Reviews in Fish Biology and Fisheries 12: 131, 2002. 2003 Kluwer Academic Publishers. Printed in the Netherlands.

The potential of sh production based on periphyton

Anne A. van Dam1,3 , Malcolm C.M. Beveridge2 , M. Ekram Azim1 & Marc C.J. Verdegem1
1 Fish

Culture and Fisheries Group, Department of Animal Sciences, Wageningen University, P.O. Box 338, 6700 AH Wageningen, The Netherlands; 2 FRS Freshwater Laboratory, Faskally, Pitlochry, Perthshire, Scotland, UK PH16 5LB; 3 Current address: Department of Environmental Resources, IHE-Delft, P.O. Box 3015, 2601 DA Delft, The Netherlands (E-mail: avd@ihe.nl; Fax: 31-15-2122921; Phone: 31-15-2151712/2151715)

Received 5 December 2001; accepted 17 October 2002

Contents
Abstract Introduction Background Objectives and scope of the review Terminology Natural and articial periphyton-based systems Natural systems with periphyton Brush-park sheries Traditional aquaculture systems in southeast Asia Aquaculture experiments Water treatment with periphyton Periphyton productivity Development of the periphyton assemblage and species composition Biomass and productivity Effects of environmental factors Fish Morphological and physiological adaptations to herbivory Periphyton ingestion by sh Periphyton as sh feed: proximate composition Assimilation efciency and food conversion ratio Potential sh production based on periphyton Conclusions and recommendations for further research Role of periphyton in aquaculture systems Ability of sh to utilize periphyton Potential of periphyton-based sh production Acknowledgements References page 1 2

10

17

21 24

26 26

Key words: sh ponds, herbivory, nutrients, periphyton, phytoplankton

Abstract Periphyton is composed of attached plant and animal organisms embedded in a mucopolysaccharide matrix. This review summarizes research on periphyton-based sh production and on periphyton productivity and ingestion by sh, and explores the potential of developing periphyton-based aquaculture. Important systems with periphyton are brush-parks in lagoon areas and freshwater ponds with maximum extrapolated sh production of 8 t ha1 y1 and 7 t ha1 y1 , respectively. Experiments with a variety of substrates and sh species have been done, sometimes with supplemental feeding. In most experiments, sh production was greater with additional substrates compared to controls without substrates. Colonization of substrates starts with the deposition of organic substances

2 and attraction of bacteria, followed by algae and invertebrates. After initial colonization, biomass density increases to a maximum when competition for light and nutrients prevents a further increase. Often, more than 50% of the periphyton ash-free dry matter is of non-algal origin. Highest biomass (dm) in natural systems ranges from 0 to 700 g m2 and in aquaculture experiments was around 100 g m2 . Highest productivity was found on bamboo in brush-parks (7.9 g C m2 d1 ) and on coral reefs (3 g C m2 d1 ). Inorganic and organic nutrients stimulate periphyton production. Grazing is the main factor determining periphyton density, while substrate type also affects productivity and biomass. Better growth was observed on natural (tree branches and bamboo) than on artical materials (plastic and PVC). Many herbivorous and omnivorous sh can utilize periphyton. Estimates of periphyton ingestion by sh range from 0.24 to 112 mg dm (g sh)1 d1 . Ingestion rates are inuenced by temperature, sh size, sh species and the nutritional quality of the periphyton. Periphyton composition is generally similar to that of natural feeds in shponds, with a higher ash content due to the entrapment of sand particles and formation of carbonates. Protein/Metabolizable Energy (P/ME) ratios of periphyton vary from 10 to 40 kJ g1 . Overall assimilation efciency of sh growing on periphyton was 2050%. The limited work on feed conversion ratios resulted in values between 2 and 3. A simple simulation model of periphyton-based sh production estimates sh production at approximately 2.8 t ha1 y1 . Together with other food resources in shponds, total sh production with the current technology level is estimated at about 5 t ha1 y1 . Because grazing pressure is determined by sh stocking rates, productivity of periphyton is currently the main factor limiting sh production. We conclude that periphyton can increase the productivity and efciency of aquaculture systems, but more research is needed for optimization. Areas for attention include the implementation and control of periphyton production (nutrient levels, substate types and conformations), the ratio of sh to periphyton biomass, options for utilizing periphyton in intensive aquaculture systems and with marine sh, and possibilities for periphyton-based shrimp culture.

Introduction Background Fish production through aquaculture is realized in a wide variety of culture systems, from extensive seasonal ponds to intensive concrete raceways or oating marine cages. In 1998, 53% of the 30 million tonnes of nsh, molluscs and shrimp produced in aquaculture were predominantly cultured in extensive to semi-intensive pond systems (mainly Chinese carps like Hypophthalmichthys molitrix, Ctenopharyngodon idella, and Aristichthys nobilis, all Cyprinidae; common carp, Cyprinus carpio, Cyprinidae; and Nile tilapia, Oreochromis niloticus, Cichlidae). Ponds are also important in terms of production value, accounting for some 47% of the total value. Carps and the tiger shrimp (Penaeus monodon, Penaeidae) are among the most important commodities (Table 1; FAO, 2001). All pond species feed low in the food chain, most being lter feeders, herbivores, or omnivores. Production in extensive pond systems is based on the natural productivity of the pond and solar energy. In semi-intensive systems, organic and chemical fertilizers and supplemental feeds are added whereas intensive systems are based predominantly on high-quality complete feeds. Only 515% of the

nitrogen added to the ponds as fertilizer is harvested as sh biomass (Edwards, 1993; Gross et al., 1999). In feed-driven systems, only 2030% of the nitrogen in the feed is retained in the sh biomass (Avnimelech and Lacher, 1979; Boyd, 1985; Jimnez-Montealegre, 2001). The nutrients that are not harvested as sh biomass either accumulate in the pond sediment, volatilize, or are discharged into the environment. From economic and environmental points of view, there is a need to examine options to make aquaculture systems more nutrient efcient. Generally, three food pathways can be distinguished in aquaculture systems: (1) direct feeding by the sh on feeds; (2) the autotrophic pathway, in which solar energy is used by primary producers (mainly algae) to convert carbon dioxide into organic matter that can be utilized by sh; and (3) the heterotrophic pathway, in which heterotrophic organisms (bacteria, protozoa, and other invertebrates) decompose organic matter that can be utilized by the sh (Schroeder, 1978). These three pathways are linked through uxes of organic and inorganic nutrients. In wastefed systems, the heterotrophic pathway can be more important than the autotrophic pathway, but stable isotope studies show that a large part of the microbial production in ponds is based on algal detritus (Schroeder, 1978; Schroeder et al., 1990). Estimates

3
Table 1. Importance of pond systems in world aquaculture production of nsh, molluscs, and shrimp in 1998. Data: FAO (2001). CM = coastal marine system; P = pond; R = raceway; C = oating cage Predominant production system

Common name Molluscs Pacic cupped oyster Japanese carpet shell Yesso scallop Blue mussel Blood cockle Mediterranean mussel Shrimp Giant tiger prawn Whiteleg shrimp Finsh Silver carp Grass carp Common carp Bighead carp Crucian carp Nile tilapia Rohu Atlantic salmon Catla Mrigal White Amur bream Rainbow trout Milksh Channel catsh Japanese eel Mud carp Total this list Total ponds (excluding Oncorhynchus mykiss) Total world (nsh, molluscs and shrimp)

Scientic name

Volume 106 MT

Value 106 US$

Crassostrea gigas Ruditapes philippinarum Pecten yessoensis Mytilus edulis Anadara granosa Mytilus galloprovincialis

3.44 1.43 0.86 0.50 0.25 0.16

11.1 4.6 2.8 1.6 0.8 0.5

3.27 1.86 1.18 0.26 0.23 0.11

6.9 4.0 2.5 0.6 0.5 0.2

CM CM CM CM CM CM

Penaeus monodon Penaeus vannamei

0.58 0.19

1.9 0.6

3.86 1.03

8.2 2.2

P P

Hypophthalmichthys molitrix Ctenopharyngodon idellus Cyprinus carpio Aristhichthys nobilis Carassius carassius Oreochromis niloticus Labeo rohita Salmo salar Catla catla Cirrhinus mrigala Parabramis pekinensis Oncorhynchus mykiss Chanos chanos Ictalurus punctatus Anguilla japonica Cirrhinus molitorella

3.31 2.89 2.47 1.58 1.04 0.79 0.75 0.69 0.63 0.56 0.45 0.44 0.37 0.26 0.21 0.16 24.01 16.24 30.86

10.7 9.4 8.0 5.1 3.4 2.6 2.4 2.2 2.0 1.8 1.5 1.4 1.2 0.8 0.7 0.5 77.8 52.6 100.0

3.09 2.66 2.83 1.45 0.83 0.89 1.94 2.20 0.55 0.47 0.54 1.36 0.55 0.42 0.82 0.16 32.56 22.09 47.08

6.6 5.6 6.0 3.1 1.8 1.9 4.1 4.7 1.2 1.0 1.1 2.9 1.2 0.9 1.7 0.3 69.2 46.9 100.0

P P P P P P P C P P P P/R P P P P

of the proportion of the standing stock of phytoplankton that accumulates as sediment in the bottom range from 20 to 50% per day (Jimnez-Montealegre, 2001). Thus, a large part of the phytoplankton production is decomposed on the pond bottom and contributes to the accumulation of nutrients in the sediment. Because many sh species are not able to harvest phytoplankton directly from the water column, an extra trophic level is involved in converting phytoplankton into sh biomass. With an estimated energy transfer efciency of 10% per trophic level (Pauly and Christensen, 1995), maximum sh yield may be no more than 1% of the energy xed by the phytoplankton consumed. Fish yields from extensive and

semi-intensive ponds could be up to ten times higher if primary production could be harvested directly by herbivorous sh. Whether phytoplankton can be harvested directly by sh depends largely on the sh species stocked. Although species like silver carp and bighead carp are capable of harvesting microalgae directly, many species used in aquaculture cannot. Even for Nile tilapia, generally regarded as a phytoplankton feeder, it seems questionable whether it can derive enough energy from exploiting phytoplankton (Dempster et al., 1993, 1995). Phytoplankton has some other disadvantages. Nighttime respiration by phytoplankton in ponds may lead to oxygen depletion during the

4 early morning hours, causing a risk of sh mortality (Madenjian et al., 1987). Decomposition of sedimented phytoplankon may result in toxic decomposition products (ammonia, nitrite) and increased oxygen demand. Phytoplankton blooms are unstable and may collapse unexpectedly, resulting in a sudden drop in dissolved oxygen concentrations and sh mortality (Delinc, 1992). On the other hand, phytoplankton serves a number of very important functions in pond aquaculture. It is a net producer of dissolved oxygen, which is indispensable for sh growth and production (Smith and Piedrahita, 1988; Teichert-Coddington and Green, 1993). It is also the most important sink of ammonianitrogen, which is excreted by sh and potentially toxic (Hargreaves, 1998; Jimnez-Montealegre, 2001). Most truly herbivorous sh species feed on larger, benthic, epilithic or periphytic algae, rather than on phytoplankton (Horn, 1989). Such algae require substrates for attachment, which are virtually absent in sh ponds. In response to the high nutrient levels that are maintained by pond fertilisation and sh excretion, high-density phytoplankton blooms usually develop. These reduce the light penetration to the pond bottom, thus preventing the development of benthic algal mats. If pond algae could be grown on substrates, more sh species may be able to harvest them, resulting in a more efcient utilization of primary production. Communities of attached algae are generally more stable than phytoplankton and the risk of collapse is much lower (Westlake et al., 1980). Some studies suggest that the production of attached algae per unit water surface area is higher than of phytoplankton (Wetzel, 1964). Horne and Goldman (1994) stated that it is mechanically more efcient to scrape or graze a two-dimensional layer of periphyton than to lter algae from a three-dimensional planktonic environment. Considering all these aspects, it might be advantageous to develop periphyton-based pond culture. Objectives and scope of the review The main objective of this review is to assess the potential of periphyton-based sh/shrimp production in aquaculture pond systems on the basis of the available literature on periphyton productivity and on periphyton utilization by sh and shrimp. We present an overview of data on natural and culture systems where sh utilize periphyton and describe the species composition of periphyton and the architecture and functionality of the periphyton assemblage. The productivity of periphyton in natural systems in relation to environmental factors, substrate types and grazing are reviewed and the potential productivity in culture sytems is estimated. We also review the quality of periphyton as a sh feed and examine the morphological and physiological adaptations of sh for utilizing periphyton. Data on periphyton grazing by sh and the effects of grazing on periphyton productivity are discussed. Based on this body of information, we estimate potential periphyton-based sh production with a simple simulation model. To conclude, we indicate knowledge gaps for developing periphytonbased aquaculture and make recommendations for further research. Terminology Throughout this paper, we will use the term periphyton to indicate the assemblage of attached aquatic plant and animal organisms on submerged substrates, including associated non-attached fauna. Several other terms are used with regard to this assemblage. The most general terms are aufwuchs (often also written with a capitalized A, from the original German word Aufwuchs) and biolm. Some authors prefer to talk about attached algae, but this disregards the many other forms that live in periphyton assemblages. Aufwuchs includes all the organisms that are attached to, or move upon, a submerged substrate, but which do not penetrate into it, whereas periphyton refers to the total assemblage of sessile or attached organisms on any substrate (Reid and Wood, 1976; Weitzel, 1979). The difference is in the unattached organisms that are often found in association with the periphyton assemblage. Sometimes, the terms euperiphyton (immobile organisms attached to the substrate by means of rhizoids, gelatinous stalks, or other mechanisms) and pseudoperiphyton or metaphyton (freeliving, mobile forms that creep among or within the periphyton) are used (Weitzel, 1979). The term biolm is preferred in other elds of application, such as wastewater treatment (Cohen, 2001), drinking water technology (Momba et al., 2000), food processing (Joseph et al., 2001) and dentistry (Rosan and Lamont, 2000) and is used mainly for attached bacteria and protozoa but not algae (OToole et al., 2000). Other terms used to indicate periphyton indicate the substrate on which it grows: epiphyton (on

