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Formation of periphyton biofilm and subsequent biofouling on different substrates in nutrient enriched brackishwater shrimp ponds
Helena Khatoon, Fatimah Yusoff , Sanjoy Banerjee, Mohamed Shariff, Japar Sidik Bujang
Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Malaysia Received 17 April 2007; received in revised form 3 October 2007; accepted 8 October 2007

Abstract Periphyton grown on substrates is known to improve water quality in aquaculture ponds. Five different substrates, (i) bamboo pipe (ii) plastic sheet (iii) polyvinylchloride (PVC) pipe (iv) fibrous scrubber, and, (v) ceramic tile were evaluated for the formation of biofilm in this experiment. The substrates were suspended 25 cm below the water surface. Each type of substrate was collected fortnightly to analyze the abundance and biomass of different periphytic algae and of the biofouling organism. The study was terminated after 60 days due to severe fouling by polychaete. Results showed that pond water nutrients were high on day 60 with mean total ammonia-N, nitrite-N and soluble reactive phosphorus concentrations of 309.6 8.6 g L 1, 26.0 2.7 g L 1 and 87.2 7.1 g L 1 respectively. During the first two weeks the substrates were colonized by 19 periphytic algae. The most abundant family was Bacillariophyta (8 genera) followed by Chlorophyta (7 genera) and Cyanophyta (4 genera). Periphyton colonization on bamboo pipe showed the highest (p b 0.05) biomass in terms of chlorophyll a amongst all the substrates used. The biomass varied from 179 to 1137 g m 2 with mean values of 1137.2 0.6, 929.6 0.6, 684.2 1.2, 179.1 0.6 and 657.0 0.6 g m 2 on bamboo pipe, PVC pipe, plastic sheet, fibrous scrubber and ceramic tile respectively for the first 15 days. From 3rd week, polychaetes began to form tubes on the substrate. By day 60, the whole surface of all substrates was covered with tightly packed polychaete tubes with mean densities of 168.0 15.4, 121.0 13.5, 72.8 9.8, 72.4 7.4 and 56.0 6.8 polychaete tubes cm 2 for bamboo, PVC, plastic, fibrous scrubber and ceramic tile respectively. This study illustrated the invasive nature of attached polychaete thus hampering the formation of periphyton biofilm on substrates which could have been used for improving water quality in enriched brackishwater shrimp ponds. 2007 Elsevier B.V. All rights reserved.
Keywords: Biofouling; Substrates; Shrimp pond; Biofilm; Periphyton; Polychaete

1. Introduction Cultivation of marine shrimp has gained popularity in recent years and call for certain amount of water exchange. Water exchange mitigates eutrophicating
Corresponding author. Tel.: +60 3 8947 2111; fax: +60 3 8948 8246. E-mail address: fatimah@ibs.upm.edu.my (F. Yusoff). 0044-8486/$ - see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.aquaculture.2007.10.040

effects of feed addition and prevents accumulation of toxic metabolites (Hopkins et al., 1993). However, frequent water exchange is laborious, expensive (Thompson et al., 2002) and may increase the risk of introducing disease causing agents. In addition, the discharge of large amount of wastewater from ponds may cause eutrophication in rivers and coastal waters (Ziemann et al., 1992).

