International Biodeterioration & Biodegradation 65 (2011) 253e257

Contents lists available at ScienceDirect

International Biodeterioration & Biodegradation

journal homepage: www.elsevier.com/locate/ibiod

Bioremediation of aquaculture wastewater using macroalgae and Artemia

E. Marinho-Soriano*, C.A.A. Azevedo, T.G. Trigueiro, D.C. Pereira, M.A.A. Carneiro, M.R. Camara
Universidade Federal do Rio Grande do Norte-Depto. de Oceanograa e Limnologia, Via Costeira, Praia de Me Luiza, s/n e Natal, RN 59014-100, Brazil

a r t i c l e i n f o
Article history: Received 2 September 2010 Received in revised form 30 September 2010 Accepted 3 October 2010 Available online 2 November 2010 Keywords: Bioremediation Nutrients Removal efciency Macroalgae Artemia

a b s t r a c t
Aquaculture produces a signicant amount of wastes, consisting of metabolic by-products, leading to general deterioration of water quality. An alternative for reducing nutrient excess is the use of biolters that can effectively remove nutrients from the water. A laboratory study was conducted to test the efcacy of the macroalgae Gracilaria caudata and the microcrustacean Artemia franciscana to remove nutrients from aquaculture efuents. The experiment consisted of three treatments: macroalgae (1) macroalgae and Artemia (2), and only Artemia (3). The results indicated that the treatment 2 was the most efcient in removing the nitrogenated forms. The maximum reduction values were: NH4 29.8%; NO2 100%; NO3 72.4% and DIN 44.5%. In contrast, the nutrients concentration was signicantly higher at the end of the experiment in the treatment 3 than in the other treatments (ANOVA: p < 0,001). PO4 showed a signicant increase during the experiment, especially in treatments 2 and 3 (ANOVA: p < 0.001). The results showed that G. caudata and A. franciscana performed well in absorbing nitrogenated forms. This indicates that the use of these organisms as biolters has the potential for being an ecologically correct practice and may contribute to improving the water quality of coastal regions. 2010 Elsevier Ltd. All rights reserved.

1. Introduction The eutrophication of coastal waters caused by anthropic activities has become a common problem in many countries. This process contributes to an increase in primary productivity and often generates undesirable changes in the structure and function of aquatic ecosystems. In recent decades, eutrophication caused by intensive aquaculture has become a constant concern for government authorities, owing to the large amount of residues produced by this activity. These wastes (mostly fecal material and uneaten food) increase water nutrients, in particular nitrogen and phosphorous (Beveridge 1996). One of the main environmental concerns in aquatic animal cultivation is the direct discharge of wastewaters into coastal waters. Wastewater pollutants promote organic enrichment, sedimentation and toxicity. This set of factors results in the deterioration of coastal ecosystems and consequent loss of biodiversity (Chopin et al. 2001). Although water is the major recipient of dissolved residues from aquaculture, a considerable portion of the solid material is retained inside the ponds, discharge canals or in the vicinity of the farms (Funge-Smith and Briggs 1998). Both the culture medium and coastal habitats where the activity is practiced

* Corresponding author. Tel.: 55 84 3342 4950; fax: 55 84 3342 4951. E-mail address: eliane@ufrnet.br (E. Marinho-Soriano). 0964-8305/$ e see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.ibiod.2010.10.001

show high rates of biological activity and organic matter decomposition. Recently, particular emphasis has been directed on development sustainable approaches to coastal aquaculture. In the sense, the promotion of ecological practices to improve the ecosystem health have been closely encouraged (Costa-Pierce 2002). These issues include water recycling, efuent management and biological treatment by integrated culture. The use of lter feeding organisms as nutrient (inorganic and organic) extractors has proven to be a valid alternative for nutrient bioremediation. The most frequently tested organisms are mollusks, which lter organic particles and phytoplankton and macroalgae, which absorb dissolved inorganic nutrients (Chopin et al. 2001). Recently, the inclusion of other groups of lter feeding animals belonging to the Porifera and Annellida group has been investigated (Milanese et al. 2003; Giangrande et al. 2005). The ability of macroalgae to respond to availability of anthropogenic nutrient (nitrogen and phosphorus) input, make them an efcient instrument for bioremediation (Neori et al. 2004). In addition to removing nutrients from residual wastewater, macroalgae have several applications such as the production of phycocolloids and bioactive substances. Filter feeding animals, in turn, also play a signicant role in wastewater treatment. These organisms lter organic particles, avoiding an increase in nutrients in the water column. Most of the studies published on lter feeding animals have been conducted with bivalves (Qian et al. 1996; Jones et al. 2001, Elliott et al. 2008). Studies with sponges (Stabilli et al.

