Aquaculture 507 (2019) 377–384

Contents lists available at ScienceDirect

Aquaculture
journal homepage: www.elsevier.com/locate/aquaculture

Bioremediation of organic sludge from a marine recirculating aquaculture T

system using the polychaete Abarenicola pusilla (Quatrefages, 1866)
Silvia Gómeza,⁎, Carlos Felipe Hurtadoa, Jaime Orellanab
a
Escuela de Ciencias del Mar, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile
b
Erwin Sander Elektroapparatebau GmbH, 31311 Uetze-Eltze, Germany

ARTICLE INFO ABSTRACT

Keywords: Organic sludge, the principal residual product from recirculating aquaculture systems (RAS), is one of the main
Recirculating aquaculture systems environmental and logistical challenges associated with aquaculture production. Polychaetes have been de-
Bioremediation scribed as potential remediating organisms of aquaculture sludge due to its ability to survive in organic en-
Polychaete richment conditions and its capability to assimilate particulate organic waste from intensive aquaculture. The
Organic matter
aim of this study was to evaluate the removal performance of Abarenicola pusilla (Quatrefages, 1866) when fed
Integrated aquaculture
with organic sludge from a marine RAS. Experiments consisted of the addition of fish sludge (0.5, 2.0, 4.0 and
10.0% respect total inert sediment) at different densities of A. pusilla (60, 75, 150 and 200 organisms m−2), with
3 replicates each, and during 45 days. The highest removal rate of total organic matter (85.37%;
35.77 ± 0.05 g m−2 day−1), total N (33.63 ± 0.003 mg day−1) and total C (236.78 ± 0.003 mg day−1) were
obtained with 10% of sludge and 200 organisms m−2. Assimilation was corroborated by obtaining significant
increase (p < .05) in total carbon (24.75%), organic carbon (24.68%) and lipid content (0.65%) in the biomass
of organisms. Highest specific growth rate achieved by A. pusilla was 3.06% day−1 and the highest survival was
91.67% over the 45 days trial period. These results indicated that A. pusilla could be used as remediating species
of organic compounds contained in marine aquaculture sludge. The results also suggest that A. pusilla is a
potential candidate for nutrient recycling from marine aquaculture systems on land. Further studies should
determine the possible changes in the fatty acid profile of A. pusilla in order to use it in aquaculture feeds.

1. Introduction components by organisms (Bureau and Hua, 2010; Cubillo et al., 2016)
and consists mainly of organic compounds of carbon (C), nitrogen (N)
Global aquaculture production has achieved an average annual and phosphorus (P) present in the diet such as proteins, lipids, carbo-
growth of 8% since 1980, becoming the food industry with greater hydrates, vitamins, minerals or pigments (Herath and Satoh, 2015). The
development in recent decades (White et al., 2017). Recirculating POM generation rate in RAS cultures can reach until 52% respect to the
aquaculture systems (RAS) emerged to respond to the global demand total food added to the system (Mongirdas et al., 2017).
increase for aquaculture products and the new environmental regula- Aquaculture sludge results of the POM accumulation extracted from
tions (Guerdat et al., 2013; Yogev et al., 2017). The reduction, disposal RAS water. This compound presents a high concentration in organic
and reuse of wastes generated during the culture cycle is one of the solids, nutrients and water content (Zhang et al., 2013; Madariaga and
main current challenges associated with production in RAS (Martins Marín, 2017; Mongirdas et al., 2017). Currently, in a RAS it is produced
et al., 2010; van Rijn, 2013). These wastes, eliminated as effluents, are between 11 and 40% of sludge respect to the total food supplied to
the result of metabolic excretion of the culture organisms after feeding organisms (Luo et al., 2015). Its accumulation is a logistical problem in
process and correspond mainly to particulate organic matter (POM), aquaculture facilities and its discharge in water bodies causes organic
dissolved organic matter (DOM), total suspended solids (TSS) and dis- enrichment, ecological changes and deterioration of water quality (van
solved metabolites (Piedrahita, 2003). Rijn, 2013; Edwards, 2015; Gunning et al., 2016; Akizuki and Toda,
Particulate organic matter (POM) is the main form of waste gener- 2018).
ated in RAS (Chen et al., 1997). POM includes solids released into the Recently, sludge treatment have been coupled with extractive spe-
water as a product of not ingested, not digested or not absorbed cies in marine aquaculture systems (Marques et al., 2017; Pajand et al.,