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An ecofriendly, low-cost technology whereby substrate based aquaculture using periphyton as biofilm has generated much interest in maintaining better water quality. It sequesters excess nutrients (van Dam et al., 2002) and improves health of cultured organisms (Shankar and Mohan, 2001). Periphyton growing on substrates in freshwater ponds also serves as an additional food source for cultured organisms (Azim and Wahab, 2005). Based on previous reports by several researchers who has used periphyton in improving production and water quality mostly in freshwater fish ponds (Wahab et al., 1999; Azim, 2001; van Dam et al., 2002 and Milstein, 2005), our objective was to find a suitable substrate for periphyton colonization to improve production and decrease ammonia and nitrite especially in brackishwater shrimp ponds. However, in marine aquaculture ponds, biofouling occurs especially on substrate in the pond such as paddle wheel, feeding trays, inlet pipes and jetty unlike in freshwater where it is not generally considered a problem (Billard, 1978; Beveridge, 1987). In the marine system the formation of biofilm can be a precursor to subsequent fouling by macrofoulers (Chambers et al., 2006). Biofouling in marine water generally starts when substrate such as metal, timber, stone or plastic is immersed in seawater. Microbial biofilm is formed on these substrate, followed by the growth of other organisms such as algae, marine fungi and protozoans. Afterwhich, larger marine invertebrates such as barnacles and polychaetes graze on them and make a permanent settlement on the substrates (Davis, 1995). According to Davis (1995), high accumulation of organic materials on the pond bottom makes the water environment of mariculture ponds to be nutrient enriched which is also conducive to rapid fouling development. Substrate type has a distinct effect on periphyton colonization. Substrates introduced into a body of water for biotic colonization have been used extensively to characterize algal communities inhabiting aquatic ecosystems. They have been used for studying colonization, succession, productivity dynamics and pollution assessment (Tuchman and Stevenson, 1980). In aquaculture they have been used for improving water quality and production of the cultured species and various types of materials such as halved plastic bottles, bamboo, fire wood, water hyacinth, bamboo mat, nylon netting (Huchette and Beveridge, 2005), PVC pipes (Keshavanath et al., 2001), plastic sheets (Shrestha and Knud-Hansen, 1994; Tidwell et al., 1998) and custom designed materials like Aquamats (Bratvold and Browdy, 2001) have been used as substrates. The present experiment was part of a series of experiments conducted in assessing the role of periphyton biofilm in improving water quality of brackish-

water shrimp ponds. This experiment was designed to find suitable substrates for periphyton biofilm formation. Our study reports the subsequent biofouling by polychaete on different substrates in the course of periphyton biofilm formation on these substrates in tropical marine shrimp culture ponds.
2. Materials and methods 2.1. Study area Three 0.3ha intensive shrimp (Penaeus monodon) culture ponds located at Banting, Malaysia (2 48 N and 101 24 E) were selected for this experiment. 2.2. Experimental design The shrimp growout ponds were prepared as described by Shariff et al. (2001). The ponds were filled with filtered (400 500 m mesh net) seawater and kept for one week allowing phytoplankton to grow. Triple super phosphate (TSP) was applied at the rate of 9 kg ha 1 at the beginning of the culture period to enhance algal growth. The water depth was then adjusted to 1m prior to stocking. The ponds were stocked with postlarvae (PL 43) at a stocking density of 30m 2. Shrimps were fed with commercial pellets twice a day at the rate of 610% of the body weight during the first month and four times a day at the same rate thereafter. Twenty two pieces of five different types of substrates (i) bamboo pipe (ii) plastic sheet (iii) polyvinylchloride (PVC) pipe (iv) fibrous scrubber, and, (v) ceramic tile were suspended by plastic rope 25 cm below the water surface one day before the introduction of shrimp PL in all the culture ponds. The substrates were hung at a depth of 25 cm because in our experience, the compensation depth of shrimp ponds in this area ranges from 2030 cm. To place the substrates above this level might jeopardize the experiment as the water level in the ponds fluctuates drastically from time to time when farmers flush their ponds to overcome water quality problems. Furthermore, it was observed that the periphyton grew on the pond substrates which were found deeper than ours, indicating that light in general was available below 25 cm. The substrates had a surface area of 144 cm2 each. Three pieces of each type of substrates were collected fortnightly to analyze the abundance of different periphytic algae and biomass. Biofouling organisms attached to the substrates were also determined. The experiment was originally scheduled for 90 days but was terminated after 60 days due to severe fouling of polychaete on the substrates. During the 60 days culture period 30% water in ponds was replaced by fresh seawater. 2.3. Physical and chemical factors of pond water Physical and chemical factors such as temperature, dissolved oxygen, pH and salinity were measured every 2weeks using Hydrolab Surveyor 3 (Hydrolab Corp., Austin, Texas, USA) between 09.00 and 10.00 h. Water transparency