5 plants), epipelon (on sediment), epixylon (on wood), epilithon (on rocks). Natural and articial periphyton-based systems Natural systems with periphyton Some of the earlier research on periphyton was carried out in lakes, and it was shown that periphyton can have an important share (42.4% for the lake studied) in the total annual production, especially in shallow lakes with large littoral zones (Wetzel, 1964). In ve oligotrophic lakes, periphyton contributed 4397% of the total productivity in the shallow (23 m) zone (Loeb et al., 1983). In the littoral zone, periphyton can grow on rocks and sediments but also as epiphyton on macrophytes. In natural, unpolluted streams periphyton density is highest in the mid-waters, where currents are moderate and erosion and deposition are balanced. Nutrients, imported from upstream, are absorbed by periphyton attached in locations with sufcient light for photosynthesis, such as rocks or the stream bed. Upstream, the current is stronger, erosional processes and allochthonous inputs are more important, nutrients are scarce and shredders dominate the food chain. In the lower reaches, the currents are slow and deposition processes are dominant. In this nutrient-rich environment phytoplankton thrives (Welcomme, 1985). An example of this trophic gradient was shown in a study of a grassland stream in New Zealand, where nitrate concentrations increased along a downstream gradient which was reected in the species composition and biomass of the periphyton (Biggs et al., 1998a, b). The key feature of periphyton in running water environments is its ability to utilize scarce nutrients in a xed position favourable for photosynthesis. Once trapped, nutrients can be recycled within the periphyton assemblage. A model of periphyton biomass with nutrient concentration and water velocity as main driving variables gave good results when compared to eld data from a river in Argentina (Saravia et al., 1998). In marine habitats, periphyton is also found in littoral zones (mangrove forests, estuaries) and on coral reefs. On coral reefs, the accumulation of organic material is facilitated by the combination of a high primary productivity of the attached algae with nitrogen xing by cyanophytes, the capture of N from the surrounding ocean and the recycling of nutrients within the reef. This explains how a relatively high sh biomass can be sustained in oligotrophic water. An important part of the primary production is transferred to the coral host in the form of organic exudates. The main limiting factor is the surface area available for photosynthesis by attached primary producers (Longhurst and Pauly, 1987). A range of herbivores, including sh, echinoids, and other invertebrates, graze on coral reef algae. Exclusion experiments with cages have shown that intense grazing by sh or sea urchins leads to reefs with a less diverse (in terms of species), lower algal biomass dominated by the smaller turfs and crustose corallines than ungrazed reefs (Ogden and Lobel, 1978; Hatcher, 1983; Steneck, 1988). Primary productivity in coral reefs is very high, but values probably depend a lot on the part of the reef where the measurement was done (depth, exposure to currents). Algal productivity is generally believed to increase when the standing crop is reduced by grazing, because of reduced self-shading, enhanced nutrient exchange with the water and maintenance of the plants in the exponential growth phase (Ogden and Lobel, 1978; Hatcher, 1983). Brush-park sheries Brush-park sheries are practiced in a large number of countries and areas: West Africa, Madagascar, Sri Lanka, Mexico, Bangladesh, Cambodia, China, and Ecuador (Kapetsky, 1981). It is a traditional technology that shares features of both capture sheries and aquaculture. Research on brush-park sheries in the United States was done as early as the 1930s (Rodeheffer, 1940; in Pardue, 1973). Current examples are the katha shery in Bangladesh (also called jhag, katta or jhata; Wahab and Kibria, 1994), samarahs in Cambodia (Shankar et al., 1998), and athkotu in Sri Lanka (Senanayake, 1981). Kathas are constructed from the branches of trees such as hizol (Barringtonia sp.), jamboline (Eugenia sp.) or acacia (Streblus sp.). Branches are piled up between a number of bamboo poles xed in the bottom to maintain the structure and delimit the area of the katha. Kathas are usually built in secondary rivers or canals in oodplain lakes. Water hyacinth may be used to cover the katha. The whole structure can be 69 m long, 26 m wide and approximately 1.25 m deep. Kathas are usually operated for 57 months each year, during which period they are shed 3 4 times, principally between September and March when water levels recede and the water becomes cool.

6 For shing, the katha is encircled with a net and all branches are removed. Fishing the whole katha may take several days and usually involves 45 persons using scoop nets. Harvests range from 100 to 1000 kg, depending on the size of the katha. Similar sheries are the kua shery where branches are placed in natural or excavated depressions at the beginning of the rainy season (Wahab and Kibria, 1994) and the juk shery in Kaptai Lake (Ahmed and Hambrey, 1999). Much better studied are the acadjas in West African coastal lagoons. An acadja is an articial reef made of tree branches (mangrove poles or sometimes palm trunks) and installed in water of about 11.5 m deep. It attracts sh and is colonized by periphyton which serves as food for the sh. Most important species in the brush-park sheries of Benin are the blackchin tilapia (Sarotherodon melanotheron, Cichlidae) and bagrid catsh (Chrysichthys nigrodigitatus, Bagridae), but many other species are reported from other countries. After encircling the acadja with a net and removing the branches, the sh are removed, if necessary using traps or baskets. In some areas, shing is done by hook and line. Apart from attracting sh from outside, sh also reproduce inside the acadja system. Production gures reported are high, from 420 t ha1 y1 (Welcomme, 1972; Hem and Avit, 1994). Because of their protability, acadjas proliferated in West-Africa which led to resource use conicts: competition with navigation for space in the lagoon, competition with capture sheries for wild sh stock (although it is also claimed that brush-parks may be benecial to capture sheries because sh disperse to adjacent open waters; Welcomme, 1972). There are also negative environmental impacts, such as increased silting in the lagoons due to accelerated sedimentation around the brush-parks, organic pollution caused by the decaying branches in the water, increased erosion as a result of deforested catchment areas and a net export of nutrients in areas of intense acadja harvesting (Durand and Hem, 1996; Weinzierl and Vennemann, 2001). An experiment with the so-called acadja-enclos was reported by Hem and Avit (1994). They compared three 625 m2 enclosures in the Ebri lagoon in Ivory Coast (salinity 09 ): one empty, one with a 100 m2 acadja of Echinochloa pyramidalis (a oating macrophyte), and one with a 100 m2 traditional acadja made of the usual tree branches. Fish recruited to these systems naturally by swimming through the 14mm mesh surrounding nets. After 12 months, total biomasses of 11.7, 18.2, and 80.5 kg, respectively were harvested from the three enclosures. Blackchin tilapia was the dominant sh species in the enclosure with tree branches. Subsequent trials with different sizes of acadja-enclos (2002500 m2 ) yielded on average 1.8 t ha1 . Because of the high requirements for wood, additional trials with bamboo sticks (10 sticks m2 , appoximately 6 cm diameter) were done, leading to average yields of 8.3 t ha1 . The authors expect even higher yields if a scheme of successive selective harvesting would be employed. Similar experiments in Sri Lanka using different mangrove and non-mangrove tree species to construct brush-parks of 4-m diameter resulted in comparable yields (extrapolated: 2.312.9 t ha1 y1 , assuming 10 productive months per year and depending on substrate species) of mainly green chromide (Etroplus suratensis, Cyprinidae), streaked spinefoot (Siganus javus, Siganidae), dory snapper (Lutjanus fulviammus; Lutjanidae), prawns (Penaeus spp., Metapenaeus dobsoni and Macrobrachium spp.) and ornamental sh (Costa and Wijeyaratne, 1994). Traditional aquaculture systems in southeast Asia In the Philippines, Indonesia, and Taiwan, the traditional culture system for milksh (Chanos chanos, Chanidae) in coastal ponds was based on mats of benthic algae, protozoa, and detritus (in the Philippines called lablab) stimulated by organic fertilization. In Indonesia, mangrove leaves (notably Avicennia sp.) and twigs were used, whereas in the Philippines green manures or copra slime were applied. Inorganic fertilization was rare. Supplementary feeding with rice straw, rice bran, oil cake, wheat starch, water hyacinth, or other macrophytes was sometimes applied. In the shallow ponds (0.30.7 m water depth), a thick mat of algae developed, consisting of cyanobacteria (e.g., Oscillatoria, Lyngbya, Phormidium, Spirulina, Microcoleus, Chroococcus, Gomphosphaeria) and diatoms (e.g., Navicula, Pleurosigma, Nastogloia, Stauroneis, Amphiora, Nitzschia and Gyrosigma). Other benthic ora and fauna as well as lamentous green algae were also ingested. Apart from the target species, about 20% of the harvest in Java could consist of prawns (Huet, 1986). Benitez (1984) distinguished between oating lablab that contained 15% protein (ash-free dry matter basis), and benthic lablab with only 6% protein, and reports an observation by sh farmers

7 that milksh consuming lamentous green algae had a slower growth rate than sh eating lablab consisting of unicellular algae and diatoms. Nowadays, many of these traditional systems have been replaced by deeper milksh ponds with higher stocking density and supplemental feeding. Another traditional culture system is rice-sh culture. Rice elds harbour lots of phytoplankton and lamentous higher algae, but studies on rice eld ecology do not reveal much about periphyton (Roger, 1996). Not much is known about the natural feeds consumed by sh in these systems. Periphytic detrital aggregate was the most important item in the food of Nile tilapia and common carp in rice elds in northeast Thailand (Chapman and Fernando, 1994). Fish feeding on periphyton on rice plants could be so vigorous that the rice plants were observed to be shaking (Chapman, 1991). Similar observations are reported from rice-sh studies in Vietnam (Rothuis et al., 1999) and in Bangladesh (Gupta et al., 1998). Aquaculture experiments Pardue (1973) reports two experiments from Alabama with bluegill (Lepomis macrochirus, Centrarchidae) grown in plastic circular tanks (stocking density 2 m2 , 3 m diameter, water volume 5,400 l) equipped with yellow pine boards with surface areas equivalent to 20, 40, 60, 80 and 100% of the tank surface. Fish production increased linearly with increasing surface area, and the highest mean yield with 100% surface area added was 384 kg ha1 of bluegill in 180 days (y = 243.34 + 1.408x, in which y = sh yield [kg] and x = added substrate [%]). In a subsequent experiment with 40 and 100% additional surface area and three levels of fertilization, there was no difference between the two levels of added surface but a strong effect of fertilization with the highest yields (mean: 430 kg ha1 ) achieved under complete fertilization (8-8-2 NPK at 112 kg ha1 , applied 6 times during the 180 days culture period). The increase in bluegill production was linked to the increase in macroinvertebrates on the substrates, notably Diptera, Hemiptera, Odonata, and Plecoptera. Cohen et al. (1983) added substrates to 200-m2 ponds for freshwater prawn (Macrobrachium rosenbergii) in Israel in an attempt to mimic the natural conditions for prawn growth by increasing the available surface area. The substrates consisted of two horizontal layers of plastic 2-cm mesh nets with corrugated plastic pipes attached to them (8-cm diameter, 1520 cm long). Each pond was stocked with 2,000 juvenile prawns (2 g) and 15 common carp (100 g). The ponds were fertilized with chicken manure and a 25% pellet feed was given during the night. Selective harvesting of large prawns was done to prevent suppression of growth caused by territorial behaviour. The total marketable yield from ponds with substrates was 2,850 kg ha1 in six months, against 2,500 in ponds without substrates. The average nal weight of the prawns was also higher with substrates (40.3 g versus 35.8 g). There was no difference in survival. Another set of experiments with Macrobrachium rosenbergii was done at Kentucky State University. In a rst experiment, substrates consisting of 3 horizontal levels of plastic mesh sheet, suspended 30 cm apart in a PVC pipe frame were placed in 400 m2 ponds. The substrates increased available surface area by about 20%. The substrates increased the yield of prawns stocked at 0.33 g individual weight and 59,280 individuals ha1 from 1,060 kg ha1 (without substrates) to 1,268 kg ha1 in 106 days. Mean size at harvest was also bigger (37 g against 30 g without substrates) and the number of mature females increased with substrates (Tidwell et al., 1998). In a similar experiment with two stocking densities and substrates that added 80% of available surface area to the ponds, the substrates produced an increase in yield from 1,243 to 1,469 kg ha1 in 95 days (averages for stocking densities 60,000 and 120,000 ha1 , size at stocking = 0.24 g) and a decrease in food conversion ratio from 2.9 to 2.4 (Tidwell et al., 1999). In a third experiment, the density of the substrates was varied at a xed prawn density of 74,000 ha1 with a stocking weight of 0.24 g. Substrates consisted of 120 cm wide, 7.03.5 cm mesh polyethylene panels suspended horizontally across the ponds, 30 cm above the bottom with 30 cm between the layers when multiple layers were installed. Three treatments were created, corresponding to surface area increases of 0, 40 and 80%. There was a positive linear response of yield on substrate density: Y (kg/ha) = 1466.3 + 4.4604 X (% increase in surface). Feed conversion ratio was inversely related to increase in surface area: Y (FCR) = 2.784 0.0052 X (% increase in surface). There was no signicant difference in individual weight at harvest (Tidwell, 2000). Bender et al. (1989) produced a microbial mat by applying 32 g dm m2 of native (Dominican Republic) grass clippings to small (3014 cm), shallow (4.5 cm) laboratory ponds and inoculating with Oscillatoria sp.

8 The experiment was repeated with larger (135 m) concrete ponds and a water depth of 20 cm. Apart from the microbial layer growing on the grass substrate, a detrital gelatinous deposit developed on the pond sediment. Weight gain of 0.53.5 g Nile tilapia in a 2-week growth trial was higher with the silage-microbial mat than with a commercial catsh feed applied at 3% BW day1 . For the microbial mat to induce sh growth, it had to be offered in the same pond where it was grown. Microbial mats or detrital material grown in one pond and offered to sh in another pond (either with or without sediment) did not induce growth in the sh. It was concluded that the integration of the detrital materials with the biomass is crucial for the productivity of such a system. Phillips et al. (1994) obtained signicant differences in nal individual weight of Nile tilapia between ponds with and without similar microbial mats, but no data on mortality and total yield were given. At the Asian Institute of Technology in Bangkok, Shrestha and Knud-Hansen (1994) carried out two experiments in concrete tanks (2.521.1 m) using vertically suspended corrugated plastic sheets (7.7 m2 extra surface area per tank) as substrates with inorganic fertilization (2.1 g N m2 week1 ). Sexreversed all-male Nile tilapia were stocked (20 g sh1 , 3 sh m2 ) for 56 days. Microscopic examination of gut contents and periphyton, as well as observation of feeding behaviour showed that the sh were feeding on the periphyton. In the rst experiment, net sh yields were not signicantly different between tanks with and without substrates (1.05 and 0.88 g m2 d1 , respectively). Although there was a difference in mean periphyton density between tanks with and without sh (0.78 and 0.93 mg dm cm2 , respectively), the difference was not statistically signicant. The second experiment compared the plastic substrate with a similar surface area of bamboo poles. Fish yield was higher with bamboo poles than with plastic (3.43 and 2.51 g m2 d1 , respectively) and on this occasion, there was signicantly less periphyton in tanks with sh (0.80 against 1.71 mg dm cm2 on plastic sheets; bamboo substrates were only tested with sh). From the data on dry matter and ash-free dry matter, it can be seen that the ash content of the periphyton (appr. 50%) was consistently much higher than in the suspended solids (consisting mainly of phytoplankton; ash appr. 20%), with the exception of periphyton on bamboo (appr. 20%). Shankar and co-workers (Shankar et al., 1998; Ramesh et al., 1999; Umesh et al., 1999) used bio-degradable substrates to stimulate sh production. In two preliminary experiments, production of common carp, Mozambique tilapia (Oreochromis mossambicus, Cichlidae) and rohu (Labeo rohita, Cyprinidae) in concrete tanks was 4550% higher with sugarcane bagasse as a substrate, compared to control tanks without substrates. A subsequent bigger trial compared dried sugarcane bagasse, paddy straw, and dried water hyacinth (Eichhornia sp.) leaves, suspended as 6090 cm bundles in 25 m2 concrete tanks with soil bottoms. Substrate density was 12.5 kg tank1 (as the authors stressed the biodegradability of the substrates, substrate density was expressed as mass per surface area) and the tanks were fertilized with cow manure and urea. A mixture of common carp (2.1 g) and rohu (1.5 g) were stocked at 13 and 12 per tank, respectively. The experiment lasted 133 days and on day 70, 7.5 kg of fresh substrates were suspended. Both rohu and common carp grew best in the treatment with sugarcane bagasse, yielding 3,088 g tank1, against 2,873, 2,403 and 1,865 g with paddy straw, Eichhornia and in tanks without substrate, respectively. In all substrate treatments, sh survival on average was higher than in the control (85.793.7%, versus 81.3%). The increased sh production could, in the absence of major differences in dissolved oxygen and ammonia concentrations, be attributed to the periphyton on the substrates, as shown by total plate counts of bacteria in the water and on the substrates, and by phytoplankton and zooplankton enumeration. The superior sh production with bagasse was attributed to its higher bre content and surface area, favouring better bacterial growth and subsequent sh production than the other two substrates. Zooplankton density in the water of the bagasse treatment was higher than in the other treatments. The authors concluded that biodegradable substrates led to better results than less degradable substrates like bamboo or non-biodegradable substrates like PVC and plastic. Bratvold and Browdy (2001) studied changes in water quality and microbial community activity due to AquaMatsTM substrates (3.4 m2 per m3 tank) added to polyethylene tanks stocked with Litopenaeus vannamei postlarvae. Shrimps were fed with a commercial feed. Compared to tanks with only sand sediment or tanks without sediment, tanks with substrates and sand sediment had a higher pH and higher total photosynthesis, lower abundance of pelagic bacteria and phytoplankton, lower turbidity, lower ammonia and orthophosphate and higher nitrication. Shrimp production was signicantly higher