⁎
Corresponding author at: Avda. Altamirano 1480, Valparaíso, Chile.
E-mail addresses: [email protected] (S. Gómez), [email protected] (C.F. Hurtado), [email protected] (J. Orellana).

https://doi.org/10.1016/j.aquaculture.2019.04.033
Received 30 January 2019; Received in revised form 8 April 2019; Accepted 9 April 2019
Available online 10 April 2019
0044-8486/ © 2019 Elsevier B.V. All rights reserved.
S. Gómez, et al. Aquaculture 507 (2019) 377–384

2017). Due to its ability to assimilate particulate organic waste from (Austral University, Valdivia, Chile) to the Aquaculture Experimental
intensive aquaculture, polychaetes are one of the main candidates for Laboratory (PUCV, Valparaíso, Chile). A purge was made to eliminate
remediation in aquaculture effluents (Palmer, 2010; Robinson et al., possible stomach contents (Palmer, 2010) and a period of 48 h was
2015). Integrated culture systems with polychaetes can increase aqua- established for acclimatization (Weston et al., 2000; Nelson et al.,
culture sustainability decreasing environmental impacts (Pombo et al., 2012). Initial wet weight was determined with a precision scale (Jad-
2018). ever SNUG II-300) and dry weight was determined after dehydration
Species of family Nereididae are the most commonly studied, (333.15 K until constant value) (Mason et al., 1983; Bischoff, 2007).
reaching significant organic matter removal from aquaculture sludge in Growth performance was calculated using the specific growth rate (μ)
several studies (Honda and Kikuchi, 2002; Bischoff, 2007; Palmer, according to Jørgensen et al. (1993). Absolute growth rate was calcu-
2010; Brown et al., 2011; Fang et al., 2016; Marques et al., 2017; lated according to Hopkins (1992). Mortality was recorded daily, col-
Pajand et al., 2017). Polychaetes of family Arenicolidae are sub- lecting the dead organisms from the sediment surface. Survival was
superficial sedimentivorous species capable of assimilating organic calculated like the proportion of alive organisms at the end of the trials,
matter (mainly detritus) available in the sediment and interstitial water expressed as percentage (Pechenik and Cerulli, 1991; Bischoff, 2007).
(Hylleberg, 1975; Alyakrinskaya, 2003). Species of this family have also
shown its capability as organic bioremediators. Arenicola marina re-
moved organic components present in fish sludge from marine RAS 2.2. System design
aquaculture (Olive et al., 2006; Guerrero and Cremades, 2012;
Lamprianidou, 2015). Yearsley et al. (2011) indicated in Arenicola During the trials, the polychaetes were randomly implemented in 16
loveni loveni significant growth when it was implemented in abalone experimental units (organized in 4 independent series) with a surface of
effluent culture (Haliotis midae). 0.134 m2. In each series, 3 culture units correspond to replicates and
Polychaetes are an important source of proteins and lipids (Wang one control without polychaetes. The prototype system was carried out
et al., 2019). Some species are used as ingredient in fish broodstock and with biofiltration and water recirculation. The biofilters (moving bed
shrimp food, improving the diets supplied during the maturation stage biofilm reactor) were sized according to the ammonia excretion rate
(Palmer et al., 2014). The integration of polychaetes in multitrophic achieved by A. pusilla in previous experiments (Drennan II et al., 2006;
cultures offers a potential of recycling proteins and polyunsaturated Ebeling and Timmons, 2012; Gómez et al., 2018). The water was re-
fatty acids present in aquaculture sludge (Bischoff et al., 2009; Olive circulated with a submersible pump (Grech CHJ – 6000; 54 l h−1) to
et al., 2009; Brown et al., 2011; Callier et al., 2014; Fang et al., 2016; allow the total water change in the units every hour (Bischoff, 2007).
Marques et al., 2017; Pajand et al., 2017; Marques et al., 2018). Peri- To provide the habitat to the organisms, a 10 cm layer of sand
nereis nuntia vallata was able to assimilate organic N contained in P. (63–500 μm; Caval substrate n° 133,631) filled each unit (Crane and
olivaceus feces (Honda and Kikuchi, 2002). Nereis diversicolor showed a Merz, 2012). The sand was previously incinerated (823.15 K, 24 h) to
significant increase in its lipid content fed with salmon sludge eliminate possible organic matter content (APHA, 1998; Palmer, 2010).
(Seekamp, 2017). These valuable components can be reintroduced in In the bottom of the units, a 5 cm layer of gravel and a subsurface
aquaculture feeds helping to reduce the demand for fishmeal and fish drainage pipe were installed to ensure water recirculation (Honda and
oil for aquaculture industry (Pajand et al., 2017; Wang et al., 2019). Kikuchi, 2002; Palmer, 2010). A textile fiber was incorporated to avoid
Preliminary studies suggest that the polychaete Abarenicola pusilla mixing between both substrates. The introduction of the sludge was
(Quatrefages, 1866) can be used as extractive species (Gómez et al., done in the subsurface layers after the substrate implementation (Olive
2018). This study aimed to evaluate its performance to remove and et al., 2006).
assimilate organic components present in aquaculture sludge from a Two types of experiments were carried out. One, implementing
marine RAS. The removal of total organic matter (TOM), total C and different sludge concentrations in each series of filtration units (0.5,
total N were analyzed during the experiments. Growth rate, protein and 2.0, 4.0 and 10.0% of sludge with respect to the total inert sediment) at
lipid content were also evaluated in the organisms. a density of 60 organisms m−2 (24 polychaetes per series) during
45 days. The other, incorporating the highest amount of sludge during
2. Materials and methods the same period and implementing different densities of polychaetes
(60, 75, 150 and 200 org m−2; which means 24, 30, 60 and 78 org
2.1. Abarenicola pusilla series−1 respectively). The details of the experimental design are shown
in Table 1.
Animals (n = 200) were transported alive in humid plant substrate
at 291.15 K from the Marine and Limnological Sciences Laboratory