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was determined using a Secchi disk. For laboratory analyses water sample was collected from a depth of 0.5m using a Van Dorn water sampler (Wildlife Supply Company, Saginaw, Michigan, USA) and analyzed for total ammonia nitrogen (TAN), nitrite nitrogen (NO2-N) and soluble reactive phosphorus (SRP) according to Parsons et al. (1984). 2.4. Periphyton identification and biomass At the beginning of the experiment, since algae covered 95% of the total surface area of the substrates, therefore only the identification of periphytic algae was taken into consideration at this stage. Both fresh and permanent mount slides were prepared according to Verlencar and Desai (2004) for identification using light microscope (400; Nikon Eclipse E600, Japan). In addition, scanning electron microscopy (SEM) was done for common periphyton such as Navicula, Cymbella and Amphora. Identification to genus level was done according to Ward and Whipple (1959), Prescott (1962), Belcher and Swale (1976) and Bellinger (1992). Biomass on the substrate, in terms of chlorophyll a was determined fortnightly following standard methods (APHA, 1992). The polychaete density was determined manually by counting five 1 cm2 randomly selected portion on each substrate and taking the average count. 2.5. Data analysis The data were analyzed using one-way analysis of variance (ANOVA). Significant differences amongst the different substrate were determined using Duncan Multiple Range Test at 0.05 level of probability. All statistical analyses were done using the Statistical Analysis System computer package (SAS, 9.1) (SAS, 2002). 3. Results 3.1. Water quality parameters Significant changes in water quality parameters were observed in all the experimental ponds as the culture

progressed (Table 1). Mean TAN gradually increased from 135.8 4.5 to 309.6 8.6 g L 1 (Table 1). There was a decrease of nutrients on day 45 due to exchange of pond water. Pond water nutrients were consistently high with mean TAN, NO2-N, SRP of 309.6 g L 1, 26.0 g L 1 and 87.2 g L 1 respectively on day 60. The range of salinity, dissolved oxygen and Secchi disk transparency was 26.4 3 to 31.1 0.3ppt; 4.8 0.1 to 5.6 0.2mg L 1 and 19.3 5.0 to 23.3 1.5 cm, respectively. 3.2. Biofilm and biofouling Diatoms consisting of Amphora, Navicula and Cymbella were found to be most abundant on all the substrates followed by bluegreen algae consisting of Oscillatoria while green algae were the lowest (Table 2). Zooplankters were rarely seen in the biofilm community and hence not taken into consideration for identification. Chlorophyll a on bamboo pipe was highest (1137 0.6) on day 15 and subsequently decreased to 0 0 g m 2 on day 60. Similar trend was shown by PVC, plastic sheet, fibrous scrubber and ceramic tile (Table 3). Amongst the different substrates, mean biomass on day 15 was highest on bamboo (1137 g m 2) followed by PVC (929 g m 2), plastic sheet (684 g m 2), ceramic tile (657 g m 2) and fibrous scrubber (179 g m 2) respectively. Thus, chlorophyll a showed a decreasing trend on all the substrates as the culture progressed due to biofouling with polychaetes which began to appear 3weeks after the culture period. Biofouling with polychaete was first noticed from the 3rd week and gradually increased in density occupying the whole surface area of the substrates. On day 30, bamboo pipe had the highest density of 17.6 2.7 followed by PVC pipe (14.4 1.2), plastic sheet (12.8 1.0), fibrous scrubber (12.4 1.0) and ceramic tile (11.6 0.6) polychaete tubes cm 2 respectively (Table 4). On day 60, the whole surface of the substrates was covered with densely packed multilayered polychaete tubes (Fig. 1) with densities of 168.0 15.4, 121.8 13.5, 72.8 9.8, 72.4 7.4 and 56.0 7.0 polychaete tubes cm 2 for bamboo, PVC, plastic, fibrous scrubber and ceramic tile respectively. The polychaete was identified from the Serpulidae family

Table 1 Water quality parameters in marine shrimp culture ponds Parameters Culture period 0 day Temperature (C) Transparency (cm) Salinity (ppt) pH Dissolved oxygen (mg L 1) Total ammonia-N (g L 1) Nitrite-N (g L 1) Soluble reactive P (g L 1) 30.1 0.1 20.3ab 1.8 26.4c 0.3 7.67.8 4.8b 0.0 135.8c 4.5 4.2b 1.0 6.5d 6.0
c

15 days 30.4 0.1 20.7ab 1.47 26.9c 0.4 7.47.4 4.9b 0.5 169.5bc 9.3 13.4b 0.3 48.0c 6.9
c

30 days 31.5 0.1 23.3a 1.5 30.3b 0.6 7.37.5 4.8b 0.4 245.0ab 11.7 47.6a 2.4 153.2a 9.0
b

45 days 32.5 0.1 22.7ab 1.8 30.4b 0.6 7.47.5 5.6a 0.2 65.0c 2.5 10.4b 6.8 24.6bc 8.1
a

60 days 33.3a 0.1 19.3b 5.0 31.1a 0.4 7.27.4 5.4a 0.9 309.6a 8.6 26.0ab 2.7 87.2b 7.1

Means standard errors in a row with different superscripts are significantly different at p b 0.05.