254

E. Marinho-Soriano et al. / International Biodeterioration & Biodegradation 65 (2011) 253e257

2006, 2008) and polychaetes (Licciano et al. 2005) are scarce and tend to focus more specically on the clearance rate and bacterial removal from the water column. Artemia is small non-selective crustacean lter feeders that consume organic particles, microalgae and bacteria (Lenz and Browne 1991). Assimilating the particulate matter as nourishment, Artemia is capable of decreasing turbidity and removing nutrients from water. Although they occur naturally in hypersaline environments, these brine shrimp are physiologically prepared to live in a wide range of salinity (5 to 250 PSU), temperatures (5e40  C) and ionic composition (chloride, sulphate or carbonate waters) (Lenz and Browne 1991). Through its ability in ltering particles to feed it self, Artemia can help clean the water by removing phytoplankton, debris and inorganic particles. This contributes to reducing water turbidity and increasing light availability to photosynthesizing organisms. Their ltering capacity can also be used in the biomanipulation of eutrophic waters and conversion of algal blooms in animal protein (Lavens and Sorgeloos 1996). The species Artemia franciscana is found throughout the year in the state of Rio Grande do Norte, in northeast Brazil and has become an important asset for the development of the local aquaculture industry (Camara et al. 2004). The macroalga Gracilaria caudata occurs naturally along the Brazilian coast and is collected by coastal populations for agar extraction. The present study examined in laboratory-scale the bioremediation potential of the macroalga G. caudata and the microcrustacean A. franciscana in an attempt at nding mitigating solutions to the growing eutrophication of coastal waters. 2. Material and methods Specimens of G. caudata were collected at Me Luiza beach (5 470 Se35110 W) located in the northeast of Brazil. After collection, the algae were taken to the laboratory where they were washed with seawater to remove sediment and epiphytes. Algae with reproduction structures, signs of depigmentation and necrosis were discarded. After these procedures the algae were kept in an aquarium containing 50 L of seawater from the collection site, under constant aeration, light (250 mmol m2 s1) and photoperiod of 12:12 h light:dark cycle. These conditions were maintained for 7 days until the start of the experiment. Adult individuals of the species A. franciscana were collected at a saltworks in the state of Rio Grande do Norte (05 050 Se36 160 W). These organisms underwent a 7-day acclimatization process. The initial salinity of 94 PSU at the collection site was reduced to 35 PSU, equivalent to the concentration of the water used in the experiment. This water was obtained from a shrimp farm (PRIMAR) located in Rio Grande do Norte (06 110 Se35 050 W). The experimental system was composed of nine 10-liter aquariums. The experiment consisted of three treatments in triplicate. In the rst treatment (1) the macroalgae G. caudata (20 g) was used, in the second treatment (2) G. caudata (20 g) plus 100 adult individuals of A. franciscana and in the third treatment (3) only A. franciscana (100 adult individuals). The experiment lasted 72 h, with monitoring of environmental parameters (turbidity, pH, salinity, dissolved oxygen, temperature and nutrients) every 24 h. The experiment was conducted at constant temperature (28  C), salinity (35 PSU), aeration, light exposure (250 mmol m2 s1) and 12:12 h (light/dark) photoperiod. The dissolved nutrients in the water (NH4, NO2, NO3 and PO4) were analyzed according to Strickland and Parsons (1972). All data are expressed as means SD. Differences between treatments (1, 2 and 3) and time (0 h, 24 h, 48 h and 72 h) were assessed using two-way analysis of variance (ANOVA).