9 (1.69 vs. 0.981.07 kg m2 in 100 days) and food conversion ratio (feed given/shrimp produced) was signicantly lower (1.5 vs. 1.92.1) in the tanks with substrates compared to the tanks without substrates. Keshavanath et al. (2001b) reported an experiment with different types of substrates for enhancing the production of mahseer (Tor khudree, Cyprinidae). Bamboo poles, PVC pipes and sugarcane bagasse substrates were placed in 25 m2 concrete tanks with mud bottoms and ngerlings of about 3 g were stocked at densities of 1, 1.5, and 2 sh per m2 . After 90 days, the highest net production with bamboo substrate was 447 kg ha1 at the highest sh density, against 399 kg ha1 with the PVC pipes. In the bagasse treatment, all sh died due to low oxygen concentrations. In subsequent experiments (Keshavanath et al., 2002) using the same tank systems, the effects of periphyton, supplemental feeding and their combination were investigated using mahseer (3.5 g) or the fringed lipped peninsula carp (Labeo mbriatus, Cyprinidae) (0.73 g), both stocked at 25 per tank. Two densities of bamboo poles (98 or 196 poles per 25 m2 ) were compared with tanks without substrates. Periphyton alone and feeding alone led to comparable sh yields that were signicantly higher (by 3075%) than the yields obtained without periphyton or feed. There was a signicant effect of substrate density on sh survival in both species. The combination of periphyton and feeding resulted in even higher yield increases (5487%). The higher substrate density improved yield only slightly without feeding and not at all with feeding, suggesting that at the sh stocking density used the carrying capacity of the periphyton was never exceeded. A similar experiment with red tilapia (Oreochromis mossambicus O. niloticus hybrid) gave similar results, with even more pronounced differences between tanks with and without substrates. Highest tilapia yields were 1,834 g per 25 m2 without, and 2,142 g per 25 m2 with feeding in 75 days (Keshavanath, unpublished results). Sugarcane bagasse was again used as a substrate in a farm trial. At densities of 156 bagasse bundles (about 28 kg) per 100 m2 , total sh yield of catla (Catla catla, Cyprinidae), rohu, and common carp was 13,104 and 14,842 g per 100 m2 in 180 days without and with feed, respectively, compared to 8,076 g in the control without feed or periphyton (Keshavanath et al., 2001a). At the Bangladesh Agricultural University in Mymensingh, Bangladesh, research on periphyton started with a series of monoculture experiments in which the performance of several sh species with periphyton was assessed. An experiment in six 75 m2 ponds compared the production of 2.1 g orangen labeo (Morulius calbasu, Cyprinidae) stocked at 1 m2 with and without substrates made of kanchi (bamboo trimmings). While no differences in water quality were observed, survival (8790% versus 72 77%) and sh growth were higher with substrates than without, resulting in a net yield of 713 kg ha1 versus 399 kg ha1 in 120 days. These relatively low yields could be explained by the sub-optimal water temperatures that prevailed during the experiment (23.632.7 C) (Wahab et al., 1999). Rohu (10 g at 1 m2 ) and kuria labeo (Labeo gonius, Cyprinidae; 4 g at 1 m2 ) were then grown with bamboo substrates at 9 poles m2 (but leaving the pond perimeter free; total substrate area was about 75 m2 ). Net rohu yield with substrate was signicantly greater (1,901 kg ha1 in 120 days) than without substrate (1,073 kg ha1 ). For kuria labeo (separate experiment), yields were not signicantly different (794 and 788 kg ha1 , respectively, in 120 days). The kuria labeo were never observed to feed actively on the periphyton whereas rohu could be clearly seen eating the periphyton (Azim et al., 2001a). It was concluded that rohu and orangen labeo are more suitable candidates for periphytonbased aquaculture than kuria labeo. To utilize the other food sources in the pond (plankton, detritus) as well, polyculture of rohu and catla was investigated. With different stocking ratios and bamboo substrate, the growth and total yields from any combination of rohu and catla were higher by 3 40% (individual weight) and 50300% (total yield), than those from rohu or catla in monoculture. The combination of 60% rohu and 40% catla was optimal, resulting in a net yield of 586 kg ha1 70 d1 (Azim et al., 2002a). Using this stocking ratio, another trial with and without bamboo substrates was carried out to verify the effect of substrates with this species combination. Survival, growth rate, individual weight at harvest as well as net yield of both rohu and catla, were signicantly higher in the ponds with bamboo substrates. In ponds with substrate, not only the net yield of periphyton-feeding rohu was higher (by 160%), but also that of surface-feeding catla (220%). The combined net production of the two species in ponds with substrates was 180% higher (1,652 kg ha1 90 d1 on average) than that of control ponds (577 kg ha1 90 d1 ). In the same experiment, it was found that the addition of 15% orange-n labeo to the

10 optimum mix of catla and rohu further increased total production by 40% (2,306 kg ha1 90 d1 ) (Azim et al., 2001b). Water treatment with periphyton Some investigators have attempted to use sh and periphyton for removing nutrients from wastewater. The nutrients acccumulated in the periphyton are removed by harvesting the periphyton. This can be done manually or mechanically, but sh can also be used. Drenner et al. (1997) used Mozambique tilapia and central stoneroller (Campostoma anomalum, Cyprinidae) for harvesting periphyton grown in circular tanks fed with mock wastewater treatment plant efuent. Maximum removal rates were 48.4 mg total phosphorous (removed in sediments and sh biomass) per m2 water surface per day. In their study with grass silage, Bender et al. (1989), using colony counts of nitrogen-xing bacteria, microscopic identication of species and characterization of chemotactic response on agar plates, showed that a chemotactic response to the lactic and acetic acid in the silage from bacteria in the sediments was the rst step in the colonization of the grass substrates. The bacteria bloomed in the water and produced slimy exudates that annealed the mirobes to the silage after which cyanobacteria invaded the substrates and caused a further increase in biomass. Using microprobes, it was shown that within the mat, different heterogeneous micro-environments existed with oxic and anoxic zones. Periphyton organisms have various ways of attaching to the substrate: stalks with sticky ends (e.g. ciliates), sticky capsules (bacteria and bluegreen algae), cushions of laments (seaweeds, algae and aquatic mosses), muscular suction pads (snails), glue (barnacles) or simply clinging to the substrate (e.g., insect larvae). Attachment to sediment can be achieved by rooting (especially higher plants), rhizoids (seaweeds on corals), and with a muscular foot (clams) (Reid and Wood, 1976). During the development of the periphyton layer, conditions for growth of the various algal species change drastically. As the density of organisms increases, there is more competition for substrate surface area and this affects the composition of the periphyton community. The organisms also compete for carbon dioxide, nutrients and light. This explains the development of the periphyton layer away from the substrate, resulting in something that can be compared to the canopy of a terrestrial forest (Hoagland et al., 1982; Figure 1). Another strategy for ensuring optimal nutrient and light conditions is shown by pennate diatom and cyanobacterial cells that can move around the substrate. They glide by excreting a polysaccharide mucilage that sticks to the substrate (diatoms) or by using contractile brils in their cell walls (cyanobacteria). In this way, they can move away from areas where light or nutrients have become limiting (Horne and Goldman, 1994). It is probably also a way to escape being covered by sediment deposits (Hutchinson, 1975). Some diatoms remain at the base of the periphyton assemblage throughout its development, withstanding extremely low light conditions, while other species move around the periphyton layer looking for the best conditions available (Johnson et al., 1997).

Periphyton productivity Development of the periphyton assemblage and species composition Development of a periphyton layer on a clean surface generally starts with the deposition by electrostatic forces of a coating of dissolved organic substances (mainly mucopolysaccharides), to which bacteria are attracted by hydrophobic reactions (Hoagland et al., 1982; Cowling et al., 2000). The presence of freeoating organic micro-particles in eutrophic waters stimulates this process. Bacteria actively attach using mucilaginous strands. This can take a week, but in some studies this was observed within days and even within a matter of hours. It is not clear whether bacterial colonization is a prerequisite for subsequent attachment of other organisms, or what the exact role of the bacteria is in this process (Hoagland et al., 1982). In the days that follow, algae start to grow. Low-prole diatoms appear rst, followed by small pennate diatoms, short-stalked and longer stalked species and then by diatoms with rosettes and mucilage pads. In the nal stages of development, species of green algae with upright laments or long strands can grow (Hoagland et al., 1982; Horne and Goldman, 1994). On plants, the periphyton community is attached on gelatinous stalks of algal and bacterial mucus interspersed with deposits of calcium carbonate (Wetzel, 1975).

11

Figure 1. Range of vertical structure in the periphyton community. Drawn to scale, 400. From: Hoagland et al., 1982. Reproduced with permission, Botanical Society of America.

Coral reef algae can be classied as algal turf (110 mm), macroalgae or eshy algal pavement (larger than 10 mm) and crustose algae (encrusting noncalcied algae or calcareous, crustose corallines) (Steneck, 1988). The epilithic algal community (EAC) of coral reefs generally consists of a mixture of turf and crustose algae (Klumpp and McKinnon, 1992). Species diversity within the periphyton assemblage can be high. Planas et al. (1989) found on average 41 different species in periphyton on ceramic tiles. Wahab et al. (1999) encountered 12 genera of Bacillariophyceae, 25 of Chlorophycaea, 10 of Cyanophyceae, 4 of Eugleno-

phyceae, 1 of Rhodophyceae and 5 of zooplankton in periphyton, as well as a variety of macrobenthic organisms, notably chironomid larvae on scrap bamboo substrates in shponds in Bangladesh. Lam and Lei (1999) found 81 algal species in periphyton on glass slides in the Lam Tsuen River, Hong Kong. KonanBrou and Guiral (1994) found 24 species of algae in the periphyton community on bamboo substrates in acadjas in Cote dIvoire. Often, one or a few species of algae dominate the assemblage. The EAC of coral reefs is often dominated by lamentous Chlorophytes and Rhodophytes (Klumpp and McKinnon, 1992). On plastic substrates

12 in tilapia cages in Bangladesh, lamentous Chlorophyceae and Myxophyceae dominated the periphyton before sh stocking, whereas after stocking of the sh diatoms became more important. Diatoms could be attached directly to the substrate but were also found as epiphytes on larger lamentous algae. In addition, freshwater oligochaetes, Protozoa, Rotifera, and coelenterate Hydrozoa were observed (Huchette et al., 2000). Dominant species in the periphyton of the acadjas were the lamentous algae Rhizoclonium riparium (Chlorophyceae) and Lyngbia (Cyanobacteria) in the surface layers, and Audouinella daviesii (Rhodophyceae) in the deeper layers. Diatoms of the genera Nitzschia and Melosira grew on the lamentous algae (Guiral et al., 1993; Konan-Brou and Guiral, 1994). In eutrophic Lake Valencia, Venezuela, the basis of the periphytic detrital aggregate (PDA) growing on macrophytes (Potamogeton sp.) consisted of lamentous cyanophytes, which were also the dominant phytoplankton in the lake. Diatoms, bacteria, and amorphous detritus were the main components of the periphyton attached to the cyanophyte matrix (Bowen, 1979). The composition, biomass, and productivity of the periphyton community vary with season, year, location, and grazing pressure. On coral reefs, the structure of the community is often different depending on whether it is located on the reef ats, the inner or outer shelf, windward or leeward side, and shallow or deep parts and slopes (Klumpp and McKinnon, 1992). Within a single water body, there can be a considerable overlap in species between phytoplankton and peripyton (Havens et al., 1996). In shponds in Bangladesh, 23 of the 39 genera found in the periphyton were common to the phytoplankton (Azim et al., 2001a). The algal contribution to the dry matter can be estimated from the ratio of ash free dry matter to pigment, called the autotrophic index (AI, mg ash free dry matter/mg chlorophyll a on a given surface area; APHA, 1998). Huchette et al. (2000) reported that AIs were between 150 and 300 in ungrazed conditions and remained stable around 300 when grazed. Azim et al. (2002b) reported AI values ranging from 189 to 346 in freshwater fertilized ponds without sh, depending on substrate. The values decreased with time, indicating that ash-free dry matter of non-algal origin dominated in young periphyton. In general, 1 mg chlorophyll a is equivalent to 6585 mg algae (Dempster et al., 1993; APHA, 1998), so more than 50% of the periphyton ash-free dry matter is not of algal origin. Biomass and productivity Table 2 gives an overview of the standing crop of periphyton found on different substrates in various kinds of natural and culture systems. Because of the wide variation in methods used, environments, and species composition it is difcult to compare the biomass gures directly. The biomasses found in natural systems (streams, lakes, and coral reefs) can be greater than those found in brush-parks and culture systems (tanks and ponds). In a comparison of periphyton biomass in a wide range of natural systems, biomasses of up to 2350 mg m2 chlorophyll a were reported (Westlake et al., 1980). The main reasons for this are probably the higher sh densities and associated grazing intensity in culture systems, although biomasses in culture systems without sh were still lower than those in, for example, coral reefs. Likely, there is a strong effect of the algal species composition, with especially high biomasses attained by the eshy macroalgae that make up the algal assemblage on lightly grazed coral reefs. Furthermore, substrate type has a strong effect on the density of periphyton, as shown by the differences between different substrate types in experiments in India and Bangladesh (Azim et al., 2002b; Keshavanath et al., 2001b). Table 3 shows data on periphyton productivity. Again, comparisons are difcult, but the productivity of the acadja systems was the highest (7.9 g C m2 d1 ). In this study, not only area-specic but also chlorophyll-specic productivity was higher in periphyton than in phytoplankton (highest contrast measured on one day was 22.5 and 5.9 mg C (mg chla)1 h1 for periphyton and phytoplankton, respectively) (Guiral et al., 1993). Coral reefs are also very productive, with net productivity of up to 3 g m2 d1 (see Table 3). Productivity measurements were much lower in temperate lakes. An intermediate estimate (1.7 g C m2 d1 ) was made in a pond in Bangladesh (Azim et al., 2002b), based on the periphyton biomass after the rst two weeks of colonization on clean bamboo substrates. The other measurements were made using UV-transparent respiration chambers. Effects of environmental factors Nutrients Inorganic nutrients can have a strong effect on periphyton biomass, as shown by numerous enrichment studies in both natural and articial systems

Table 2. Periphyton biomass (standing crop) in various natural and articial systems. Biomass is expressed as g ash-free dry matter (AFDM), or as mg chlorophyll a (Chl-a) per m2 of substrate area AFDM (g m2 ) Chl-a (mg m2 ) Remarks Reference

System type

Substrate type

Stream/river 0.820 733 76.8397 0.433.1 350 550 132683 6001200 2.5154 4.1167a 125228 60300a 2100

Natural stones Cellulose-acetate Wood

048.4 7.146 1342

Glacial stream, Antarctica Amazon river, depending on zone Billabong (lentic system), on Eucalyptus camaldulensis

Izaguirre and Pizarro, 1998 Putz, 1997 Scholz and Boon, 1993 Lohman et al., 1992 Bafco and Pedrozo, 1996 Cattaneo, 1987 Meulemans and Heinis, 1983

Lake/reservoir

Sediment X-ray plates

Natural stones Dead reed stems

Nutrient-poor vs. enriched sites, Northern Ozarks, Missouri (USA) Oligotrophic lake with trout farm, Patagonia, Argentina. Higher values near sh farm 10 lakes of varying trophy, Quebec, Canada Oligo-mesotrophic lake, Netherlands. Low biomass in summer, high in winter Depending on algal type, Bonaire (Netherlands Antilles) Macroalgae, Great Barrier Reef Mainly crustose and turf algae, Great Barrier Reef. Depending on depth and location Ebri e Lagoon, Cote dIvoire Depending on depth and salinity/season, Ebri e Lagoon, Cote dIvoire 5 m2 concrete tanks, Asian Institute of Technology, Bangkok 75 m2 shponds Mymensingh, Bangladesh 25 m2 concrete tanks, Mangalore, India. Higher values without sh Biolm on biological reactors with layer of substrate, value depending on substrate

Coral reef 400700 160240b 0.238 7.225 3.39.5 19113 718 70100

Coral rock Coral rock Coral rock

Bruggeman, 1995 Hatcher and Larkum, 1983 Klumpp and McKinnon, 1992 Konan-Brou and Guiral, 1994 Ar et al., 1997 Shrestha and Knud-Hansen, 1994 Azim et al., 2002b Keshavanath et al., 2001b, 2002 Apil anez et al., 1998

Brush-park

Bamboo

Bamboo

Culture system

Plastic sheets Bamboo, kanchi, hizol Bagasse, bamboo, PVC

Water treatment system

Glass, sand disk, Active carbon

a Total pigment (chlorophyll a + pheophyton). b Converted from g carbon by assuming 48% C in afdm.