Table 1
Design of the experiments achieved in the remediation system.
Test sludge

Sludge (g per unit) 125.03 ± 0.06 500.69 ± 0.67 1000.79 ± 0.51 2503.62 ± 2.73
(g per g org) 16.80 ± 1.36 64.10 ± 9.52 116.13 ± 5.10 261.38 ± 46.91
Polychaetes per unit 8.0 8.0 8.0 8.0
Density (org m−2) 60.0 60.0 60.0 60.0
(g org m−2) 112.1 ± 8.64 118.8 ± 16.37 129.45 ± 5.66 147.00 ± 27.69

Test density

Sludge (g per unit) 2503.62 ± 2.73 2503.62 ± 2.73 2503.62 ± 2.73 2503.62 ± 2.73
(g per g org) 325.01 ± 57.84 264.84 ± 33.14 121.01 ± 13.69 95.33 ± 5.47
Polychaetes per unit 8.0 10.0 20.0 26.0
Density (org m−2) 60.0 75.0 150.0 200.0
(g org m−2) 117.9 ± 19.7 143.35 ± 18.34 312.90 ± 33.25 404.97 ± 22.89

Values are mean ± SD of 3 determinations. Weight is expressed as wet weight (g).

378
S. Gómez, et al. Aquaculture 507 (2019) 377–384

Table 2 A. pusilla = final isotopic composition of organisms; ƌ13C initial = in-

Aquaculture sludge characterization. itial isotopic composition of organisms; ƌ13C standard = isotopic
pH 6.65 ± 0.03 composition of Abarenicola in marine environments (LeDuc et al., 2006;
T (K) 292.85 ± 0.14 Silva et al., 2011). Total lipid content was determined as ethereal ex-
O2 (% saturation) 14.60 ± 0.42 tract by extraction with organic solvent (Soxhlet technique, AOAC
O2 (mg l−1) 1.09 ± 0.18 963.15). Protein content was determined by Kjeldahl method (AOAC
Ammonia (NH3-N) (mg l−1) 10.70 ± 0.08
Nitrite (NO2−-N) (mg l−1) 0.27 ± 0.02
960.52) (Vergara et al., 1999).
Nitrate (NO3−-N) (mg l−1) 0.7 ± 0.12
Salinity (g l−1) 36.00 ± 0.50 2.6. Statistical analysis
Water content (%) 96.18 ± 0.23
TOM content (%) 82.17 ± 0.12
The analyses were carried out using SPSS 19 software. All data were
N total (%) 4.1 ± 0.05
C total (%) 29.4 ± 0.32 tested for normality and homogeneity of variance using the Shapiro-
C/N ratio 7.16 ± 0.18 Wilk test. The removal and assimilation of total N and total C, protein
and lipid content were determined with independent samples t-test
Values are mean ± SD of 5 determinations. TOM: Total organic (p < .05). Data of TOM removal, survival and growth performance of A.
matter. pusilla were determined by one-way ANOVA analysis. The significant
differences were analyzed using the Tukey test (p < .05).
2.3. Aquaculture sludge
3. Results
The aquaculture sludge was obtained from a marine RAS with
Seriola lalandi (Valenciennes, 1833) (average weight = 3.1. Aquaculture sludge
3000.2 ± 110.4 g, culture density = 10.85 kg m−3). The sludge was
collected from a radial drum filter with rotating screen (60 μm, HEX F1- The results of the sludge analysis are shown in Table 2. The sludge
1, Germany) coupled to a cylindrical sedimentation tank with conical generation rate in the RAS was 10.12 ± 0.19 kg of sludge per kg of food
bottom (Tal et al., 2009). The sludge was used less of 48 h after its supplied daily to the fish (mean ± SD, n = 5), approximately 400 g of
removal to reduce microbiological action (Mongirdas et al., 2017). dry sludge per kg of feed supplied. The range of majority particle sizes
Ammonia (NH3-N) (Nessler method 8038), nitrite (NO2−-N) (diazoti- (49%) was found between 1000 and 3000 μm.
zation method 8507) and nitrate (NO3−-N) (cadmium reduction
method 8192) concentration were determined using a spectro- 3.2. Abarenicola pusilla
photometer (Hach DR-3900). Before these analyzes, the sludge was
filtered (0.2 μm) to reduce suspended solids, microorganisms and col- The initial wet weight of A. pusilla was 2.11 ± 0.25 g (test sludge,
loidal particles (Aruety et al., 2016; Mayhead et al., 2018). Tempera- n = 48) and 2.00 ± 0.07 g (test density, n = 96) with a water content
ture, dissolved oxygen and pH were measured with a portable multi- of 86.93 ± 3.97% (n = 15). The detail of the growth and survival