When differences were detected, means were compared using StudenteNewmaneKeuls test (Zar 2009). Normality and homogeneity of variances in the data before analysis were examined using ShapiroeWilk and Levene tests, respectively. The statistical tests were conducted using SigmaStat 3.1 software. 3. Results During the experiment, the environmental variables water temperature, dissolved oxygen, pH and salinity did not vary signicantly (p > 0.05). With respect to turbidity, a signicant decrease of 75%, 68% and 65% in treatments 1, 2 and 3 respectively was observed in the rst 24 hours of the experiment. After this period, the values remained relatively constant until the end of the experiment (p > 0.05). Table 1 shows the effect of G. caudata and A. franciscana on nutrient concentration in the three treatments. The concentration of NH4 varied signicantly among treatments (p < 0.001). In treatment 1 a reduction of 23% was recorded in 48 h; however, after this period a slight increase in concentration (1.35 mmol L1) was observed compared to the initial value (11.84e13.19 mmol L1). The mean NH4 value recorded for treatment 1 was 12.51 mmol L1. In treatment 2, a gradual reduction of this nutrient was observed over the study period (11.84e8.31 mmol L1), reaching an absorption efciency of approximately 30%. In treatment 3, NH4 concentration varied signicantly (p < 0.05). In this treatment, NH4 concentration showed increasing values until the end of the experiment. At the end of the experiment (72 h), NH4 was at almost double the initial value (11.84e23.62 mmol L1), corresponding to an increase of approximately 99% (Fig. 1). The concentration of NO2 showed no signicant variations among the treatments (p > 0.05). In treatment 1, NO2 uctuated during the sampling periods (p < 0.05) and a complete reduction of this nutrient was recorded after 72 h (0.03e0.0 mmol L1). In treatment 2 (0.03e0.05 mmol L1) and 3 (0.03e0.1 mmol L1), a signicant increase in NO2 was observed in the rst 24 h (p < 0.05). After this period, these values were reduced and no evidence of this nutrient was detected in the nal observation hours (Fig. 2). NO3 concentration was signicantly reduced in all the treatments (p < 0.001), but no signicant differences were found among them (p > 0.05). The concentration of this nutrient showed a sharp decrease in the rst 24 h of the experiment. After this period the values remained relatively constant (1.73 mmol L1) for the three
Table 1 Mean and standard deviation (SD) of nutrients concentration (NH4, NO2, NO3, DIN and PO4) under different treatments. Nutrients Treatment Initial 24 h 48 h 72 h Treatment Initial 24 h 48 h 72 h Treatment Initial 24 h 48 h 72 h 1 11.84 0.54 10.62 1.67 9.13 2.15 13.19 0.50 2 11.84 0.54 11.31 1.56 10.60 4.13 8.31 3.54 3 11.84 0.54 16.57 2.62 17.25 5.87 23.62 7.28 0.03 0.00 0.11 0.03 0.02 0.02 0.00 0.00 6.16 0.30 2.00 0.34 1.80 0.22 1.73 0.09 18.03 0.84 18.68 2.51 19.06 5.72 25.35 7.33 0.17 0.01 0.60 0.12 1.27 0.35 1.70 0.60 0.03 0.00 0.05 0.01 0.03 0.02 0.00 0.00 6.16 0.30 2.01 0.37 1.68 0.21 1.70 0.04 18.03 0.84 13.37 1.51 12.31 4.31 10.01 3.56 0.17 0.01 0.37 0.04 0.64 0.02 0.70 0.11 0.03 0.00 0.01 0.01 0.05 0.03 0.00 0.00 6.16 0.30 1.50 0.39 1.76 0.30 1.78 0.05 18.03 0.84 12.13 1.62 10.94 2.03 14.98 0.50 0.17 0.01 0.28 0.04 0.45 006 0.41 0.07 NH4 (mmol L1) NO2 (mmol L1) NO3 (mmol L1) DIN (mmol L1) PO4 (mmol L1)

E. Marinho-Soriano et al. / International Biodeterioration & Biodegradation 65 (2011) 253e257

255

Fig. 1. Ammonium concentration (mmol L1) during the experimental period. Vertical bars represent SD for the means (n 3).

Fig. 3. Nitrate concentration (mmol L1) during the experimental period. Vertical bars represent SD for the means (n 3).

treatments until the end of the experiment (72 h). The removal percentage was greater than 70% in the three treatments tested (Fig. 3). Analysis of dissolved inorganic nitrogen (DIN), which represents the sum of NH4 NO3 NO2, showed signicant differences among the three treatments (p < 0.001). In treatment 1 there was a considerable decrease in DIN after 48 h, with a removal efciency of around 40%. After this period an increased concentration (w4 mmol L1) was observed, likely as a result of the increase in NH4 concentration. The DIN removal efciency of G. caudata in treatment 1 was 17%. In treatment 2 (G. caudata Artemia), there was a marked reduction of this nutrient until the end of the experiment (w8 mmol L1), resulting in an absorption efciency of 44.5% (Fig. 4). In contrast, in treatment 3, DIN concentration increased over time, reaching values above 25 mmol L1. In regard to PO4, signicant variations were found among the treatments (p < 0.01), as well as among the sampling periods (p < 0.001). PO4 showed a signicant increase over the course of the three treatments. However, it should be noted that the values obtained in treatment 1 (0.17e0.41 mmol L1) and 2 (0.17e0.70 mmol L1), in which the macroalga G. caudata was used, were lower than those found in treatment 3 (0.17e1.70 mmol L1) (Fig. 5).