H. Khatoon et al. / Aquaculture 273 (2007) 470477 Table 2 Relative abundance (%) of different periphyton genera found in different substrates Substrates Group Bacillariophyta (diatoms) Genus Amphora Cymbella Fragilaria Gomphonema Melosira Navicula Nitzschia Pleurosigma Chaetophora Cladophora Gonatozygon Draparnaldia Oedogonium Stigeoclonium Zygnema Aphanocapsa Gomphosphaeria Gloetrichia Oscillatoria Bamboo 17.2 20.2 0 3.0 2.1 23.0 1.35 1.5 68.3 2.0 1.2 1.0 1.4 1.0 2.05 0 8.65 3.0 2.0 1.0 17.0 23.0 100 Polyvinyl chloride 16.0 18.2 2.4 3.8 2.0 22.0 1.5 2.0 67.9 2.0 2.0 0 2.4 3.2 0 0 9.6 3.5 2.0 0 17.0 22.5 100 Plastic sheet 14.0 13.2 2.8 3.3 3.1 18.0 2.0 4.5 60.9 2.0 3.0 3.0 2.0 4.0 2.1 0 16.1 5.0 3.0 0 15.0 23.0 100 Fibrous scrubber 12.0 11.0 4.0 4.0 3.0 14.0 4.6 2.3 54.9 3.0 5.0 4.0 3.1 2.0 3.0 0 20.1 4.0 3.0 2.0 16.0 25.0 100

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Ceramic tile 11.0 5.4 3.4 2.8 4.0 17.0 5.0 4.0 52.6 3.8 4.0 4.0 4.1 3.0 4.0 1.2 24.1 5.0 2.3 0 16.0 23.3 100

Subtotal Chlorophyta (green algae)

Subtotal Cyanophyta (bluegreen algae)

Subtotal Grand total

belonging to the genus Serpula (Rouse and Pleijel, 2002) (Fig. 2).

4. Discussion Water quality parameters in ponds play a critical role as they have direct impacts on the general health status of the cultured organisms. According to van Dam et al. (2002), periphyton forms a living mat on substrates and serves as natural food source for the culture organisms
Table 3 Mean values standard errors of periphyton biomass (chlorophyll a) (g m 2) on different substrates during the experimental period Substrates Culture period (days) 15 Bamboo Polyvinylchloride pipe Plastic sheet Fibrous scrubber Ceramic tile 1137.2 0.6 929.6b 0.6 684.2c 1.2 179.1e 0.6 657.0d 0.6
a

as well as improves the water quality by sequestering the excess nutrients such as dissolved reactive phosphorous, ammonia ions, nitrates in aquaculture ponds. In freshwater fish pond system, periphyton grown on substrate facilitates fish grazing and becomes a significant contributor to primary production (Milstein et al., 2003). However, many marine organisms themselves face the constant problem of being colonized and overgrown by fouling organisms. Immobile plants and animals are generally exposed to biofouling and
Table 4 Mean values standard errors of polychaete density (polychaete tubes cm 2) in different substrates at different phases of culture period Substrates Polychaete density 15 days 30 days Bamboo Polyvinylchloride pipe Plastic sheet Fibrous scrubber Ceramic tile nd nd nd nd nd 17.6 2.7 14.4b 1.2
a

30 1094.0 2.5 407.7c 0.9 579.1b 0.6 80.7e 0.7 201.3d 0.9
a

45 398.9 1.5 138.1c 0.9 217.9b 1.1 35.9d 9.5 154.2c 30.5
a

60 nd nd nd nd nd

45 days
a

60 days

42.0 4.0 168.0a 15.4 40.6a 4.5 121.0b 13.5 72.8c 9.8 72.4c 7.4 56.0c 7.0

12.8ab 1.0 36.4ab 5.0 12.4b 1.0 36.2ab 4.0 11.6b 0.6 28.0b 3.4

nd = not detected. Means in a column with different superscripts are significantly different at p b 0.05.

nd = not detected. Means in a column with different superscripts are significantly different at p b 0.05.