parameter probe (Hach HQ40D) and salinity was determined using a data are shown in Table 3. The highest specific growth rate (μ)
refractometer (RHS-10ATC). The particle size distribution was de- achieved during the experiments was 3.06 ± 0.22% day−1
termined by passing the sludge through a sieve tower (80–5000 μm). (8.48 ± 0.74 g m−2 day−1). In the test of density, the highest specific
Water content was determined by dehydration (333.15 K until constant growth rate was reached in the filtration units implemented at 60 and
weight) (Winberg and Duncan, 1971; Bischoff, 2007). 75 organisms m−2 (2.57 ± 0.05% day−1 and 2.58 ± 0.20% day−1,
respectively). There were significant differences in growth rates for
2.4. Water quality analysis these conditions compared to the rest (p < .05). No significant differ-
ences in survival were observed between different experimental con-
A surface water layer was permanently present in the experi- ditions in both trials.
mental units (Görlitz, 2012). The water was extracted directly from
the sea by a pumping system (Vogt AM-751), filtered using sand fil- 3.3. Water quality analysis
ters (Jacuzzi TP-30) and cartridge filters (Vigaflow 20, 5 and 1 μm)
and treated with ultraviolet system (Red Base 40w UV-C T5). Water The parameters of the initial water introduced into the remediation
quality analyzes were measured daily. The physicochemical para- system were: ammonia (NH3-N: 0.16 ± 0.03 mg l−1), nitrite (NO2−-N:
meters and the concentration of nitrogenous components were de- 0.10 ± 0.01 mg l−1), nitrate (NO3−-N: 0.40 ± 0.03 mg l−1), pH:
termined as the same procedure as the sludge analyzes. Carbon di- 7.80 ± 0.01, CO2: 1 ± 0.00 mg l−1, temperature: 291.35 ± 0.09 K.
oxide (CO2) dissolved was determined using an analyzer (OxyGuard During the trials, the water temperature was maintained between
G03C2). Oxygen saturation was maintained using an air pump 290.15 and 293.15 K (291.11 ± 0.24 K) (Retraubun et al., 1996; Olive
(Sunsun HT-500). et al., 2006; Moreno et al., 2007). No pH adjustment was necessary
(7.77 ± 0.05) (Toulmond, 1975; Alyakrinskaya, 2003). Oxygen sa-
2.5. Organic components analysis turation remained at 107.59 ± 2.67% (8.39 ± 0.24 mg l−1)
(Alyakrinskaya, 2003; Bischoff, 2007) and salinity at
The TOM content in the substrate was determined once a week after 30.19 ± 0.41 g l−1 (Alyakrinskaya, 2003; Hiebert, 2014; Valenzuela-
dehydration (333.15 K until constant weight) as the biomass loss by Olea, pers. comm.). Carbon dioxide (CO2) dissolved remained below
incineration (823.15 K, 24 h) (Palmer, 2010). To determine the flow 1 mg l−1. Ammonia concentration (0.44 ± 0.24 mg l−1) was main-
and transfer of organic matter to the organisms, total nitrogen (N), total tained below the toxicity limit for other species of polychaetes
carbon (C) and the stable isotope profile (δ13C and δ15N) were de- (8 mg l−1) in laboratory tests (Bischoff, 2007; Seekamp, 2017). Sig-
termined using an isotope ratio mass spectrometer (IRMS, Thermo nificant differences were found in ammonia concentration only at the
Delta Advantage) coupled to an elemental analyzer (Flash EA2000) beginning of the trials (until day 6) in the filtration units with the
(Callier et al., 2014; Andrade et al., 2016). The analysis of assimilation highest implementation of sludge (p < .05). Nitrate concentration
by isotopes was carried out adapting the Bianchi (2007) model ac- (average: 0.76 ± 0.21 mg l−1) remained between 0.40 and 1.50 mg l−1
cording to the following formula: OC Assimilated (%) = {(ƌ13C A. pu- in all treatments. The levels of nitrite concentration presented higher
silla – ƌ13C standard)/(ƌ13C initial – ƌ13C standard)} * 100. Where: ƌ13C values at the beginning of the experiments, reaching a maximum of