4. Discussion The use of biolters in integrated aquaculture systems has been investigated to reduce the concentration of dissolved nutrients and particulates in the water column and consequent decrease in

eutrophication (Neori et al. 2004). The employ of lter feeding animals (organic extractors) in bioremediation studies has several advantages. First, because they consume organic particulates, thereby avoiding future release of nutrients from bacterial degradation of organic matter and second, because they feed on phytoplankton and bacteria, which use inorganic nutrients for their growth (Qian et al. 2001). Macroalgae as inorganic extractors remove the nutrients after the organic matter is mineralized by the bacteria. Thus, the ltration capacity of organisms may play an important role in the equilibrium of aquatic ecosystems, such as reducing phytoplankton populations, increasing water transparency and decreasing the nutrient load in the water column. Among the forms of dissolved nitrogen, ammonium is considered one of the excretion products of marine invertebrates (Bracken 2007). Our results also showed a signicant effect of A. franciscana on the concentration of NH4 in the treatments used. Indeed, NH4 levels increased considerably in the treatment where there was only Artemia (treatment 3) to nearly 100% higher than the initial concentration (11.84e23.62 mmol L1). This excessive increase in NH4 during the experiment was attributed to the metabolic residues of animals, considering that these lter feeding organisms are ammoniotelic arthropods that excrete part of their nal nitrogenated products under the form of NH3 and NH4 (Freire et al. 2008). The highest removal efciency of NH4 was in treatment 2 (29.8%), which was composed of A. franciscana and G. caudata. This suggests that particulate nitrogen was removed by the ltration activity of Artemia, whereas dissolved inorganic nitrogen was removed by G. caudata. Furthermore, the fact that the NH4 concentration in treatment 1, which contains only G. caudata, was

Fig. 2. Nitrite concentration (mmol L1) during the experimental period. Vertical bars represent SD for the means (n 3).

Fig. 4. Dissolved inorganic nitrogen concentration (mmol L1) during the experimental period. Vertical bars represent SD for the means (n 3).