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and transparency) were observed in the present study as culture period progressed and nutrients were relatively high on day 44. However, all the nutrients subsequently decreased significantly on day 45 probably due to replenishing of water in the ponds. Generally, nutrient concentrations increased gradually with the progress of culture period due to excess feed and excreta from the cultured organisms. Increase in nutrients such as ammonia, nitrite and phosphorus can accumulate within the first 45 days to high levels which are beyond the recommended safe level for shrimp culture (Boyd and Fast, 1992). In the present experiment during the 1st week of the culture period, all the substrates in the marine shrimp ponds were colonized by 19 different periphytic algae, mainly diatoms (eight genera), green algae (seven genera) and bluegreen algae (four genera). The most dominant genera were Amphora, Cymbella, Gomphonema, Navicula, Nitzschia, Cladophora, Stigeoclonium, Gomphosphaeria, Pleurosigma and Oscillatoria. Similar to the result of this study, Huet (1986) reported a large number of bluegreens such as Oscillatoria, Phormidium , Spirulina , Chroococcus , Gomphosphaeria and diatoms such as Navicula, Pleurosigma, Stauroneis, Amphora, Nitzschia and Gyrosigma on artificial substrate in shallow freshwater ponds. In addition, Wahab et al. (1999) reported 53 genera of periphyton collected from scrap bamboo in freshwater fishponds in Bangladesh.

Fig. 1. Total colonization by polychaete on substrate (a) bamboo; (b) ceramic tile, forming multilayered calcareous tubes on the substrates.

consequent loss of species and community assemblages (Abdul Azis et al., 2001). In the present experiment, substrate material was selected based on its availability and low price. Studies by Danilov and Ekelund (2001); Keshavanath et al. (2001) and Azim et al. (2001) showed that glass tubes, bamboo and PVC have higher densities and more diverse periphyton community than sugarcane bagasse and wood. Moreover, survey of different shrimp farms in Malaysia showed that existing substrates in the ponds such as bamboo, PVC and plastic sheet had higher periphyton colonization (Khatoon, 2006). Gradual changes in pond water quality parameters (temperature, salinity, dissolved oxygen, conductivity

Fig. 2. Biofouling polychaete Serpula sp.

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Periphyton biomass increased upto day 15 along with gradual increase of nutrients in the present study. As a result, all the substrates were colonized by a large number of periphyton of different genera. According to Marks and Lowe (1993), periphyton bio-volume and cell densities increase significantly due to nitrogen and phosphorous enrichment. Phosphorous enrichment increased periphyton accumulation only when nitrogen levels were high indicating a significant interactive effect between nitrogen and phosphorous. In fact, inorganic nutrients can have a strong effect on periphyton biomass, as shown by numerous enrichment studies in both natural and artificial systems (Ghosh and Guar, 1994; Aizaki and Sakamoto, 1998). The highest periphyton biomass in terms of chlorophyll a (1137.2 g m 2) was found on bamboo substrate and lowest on fibrous scrubber (179.1 g m 2). The present findings are similar to that reported in freshwater ponds by Wahab and Azim (2001). They reported that highest periphyton biomass, in terms of dry matter and chlorophyll a was found in hizol (Barringtonia sp.) branches and bamboo substrates. Keshavanath et al. (2001) and Azim et al. (2001) stated from their studies in India and Bangladesh that bamboo resulted in maximum densities of periphyton than PVC pipes or sugarcane bagasse bundles. In the present study, after the 2nd week of culture, polychaete started to form tubes on all the substrates that prevented further growth of periphyton. On day 30, 70% of the periphyton coverage was replaced by the growth of calcareous tube polychaetes on all the substrates, and by day 60 the substrates were fully covered by them. The identified polycheate, Serpula sp., are sedentary tube dwelling polychaetes having worldwide distribution in marine waters associated with hard substrates. They are often found epizooically on crustacean carapaces, the shells of molluscs and on algae (Rouse and Pleijel, 2002). The colour of the calcareous tubes varied from white to orange as have been reported by Rouse and Pleijel (2002). In the present study, the polychaete was found at shallow depth which has been corroborated by Zibrowius (1977) who states that majority of Serpulidae have been described from shallow or intertidal waters. Many groups of plants and animals are recognized as foulers and the most important members of the fouling community are the diatoms, protozoa, hydrozoa, nematode, annelida and crustacea (Abdul Azis et al., 2001). According to Fusetani (2004), larvae of many benthic invertebrates attach themselves to substrates where they metamorphose into juveniles when they encounter suitable substrates. Moreover, the role of substratum is of paramount importance in the formation of primary