379
S. Gómez, et al. Aquaculture 507 (2019) 377–384

Table 3
Growth rates and survival of A. pusilla.
Test sludge

Sludge (g) C1 C2 C3 C4
AGR (g m−2 day−1) 7.14 ± 0.42† 7.41 ± 0.18† 8.48 ± 0.74† 7.00 ± 1.09†
SGR (μ) (% day−1) 3.02 ± 0.23† 3.00 ± 0.26† 3.06 ± 0.22† 2.58 ± 0.51†
Survival (%) 91.67 ± 14.43† 91.67 ± 14.43† 83.34 ± 14.43† 66.67 ± 14.43†

Test density

Density (org m−2) 60.0 75.0 150.0 200.0

AGR (g m−2 day−1) 6.39 ± 1.10a 6.89 ± 0.46a 8.57 ± 0.06b 8.72 ± 0.42b
SGR (μ) (% day−1) 2.57 ± 0.05a 2.58 ± 0.20a 1.80 ± 0.13b 1.54 ± 0.03b
Survival (%) 83.34 ± 28.67† 80.00 ± 20.00† 90.00 ± 10.00† 76.92 ± 7.69†

AGR: Absolute growth rate. Values are mean ± SD for 45 days of trials. n = 48 (test sludge), n = 96 (test density). Sludge added: C1 = 500.11 g; C2 = 2002.76 g;
C3 = 4003.16 g; 10,014.50 g per experimental series. Values with different letters are significantly different from each other (p < .05). Values with “†” are not
different from each other.

0.42 mg l−1. After that, the average concentration remained at

0.04 ± 0.07 mg l−1.