256

E. Marinho-Soriano et al. / International Biodeterioration & Biodegradation 65 (2011) 253e257

and presumed that this increase was associated to the regeneration of this nutrient in the system by the mussel. Likewise, Jones et al. (2001) used oysters and macroalgae in the treatment of wastewater from shrimp ponds observed that PO4 concentration was higher in the treatments involving only oysters, a nding attributed to the excretion of these bivalves. Earlier studies showed that marine bivalves produce wastes as a result of incomplete digestion and metabolism. Thus, several forms of carbon, nitrogen and phosphorous may be found in organism excretions (Dame 1996). In the present study, we observed that in treatments 1 and 2, which contained G. caudata, PO4 concentration was lower than in the treatment that used only A. franciscana. This suggests that macroalgae contributed to the PO4 reduction in the water column. 5. Conclusion This study demonstrated that the macroalga G. caudata and the crustacean A. franciscana are potential candidates for nitrogen bioremediation. This ability was conrmed by the signicantly decreases of nitrogen in treatment 2 (NH4 29.8%; NO2 100%; NO3 72.4% and DIN 44.5%), which encourage the employment of these organisms as biolters. In conclusion, this study conrms the bioltration capacity of G. caudata (Marinho-Soriano et al. 2009) and points to A. franciscana as a new candidate to be integrated into bioremediation processes. The interaction of these two organisms represents an efcient and environmentally valid alternative for improving the environmental conditions of coastal areas susceptible to discharges of aquaculture wastewaters. Acknowledgements This study was supported by the Ministrio da Cincia e Tecnologia (MCT) and Conselho Nacional de Desenvolvimento Cientco e Tecnolgico (CNPq). We also thank two reviewers for their comments. References
Beveridge, M.C.M., 1996. Cage aquaculture, 2nd ed. Fishing News Books, Blackwell Science Ltd, Oxford, UK. Bracken, M.E.S., 2007. Excretion. In: Denny, M.W., Gaines, S.D. (Eds.), Encyclopedia of tidepools and rocky shores. University of California Press, California, USA, pp. 215e217. Camara, M.R., Monteiro, P.A., Reis, L.G., Costa, M.F., 2004. Farming Artemia in a multi-cycle culture system in Northeastern Brazil. World Aquaculture 35, 40e42. Chopin, T., Buschmann, A.H., Troell, M., Kautsky, N., Neori, A., Yarish, C., Neefs, C., 2001. Integrating seaweeds into marine aquaculture systems: a key toward sustainability. Journal of Phycology 37, 975e986. Costa-Pierce, B.A., 2002. Ecological aquaculture: the evolution of the blue revolution. Blackwell Science, Oxford, UK. Dame, R.F., 1996. Ecology of marine bivalves: an ecosystem approach. CRC Press, Boca Raton, New York. Deviller, G., Aliaume, C., Franco-Nava, M.A., Casellas, C., Blancheton, J.P., 2004. High rate algal pond treatment for water reuse in an integrated marine sh recirculating system: effect on water quality and seabass growth. Aquaculture 235, 331e344. Elliott, P., Aldridge, D.C., Moggridge, G.D., 2008. Zebra mussel ltration and its potential uses in industrial water treatment. Water Research 42, 1664e1674. Freire, C.A., Onken, H., McNamara, J.C., 2008. A structureefunction analysis of ion transport in crustacean gills and excretory organs. Comparative Biochemistry and Physiology 151, 272e304. Funge-Smith, S.J., Briggs, M.R.P., 1998. Nutrient budgets in intensive shrimp ponds: Implications for sustainability. Aquaculture 164, 117e133. Giangrande, A., Cavallo, A., Licciano, M., Mola, E., Pierri, C., Trianni, L., 2005. Utilization of the lter feeder Sabella spallanzani as bioremediator in aquaculture. Aquaculture International 13, 129e136. Hernndez, I., Fernndez-Engo, M., Prez-Llorns, J., Vergara, J., 2005. Integrated outdoor culture of two estuarine macroalgae as biolters for dissolved nutrients from Sparus auratus waste waters. Journal of Applied Phycoly 17, 557e567. Hwang, S.J., Kim, H.S., Shin, J.K., Oh, J.M., Kong, D.S., 2004. Grazing effects of a freshwater bivalve (Corbicula leana Prime) and large zooplankton on phytoplankton communities in two Korean lakes. Hydrobiologia 515, 161e179.

Fig. 5. Orthophosphate concentration (mmol L1) during the experimental period. Vertical bars represent SD for the means (n 3).

inferior to treatment 2 demonstrates that Artemia effectively removes the particulate nitrogen from the water. The experiment also showed high removal efciency for nitrite and nitrate (Figs. 2 and 3). In the treatment where there was only G. caudata, there was a considerable reduction in NO2 after 24 h (w57%), while in the treatments 2 and 3, an increase in NO2 was recorded in the rst 24 h. At the end of the experiments (72 h), NO2 was totally removed from the water column in the three treatments. In an integrated system, the reduction in NO2 is important because the accumulation of this nutrient in water may be toxic to aquatic organisms. The removal of NO3 in the three treatments was approximately 70% and was generally higher than the values previously reported in literature (see Neori et al. 2004). Dissolved integrated nitrogen (DIN) consists mainly of ammonium (NH4) and oxidized forms (NO2 and NO3). In this study, NH4 was the predominant form of nitrogen, followed by NO3 and NO2. Treatment 2 was also quite efcient in removing DIN (18.03e10.01 mmol L1), as evidenced by the high removal rate (44.5%). Treatment 1 showed removal capacity of 17%, whereas in experiment 3 DIN concentration increased considerably (18.03e25.35 mmol L1) after 72 hours (40%). The comparison among the three treatments showed higher removal capacity for treatment 2 (G. caudata Artemia). In general, the removal efciency of DIN (44.5%) in this treatment was higher than the values described in the literature. For example, Hernandez et al. (2005) showed that the macroalga Gracilariopsis longissima cultivated with the sh Sparus aurata, removed only 17% of DIN. These values were similar to those found in the present study for treatment 1, where there was only G. caudata. In another study, Deviller et al. (2004), using Ulva, Enteromorpha and Cladophora, obtained a removal rate of 25% for this nutrient. Filter feeding organisms excrete NH4 and PO4 (Lauritsen and Mozley 1989), which are inorganic nutrients readily available to inorganic extracting species such as macroalgae. In the present study, it is likely that the metabolism of Artemia along with the solid matter in suspension accounted for the increase in PO4 concentration during the experiment. Indeed, the increase in PO4 concentration in treatments 2 and 3 may have occurred due to the rapid release of this nutrient from excrements of Artemia during the experimental period. Motohashi and Matsudaira (1969), showed that during excessive grazing of zooplankton PO4 tends to be released more rapidly into the medium. This occurs because the fecal pellets produced by the herbivorous plankton contain organic phosphorous in a labile state which are transformed more rapidly by phosphatases. Hwang et al. (2004), studying the mussel Corbicula leana, observed that the concentration of phosphate increased between 30 and 50% compared to the control treatment