film and settlement of organisms and many organisms prefer slimed surfaces (Crisp and Meadows, 1963). In the present experiment, the slimy periphyton mat growing on substrates had thus provided a suitable surface for the polychaetes to thrive on. Serpula is suspension feeder and feeds suspended material from the water column. According to Matias et al. (2002), there is usually high accumulation of organic materials on the pond bottom due to uneaten feeds, feces and plankton die-offs in intensive pond culture systems. The availability of high nutrient concentrations such as nitrogen and phosphorus would accelerate phytoplankton growth rates in aquatic ecosystems, resulting in eutrophication (Yusoff et al., 2001). This condition in turn makes the environment of mariculture ponds to be nutrient-rich which is conducive to rapid fouling development (Davis, 1995). Therefore, the organically enriched pond water was probably conducive for the growth of Serpula as they could filter the particle-laden water for their food. Temperature of the water also plays a very significant role in the settlement of organisms. Being located in the tropics, the temperature of the pond water was also high (32 to 33C). According to Melo and Bott (1997), the growth and development of the biofouling organisms are dependent on the nutrient availability, water flow rate, temperature and pH. In warmer seas of the world, biofouling takes place at all times of the year, whereas in the cool waters it is more pronounced only during the warmer months (Nair and Thampy, 1980). The other factor which may have hastened the rate of polychaete proliferation is salinity. According to Beveridge (1987), the rate of fouling decreases with decreasing salinity. In the present experiment no fouling was observed in the beginning when the salinity was low (26ppt). But as salinity increased (30ppt), polychaetes started to colonize and ultimately covered the whole surface area of the substrates. Therefore, slimy surface, temperature, nutrient and salinity may have contributed to the rapid fouling by polychaetes in the present experiment. The authors have observed fouling by barnacles in shrimp ponds lined by plastic sheet. At the end of the harvest, it is a laborious work to remove the barnacles from the plastic lining. 5. Conclusion This study illustrated that periphyton biofilm showed good growth for the first 2weeks of culture period in the shrimp ponds. The best periphyton growth was found on bamboo pipe followed by PVC pipe, plastic sheet, fibrous scrubber and ceramic tile. In shrimp ponds, this periphyton biofilm can serve as feed for the