13

14
Table 3. Net productivity of periphyton in various natural and culture systems. Productivity is expressed as g C m2 d1 and was converted to these units assuming 50% C in dm and 12 hours active feeding per day System type Substrate type Net productivity Remarks (g C m2 d1 ) Amazon river, depending on zone Reference

River Lake

Cellulose-acetate 0.0821.04 0.020.20 Reed stems 0.140.72 Debris and plants 0.73

Putz, 1997

Littoral zone, 5 oligotrophic lakes, California/Nevada Loeb et al., 1983 USA Littoral zone, oligo-mesotrophic lake, Netherlands Meulemans and Heinis, 1983 Whole-lake estimate, large shallow lake Wetzel, 1964 Great Barrier Reef. Lower on slopes, higher on ats Great Barrier Reef. Damselsh territories Papua New Guinea, turf algae Virgin Islands Several Pacic benthic reef communities Fringing coral reef, Bonaire Klumpp and McKinnon, 1992 Klumpp and Polunin, 1989 Polunin, 1988 Carpenter, 1986 Marsh, 1976 Van Rooij et al., 1998 Guiral et al., 1993 Estimated from increase in total biomass on clean substrates Azim et al., 2002b Bender et al., 1989

Coral reef

Coral rock Coral rock Coral rock Coral rock Coral rock Coral rock Bamboo Bamboo Grass silage

0.61.1 1.792.15 1.982.00 1.83.1 2.164.80 0.642.00 7.9 1.7 7.5

Brush-park Fishpond Fishpond

(e.g., Aizaki and Sakamoto, 1988; Lohman et al., 1992; Ghosh and Gaur, 1994). Periphyton biomass and productivity can thus be used as indicators of eutrophication in natural waters (e.g., Mattila and Raeisaenen, 1998). In a fertilization experiment in ponds, a quadratic relationship between periphyton biomass and fertilization level was established, with a maximum periphyton biomass (mean biomass over 6 weeks) of 3.3 mg cm2 dry matter realized with fertilization rates of 4,500, 150, and 150 kg ha1 of cow manure, urea, and TSP, respectively (equivalent to 1.5 times the standard rate for shponds in Bangladesh). Phytoplankton biomass increased linearly with increasing fertilization rate up to 2 times the standard rate (Azim et al., 2001c). Investigative studies of nutrient limitation show mixed results. In most freshwater studies, phosphorous was identied as the limiting nutrient (e.g., Ghosh and Gaur, 1994; Vymazal et al., 1994), but nitrogen (Barnese and Schelske, 1994), carbon (Sherman and Fairchild, 1989) and silica can also be limiting, depending on the algal species and on other environmental factors such as hardness and acidity. High Si:P and N:P ratios favoured diatoms, and low N:P and Si:P ratios favoured cyanophytes in a reservoir in Patagonia (Bafco and Pedroso, 1996). Similarly, high Si:N or Si:P ratios favoured diatoms, low

N:P ratios favoured cyanophytes and high N:P ratios favoured chlorophytes in periphyton of the Baltic Sea (Sommer, 1996). Whether or not nutrient enrichment stimulates periphyton productivity also depends on the type of substrate. Benthic periphyton has an advantage over phytoplankton because it is closer to the nutrient-rich sediment and the interstitial pore water, or in the case of epiphytes to macrophyte nutrients. In a series of whole-lake experiments, it was shown that periphyton on sediments utilized the nutrients in the sediment pore water and therefore, responded much less to enrichment than periphyton growing on wood in the same lake (Blumenshine et al., 1997). Epilithic algae are more likely to become nutrient-limited because they have to absorb nutrients from the water (SandJensen and Borum, 1991). Lower nutrient concentrations do not necessarily mean lower biomass and productivity. In an experiment with an articially created upstream-downstream gradient, there were large differences in nutrient concentrations between upstream and downstream parts of the stream, but differences in periphyton biomass were poorly related to gradient. More nutrient recycling took place downstream (Mulholland et al., 1995). Similarly, in a modelling study of nutrient enrichment in seven New Zealand streams, biomass

15 levels in the area studied were lower than predicted using the calibrated model. This was ascribed to differences in other factors such as increased grazing, shading, or differences in substrate characteristics (Welch et al., 1992). Apart from the impact of enrichment on periphyton, periphyton has an effect on the nutrient concentration in the overlying water. Periphyton lowered the phosphorous of the overlying water (Hansson, 1989; Bratvold and Browdy, 2001) and sediment (Hansson, 1989). By lowering the nutrient concentration of the water, periphyton can affect the growth of the phytoplankton, as was shown in a study in Swedish lakes (Hansson, 1990). In aquaculture experiments with periphyton, ammonia concentrations in tanks with periphyton were lower than in control tanks, indicating a stimulating effect of the periphyton on nitrication (Langis et al., 1988; Ramesh et al., 1999; Bratvold and Browdy, 2001). Organic nutrients are also important for the heterotrophic components of the periphyton. The activity of ectoenzymes in a Mediterranean river was higher during periods of high dissolved organic matter concentrations (Roman and Sabater, 2000). Such enzymes are retained in the periphyton layer by the extracellular polysaccharide matrix (Thompson and Sinsabaugh, 2000). Similarly, the organic load caused by decomposing salmon carcasses led to increased stream periphyton growth (Fisher-Wold and Hershey, 1999). Pulses of highly available carbon created a change in the composition of the biolm from chemoautotrophic to heterotrophic organisms, and biolms adapted their metabolism to the prevailing environmental conditions (Battin et al., 1999; Butturini et al., 2000). The algae from the periphyton are important suppliers of organic matter to the heterotrophs. In river biolms, maximum enzyme activity was seen with an algal biomas that was two to three times as high as the bacterial biomass. Bacteria are likely to utilize the algal exudates and lysis products, as well as photosynthetically produced oxygen, whereas algae utilize the inorganic carbon produced by the heterotrophs (Kuehl et al., 1996; Roman and Sabater, 2000). Organic matter quality affects the rate at which it is processed, as shown by differences in turnover times between two rivers with different sources of organic matter (Roman, 2000). Dissolved organic matter may play a role in determining the structure of the periphyton. Periphyton communities treated experimentally with dissolved organic carbon contained less mucilage than untreated controls (Wetzel et al., 1997). Grazing Grazing is the most important determinant of periphyton biomass. On coral reefs, algal communities can be grazed down completely by sh or echinoids (e.g. Hixon and Brostoff, 1981; Hay, 1981). Exclusion and removal experiments on coral reefs showed that algal standing crop could increase 1.5 to 15-fold when grazers were excluded (Hatcher and Larkum, 1983). Not all components of the periphyton assemblage are equally susceptible to grazing. Diatoms belonging to the overstory of the periphyton layer were removed by grazing snails while more prostrate basal cells (e.g., Stigeoclonium sp.) were unaffected by grazing (McCormick and Stevenson, 1991; Hill et al., 1992; Steinman et al., 1992). Generally, periphyton algal diversity is lower when grazed (Jacoby, 1987; Horn, 1989; Swamikannu and Hoagland, 1989). Most studies of grazing have been done with invertebrate grazers such as snails and insect larvae while studies with sh are much less common. Huchette et al. (2000) compared the species composition of grazed (four weeks) and ungrazed periphyton communities on plastic substrates in tilapia cages in a Bangladesh river. Four weeks after stocking with sh, the lamentous algae were reduced to short colony lengths and other species such as Ankistrodesmus became more important in the periphyton. Grazing also resulted in a size reduction of epiphytic diatoms. Grazing tilapia preferred the larger-sized diatoms (Melosira spp., Cycotella spp.) as shown by a larger proportion of these species in the stomachs. The sh were not grazing on the periphyton only, as shown by a higher diversity of diatom species in the sh stomachs than in the periphyton and the presence of nanoplankton in the sh stomachs. Grazing resulted in a much lower standing biomass of periphyton than in the ungrazed treatment. Similarly, periphyton biomass was 60% lower on nets in cages stocked with Kariba tilapia (Oreochromis mortimeri, Cichlidae), redbreast tilapia (Tilapia rendalli, Cichlidae) and Nile tilapia than in unstocked cages (Norberg, 1999). However, in a study with redear sunsh (Lepomis microlophus, Centrarchidae) and snails, it was shown that the sh had a positive effect on periphyton biomass by reducing the grazing on the periphyton by snails. Nutrient concentrations in the water were also higher with sh, but this did not affect the periphyton much (McCollum et al., 1998). In streams in the Ozark

16 Mountains (Missouri, USA), stones were covered by cyanobacteria (Calothrix sp.) when exposed to grazing by sh and invertebrates. Once protected from grazing, diatoms would overgrow the cyanobacteria within four to ten days. Grazing minnows could strip the diatom layer in a matter of minutes, after which regeneration of the cyanobacteria happened in 11 days (Power et al., 1988). Predators of grazing sh can also affect the density of periphyton layers, as shown by a study in streams in Panama where bands of high periphyton biomass occurred in the top water layer (<20 cm deep) which is avoided by the sh for fear of predation (Power et al., 1989). Although grazing diminishes biomass, algal productivity is enhanced by grazing because selfshading and competition for nutrients are reduced (Hatcher, 1983; Carpenter, 1986; McCormick and Stevenson, 1989). Grazing removes dead or senescent algae and keeps the algal assemblage in a productive, early-successional stage. Coral reef algae grazed by sea urchins had a lower biomass than ungrazed algae but production was not reduced, indicating a positive effect of grazing on the productivity of the algae (Carpenter, 1986). Norberg (1999), using transparent incubation chambers, measured a fourfold increase in periphyton specic productivity in grazed periphyton compared to ungrazed controls. Apart from the direct effect of reduced competition, there is also the indirect effect of excreted nutrients that can benet the periphyton. Rudd (Scardinius erythrophthalmus, Cyprinidae) grazing on macrophytes increased the phosphorous content in water and periphyton, as shown by a 33 PO4 tracer experiment (Hansson et al., 1987). Some algae have developed defenses against grazing. Seaweeds have chemical defenses against grazing (chemical compounds that determine palatability for certain species of grazers) that may be changed by e.g. desiccation or exposure to light (Cronin and Hay, 1996). The foliaceous red alga Iridaea cordata is protected from predation by some herbivores (isopod Idotea) by its cuticle. Other herbivores (chiton, sea urchin, limpet) eat Iridaea in all its stages, and yet others (Lacuna, snail), while not deterred by the cuticle, do not eat infertile Iridaea anyway (Gaines, 1985). Light The importance of self-shading in mature perihyton assemblages has already been mentioned and illustrates the importance of light for the productivity of periphyton. Periphyton standing stock is reduced at greater depths because of reduced light incidence (Konan-Brou and Guiral, 1994; Azim et al., 2002a). Irradiance induced more differences in epilithic biomass than differences in hydrology in laboratory streams, with higher levels of irradiance leading to a higher biomass and changes in the taxonomic composition of the epilithic assemblage (DeNicola and McIntire, 1999). Light is less important for the non-algal components of the periphyton, although non-algal periphyton probably benets from organic exudates produced by the algal components (Kuehl et al., 1996; Roman and Sabater, 2000). Photoadapation was observed in coral reef algae, such that changes in irradiance were compensated by changes in the photosynthetic efciency and maximum rate of photosynthesis (Klumpp and McKinnon, 1992). Periphyton productivity was not suppressed at high light intensities, which normally inhibit phytoplankton productivity (Loeb et al., 1983). The effects of changes in irradiance on acadjas were investigated by Ar et al. (1997). Ninety-ve percent of the variation in periphyton dry matter in an acadja in the Ebri lagoon was due to three variables: photosynthetically active radiation, phytoplankton chlorophyll a, and the discharge volume of the river into the lagoon. In the rainy season, river discharge and cloud cover increase, leading to a reduced salinity in the lagoon and a decrease in radiation and photosynthesis. These environmental changes lead to a decrease in phytoplankton settling on the substrates and an overall reduction in periphyton production and biomass. Substrate type The seasonal changes of epiphyton in natural systems are affected by the seasonal changes in the vegetation upon which it grows (Horne and Goldman, 1994). Apart from aquatic vegetation, inorganic substrates such as rocks and sand are important in natural systems. In aquaculture, a variety of materials can be used: PVC pipes (Keshavanath et al., 2001b), plastic sheets (Shrestha and Knud-Hansen, 1994; Tidwell et al., 1998) and custom-designed materials such as AquamatsTM (Bratvold and Browdy, 2001). Living substrates are not common in aquaculture but occur in integrated cultures of (semi-) aquatic crops like rice with sh, where periphyton grows on plant stems (Chapman, 1991; Chapman and Fernando, 1994). Dead organic materials such as bamboo poles, tree