3.4. Organic components analysis

3.4.1. Total organic matter

The percentage of TOM removal achieved by A. pusilla after 45 days
in the different treatments is shown in Figs. 1 and 2. The highest TOM
removal was obtained with 200 organisms m−2 (85.37 ± 7.65%;
35.77 ± 0.05 g m−2 day−1). Significant differences were found be-
tween the series with 60 and 75 organisms m−2 compared to 150 and
200 organisms m−2 (p < .05). All series show differences with their
respective controls. Table 4 shows the removal rate obtained per unit
area and time period for both experiments. According to the TOM re-
moval obtained, the highest conversion factor obtained by the organ-
isms was 4.22 g TOM per g polychaete. Fig. 2. TOM removal rate achieved by A. pusilla during the trials (experiment
2). A. pusilla performance is shown in grey columns. Black columns represent
3.4.2. Total N and total C control units (mean values ± SD). It is expressed as percentage of removal (%)
Significant total N (33.63 ± 0.003 mg day−1) and total C at 45 days. Sludge added: 10.014,50 g in each experimental series. Significant
differences (p < 0.05) are expressed with (*).
(236.78 ± 0.003 mg day−1) removal rates were obtained in the units at
200 organisms m−2 (p < .05). This meant a decrease of 91.85% (N) and
90.55% (C) in 45 days. The biomass of the organisms presented an in- displacement of the isotope ƌ13C (1.17 ppm, p < .05), indicating OC
crease of 6.92 ± 1.19% (total N) and 24.75 ± 4.18% (total C) about its assimilation (24.68%). No significant assimilation was found for ƌ15N
initial content. The assimilation of total C was significant (p < .05). isotope (p < .05).
In addition, the biomass of A. pusilla showed a significant
3.4.3. Lipid and proteins content
Mean values of lipid content (14.43 ± 1.18%) and protein content
(17.70 ± 0.15%) were obtained in the aquaculture sludge (n = 5). To
determine the assimilation of these compounds by the organisms, their
content in the biomass were analyzed before and after the experiments
(Bischoff et al., 2009). A. pusilla achieved a significant increase
(p < .05) in its lipid content (dry basis) during the trials
(0.65 ± 0.07%) and a significant decrease in protein content
(2.98 ± 0.01%).

4. Discussion

The results of this study demonstrated that A. pusilla is an efficient

remediating species for reduction of organic waste presented in RAS
aquaculture sludge.

4.1. TOM removal rate

Fig. 1. TOM removal rate achieved by A. pusilla during the trials (experiment
1). A. pusilla performance is shown in grey columns. Black columns represent The highest removal efficiency of TOM achieved by A. pusilla
control units (mean values ± SD). It is expressed as percentage of removal (%) (85.37%, 27.4 mg g polychaete−1 day−1, 45-day trial) was obtained in
at 45 days. Sludge added: C1 = 500,11 g; C2 = 2.002,76 g; C3 = 4.003,16 g; the filtration units with 200 org m−2. Other studies in bioremediation
C4 = 10.014,50 g. Significant differences (p < 0.05) are expressed with (*). with polychaetes reported lower TOM removal rates. Fang et al. (2016)

380
S. Gómez, et al. Aquaculture 507 (2019) 377–384

Table 4
TOM removal rate achieved by A. pusilla.
Test sludge

Sludge (g) C1 C2 C3 C4
Density (org m−2) 60.0 60.0 60.0 60.0
TOM RR (g m−2 day−1) 0.89 ± 0.06a 3.80 ± 0.12a 6.67 ± 0.10 12.35 ± 0.09†

Test density

Sludge (g) C4 C4 C4 C4
Density (org m−2) 60.0 75.0 150.0 200.0
TOM RR (g m−2 day−1) 13.65 ± 0.04a 18.53 ± 0.05a 33.86 ± 0.05† 35.77 ± 0.05†

TOM RR: Total organic matter removal rate. Values are mean ± SD of 3 determinations. Sludge added: C1 = 500.11 g; C2 = 2002.76 g; C3 = 4003.16 g;
C4 = 10,014.50 g implemented to each experimental series. Values with different letters are significantly different from each other (p < .05). Values with “†” are not
different from each other.