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H. Khatoon et al. / Aquaculture 273 (2007) 470477 Beveridge, M.C.M., 1987. Cage Aquaculture. Fishing News Books, Farhnam. 352 pp. Billard, R., 1978. La production de carpes en cages flottantes sur te lac Suwa au Japon. Bull. Fr. Piscic. 269, 176186. Boyd, C.E., Fast, A.W., 1992. Pond monitoring and management. In: Fast, A.W., Lester, L.J. (Eds.), Marine Shrimp Culture: Principles and Practices. Elsevier Science Publishers, Amsterdam, pp. 497513. Bratvold, D., 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. Chambers, L.D., Stokes, K.R., Walsh, F.C., Wood, R.J.K., 2006. Modern approaches to marine antifouling coatings. Surf. Coat. Technol. 201, 36423652. Crisp, D.J., Meadows, P.S., 1963. Adsorbed layers: the stimulus to settlement in barnacles. Proc. R. Soc. London Ser. B. 158, 364387. Danilov, R.A., Ekelund, N.G.A., 2001. Comparison of usefulness of three types of artificial substrata (glass, wood and plastic) when studying settlement patterns of periphyton in lakes of different trophic status. J. Microbiol. Methods 45, 167170. Davis, A., 1995. Marine Biofouling: a sticky problem. http//www. biosciences.bham.ac.uk/external/biofoulnet/WhatisBiofouling/NERC. Fusetani, N., 2004. Biofouling and antifouling. Nat. Prod. Rep. 21 (9), 410. Ghosh, M., Guar, J.P., 1994. Algal periphyton of an unshaded stream in relation to in-situ nutrient enrichment and current velocity. Aquat. Bot. 47, 185189. Hopkins, J.S., Hamilton, R.D., Sandifer, P.A., Browdy, C.L., Stokes, A.D., 1993. Effect of water exchange rate on production, water quality, effluent characteristics, and nitrogen budgets of intensive shrimp ponds. J. World Aquac. Soc. 24, 304320. Huchette, S.M.H., Beveridge, M.C.M., 2005. Periphyton-based cage aquaculture. In: Azim, M.E., Verdegem, M.C.J., van Dam, A.A., Beveridge, M.C.M. (Eds.), Periphyton: Ecology, Exploitation and Management. CABI Publishing, UK, pp. 237245. Huet, M., 1986. Textbook of Fish Culture, 2nd Edition. Fishing News Books Ltd., Farnham, Surrey. 438 pp. Keshavanath, P., Gangadhar, B., Ramesh, T.J., Beveridge, M.C.M., van Dam, A.A., Verdegem, M.C.J., 2001. On-farm evaluation of Indian major carp production with sugarcane bagasse as substrate for periphyton. Asian Fish. Sci. 14, 367376. Khatoon, H., 2006. Use of Selected Periphyton Species to Improve the Water Quality and Shrimp Postlarval Production. Ph.D. Thesis. Universiti Putra Malaysia, Malaysia, 180 pp. Marks, J.C., Lowe, R.L., 1993. Interactive effects of nutrient availability and light levels on the periphyton composition of a large oligotrophic lake. Can. J. Fish. Aquat. Sci. 50, 12701278. Matias, H.B., Yusoff, F.M., Shariff, M., Azhar, O., 2002. Effects of commercial microbial products on water quality in tropical shrimp culture ponds. Asian Fish. Sci. 15, 239248. Melo, L.F., Bott, T.R., 1997. Biofilm in water systems. Exp. Therm. Fluid Sci. 14, 375381. Milstein, A., 2005. Effect of periphyton on water quality. In: Azim, M.E., Verdegem, M.C.J., van Dam, A.A., Beveridge, M.C.M. (Eds.), Periphyton: Ecology, Exploitation and Management. CABI Publishing, UK, pp. 179190. Milstein, A., Azim, M.E., Wahab, M.A., Verdegem, M.C.J., 2003. The effects of periphyton, fish and fertilizer dose on biological processes affecting water quality in earthen fish ponds. Environ. Biol. Fish. 68, 247260. Nair, N.B., Thampy, D.M., 1980. A Textbook of Marine Ecology. The Macmillan Company of India, New Delhi. 352 pp.

invertebrates which in turn can be consumed by the benthic shrimp. In addition, substrate based periphyton system is a significant contributor to primary production by providing food for the cultured organisms and also sequesters the excess dissolved nutrients in freshwater fish ponds (Milstein et al., 2003). In the present field study, factors such as nutrient-rich environment, temperature and salinity were conducive for the fast proliferation of inedible calcareous-tubed polychaetes on substrates. This hindered the effectiveness of periphyton biofilm as an additional food source and in improving water quality in brackishwater shrimp ponds compared to freshwater ponds. Studies on the prevention of biofouling should be pursued in brackishwater shrimp ponds so that biofilms can serve effectively as source of supplementary food for the cultured organisms and maintenance of water quality. Acknowledgements This study was supported by the Malaysian Government Research Grant number IRPA-EAR-01-02-040392. Third World Organization for Women in Science (TWOWS) which supported the first author in carrying out postgraduate studies in Malaysia is greatly acknowledged. We thank Mr Perumal and Ms Hazel for their technical assistance. References
Abdul Azis, P.K., Al-Tisan, I., Sasikumar, N., 2001. Biofouling potential and environmental factors of seawater at a desalination plant intake. Desalination 135, 6982. Aizaki, M., Sakamoto, K., 1998. Relationship between water quality and periphyton biomass in several streams in Japan. Proc. Int. Assoc. Theor. Appl. Limnol. 23, 15111517. APHA, 1992. Standard Methods for the Examination of Water and Wastewater. American Public Health Association, Washington DC. 1268 pp. Azim, M.E., Wahab, M.A., 2005. Periphyton-based pond polyculture. In: Azim, M.E., Verdegem, M.C.J., van Dam, A.A., Beveridge, M.C.M. (Eds.), Periphyton: Ecology, Exploitation and Management. CABI Publishing, UK, pp. 207222. Azim, M.E., 2001. The potential of periphyton-based aquaculture production systems. PhD dissertation, Wageningen University, 219 pp. Azim, M.E., Wahab, M.A., van Dam, A.A., Beveridge, M.C.M., Verdegem, M.C.J., 2001. The potential of periphyton-based culture of two Indian major carps, rohu Labeo rohita (Hamilton) and gonia Labeo gonis (Lennaeu). Aquac. Res. 32, 209216. Belcher, H., Swale, S., 1976. A Beginner's Guide to Freshwater Algae. Institute of Terrestrial Ecology. Natural Environmental Research Council, London. 248 pp. Bellinger, E.G., 1992. A Key to Common Algae. Freshwater, Estuarine and Some Coastal Species. The Institute of Water and Environmental Management, London. 659 pp.

