceed 48 h (Thors, 1997) to ensure destruction of the bilharzia parasites if they are in the form of cercariae. However, if the hosting snails are present, the bilharzia parasite can survive in the snail for a long time. Cutting the water hyacinths before processing is necessary for anaerobic digestion. For composting, cutting might not be necessary because whole plants will enhance aeration. It is favourable if the preparation of the fresh water hyacinths can be done during the dry season. During

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that season people are not engaged in fieldwork, and consequently have more time for working with the water hyacinths. Since not all water hyacinths can be expected to be used directly on the lakeshore, it is also necessary to find a way to transport the harvested material. The high water content of the fresh water hyacinths makes drying them directly on the beach after harvesting of interest in order to decrease the labour needed for transport. Fresh water hyacinths can be transported in buckets or baskets. When it comes to transport of the product, the nitrogen concentration in the sludge from an anaerobic process is, due to its high water content, very low and the sludge is difficult to transport. Buckets could be used but the large amount that must be transported to provide nutrients, in the form of sludge, for the crop would make it necessary to develop a new transport system. The heavy workloads otherwise might lead to a risk of the sludge being dumped close to the processing plant. If so this would probably lead to even greater eutrophication of the lake and an increase in the amount of water hyacinths. The compost product can be transported in baskets. The amount of compost product to be transported to the field is twice as much as when transporting dried water hyacinths. Once in the field the compost product has a texture very similar to the soil and is therefore easy to work into the soil. Use of dried water hyacinths as green manure is the alternative with the lowest transport requirement. The product is also easy to handle. Drying gives the option to store the plants so they can be harvested when there is time, during the dry season. They can then be used when needed by the farmer, during the rainy season. Sun drying for a few days decreases the weight of the water hyacinths by about half. A dry matter content of 87% is needed to stop microbial activity decomposing the product (Apori, 1994). The advantage of the anaerobic process is that it results in two useful products: a nutrient-rich sludge that can be used for soil improvement and a gas that can be used for cooking, heating, lighting and electricity production, thus decreasing the need for firewood. One restriction with anaerobic digestion is the large need for water, but as water hyacinths grow in water that should not be a problem. Implementation of a biogas program requires, according to experiences from China and India, strong governmental support to be successful. It also requires quite large initial investments and technological skills. Small-scale digester designs exist that, with proper implementation, could work very well in these areas (Gunnersson and Stuckey, 1986). Composting of water hyacinths is a possible treatment method. Compared with drying, the nitrogen loss is larger but the compost product obtained is stable, and if a temperature of above 55 °C is reached for a day or two essentially all pathogens are destroyed (Haug, 1993). To ensure kill-off, the high temperature must be reached throughout the compost pile. The high water content of the fresh water hyacinths probably does not cause any problems when composting in hot, evaporative climates. On the other

hand, regardless of the high water content of the substrate, the compost might need watering, especially during the dry season. If possible, the compost should be placed close to a source of water that does not dry out during the dry season since that is when the water demand is highest. If the water hyacinths are cut into smaller pieces before composting, addition of a material providing structure might be needed (Dalzell et al., 1979). As water hyacinths have a relatively high ash content, 25.7% of DM (Abdelhamid and Gabr, 1991), adding extra ash will probably not be needed. The water hyacinths are valuable in the compost due to their high content of nitrogen. Compost of water hyacinths will off-set the cost of cleaning the irrigation system of this weed and will prevent the health hazard arising from leaving the plant material on the beach (Elserafy et al., 1980). Water hyacinth compost has been shown to have positive effects on crop growth (Sharma and Mittra, 1991; Abdel-Sabour and Abo El-Seoud, 1996; Thakuria et al., 1991). When using the water hyacinth compost as organic fertiliser, it may be advisable not to add the compost too near in time to sowing or transplanting, since it has been concluded that yield may be enhanced by incorporating water hyacinth compost up to 10 days before transplanting rice to the field (Sharma and Mittra, 1990). Due to its positive charge, ammonium can bind to the negatively charged soil particles and the risk is low that it leaches away. The negatively charged nitrate does not bind to the soil particles but remains in the soil fluids and is thus available for leaching (Claesson and Steineck, 1991). Whether ammonium or nitrate is to be preferred as a fertiliser on a certain soil depends on the type of crop grown, the climate, soil conditions and the agricultural practices in the area (Stout, 1983). A large proportion of the nitrogen in anaerobic sludge is in ammonium (NHþ ) form (Stout, 4 1983). In the compost product only a small amount of the nitrogen is in the form of nitrate, and thereby directly available to the plants. Most of the nitrogen is bound in humus and must be mineralised to be plant available. In the dried water hyacinths, all nitrogen is in organic form and the release of nutrients might therefore be delayed with little becoming available to the crop during the first growing season. In the long run, adding organic material will, as mentioned before, increase both the nutrient content and the water-holding capacity of the soil. When comparing the contents of phosphorus and potassium in the fresh water hyacinths with the water hyacinths dried for 3 and 13 days (Gunnarsson and Mattsson, 1997), it can be seen that the two nutrients behave differently. While the phosphorus content stays relatively constant during the drying process, the potassium content has decreased to a value that is half of the original. Phosphorus and potassium can only be lost through leaching (van der Werff et al., 1995). The fact that the dried samples lost potassium indicates that the losses of nitrogen also were mostly due to leaching. The literature also confirms that potassium is lost to a greater extent than phosphorus ´ (Ulen, 1991). The amount of nutrients lost through leach-