17 branches, jute sticks, or bundles of sugarcane bagasse are likely to become more important in pond aquaculture. In periphyton research, a variety of other materials is used including ceramic tiles, glass bre lters, and glass slides. For sampling microbial periphyton communities, and especially protozoa, polyurethane foam has been recommended (Cairns et al., 1979). Substrate type has a distinct effect on periphyton growth. A comparison of periphyton growing on natural (leaves) and articial (glass slides) substrates in a Malaysian river, totals of 37 and 35 species, respectively, were found of which 25 were common to both substrate types (Nather Khan et al., 1987). In a study in Swedish lakes, glass tubes supported greater numbers and a more diverse periphyton community than wood substrates, whereas plastic substrates supported only a layer of bacteria (Danilov and Ekelund, 2001). In studies in India and Bangladesh, bamboo resulted in greater maximum densities of periphyton than PVC pipes or sugarcane bagasse bundles (Keshavanath et al., 2001b; Azim et al., 2002b). The reasons for these differences are unknown, but they may be attributable to leaching of nutrients or toxic substances from the substrates, or differences in surface roughness. Similarly, the density of biolms of Salmonella sp. on plastic, cement, and steel surfaces was found to differ by an order of magnitude, probably due to differences in hydrophobicity of the materials (Joseph et al., 2001). Most microorganisms are hydrophilic and probably adsorb more strongly to hydrophobic surfaces (Cowling et al. 2000). Generally, plastic and PVC also performed worse in terms of sh production than bamboo or tree branches (Shrestha and Knud-Hansen, 1994; Keshavanath et al., 2001b). Interactions Grazing has an overriding effect on biomass while the effect of nutrients is not discernible or much less apparent due to the ability of the periphyton to recycle nutrients and to utilize nutrients from the substrate (Steinman et al., 1992; Hill et al., 1992; Pan and Lowe, 1994). Greater nutrient levels lead to greater grazer biomass, indicating that the nutrients are effectively passed on to higher trophic levels. Low nutrient concentrations limit periphyton biomass but when nutrient supply increases, competition for light, both within the periphyton and with other primary producers, becomes more important. In lakes, phytoplankton can contribute to shading at high nutrient concentrations (Hansson, 1992). Fish Morphological and physiological adaptations to herbivory Although animal components constitute an important part of periphyton layers, periphyton is best utilized by shes that are adapted to herbivorous diets. These adaptations reect their evolution from carnivores to herbivores. Specialization towards a plant diet required adaptation of the teeth (from grasping to cropping teeth), a grinding apparatus (a pharyngeal mill or a gizzard-like stomach), a less acidic stomach (because plant cell walls are destroyed mechanically and acidity possibly interferes with the ingestion of carbonates from coral substrates or sand), and a longer gut length to increase the residence time, thus ensuring better digestion (Horn, 1989). Fishes generally possess the enzymes to digest the contents of plant cells, but lack the enzymes capable of disrupting cell walls by digesting the beta-linked polymers, such as cellulase (Lobel, 1981). Cellulase activity in sh guts can probably be ascribed to microorganisms colonizing the sh gut contents (Stickney and Shumway, 1974). Many important pond aquaculture species possess some of these adaptations to herbivory and are probably able to utilize periphyton. However, some other species that seem to be adapted to a herbivorous diet do not utilize periphyton well (e.g., catla; Azim et al., 2002a). The reason for this is not clear, but it seems likely that they lack the ability to crop the attached vegetative part of the periphyton. Detritivorous tilapias are exceptional in the sense that they use low stomach pH (as low as 1.4) to digest cell walls (Beveridge and Baird, 2000). Periphyton ingestion by sh Most of the quantitative data on periphyton grazing is from coral reef studies and deals with species that are not of direct interest for aquaculture. For whitespotted devil (Plectroglyphidodon lacrymatus, Pomacentridae) of 62.7 mm mean standard length, a grazing rate of 304.3 g C (g sh)1 y1 was measured on shallow fringing reefs in Papua New Guinea (Polunin and Brothers, 1989). For the same species in a coastal lagoon in Papua New Guinea, mean consumption was estimated at 1.57 g dm d1 for a 14 g sh (Polunin, 1988). For another damselsh species, the Australian gregory (Stegastes apicalis, Pomacentridae) on the Great Barrier Reef, ingestion rates of 7731433 mg C sh1 d1 were measured (mean body weight 63

18 g; Klumpp and Polunin, 1989). For herbivorous reef sh, Van Rooij et al. (1998) estimated an allometric relationship between carbon intake (g d1 ) and body weight (g) from several literature sources: C intake = 0.0342W0.816 (r2 = 0.946, n = 13, W = 10100 g). In a study in a West-African lagoon, Pauly (1976) found that a 20 g blackchin tilapia consumed 1.5 g detritus dm day1 . Dempster et al. (1993) measured the ingestion of algae by Nile tilapia ngerlings (47 mm mean standard length), both of planktonic Microcystis aeruginosa and from a periphytic assemblage consisting mainly of Oscillatoria sp. Mean ingestion rate when only periphyton was offered was about 1.8 mg dm (g sh)1 h1 (measured during a 4-hour period). In a second experiment with periphyton densities ranging from 10 to 40 g m2 , ingestion rate was independent of periphyton density (mean: 3.68 mg dm g1 h1 ). In a study with indigenous carps in Bangladesh, periphyton grazing rates of 0.51.2 (kuria labeo), 0.20.79 (rohu) and 0.21.1 mg dm sh1 h1 (orange-n labeo) were measured (Rahmatullah et al., 2001). Azim et al. (2003) measured periphyton consumption in three size groups (7, 25 and 150 g) of Nile tilapia in the laboratory and found ingestion rates of 0.90, 0.18 and 0.02 mg dry matter g1 h1 , respectively. Ingestion rate of Nile tilapia increased when periphyton density on the glass plates increased. A comparison of these observations after conversion to the same unit (mg dm [g sh]1 d1 ; Table 4) shows that ingestion rates estimated for coral reef sh are generally higher than those measured in the laboratory. Information on grazing by shes can be obtained from observations of ingestion (number of bites during a dened time period) in relation to gut fullness, through exclusion experiments in the natural habitat (using cages to exclude sh from certain areas, after which grazing rates follow from biomass differences between areas with and without cages) or by measuring biomass differences on articial substrates in the laboratory before and after grazing. Advantage of the observation and exclusion methods is that the measurements are relatively unaffected by handling stress, while laboratory studies often involve short study periods that are hard to extrapolate to longer periods of time. Laboratory conditions (solitary sh, sometimes unialgal diets, various sources of stress) strongly affect the natural feeding behaviour of sh, which probably leads to underestimations of periphyton consumption with these experiments (Horn, 1989; Azim, unpublished results). Apart from sh weight, temperature is important. A linear relationship between the number of bites per hour and temperature was observed in damselsh (whitespotted devil), with the mean number of bites doubling when water temperature increased from 26 to 32 C (Polunin and Brothers, 1989). There are also differences between species. For some species, periphyton is simply not a preferred food. Yields of the indigenous kuria labeo in ponds with periphyton in Bangladesh were not different from control ponds without substrates, suggesting that this labeo did not eat the periphyton (Azim et al., 2001a). Similarly, catla were not observed to feed on periphyton growing on bamboo substrates in ponds (Azim et al., 2002a). Common carp were observed to lose weight in tanks with periphyton substrates while Nile tilapia gained weight by utilizing the periphyton (Van Dam, unpublished). Periphyton nutritional quality also plays a role in determining ingestion rates. Fish feeding on algae, macrophytes or detritus ingest larger quantities to compensate for the low energy content. To compensate for low protein content, they selectively feed on the components with the highest protein content, thus maximizing the dietary protein to metabolizable energy (P/ME) ratio (Horn, 1989; Bowen et al., 1995). Stoplight parrotshes (Sparisoma viride, Scaridae) on Carribean reefs spend 7090% of the day foraging, ingesting huge amounts (sometimes more than 60% of their own body mass per day) of lowquality food (P/ME ratios ranging from 58 mg kJ1 ) (Bruggemann, 1995). Periphyton as sh feed: proximate composition The composition of natural food for pond shes has been reviewed by Hepher (1988). Except for macrovegetation and some algae, most natural food types are rich in protein. Fluctuations in composition are mainly caused by differences in ash content. The composition of detritus is highly variable and depends a lot on the source material. Generally, periphyton composition is similar to other natural food types. One difference is the high organic matter content of the periphyton assemblage due to the mucopolysaccharide matrix. Nielsen et al. (1997) determined the composition of biolms and found that the extracellular polymeric substances accounted for 5080% of the total organic matter. Protein was the largest fraction of the extracellular substance, despite the emphasis that is ususally put on polysaccharides. Periphyton can also

19
Table 4. Comparison of periphyton ingestion rates from different experiments. Data were converted to mg dm (g sh)1 d1 Species Location Fish size (g fw) Grazing rate Source (mg dm g1 d1 ) 75 98.8a,b 24.545.5b 112.1 21.644.2e 10.8e 2.16 0.24 614.4e 2.413.2e 2.49.5e Pauly, 1976 Polunin and Brothers, 1989 Klumpp and Polunin, 1989 Polunin, 1988 Dempster et al., 1993 Azim et al., 2003 Azim et al., 2003 Azim et al., 2003 Rahmatullah et al., 2001 Rahmatullah et al., 2001 Rahmatullah et al., 2001

Sarotherodon melanothron Plectroglyphidodon lacrymatus Stegastes apicalis Plectroglyphidodon lacrymatus Oreochromis niloticus Oreochromis niloticus Oreochromis niloticus Oreochromis niloticus Labeo gonius Labeo calbasu Labeo rohita

Lagoon, Ghana 20 Shallow fringing reef, Papua New Guinea 16.9c Great Barrier Reef, Australia 63 Coastal lagoon, Papua New Guinea 14 Laboratory, periphyton on glass plates 3.7d Laboratory, periphyton on glass plates 7 Laboratory, periphyton on glass plates 25 Laboratory, periphyton on glass plates 150 Laboratory, periphyton on glass plates 10100 Laboratory, periphyton on glass plates 10100 Laboratory, periphyton on glass plates 10100

Assumptions: a Assuming 365 days in one year. b Assuming 50% C in dm. c Calculated with L-W relationship from same publication. d Calculated with L-W relationship for O. niloticus from Fishbase (Froese and Pauly 2001). e Assuming diurnal feeding (12 h day1 ).

trap exogenous organic matter from the water column, which may account for a large part of the bio-mass (Sansone et al., 1998). Organic compounds adsorb to particulate and colloidal calcium carbonate that is formed during photosynthesis (Wetzel, 1975). Table 5 compares the composition of a number of natural pond foods with that of periphyton. Characteristics of natural foods are the low dry matter content, and variable protein, energy and ash contents. Periphyton grazed from coral reefs, mixed with sediment material, had the lowest protein and energy content, whereas bacteria, zooplankton and insects had the highest nutritional quality. Generally, periphyton has a high ash content compared to other natural feeds. Partly, this is due to the high content of carbonates that are formed (Wetzel, 1975) and the entrapment of inorganic particles. We estimated the P/ME-ratios of the various food types, assuming that metabolizable energy is 60% of gross energy (see Table 5). P/ME-ratios ranged from about 6 in periphyton ingested by parrotsh and in calcareous red algae to more than 60 in bacteria, with the majority of values between 10 and 40 kJ g1 . Values estimated for periphyton grown on different substrate types in aquaculture ponds ranged from 15.6 to 25.5 kJ g1 . There seemed to be an effect of substrate type on the nutritional quality of the periphyton, with bamboo substrates giving better quality periphyton than jutestick and branches of the hizol tree (Azim et al., 2002b). P/ME ratios for natural feeds range typically range from 1 to 30 mg kJ1 (Bowen et al., 1995).

There is little information on other indicators of periphyton nutritional quality. Phillips et al. (1994) analyzed the amino acid prole of proteins from a microbial mat grown on grass silage. A comparison with the essential amino acid requirements of several sh species shows that the microbial mat was only decient in valine. Assimilation efciency and food conversion ratio Montgomery and Gerking (1980) measured assimilation efciency in cortez damselsh (Eupomacentrus rectifraenum, Pomacentridae) and giant damselsh (Microspathodon dorsalis, Pomacentridae) from Baja California, Mexico feeding on various species of marine algae. There was a large difference between the proximate composition of the periphyton mat (81.6% ash, 1.7% protein in dm) and the stomach contents of cortez damselsh (50.2% ash, 26.1% protein), which was attributed to selective feeding. The giant damselsh fed non-selectively on algal turf consisting solely of the red alga Polysiphonia. The assimilation efciency on a biomass basis was 2024% and for protein 5767%. A digestibility study with periphyton mats grown on grass silage showed a dry matter digestibility of 62% and 60%, and protein digestibility of 81% and 75%, for Nile tilapia and silver carp, respectively. Drying signicantly lowered the digestibility of the material to less than 50% on a dry matter basis (Ekpo and Bender, 1989; Bender et al., 1989). Table 6 summarizes data on assimilation efciency (AE, or digestibility) of sh feeding on periphyton and some

20

Table 5. Comparison of periphyton nutritional quality with other types of natural foods. P/ME-ratios were estimated by us assuming that metabolizable energy is 60% of gross energy for all food types Subtrate type Dry matter (%) 80.4 53.8 17.631.3 14.6 41.564.3 34.768.8 39.5 9.228.2 20.122.5 7.5 27.3 59.9 55.7 51.9 15.5 14.6 52.1 10.2 8.3 7.7 1.2 1.7 0.352.6 2230 1928 24.8 12.7 2.05.5 3.0 9.2 2.8 7.7 4.5 4.8 2.1 1.7 2.1 2.5 3.79.9 4.5 19.026.4 4.918.6 7.8 1.7 65.3 45.7 32.7 14.9 40.1 39.1 40.5 Protein (% dm) Lipids (% dm) Carbohydrates (% dm) Ash (% dm) Energy (kJ g1 ) P/ME-ratioa (mg kJ1 ) Source

Food type

Bacteria (Pseudomonas sp.) Bacteria Algae (Spirulina sp.) Algae Macrophytes Zooplankton Insects Molluscs Detritus Average natural food Green algae Brown algae Red algae Calcareous red algae Algal mat Periphyton + sediment Periphyton Periphyton Periphyton Periphyton 14.121.7 15.8 7.335.0 14.826.0 32.2 91.5 14.2

Branitic boulders Granitic boulders Granitic boulders Granitic boulders Granitic boulders Coral rock Bamboo Kanchi Hizol Jute stick

16.9 20.5 5.4 19.7 11.1 19.6 26.946.7 9.315.8 13.9 16.3 5.119.6 20.123.8 3.711.8 20.523.6 32.9 16.3 12.4 19.7 7.7 25.4 13.8 31.3 11.7 38.3 10.4 81.6 3.3 81.6 1.71 66.695.6 0.94.2 1529 1720 1429 19 41 20.4 31 13.6

12.4 11.7 12.4 6.1 6.1 6.510.4 29.132.6 23.425.4 20.3 15.5

Matty and Smith, 1978 Hepher, 1988 Matty and Smith, 1978 Hepher, 1988 Hepher, 1988 Hepher, 1988 Hepher, 1988 Hepher, 1988 Hepher, 1988 Hepher, 1988 Montgomery and Gerking, 1980 Montgomery and Gerking, 1980 Montgomery and Gerking, 1980 Montgomery and Gerking, 1980 Montgomery and Gerking, 1980 Bruggemann, 1995 Azim et al., 2002b; 2002c Azim et al., 2002b; 2002c Azim et al., 2002b Azim et al., 2002c

21
Table 6. Assimilation efciency (AE, or digestibility) in sh feeding on periphyton and other natural foods Food type Fish species Total Organic AE (%) Carbo- Lipids Source matter protein hydrate 66 6371 65 45 Targett et al., 1995 Targett et al., 1995 Targett et al., 1995 Targett et al., 1995 Appler and Jauncey, 1983 Appler, 1985 Appler, 1985 5062 Getachew, 1988 56.4 Montgomery and Gerking, 1980 46.3 Montgomery and Gerking, 1980 Ekpo and Bender, 1989 Ekpo and Bender, 1989

Thalassa testudineum Lobophora variegata Lobophora variegata Padina gymnospora Cladophora glomerata Hydrodictyon reticulatum Hydrodictyon reticulatum Bluegreen algae Marine algae Marine algae Algal mat on silage Algal mat silage

Sparisoma radians S. radians S. chrysopterum S. chrysopterum Oreochromis niloticus O. niloticus Tilapia zillii O. niloticus Eupomacentrus rectifraenum Microspathodon dorsalis O. niloticus Hypophthalmichthys molitrix

50 4049 48 25

24.2 20.1 62.1 59.7

75 66 72.3 70.9 75.7 6472 67 57.4 80.7 75.0

3450 44 37.1

other natural foods. Overall AE ranged from 20 to 50%, but protein AE was generally higher (5780%). Very little work on feed conversion efciency of periphyton into sh biomass has been done. In some studies, algae were fed to sh to determine the feed conversion ratio (FCR, reported here as g dm feed divided by g fresh sh gain). Stanley and Jones (1976) fed the cyanobacterium Spirulina platensis to bigmouth buffalo (Ictiobus cyprinellus, Catostomidae; 53.1 g initial weight, 29.1 g kg1 d1 for 28 days, 2125 C) and blue tilapia (Oreochromis aureus, Cichlidae; 23.8 g initial weight, 28.4 g kg1 d1 for 36 days), and obtained FCRs of 2.0 for both species. Grass carp fed at 2127 g kg1 d1 did not perform well with FCR values of 9.711.4. The FCR for Nile tilapia fed on a diet containing Cladophora glomera meal as the sole protein source was 2.33 (Appler and Jauncey, 1983). Appler (1985) formulated diets incorporating varyious amounts of the green lamentous alga Hydrodictyon reticulatum. The diet containing the alga as the only protein source gave FCRs of 3.6 for Nile tilapia and 3.9 for redbelly tilapia (Tilapia zillii, Cichlidae) (initial weight 1 g, 50 days culture period, 26 C, feeding level 5% body weight per day). Azim et al. (2003) fed 78 g (individual weight) Nile tilapia with periphyton grown on glass substrates in a greenhouse in The Netherlands. Despite the high ash content of the periphyton (55% on dry matter basis), the sh grew and the FCR was 2.81.