indicated a TOM removal up to 50% in Perinereis aibuhitensis fed with Fang et al. (2016) indicated in P. aibuhitensis the removal of
Paralichthys olivaceus sludge in a 35-day trial (13.43 mg TOM g poly- 0.33 ± 0.05 mg N g polychaete−1 day−1. MacDonald et al. (2013) in-
chaete−1 day−1). Marques et al. (2017) reported a removal of 70% of dicated in Holothuria forskali fed with D. labrax sludge a removal rate of
TOM by N. diversicolor in Solea senegalensis sludge from an intensive 0.184 mg N g organism−1 day−1. Other remediation studies do not re-
RAS (150-day trial) and Palmer (2010) indicated that Perinereis nuntia port significant results of total N removal by polychaete species
and Perinereis helleri removed around 50% of the TOM contained in (Palmer, 2010).
aquaculture sludge from Penaeus monodon (16-week trial). Significant total C removal was also achieved by A. pusilla
TOM removal rate is determined by the polychaete species, the (8.99 ± 0.03 mg C g polychaete−1 day−1). Fang et al. (2016) indicated
aquaculture sludge characteristics and the design of the remediation in P. aibuhitensis fed with P. olivaceus sludge a lower removal rate
system (Bischoff, 2007; Nelson et al., 2012; Seekamp, 2017). In Palmer (2.99 ± 0.47 mg C g−1 day−1). These differences could be due to the
(2010) the differences during the experiences in the physicochemical lower content of total N and C in the aquaculture sludge respect to this
parameters of surface water and sediment in the remediation systems, work. Other factors could be the remediation system design, the phy-
implemented outdoor, could decrease the TOM removal rate by poly- sicochemical conditions in superficial water or the efficiency of the
chaetes. The sludge used in the study of Fang et al. (2016) was obtained polychaete species itself (Bischoff, 2007).
from the seabed under aquaculture cages. This could be related to the A. pusilla achieved a total C assimilation of 24.75% (45-day trial)
lower TOM removal efficiency due to sludge degradation processes respect to the contained C in the sludge, more than the indicated by
prior to the implementation in the laboratory tests (Mongirdas et al., Fang et al. (2016) in P. aibuhitensis (20.5%, 35-day trial) fed with P.
2017). olivaceus sludge. The organic carbon (OC) assimilation by A. pusilla
Palmer (2010) achieved the highest TOM removal rate im- (1.17 ppm, 45-day trial) was also significant. This result is in ac-
plementing the highest density of organisms, agreeing with the present cordance with Callier et al. (2014), where N. diversicolor fed with D.
study. Increasing this condition, could be possible to obtain better re- labrax sludge obtained approximately 1 ppm (30-day trial). Otherwise,
moval results (Palmer, 2010). As well, a higher TOM removal rate could the sea cucumber Apostichopus japonicus reported lower assimilation
be expected with a sludge richer in organic components (Honda and (1.6 ppm in 238-days trial) under aquaculture cages in the sea. This fact
Kikuchi, 2002; Brown et al., 2011) or making a mixed diet with fish could be due to the ingestion of other organic components not corre-
feed (Seekamp, 2017). In this study, better results of TOM removal rate sponding to the aquaculture sludge (Yokoyama, 2013).
were obtained increasing the sludge in the units with A. pusilla. For this,
the addition of 10% of sludge is recommended. The best rate of or-
4.3. Protein and lipids
ganisms to be implemented in this system was 150 org m−2 (no dif-
ferences in TOM removal rate were obtained about 200 org m−2). It
A. pusilla presented a significant reduction in its protein content
would be interesting to carry out more experiments to identify the
after the trials, agreeing with other remediation studies with poly-
carrying capacity of the remediation system (increasing the density of
chaetes (Suckow et al., 2010; Seekamp, 2017). This fact could be as-
polychaetes and/or the proportion of sludge).
sociated with high levels of ammonia in the surface water (Palmer,
The TOM removal rate obtained in the control units could be due to
2010). However, the average concentration of ammonia in this study
the mineralization of organic components by heterotrophic micro-
was maintained lower than the limit of toxicity indicated for other
organisms (Batista et al., 2003; Olive et al., 2009; Palmer, 2010; Palmer
species of polychaetes (Bischoff, 2007). On the other hand, the greater
et al., 2018). Due to the bioturbation activities, is possible as well that
availability of nutrients in the remediation systems compared with
an increase in the mineralization rate was obtained in the units with A.
natural environments could result in a lower energy investment for the
pusilla (Hylleberg, 1975; Görlitz, 2012; Ito et al., 2016). On the other
search for food and in a less muscle formation, component with high
hand, the increases of TOM obtained in some control units could be due
percentage of proteins (Seekamp, 2017). Also, the decrease in protein
to the degradation of the organic components in the sediment
percentage in the biomass of A. pusilla could be due to a reduction in its
(Nimptsch et al., 2015; Srithongouthai and Tada, 2017).
blood hemoglobin concentration because of the high oxygen saturation
in the surface water (Wells, 1966; Alyakrinskaya, 2003). The protein
4.2. Total N and total C content in the aquaculture sludge could influence as well in the as-
similation by polychaetes. Brown et al. (2011) indicated in N. virens a
A significant total N removal (1.27 ± 0.01 mg N g poly- significant increase in its protein content fed with a richer sludge (50%-
chaete−1 day−1) was obtained by A. pusilla. Other studies indicated protein content). More studies would be needed to verify if A. pusilla is
lower N removal rates. Honda and Kikuchi (2002) reported that P. able to increase its protein content in another type of study.
nuntia vallata was able to eliminate 0.65 ± 1.17 mg N g poly- A significant increase in lipid concentration was obtained by A.
chaete−1 day−1 respect to the total content in the aquaculture sludge. pusilla during the trials. Polychaetes accumulate lipids when increase