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ing is higher if the plant material is dry at the time of pre´ cipitation (Ulen, 1984). 5. Concluding remarks Applying the water hyacinths directly without any other processing than sun drying, seems to be the best alternative in small-scale use, due to the relatively small losses of nutrients and workload required (Gunnarsson and Mattsson, 1997). This option also does not require any large investments or new technology. If the fresh water hyacinths could be applied as mulch on the fields, the labour need for weeding could be used for handling the water hyacinths instead. But transport of fresh water hyacinths means transporting a lot of water. Drying seems to be a reasonable treatment since it will both decrease the labour required for transport and the risk of water hyacinths emerging in the field, as well as improving their hygienic status. It can be assumed that as long as drying is carried out during the dry season, reaching a high enough dry matter content is possible (Gunnarsson and Mattsson, 1997). The limitation might instead be finding time for harvesting and the availability of land to dry the water hyacinths on. To meet the ever-growing energy demand, biogas production could be one option but it requires investments and technological skills that would impose great problems in developing countries, where the water hyacinth is often found. The sludge, if used as fertiliser, would also be difficult to transport. It might be possible to use the containers that are used for carrying water, but the low dry matter content makes this a heavy task. According to calculations by Gunnarsson and Mattsson (1997) 88 ton of sludge must be transported to equal 37 kg of plant available N/ha. The advantage with anaerobic digestion is that the gas produced has multiple uses, such as cooking and lighting, but this does not outweigh the investments and technology needed. For optimal operation, the anaerobic digester should be fed continuously. Anaerobic digestion would have the same labour requirements during the dry and rainy season, and might therefore interfere with the agricultural work. Composting as an alternative treatment has the advantage of producing a product that is easy to work into the soil compared with dried water hyacinths, because of the decomposed structure. But the structure of the compost will also make it more difficult to transport and will, because of the relatively high nitrogen losses, require more transport work compared to dried water hyacinths. Also considerable work is required for taking care of the compost. To provide 37 kg plant available nitrogen, at Lake Victoria, 132 ton of fresh water hyacinths must be composted (Gunnarsson and Mattsson, 1997). That the compost product already contains a well-working flora of microorganisms will be favourable for the soil. The relatively slow mineralisation rate of organic nitrogen prevents leakage of nutrients from the soil. The nutri-

ents will probably be stored in the soil for the next growing season. In the green manure all nutrients are in organic form; in compost, only small amounts of the nutrients are found in the mineralised form. It will therefore take some time before the nutrients are available to the crop. In sludge much of the nitrogen is in the form of ammonium and can be directly available to plants. In warm climates there is a big risk of gaseous losses of nitrogen as ammonia. Therefore green manure or compost might be preferable. For other uses of the water hyacinth, such as incineration on a large scale, it must be possible to reduce the moisture content, ‘‘with relative ease’’, to 15% or less (Thomas and Eden, 1990). The highest dry matter content reached in trials by Gunnarsson and Mattsson (1997) was 87%, in roots after one week of drying on a fence. Storing or incineration might be possible if drying can be accomplished during the dry season. Burning the dried water hyacinths directly or as briquettes might therefore be a feasible solution that would not only decrease the water hyacinth problem but also provide energy and thereby decrease the deforestation. Briquetting would produce a product that is more similar to the charcoal used today, but the processing requires energy input and investments in machinery. The ashes could then be spread in the fields to provide minerals, mainly phosphorus and potassium (and larvae protection). The ash spreading would require a relatively low labour input, but the effects and application rate must be investigated. An interesting feature of the water hyacinths is their ability to accumulate nutrients and metal ions from surrounding water. If water hyacinths are introduced to the ponds for biological cleaning of the water, together with other adjustments, it might improve the degree of cleaning. This would, in a longer perspective, improve the water quality in the recipient water by decreasing the input of nutrients and pollutants. Reduced nutrient input might in turn decrease the growth rate of the water hyacinths in the recipient water and thereby further improve the quality of the water and help to control the further spread of the hyacinths. Water hyacinths pose a big, and increasing, problem in many places; mechanical control alone might not be sufficient. A more productive way to finally control the growth can be to make use of the plant by using one or several of the techniques described in this paper. Acknowledgements The authors would like to thank the Swedish Development Agency (SIDA) for financing this study. We would ˚ also like to thank Assoc Prof. Hakan Jonsson, Prof. Girma ¨ Gebresenbet, the District Commissioner of Homa Bay, Kenya, Dr. K.V. Seshu Reddy, Scientist-in-Charge at Mbi´ ta Point Field Station, Britta Widen, Stephan Noll, Dr. Lennart Bengtsson and everybody at the SNFIOH Women Centre in Homa Bay, Kenya for helping in the preparation of the manuscript.

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