Potential sh production based on periphyton To estimate the potential sh production from periphyton-based pond aquaculture systems, we constructed a simple dynamic simulation model and incorporated values found in the literature for periphyton productivity, maximum periphyton biomass, ingestion rate and conversion efciency. The model (Figure 2) consists of two state variables, periphyton biomass (PB; g) and sh biomass (FB; g), and two rate variables, net periphyton growth rate (PBgrowth; g d1 ) and periphyton conversion rate to sh (PBconversion; g d1 ). PBgrowth is dened with the equation: PBgrowth = K PB (1 PB/PBmax ) (1)

in which K is the relative growth rate of periphyton (d1 ) and PBmax is the maximum periphyton biomass (g). Equation (1) results in a curve where the periphyton biomass increases with time to a plateau (PBmax ) where PBgrowth = 0 (see broken line a in Figure 3). This is the theoretical periphyton density where self-shading and/or nutrient limitation prevent a further increase in density, and productivity and mortality are balanced. In the model, maximum periphyton growth occurs when PB = 0.5 PBmax . PBconversion is calculated by assuming that the sh consume periphyton at a constant rate (q, in d1 ) equivalent to a percentage of their biomass: PBconversion = q FB (2)

The grazing by the sh reduces the periphyton biomass, keeping it from reaching its maximum

22

Figure 2. Relational diagram of simple simulation model for periphyton-based sh production. K and PBmax are parameters of the periphyton growth rate equation. PB is the actual periphyton biomass (g). PBdens is the periphyton density in g per m2 of substrate surface area. SAI is the surface area index (m2 of substrate area per m2 of pond area). FB is the sh biomass. PBconversion is the rate at which periphyton is consumed by sh, which is calculated as a xed percentage (q) of sh biomass. FCR is the conversion ratio of periphyton to sh biomass. FBdens is the sh biomass density in g sh per m2 pond surface area. For further explanation, see text.

biomass and maintaining its productivity. In the model, it was assumed that the periphyton had a minimum biomass (PBmin ) before sh were stocked and that sh grazing would cease whenever the biomass of the periphyton would be lower than that minimum biomass. This simple model was implemented in Stella 6.0 and run with a time-step of 0.05 day. Parameter values were derived from the present review. We assumed a maximum periphyton density of 100 g dm m2 (Table 2). PBmax was derived from this value by multiplying with the substrate area. Minimum density was assumed to be 10 g m2 . Net periphyton productivity was set at 1.5 g C m2 d1 or 3 g dm

m2 d1 (assuming 50% C in dm) (see Table 3), which converts into a value of K = 0.12 (this follows from rewriting equation (1) and substituting PB = 0.5PBmax ). Ingestion rate was assumed to be 5% of sh biomass d1 (q = 0.05; based on Bowen et al., 1995 and Van Rooij et al., 1998) and the conversion efciency of periphyton dry matter to sh biomass was assumed to be 50% (FCR = 2). Fish stocking density was assumed to be 1 m2 and initial sh weight was 5 g (resulting in an initial sh density of 5 g m2 , which is typical for pond culture). Substrate density was assumed to be 1 m2 per m2 of pond surface (in the model, this is called the surface area index or SAI).

23

Figure 3. Simulation of periphyton and sh production with 1 sh m2 (5 g individual weight) and periphyton productivity of 3 g dm m2 d1 . For model description and simulation conditions, see text. A: sh and periphyton biomass. Broken lines indicate simulated periphyton biomass without sh (a) and with higher (1.5 m2 ) sh stocking density. B: rates of periphyton production and consumption. C: ratio of sh:periphyton biomass throughout the simulation. Straight dotted lines indicate the time of maximum periphyton productivity (1) and the moment when periphyton production and consumption by sh are equal (2). For further explanation, see text.

24 The results of the simulation are shown in Figure 3. All results are reported per m2 of pond surface (note that because SAI = 1, periphyton densities per m2 substrate area are the same as per m2 pond surface area). Simulated periphyton biomass increased from 10 g m2 initially to a maximum of 90 g m2 after 52 days. Before day 52, periphyton productivity was greater than the consumption of the periphyton by sh. Maximum periphyton productivity was achieved on day 22. After day 58, sh grazing exceeded periphyton productivity as a result of the increased sh biomass (about 20 g m2 on day 58) and periphyton biomass decreased steadily until reaching a value of about 50 g m2 on day 120. The model computes a net sh production of 948 kg ha1 in 120 days, and with three crops per year the production of sh due to periphyton would be 2,844 kg ha1 y1 . The model does not account for the many interactions that would occur in a periphyton-based shpond. In reality, other food resources in the pond would contribute to sh production, and shes may switch between different food resources depending on the density of the resource and feeding preferences. The model demonstrates that the ratio of sh to periphyton density is very important. In the beginning of the culture period, sh density is low, the periphyton remains under-exploited and productivity is limited. Towards the end of the culture period, sh density is much higher which results in increased grazing pressure and lower periphyton density. The highest periphyton productivity occurred between days 20 and 60, when the ratio of sh to periphyton biomass was 0.180.24. When designing periphyton-based culture systems, this should be taken into account. Multiple cropping and partial harvesting of sh may be used to keep the sh:periphyton biomass ratio at optimal levels. and heterotrophic components of the periphyton assemblage, which probably results in less accumulation of detritus of periphyton origin on the bottom of the ponds in comparison to a phytoplanktondominated system. In addition, suspended organic material is trapped and processed by the periphyton layer. Thus, in a periphyton-based system the biomass produced remains in the aerobic layers of the pond where decomposition is faster and where it is more accessible to grazing by sh. In this way, periphyton probably contributes to making pond systems more nutrient efcient. Adding substrates to experimental ponds in Bangladesh led to an 5464% increase in nitrogen use efciency (N in sh/N in inputs) but overall nitrogen efciency remained quite low (8.7% with bamboo substrates) (Azim et al., 2002c). Several studies show a positive effect of periphyton on nitrication, leading to lower ammonia concentrations. Maximum periphyton production can be achieved at lower nutrient levels than those needed for phytoplankton. Nutrient concentrations in the water column may be poor indicators of conditions for periphyton close to the substrate, where steep nutrient gradients exist. The methodology needed to study these processes is still developing (Sand-Jensen and Borum, 1991; Bott et al., 1997). Periphyton does indeed seem to be more stable than phytoplankton, as a result of which the risk of community collapse and water quality deterioration is much smaller. However, for improved management and manipulation of periphyton layers in shponds, more knowledge about the basic processes in the periphyton assemblage is needed. Ability of sh to utilize periphyton Although not all sh species are suitable for culture in periphyton-based systems, success in such systems is not limited to specialist (macro)herbivores. More general detritus and benthos feeders can also thrive on periphyton. Some species (e.g., catla and common carp) are not able to utilize periphyton directly but may play an important role in periphyton-based polyculture systems. Not enough is known about the mechanisms for exploiting periphyton and the selection of food from periphyton assemblages by sh, or about the relationship between periphyton density and the ability of different sh species to harvest periphyton (functional response types). Experiments to investigate this are extremely difcult to design, as periphyton density is affected by environmental

Conclusions and recommendations for further research Role of periphyton in aquaculture systems Periphyton is a complex mixture of autotrophic and heterotrophic organisms and cannot simply be regarded as an attached equivalent of phytoplankton, although it certainly performs similar functions in ponds, such as oxygen production and the uptake of inorganic nutrients. There is an intense exchange of inorganic and organic nutrients between autotrophic

25

Figure 4. Number of substrate poles needed to install periphyton substrates in a 200 m2 pond at different pole diameters and SAIs.

factors and changes rapidly with time. Because of this, observation periods are short which leads to large extrapolation errors. In addition, invertebrate grazers confound the effect of sh grazing. Nevertheless, more knowledge about these basic processes is needed if progress in system design is to be made. Potential of periphyton-based sh production A considerable amount of experimental work on periphyton-based sh production systems has been done but it is difcult to compare studies because of the wide variety in culture systems, substrate types, sh species and environments used. Nevertheless, there is enough evidence that ponds with substrates can produce more sh than identical ponds with the same fertilization/feeding regime, but without additional substrates. The main factors affecting the success of periphyton-based systems are the choice of species, the choice of substrate material, the density of substrate material in relation to pond surface area (SAI), and the density of sh in relation to periphyton density. There are clear indications that biolms and periphyton develop better on natural than on articial materials, probably because of differences in the hydrophilic characteristics of these materials. Also,

the presence of organic matter is important for the initiation of bacterial processes that are the start of layer formation. Bamboo seems to be especially suitable as it not only yields more, but also higher quality periphyton. Certainly, biodegradable substrates favour the growth of bacteria and more research is needed to look into the potential of their use in shponds. On the other hand, they do pose the danger of sh kills if the decomposition processes are so rapid that they consume all the oxygen (Keshavanath et al., 2001b). More research must be devoted to the relationship between substrate area and pond surface area. In the studies we reviewed, SAI ranged from 0.02 1.54, and the few studies that investigated different substrate densities showed that increasing SAI beyond a certain value did not lead to further increases in sh production. This critical value depends strongly on the type of substrate used. On the one hand, research should focus on the possibility of increasing SAI because periphyton production rather than the ability to harvest the periphyton seems to limit sh production. Can nutrient and light limits be shifted by manipulating the numbers and diameters of substrate material used without negatively affecting the physical space requirements of the sh (see Figure 4)? Maybe other, more efcient substrate conformations can be designed.

26 The extent to which periphyton can be used in intensive aquaculture systems is not clear. It may be possible to use specially designed substrates that leach nutrients in the right mix for optimal periphyton growth. Periphyton may be combined with feeds by stocking two sh species: a high-value, carnivorous sh that utilizes the feed and another species that utilizes the periphyton. Much more work is needed to determine optimum combinations and possibly special feed formulations for periphytonbased systems. Perhaps periphyton systems can be used for a specic part of the culture cycle, e.g. in ngerling production. The positive effect of substrates on sh survival, probably related to stress reduction, points in this direction. With regard to the culture of marine sh, very little experimental work is reported. Although some initial work on periphyton was done in coastal lagoons in West Africa, most of the other work reported was done in extensive and semi-intensive freshwater ponds. Mullets (Mugilidae) are probably very suitable for growing in marine periphyton-based systems because of their feeding habits, but hardly any work with mullets and periphyton has been done so far (Eskinazi-Lea et al., 1980). Another interesting option is to culture marine shrimp in periphyton based systems. Most of the work done on the use of substrates and freshwater prawns was geared towards providing shelter and preventing territorial behaviour. It is not clear whether periphyton attached to articial substrates can be harvested by shrimp. Substrates may enhance the formation of periphyton-detritus assemblages that can be utilized by shrimp, thus preventing the accumulation of decomposing organic matter and adverse environmental conditions on the pond bottom. Experimental work in shponds shows that periphyton substrates can roughly double the yield of sh in ponds with no supplemental feeding (Azim et al., 2001a, 2002a; Keshavanath et al., 2002). Based on our simple model and the productivity of periphyton, we estimate that the potential sh yield with periphyton is around 5 tonnes ha1 y1 with the current technology. Assuming that the sh:periphyton biomass ratio would be maintained in the optimum range, further increases in production can come from increasing periphyton productivity, e.g., by manipulating nutrient levels, identifying substrate materials that facilitate periphyton growth, or increasing the SAI (provided that no nutrient or light limitation occurs). Acknowledgements The authors would like to thank J.M. van Rooij, M.A. Wahab, P. Keshavanath and D.J. Baird for their contributions to this study. This review was written with nancial support from the European Commission (Grant INCO-DC IC18-CT97-0196) and the Interdisciplinary Research and Education Fund (INREF) of Wageningen University.

References
Ahmed, K.K. and Hambrey, J.B. (1999) Brush shelter: a recently introduced shing method in the Kaptai Reservoir sheries in Bangladesh. Naga 22, 2023. Aizaki, M. and Sakamoto, K. (1988) Relationship between water quality and periphyton biomass in several streams in Japan. Proc. Int. Ass. Theor. Appl. Limnol. 23, 15111517. APHA (1998) Standard Methods for the Examination of Water and Wastewater, 20th edn. American public health Association, Washington DC. Apilnez, I., Gutirrez, A. and Daz, M. (1998) Effect of surface materials on initial biolm development. Bioresource Technology 66, 225230. Appler, H.N. (1985) Evaluation of Hydrodictyon reticulatum as protein source in feeds for Oreochromis (Tilapia) niloticus and Tilapia zillii. J. Fish Biol. 27, 327334. Appler, H.N. and Jauncey, K. (1983) The utilization of a lamentous green alga (Cladophora glomerata L.) as a protein source in pelleted feeds for Sarotherodon (Tilapia) niloticus ngerlings. Aquaculture 30, 2130. Ar, R., Bouvy, M. and Luquet, P. (1997) Effects of a seasonal salinity change on periphyton biomass in a shallow tropical lagoon. Int. Revue Ges. Hydrobiol. 82, 8193. Avnimelech, Y. and Lacher, M. (1979) A tentative nutrient balance for intensive shponds. Bamidgeh 31, 38. Azim, M.E., Verdegem, M.C.J., Rahman, M.M., Wahab, M.A., van Dam, A.A. and Beveridge, M.C.M. (2002a) Evaluation of polyculture with Indian major carps in periphyton-based pond. Aquaculture 213, 131149. Azim, M.E., Wahab, M.A., van Dam, A.A., Beveridge, M.C.M. and Verdegem, M.C.J. (2001a) The potential of periphyton-based culture of two Indian major carps, rohu Labeo rohita (Hamilton) and gonia Labeo gonius (Linnaeus). Aquacult. Res. 32, 209216. Azim, M.E., Wahab, M.A., van Dam, A.A., Beveridge, M.C.M., Huisman, E.A. and Verdegem, M.C.J. (2001b) Optimization of stocking ratios of two Indian major carps, rohu (Labeo rohita Ham.) and catla (Catla catla Ham.) in a periphyton-based aquaculture system. Aquaculture 203, 3349. Azim, M.E., Wahab, M.A., van Dam, A.A., Beveridge, M.C.M., Milstein, A., and Verdegem, M.C.J. (2001c) Optimization of fertilization rate for maximizing periphyton production on articial substrates and the implications for periphyton-based aquaculture. Aquacult. Res. 32, 749760. Azim, M.E., Wahab, M.A., Verdegem, M.C.J., van Dam, A.A., van Rooij, J.M. and Beveridge, M.C.M. (2002b) The effects of articial substrates on freshwater pond productivity and water quality and the implications for periphyton-based aquaculture. Aquat. Living Resour. 15, 231241.