H. Khatoon et al. / Aquaculture 273 (2007) 470477 Parsons, T.R., Maita, Y., Laili, C.M., 1984. A Manual of Chemical and Biological Methods for Seawater Analysis. Pergamon Press, New York. 173 pp. Prescott, G.W., 1962. Algae of the Western Great Lakes Area. Wm. C. Brown Co., Iowa, USA. 977 pp. Rouse, G.W., Pleijel, F., 2002. Polychaetes. Oxford University Press, U.K.. 354 pp. SAS, 2002. Statistical Analysis System 9.1. SAS Institute Inc., Cary, NC, USA. Shankar, K.M., Mohan, C.V., 2001. The potential of biofilm in aquaculture. World Aquac. 32, 6263. Shariff, M., Yusoff, F.M., Devaraja, T.N., Srinivasa Rao, P.S., 2001. The effectiveness of a commercial microbial product in poorly prepared tiger shrimp, Penaeus monodon Fabricius ponds. Aquac. Res. 32, 181188. Shrestha, M.K., Knud-Hansen, C.F., 1994. Increasing attached microorganisms biomass as a management strategy for Nile tiapia (Oreochromis niloticus) production. Aquac. Eng. 13, 101108. Thompson, F.L., Abreu, P.C., Wasielesky, W., 2002. Importance of biofilm for water quality and nourishment in intensive shrimp culture. Aquaculture 203, 263278. Tidwell, J.H., Coyle, S.D., Schulmeister, G., 1998. Effects of added substrate on the production and population characteristics of freshwater prawns (Macrobrachium rosenbergii) to increasing amounts of artificial substrate in ponds. J. World Aquac. Soc. 31, 174179. Tuchman, M.L., Stevenson, R.J., 1980. Comparison of clay tile, sterilized rock, and natural substrate diatom communities in a small stream in southeastern Michigan, USA. Hydrobiol. 75, 7379.

477

van Dam, A.A., Beveridge, M.C.M., Azim, M.E., Verdegem, M.C.J., 2002. The potential of fish production based on periphyton. Rev. Fish. Biol. Fish. 12, 131. Verlencar, X.N., Desai, S., 2004. Phytoplankton Identification Manual. National Institute of Oceanography, Goa, India. 35 pp. Wahab, M.A., Azim, M.E., 2001. Growth and production of periphyton on substrates: experiences from Bangladesh. Summary of an EC-INCO-DC Funded Workshop Periphyton-based Aquaculture and its Potential in Rural Development, Dhaka. 58 pp. Wahab, M.A., Azim, M.E., Ali, M.H., Beveridge, M.C.M., Khan, S., 1999. The potential of periphyton-based culture of the native major carp calbaush, Labeo calbasu (Hamilton). Aquac. Res. 30, 409419. Ward, H.B., Whipple, G.C., 1959. Freshwater Biology. John Wiley and Sons Inc. New York. 423 pp. Yusoff, F.M., Matias, H.B., Khalid, Z.A., Phang, S-M., 2001. Culture of microalgae using interstitial water extracted from shrimp pond bottom sediments. Aquaculture 201, 263270. Zibrowius, H., 1977. Review of Serpulidae (Polychaeta) from depths exceeding 2000 meters. In: Reish, D.J., Fauchald, K. (Eds.), Essays on Polychaetous Annelids in Memory of Dr Olga Hartman. Allan Hancock Press, Los Angeles, USA, pp. 289306. Ziemann, D.A., Walsh, W.A., Saphore, E.G., Fulton-Bennet, K., 1992. A survey of water quality characteristics of effluent from Hawaiian aquaculture facilities. J. World Aquac. Soc. 23, 180191.