E. Marinho-Soriano et al. / International Biodeterioration & Biodegradation 65 (2011) 253e257 Jones, A.B., Dennison, W.C., Preston, N.P., 2001. Integrated of shrimp efuent by sedimentation, oyster ltration and macroalgas absorption: a laboratory scale study. Aquaculture 193, 155e178. Lauritsen, D.D., Mozley, S.C., 1989. Nutrient excretion by the Asiatic Clam Corbicula uminea. The North American Benthological Society 8, 132e139. Lavens, P., Sorgeloos, P., 1996. Manual on the production and use of live food for aquaculture. FAO Fisheries Technical Paper No. 361, 295 p. Licciano, M., Stabilli, L., Giangrande, A., 2005. Clearance rates of Sabella spallanzanii and Branchiomma luctuosum (Annelida: Polychaeta) on a pure culture of Vibrio alginolyticus. Water Research 39, 4375e4384. Lenz, P.H., Browne, R.A., 1991. Ecology of artemia. In: Browne, R.A., Sorgeloos, P., Trotman, C.N.A. (Eds.), Artemia biology. CRC Press, Boca Raton, Florida, USA, pp. 237e253. Marinho-Soriano, E., Panucci, R.A., Carneiro, M.A.A., Pereira, D.C., 2009. Evaluation of Gracilaria caudata J. Agardh for bioremediation of nutrients from shrimp farming wastewater. Bioresource Technology 100, 6192e6198. Milanese, M.E., Chelossi, E., Manconi, R., Sara, A., Sidri, M., Pronzato, R., 2003. The marine sponge Chondilla nucula Schmidt, 1862 as an elective candidate for bioremediation in integrated aquaculture. Biomolecular Engineering 20, 363e368. Motohashi, K., Matsudaira, C., 1969. Some problems relating to phosphate regeneration and oxygen consummation in seawater e Phosphate regenerated by

257

zooplankton during excessive grazing. Tohoku Journal of Agricultural Research 20, 81e88. Neori, A., Chopin, T., Troell, M., Buschmann, A.H., Kraemer, G.P., Halling, G., Shpigel, M., Yarish, C., 2004. Integrated aquaculture: rationale, evolution and state of the art emphasizing seaweed bioltration in modern aquaculture. Aquaculture 231, 362e391. Qian, P.Y., Wu, C.Y., Wu, M., Xie, Y.K., 1996. Integrated cultivation of the red alga Kappaphycus alvarezii and the pearl oyster Pinctada martensi. Aquaculture 147, 21e35. Qian, P.Y., Wu, M.C.S., Ni, I.H., 2001. Comparison of nutrients release among some maricultured animals. Aquaculture 200, 305e316. Stabilli, L., Licciano, M., Giangrande, A., Longo, C., Mercurio, M., Marzano, C.N., Corriero, G., 2006. Filtering activity of Spongia ofcinalis var. adriatica (Schmidt) (Porifera, Demospongiae) on bacterioplankton: implications for bioremediation of polluted seawater. Water Research 40, 3083e3090. Stabilli, L., Licciano, M., Longo, C., Corriero, G., Mercurio, M., 2008. Evaluation of microbilogical accumulation capability of the commercial sponge Spongia ofcinalis var. adriatica (Schmidt) (Porifera, Demospongiae). Water Research 42, 2499e2506. Strickland, J.D.H., Parsons, T.R., 1972. A practical handbook of seawater analysis, 2nd ed. Fishery and Research Board of Canada Bulletin, Ottawa, Canada. Zar, J.H., 2009. Biostatistical analysis, 5th ed. Prentice Hall, Upper Saddle River, New Jersey.