27
Azim, M.E., Verdegem, M.C.J., Khatoon, H., Wahab, M.A., van Dam, A.A. and Beveridge, M.C.M. (2002c) A comparison of fertilization, feeding and three periphyton substrates for increasing sh production in freshwater pond aquaculture in Bangladesh. Aquaculture 212, 227243. Azim, M.E., Verdegem, M.C.J., Mantingh, I., van Dam, A.A. and Beveridge, M.C.M. (2003). Ingestion and utilization of periphyton grown on articial substrates by Nile tilapia Oreochromis niloticus L. Aquacult. Res. (in press). Bafco, G.D. and Pedrozo, F.L. (1996) Growth factors controlling periphyton production in a temperate reservoir in Patagonia used for sh farming. Lakes Reserv.: Res. Manage. 2, 243249. Barnese, L.E. and Schelske, C.L. (1994) Effects of nitrogen, phosphorous and carbon enrichment on planktonic and periphytic algae in a softwater, oligotrohic lake in Florida, USA. Hydrobiologia 277, 159170. Battin, T.J., Butturini, A. and Sabater, F. (1999) Immobilization and metabolism of dissolved organic carbon by natural sediment biolms in a Mediterranean and temperate stream. Aquat. Microb. Ecol. 19, 297305. Bender, J.A., Vatcharapijarn, Y. and Russell, A. (1989) Fish feeds from grass clippings. Aquacult. Eng. 8, 407419. Benitez, L.V. (1984) Milksh nutrition. In: Juario, J.V., Ferraris, R.P. and Benitez, L.V. (eds.), Advances in Milksh Biology and Culture. Proceedings of the second International Milksh Aquaculture Conference, 48 October 1983, Iloilo City, Philippines. SEAFDEC/IDRC, Manila, pp. 133143. Beveridge, M.C.M. and Baird, D.J. (2000) Diet, feeding and digestive physiology. In: Beveridge, M.C.M. and McAndrew, B.J. (eds.), Tilapias: Biology and Exploitation. Kluwer Academic Publishers, Dordrecht, pp. 5987. Biggs, B.J.F., Kilroy, C. and Lowe, R.L. (1998a) Periphyton development in three valley segments of a New Zealand grassland river: Test of a habitat matrix conceptual model within a catchment. Arch. Hydrobiol. 143, 147177. Biggs, B.J.F., Stevenson, R.J. and Lowe, R.L. (1998b) A habitat matrix conceptual model for stream periphyton. Arch. Hydrobiol. 143, 2156. Blumenshine, S.C., Vadeboncoeur, Y., Lodge, D.M., Cottingham, K.L. and Knight, S.E. (1997) Benthic-pelagic links: response of benthos to water-column nutrient enrichment. J. N. Am. Benthol. Soc. 16, 466479. Bott, T.L., Brock, J.T., Baattrup-Pedersen, A., Chambers, P.A., Dodds, W.K., Himbeault, K.T., Lawrence, J.R., Planas, D., Snyder, E. and Wolfaardt, G.M. (1997) An evaluation of techniques for measuring periphyton metabolism in chambers. Can. J. Fish. Aquat. Sci. 54, 715725. Bowen, S.H. (1979) Determinants of the chemical composition of periphytic detrital aggregate in a tropical lake (Lake Valencia, Venezuela). Arch. Hydrobiol. 87, 166177. Bowen, S.H., Lutz, E.V. and Ahlgren, M.O. (1995) Dietary protein and energy as determinants of food quality: trophic strategies compared. Ecology 76, 899907. Boyd, C. (1985) Chemical budgets for channel catsh ponds. Trans. Am. Fish. Soc. 114, 291298. Bratvold, D. and Browdy, C.L. (2001) Effect of sand sediment and vertical surfaces (AquaMats) on production, water quality and microbial ecology in an intensive Litopenaeus vannamei culture system. Aquaculture 195, 8194. Bruggemann, J.H. (1995) Parrotsh Grazing on Coral Reefs: A Trophic Novelty. PhD thesis, University of Groningen, the Netherlands, 213 pp. Butturini, A., Battin, J.T. and Sabater, F. (2000) Nitrication in stream sediment biolms: the role of ammonium concentration and DOC quality. Water Research 34, 629639. Cairns Jr., J., Kuhn, D.L. and Plafkin, J.L. (1979) Protozoan colonization of articial substrates. In: Weitzel, R.L. (ed.), Methods and Measurements of Periphyton Communities: A Review. American Society for Testing and Materials, STP 690, pp. 3457. Carpenter, R.C. (1986) Partitioning herbivory and its effects on coral reef algal communities. Ecol. Monogr. 56, 345363. Cattaneo, A. (1987) Periphyton in lakes of different trophy. Can. J. Fish. Aquat. Sci. 44, 296303. Chapman, G. (1991) The Diet and Feeding Habits of Oreochromis niloticus (Linnaeus) and Cyprinus carpio (Linnaeus) in Lowland Rice Fields in Northeast Thailand. M.Sc. thesis, University of Waterloo, 131 pp. Chapman, G. and Fernando, C.H. (1994) The diets and related aspects of feeding of Nile tilapia (Oreochromis niloticus L.) and common carp (Cyprinus carpio L.) in lowland rice elds in northeast Thailand. Aquaculture 123, 281307. Cohen, D., Raanan, Z., Rappaport, U. and Arieli, Y. (1983) The production of freshwater prawn Macrobrachium rosenbergii in Israel: improved conditions for intensive monoculture. Bamidgeh 35, 3137. Cohen, Y. (2001) Bioltration the treatment of uids by microorganisms immobilized into the lter bedding material: a review. Bioresour. Technol. 77, 257274. Costa, H.H. and Wijeyaratne, M.J.S. (1994) Utilization of mangrove species in brush-park construction and their effects on Negombo Estuary shery (Sri Lanka). J. Appl. Ichthyol. 10, 96103. Cowling, M.J., Hodgkiess, T., Parr, A.C.S., Smith, M.J. and Marrs, S.J. (2000) An alternative approach to antifouling based on analogues of natural processes. The Science of the Total Environment 258, 129137. Cronin, G. and Hay, M.E. (1996) Susceptibility to herbivores depends on recent history of both the plant and the animal. Ecology 77, 15311543. Danilov, R.A. and Ekelund, N.G.A. (2001) Comparison of usefulness of three types of articial substrata (glass, wood and plastic) when studying settlement patterns of periphyton in lakes of different trophic status. J. Microbiol. Methods 45, 167 170. Delinc G., (1992) The Ecology of the Fish Pond Ecosystem with Special Reference to Africa. Kluwer Academic Publishers, Dordrecht, the Netherlands, 230 pp. Dempster, P.W., Baird, D.J. and Beveridge, M.C.M. (1995) Can sh survive by lter feeding on microparticles? Energy balance in tilapia grazing on algal suspensions. J. Fish Biol. 47, 717. Dempster, P.W., Beveridge, M.C.M. and Baird, D.J. (1993) Herbivory in the tilapia Oreochromis niloticus: a comparison of feeding rates on phytoplankton and periphyton. J. Fish Biol. 43, 385392. DeNicola, D.M. and McIntire, C.D. (1999) Effects of substrate relief on the distribution of periphyton in laboratory streams. II. Interactions with irradiance. J. Phycol. 26, 634641. Drenner, R.W., Day, D.J., Basham, S.J., Smith, J.D. and Jensen, S.I. (1997) Ecological water treatment system for removal of phosphorous and nitrogen from polluted water. Ecol. Appl. 7, 381390. Durand, J.R. and Hem, S. (1996) The integration of extensive aquaculture (acadja-enclos) into the lagoon village environment in Cte dIvoire. ICLARM Conf. Proc. 41, 3341.

28
Edwards, P. (1993) Environmental issues in integrated agricultureaquaculture and wastewater fed sh culture systems. In: Pullin, R.S.V., Rosenthal, H. and Maclean, J.L. (eds.), Environment and Aquaculture in Developing Countries. ICLARM Conf. Proc., Vol. 31, pp. 139170. Ekpo, I. and Bender, J. (1989) Digestibility of a commercial sh feed, wet algae and dried algae by Tilapia nilotica and silver carp. Progr. Fish Cult. 51, 8386. Eskinazi-Lea, E., Da Costa Alves, M.L. and De Paiva Rocha, I. (1980) O periton e sua relao com o cultivo de peixes mugilideos. 1st Symposium Brsilien dAquaculture, Recife, July 1978, pp. 109119. FAO (2001) World review of sheries and aquaculture. In: The state of world sheries and aquaculture 1998. Available: http://www. fao.org/docrep/w9900e/w9900e00.htm [2001, April 23]. Fisher-Wold, A.K. and Hershey, A.E. (1999) Effects of salmon carcass decomposition on biolm growth and wood decomposition. Can. J. Fish. Aquat. Sci. 56, 767773. Froese, R. and Pauly, D. (eds.) (2001) FishBase. WorldWideWeb electronic publication, http://www.shbase.org [2001, October 9]. Gaines, S.D. (1985) Herbivory and between-habitat diversity: the differential effectiveness of defenses in a marine plant. Ecology 66, 473485. Getachew, T. (1988) Digestive efciency and nutrient composition gradient in the gut of Oreochromis niloticus L. in Lake Awasa, Ethiopia. J. Fish Biol. 33, 501509. Ghosh, M. and Gaur, J.P. (1994) Algal periphyton of an unshaded stream in relation to in-situ nutrient enrichment and current velocity. Aquat. Bot. 47, 185189. Gross, A., Boyd, C.E. and Wood, C.W. (1999) Ammonia volatilization from freshwater sh ponds. J. Environm. Qual. 28, 793797. Guiral, D., Ar, R., Da, K.P. and Konan-Brou, A.A. (1993) Communauts, biomasses et productions algales au sein dun recif articiel (acadja) en milieu lagunaire tropical. Rev. Hydrobiol. Trop. 26, 219228. Gupta, M.V., Sollows, J.D., Mazid, M.A., Rahman, A., Hussain, M.G. and Dey, M.M. (1998) Integrating aquaculture with rice farming in Bangladesh: feasibility and economic visibility, its adoption and impact. ICLARM Tech. Rep. 55, 90 pp. Hansson, L.A. (1989) The inuence of a periphytic biolayer on phosphorous exchange between substrate and water. Archiv. Hydrobiol. 115, 2126. Hansson, L.A. (1990) Quantifying the impact of periphytic algae on nutrient availability for phytoplankton. Freshwat. Biol. 24, 265 273. Hansson, L.A. (1992) Factors regulating periphytic algal biomass. Limnol. Oceanogr. 37, 322328. Hansson, L.A., Johansson, L. and Persson, L. (1987) Effects of sh grazing on nutrient release and succession of primary producers. Limnol. Oceanogr. 32, 723729. Hargreaves, J.A. (1998) Nitrogen biogeochemistry of aquaculture ponds. Aquaculture 166, 181212. Hatcher, B.G. (1983) Grazing in coral reef ecosystems. In: Barnes, D.J. (ed.), Perspectives on Coral Reefs. Brian Clouston Publishers, Manaka, Australia, pp. 164179. Hatcher, B.G. and Larkum, A.W.D. (1983) An experimental analysis of factors controlling the standing crop of the epilithic algal community on a coral reef. J. Exp. Mar. Biol. Ecol. 69, 6184. Havens, K.E., East, T.L., Meeker, R.H., Davis, W.P. and Steinman, A.D. (1996) Pytoplankton and periphyton responses to in situ experimental nutrient enrichment in a shallow subtropical lake. J. Plankton Res. 18, 551566. Hay, M.E. (1981) Spatial patterns of grazing intensity on a Caribbean barrier reef: herbivory and algal distribution. Aquat. Bot. 11, 97109. Hem, S. and Avit, J.L.B. (1994) First results on acadja-enclos as an extensive aquaculture system (West Africa). Bull. Mar. Sci. 55, 10381049. Hepher, B. (1988) Nutrition of Pond Fishes. Cambridge University Press, Cambridge, 388 pp. Hill, W.R., Boston, H.L. and Steinman, A.D. (1992) Grazers and nutrients simultaneously limit lotic primary productivity. Can. J. Fish. Aquat. Sci. 49, 504512. Hixon, M.A. and Brostoff, W.N. (1981) Fish Grazing and Community Structure of Hawaiian Reef Algae. Proceedings of the 4th International Coral Reef Symposium, 1981, Manila, Vol. 2, pp. 507514. Hoagland, K.D., Roemer, S.C. and Rosowski, J.R. (1982) Colonization and community structure of two periphyton assemblages, with emphasis on the diatoms (Bacillariophyceae). Am. J. Bot. 69, 188213. Horn, M.H. (1989) Biology of marine herbivorous shes. Oceanogr. Mar. Biol. Annu. Rev. 27, 167272. Horne, A.J. and Goldman, C.R. (1994) Limnology, 2nd edn. McGraw-Hill Book Company, USA. Huchette, S.M.H., Beveridge, M.C.M., Baird, D.J. and Ireland, M. (2000) The impacts of grazing by tilapias (Oreochromis niloticus L.) on periphyton communities growing on articial substrate in cages. Aquaculture 186, 4560. Huet, M. (1986) Textbook of Fish Culture, 2nd edn. Fishing News Books Ltd., Farnham, Surrey, 438 pp. Hutchinson, G.H. (1975) A Treatise on Limnology, Vol. III. Limnological Botany. John Wiley and Sons, New York. Izaguire, I. and Pizarro, H. (1998) Epilithic algae in a glacial stream at Hope Bay (Antarctica) Polar Biol. 19, 2431. Jacoby, J.M. (1987) Alterations in periphyton characteristics due to grazing in a Cascade foothill stream. Freshwat. Biol. 18, 495 508. Jimnez-Montealegre, R. (2001) Nitrogen Transformation and Fluxes in Fish Ponds: A Modelling Approach. PhD-thesis, Wageningen University, the Netherlands, 185 pp. Johnson, R.E., Tuchman, N.C. and Peterson, C.G. (1997) Changes in the vertical microdistribution of diatoms within a developing periphyton mat. J. N. Am. Benthol. Soc. 16, 503519. Joseph, B., Otta, S.K., Karunasagar, I. and Karunasagar, I. (2001) Biolm formation by Salmonella spp. on food contact surfaces and their sensitivity to sanitizers. Int. J. Food Microbiol. 64, 367 437. Kapetsky, J. (1981) Brush park sheries. In: Some Considerations for the Management of Coastal Lagoon and Estuarine Fisheries. FAO Fisheries Technical Paper 218. Food and Agriculture Organization, Rome, pp. 1828. Keshavanath, P., Gangadhar, B., Ramesh, T.J., Beveridge, M.C.M., van Dam, A.A. and Verdegem, M.C.J. (2001a) On-farm evaluation of Indian major carp production with sugarcane bagasse as substrate for periphyton. Asian Fisheries Science 14, 367376. Keshavanath, P., Gangadhar, B., Ramesh, T.J., van Dam, A.A., Beveridge, M.C.M. and Verdegem, M.C.J. (2002) The effect of periphyton and supplemental feeding on the production of the indigenous carps Tor khudree and Labeo mbriatus. Aquaculture 213, 207218. Keshavanath, P., Gangadhar, B., Ramesh, T.J., van Rooij, J.M., Beveridge, M.C.M., Baird, D.J., Verdegem, M.C.J. and Van Dam, A.A. (2001b) Use of articial substrates to enhance production of freshwater herbivorous sh in pond culture. Aquacult. Res. 32, 189197.

29
Klumpp, D.W. and McKinnon, A.D. (1992) Community structure, biomass and productivity of epilithic algal communities on the Great Barrier Reef: dynamics at different spatial scales. Mar. Ecol. Progr. Ser. 86, 7789. Klumpp, D.W. and Polunin, N.V.C. (1989) Partitioning among grazers of food resources within damselsh territories on a coral reef. J. Exp. Mar. Biol. Ecol. 125, 145169. Konan-Brou, A.A. and Guiral, D. (1994) Available algal biomass in tropical brackish water articial habitats. Aquaculture 119, 175 190. Kuehl, M., Glud, R.N., Ploug, H. and Ramsing, N.B. (1996) Microenvironmental control of photosynthesis and photosynthesis-coupled respiration in an epilithic cyanobacterial biolm. J. Phycol. 32, 799812. Lam, P.K. and Lei, A. (1999) Colonization of periphytic algae on articial substrates in a tropical stream. Diatom Res. 14, 307 322. Langis, R., Proulx, D., de la Noue, J. and Couture, P. (1988) Effects of bacterial biolms on intensive Daphnia culture. Aquacult. Eng. 7, 2138. Lobel, P.S. (1981) Trophic biology of herbivorous reef shes: alimentary pH and digestive capabilities. J. Fish Biol. 19, 365 397. Loeb, S.L., Reuter, J.E. and Goldman, C.R. (1983) Littoral zone production of oligotrophic lakes the contributions of phytoplankton and periphyton. In: Wetzel, R.G. (ed.), Periphyton of Rreshwater Ecosystems. Developments in Hydrobiology, Vol. 17. Dr W. Junk Publishers, The Hague, pp. 161167. Lohman, K., Jones, J.R. and Perkins, B.D. (1992) Effects of nutrient enrichment and ood frequency on periphyton biomass in northern Ozark streams. Can. J. Fish. Aquat. Sci. 49, 1198 1205. Longhurst, A.R. and Pauly, D. (1987) Ecology of Tropical Oceans. Academic Press, San Diego, 407 pp. Madenjian, C.P., Rogers, G.L. and Fast, A.W. (1987) Predicting nighttime dissolved oxygen loss in prawn ponds of Hawaii: part II. A new method. Aquacultural Engineering 6, 209225. Marsh, J.A. (1976) Energetic role of algae in reef ecosystems. Micronesia 12, 1321. Mattila, J. and Raesiaenen, R. (1998) Periphyton growth as an indicator of eutrophication; an experimental approach. Hydrobiologia 377, 1523. Matty, A.J. and Smith, P. (1978) Evaluation of a yeast, a bacterium and an alga as a protein source for rainbow trout. I. Effect of protein level on growth, gross conversion efciency and protein conversion efciency. Aquaculture 14, 235246. McCollum, E.W., Crowder, L.B. and McCollum, S.A. (1998) Complex interactions of sh, snails, and littoral zone periphyton. Ecology 79, 19801994. McCormick, P.V. and Stevenson, R.J. (1989) Effects of snail grazing on benthic algal community structure in different nutrient environments. J. N. Am. Benthol. Soc. 8, 162172. McCormick, P.V. and Stevenson, R.J. (1991) Grazer control of nutrient availability in the periphyton. Oecologia 86, 287291. Meulemans, J.T. and Heinis, F. (1983) Biomass and production of periphyton attached to dead reed stems in Lake Maarsseveen. In: Wetzel, R.G. (ed.), Periphyton of Freshwater Ecosystems. Dr W. Junk Publishers, The Hague, pp. 169173. Momba, M.N.B., Kr, R., Venter, S.N. and Cloete, T.E. (2000) Overview of biolm formation in distribution systems and its impact on the deterioration of water quality. Water S.A. 26, 5966. Montgomery, W.L. and Gerking, S.D. (1980) Marine macroalgae as foods for shes: an evaluation of potential food quality. Environ. Biol. Fish. 5, 143153. Mulholland, P.J., Marzolf, E.R., Hendricks, S.P. and Wilkerson, R.V. (1995) Longitudinal patterns of nutrient cycling and periphyton characteristics in streams: a test of upstreamdownstream linkage. J. N. Am. Benthol. Soc. 14, 357370. Nather Khan, I.S.A., Furtado, J.I. and Lim, R.P. (1987) Periphyton on articial and natural substrates in a tropical river. Arch. Hydrobiol. Beih. Ergebn. Limnol. 28, 473484. Nielsen, P.H., Jahn, A. and Palmgren, R. (1997) Conceptual model for production and composition of exopolymers in biolms. Water Sci. Technol. 36, 1119. Norberg, J. (1999) Periphyton fouling as a marginal energy source in tropical tilapia cage farming. Aquacult. Res. 30, 427430. Ogden, J.C. and Lobel, P.S. (1978) The role of herbivorous shes and urchins in coral reef communities. Environ. Biol. Fish. 3, 4963. OToole, G., Kaplan, H.B. and Kolter, R. (2000) Biolm formation as microbial development. Ann. Rev. Microbiol. 54, 4979. Pan, Y. and Lowe, R.L. (1994) Independent and interactive effects of nutrients and grazers on benthic algal community structure. Hydrobiologia 291, 201209. Pardue, G.B. (1973) Production response of the bluegill sunsh, Lepomis macrochirus Ranesque, to added attachment surface for shfood organisms. Trans. Am. Fish. Soc. 3, 622626. Pauly, D. (1976) The biology, shery and potential for aquaculture of Tilapia melanotheron in a small West African lagoon. Aquaculture 7, 3349. Pauly, D. and Christensen, V. (1995) Primary production required to sustain global sheries. Nature 374, 255257. Phillips, P., Russell, A., Bender, J. and Muoz, R. (1994) Management plan for utilization of a oating microbial mat with its assciated detrital gelatinous layer as a complete tilapia Oreochromis niloticus feed system. Bioresour. Technol. 47, 239245. Planas, D., Lapierre, L., Moreau, G. and Allard, M. (1989) Structural organization and species composition of a lotic periphyton community in response to experimental acidication. Can. J. Fish. Aquat. Sci. 46, 827835. Polunin, N.V.C. (1988) Efcient uptake of algal production by a single resident herbivorous sh on the reef. J. Exp. Mar. Biol. Ecol. 123, 6176. Polunin, N.V.C. and Brothers, E.B. (1989) Low efciency of dietary carbon and nitrogen conversion to growth in an herbivorous coral-reef sh in the wild. J. Fish Biol. 35, 869879. Power, M.E., Dudley, T.L. and Cooper, S.D. (1989) Grazing catsh, shing birds, and attached algae in a Panamanian stream. Environmental Biology of Fishes 26(4), 285294. Power, M.E., Stewart, A.J. and Matthews, W.J. (1988) Grazer control of algae in an Ozark mountain stream: effects of shortterm exclusion. Ecology 69(6), 18941898. Putz, R. (1997) Periphyton communities in Amazonian blackand whitewater habitats: community structure, biomass and productivity. Aquatic Science 59, 7493. Rahmatullah, S.M., Beveridge, M.C.M., van Dam, A.A., Wahab, M.A. and Baird, D.J. (2001) Functional feeding morphometry and grazing efciencies in indigenous periphyton-feeding sh species. Summary report. In: Keshavanath, P. and Wahab, M.A. (eds.), Periphyton-Based Aquaculture and Its Potential in Rural Development. Summaty of an EC-INCO-Funded Workshop, Ahsania Mission, Dhaka, Bangladesh, 2931 January 2001. Asian Fisheries Society, Indian Branch, Mangalore, p. 23.

30
Ramesh, M.R., Shankar, K.M., Mohan, C.V. and Varghese, T.J. (1999) comparison of three plant substrates for enhancing carp growth through bacterial biolm. Aquacult. Eng. 19, 119131. Reid, G.K. and Wood, R.D. (1976) Ecology of Inland Waters and Estuaries, 2nd edn. D. van Nostrand Company, New York, 584 pp. Rodeheffer, I.A. (1940) The use of brush shelters by sh in Douglas Lake, Michigan. Pap. Mich. Acad. Sci., Arts and Letters 25, 357 366. Roger, P.A. (1996) Biology and Management of the Floodwater Ecosystem in Riceelds. International Rice Research Institute, Manila, 250 pp. Roman, A.M. (2000) Characterization of extracellular enzyme kinetics in two Mediterranean streams. Arch. Hydrobiol. 148, 99117. Roman, A.M. and Sabater, S. (2000) Inuence of algal biomass on extracellular enzyme activity in river biolms. Microb. Ecol. 40, 1624. Rosan, B. and Lamont, R.J. (2000) Dental plaque formation. Microbes and Infection 2, 15991607. Rothuis, A.J., Vromant, N., Xuan, V.T., Richter, C.J.J. and Ollevier, F. (1999) The effect of rice seeding rate on rice and sh production, and weed abundance in direct-seeded rice-sh culture. Aquaculture 172, 255274. Sand-Jensen, K. and Borum, J. (1991) Interactions among phytoplankton, periphyton, and macrophytes in temperate freshwaters and estuaries. Aquat. Bot. 41, 137175. Sansone, U., Belli, M., Riccardi, M., Alonzi, A., Jeran, Z., Radojko, J., Smodis, B., Montanari, M. and Cavolo, F. (1998) Adhesion of water-borne particulates on freshwater biota. The Science of the Total Environment 219, 2128. Saravia, L.A., Momo, F. and Lissin, L.D.B. (1998) Modelling periphyton dynamics in running water. Ecol. Model. 114, 35 47. Scholz, O. and Boon, P.I. (1993) Biolm development and extracellular enzyme activities on wood in billabongs of south-eastern Australia. Freshwat. Biol. 30, 359368. Schroeder, G.L. (1978) Autotrophic and heterotrophic production of microorganisms in intensely-manured sh ponds, and related sh yields. Aquaculture 14, 303325. Schroeder, G.L., Wohlfarth, G., Alkon, A., Helevy, A. and Krueger, H. (1990) The dominance of algal-based food webs in sh ponds receiving chemical fertilizers plus organic manures. Aquaculture 86, 219229. Senanayake, F.R. (1981) The athkotu (brush-park) shery of Sri Lanka. ICLARM Newsletter 4, 2021. Shankar, K.M., Mohan, C.V. and Nandeesha, M.C. (1998) Promotion of substrate based microbial biolm in ponds a low cost technology to boost sh production. Naga 21, 1822. Sherman, J.W. and Fairchild, G.W. (1989) Algal periphyton community response to nutrient manipulation in softwater lakes. J. Phycol. 25(2 suppl.), 13. Shrestha, M.K. and Knud-Hansen, C.F. (1994) Increasing attached microorganism biomass as a management strategy for Nile tilapia (Oreochromis niloticus) production. Aquacult. Eng. 13, 101108. Smith, D.W. and Piedrahita, R.H. (1988) The relation between phytoplankton and dissolved oxygen in sh ponds. Aquaculture 68, 249265. Sommer, U. (1996) Nutrient competition experiments with periphyton from the Baltic Sea. Mar. Ecol. Progr. Ser. 140, 161167. Steinman, A.D., Mulholland, P.J. and Hill, W.R. (1992) Functional responses associated with growth form in stream algae. J. N. Am. Benthol. Soc. 11, 229243. Steneck, R.S. (1988) Herbivory on Coral Reefs: A Synthesis. Proceedings of the 6th International Coral Reef Symposium, Australia 1988, Vol. 1, pp. 3749. Stickney, R.R. and Shumway, S.E. (1974) Occurrence of cellulase activity in the stomachs of shes. J. Fish Biol. 6, 779790. Swamikannu, X. and Hoagland, K.D. (1989) Effects of snail grazing on the diversity and structure of a periphyton community in a eutrophic pond. Can. J. Fish. Aquat. Sci. 46: 16981704. Stanley, J.G. and Jones, J.J. (1976) Feeding algae to sh. Aquaculture 7, 219223. Targett, N.M., Boettcher, A.A., Targett, T.E. and Vrolijk, N.H. (1995) Tropical marine herbivore assimilation of phenolic-rich plants. Oecologia 103, 170179. Teichert-Coddington, D. and Green, B.W. (1993) Tilapia yield improvement through maintenance of minimal oxygen concentrations in experimental growout ponds in Honduras. Aquaculture 118, 6371. Thompson, A.J. and Sinsabaugh, R.L. (2000) Matric and particulate phosphatase and aminopeptidase activity in limnetic biolms. Aquat. Microb. Ecol. 21, 151159. Tidwell, J.H., Coyle, S.D. and Schulmeister, G. (1998) Effects of added substrate on the production and population characteristics of freshwater prawns Macrobrachium rosenbergii in ponds. J. World Aquacult. Soc. 29, 1722. Tidwell, J.H., Coyle, S.D., van Arnum, A. and Weibel, C. (2000) production response of freshwater prawns Macrobrachium rosenbergii to increasing amounts of articial substrate in ponds. J. World Aquacult. Soc. 31, 452458. Tidwell, J.H., Coyle, S.D., Weibel, C. and Evans, J. (1999) Effects of interactions of stocking density and added substrate on production and population structure of freshwater prawns Macrobrachium rosenbergii. J. World Aquacult. Soc. 30, 174 179. Umesh, N.R., Shankar, K.M. and Mohan, C.V. (1999) Enhancing growth of common carp, rohu and Mozambique tilapia through plant substrate: the role of bacterial biolm. Aquaculture International 7, 251260. Van Rooij, J.M., Videler, J.J. and Bruggeman, J.H. (1998) High biomass and production but low energy transfer efciency of Caribbean parrotsh: implications for trophic models of coral reefs. J. Fish Biol. 53(Supplement A), 154178. Vymazal, J., Craft, C.B. and Richardson, C.J. (1994) Periphyton response to nitrogen and phosphorous additions in Florida Everglades. Arch. Hydrobiol. Suppl. 103, 7597. Wahab, M.A. and Kibria, M.G. (1994). Katha and kua sheries unusual shing methods in Bangladesh. Aquaculture News 18, 24. Wahab, M.A., Azim, M.E., Ali, M.H., Beveridge, M.C.M. and Khan, S. (1999) The potential of periphyton-based culture of the native major carp calbaush, Labeo calbasu (Hamilton). Aquacult. Res. 30, 409419. Weinzierl, S. and Vennemann, K. (2001) The origin of acadja branches and transport routes in the Province Atlantique. Internet URL http://www.uni-hohenheim.de/atlas308/c. . .cts/c3_3/html/ english/btext_en_c3_3.htm. 25/01/2001. Weitzel, R.L. (1979) Periphyton measurements and applications. In: Weitzel, R.L. (ed.), Methods and Measurements of Periphyton Communities: A Review. American Society for Testing and Materials, STP 690, pp. 333.

31
Welch, E.B., Quinn, J.M. and Hickey, C.W. (1992) Periphyton biomass related to point-source nutrient enrichment in seven New Zealand streams. Water Research 26, 669675. Welcomme, R.L. (1972) An evaluation of the acadja method of shing as practised in the coastal lagoons of Dahomey (West Africa). J. Fish Biol. 4, 3955. Welcomme, R.L. (1985) River sheries. In: FAO Fish. Tech. Pap., Vol. 262. Food and Agriculture Organization, Rome. Westlake, D.F., Adams M.S., Bindloss, M.E., Ganf, G.G., Gerloff, G.C., Hammer, U.T., Javornicky, P., Koonce, J.F., Marker, A.F.H., McCracken, M.D., Moss, B., Nauwerck, A., Pyrina, I.L., Steel, J.A.P., Tilzer, M. and Walters, C.J. (1980) Primary production. In: LeCren, E.D. and Lowe-McConnell, R.H. (eds.), The Functioning of Freshwater Ecosystems (International Biological Programme 22). Cambridge University Press, Cambridge, 588 pp. Wetzel, R.G. (1964) A comparative study of the primary productivity of higher aquatic plants, periphyton and phytoplankton in a large, shallow lake. Int. Rev. Ges. Hydrobiol. 48, 161. Wetzel, R.G. (1975) Limnology. W.B. Saunders Comp., Philadelphia, 743 pp. Wetzel, R.G., Ward, A.K. and Stock, M. (1997) Effects of natural dissolved organic matter on mucilaginous matrices of biolm communities. Arch. Hydrobiol. 139, 289299.