381
S. Gómez, et al. Aquaculture 507 (2019) 377–384

the quality of the available food in the system, offering the possibility of land. A. pusilla achieved a significant removal of organic components
reusing these nutrients contained in the sludge (Bischoff et al., 2009; and obtained a production of about 9 g m−2 day−1 (almost 400 g m−2
Brown et al., 2011). The sludge characteristics, the efficiency of the in 45 days). These results demonstrated that A. pusilla is able to con-
species or the stability in the experimental conditions could influenced sume aquaculture sludge as the only source of food. The assimilation of
in the lipid assimilation by polychaetes (Bischoff et al., 2009; Olive organic compounds achieved by A. pusilla suggests that it is an excellent
et al., 2009; Brown et al., 2011). For instance, Seekamp (2017) reported potential candidate for an integrated culture. Further studies could be
differences in lipid assimilation in N. diversicolor fed with commercial interesting to determine possible changes in its fatty acid profile and the
fish feed (1.6%, 30-day trial) comparing with Salmo salar sludge (1.2%, use of its biomass as a valuable product in aquaculture feeds.
30-day trial). Other studies feeding A. pusilla with a mixed diet of fish
feed and aquaculture sludge could result in a greater assimilation of 6. CRediT authorship contribution statement
lipids. New analyzes of fatty acid profiles would be needed to verify the
presence of important components in A. pusilla for its use in aquaculture Dra. Silvia Gómez: Conceptualization, Methodology, Validation,
feeds. Formal analysis, Investigation, Writing – original draft, Project ad-
ministration. Dr. Carlos Felipe Hurtado: Conceptualization,
4.4. Growth of A. pusilla Methodology, Project administration, Supervision, Writing – Review
and editing, Funding acquisition. Dr. Jaime Orellana:
The highest specific growth rate achieved by A. pusilla (3.06% Conceptualization, Methodology, Project administration, Supervision,
day−1, 45-day trial) was obtained in the filtration units with lower Writing – Review and editing, Funding acquisition.
density of organisms, results that agree with Palmer (2010). Other
studies with polychaetes reported similar or lower growth rates. For Acknowledgements
instance, Honda and Kikuchi (2002) obtained 1.66% day−1 in Perinereis
nuntia vallata fed on aquaculture sludge from Paralichthys olivaceus in a The authors are grateful for the financial support provided by the
15-day trial. Pajand et al. (2017) indicated a SGR of 3.4% day−1 in N. Aquaculture PhD programme of the Pontifical Catholic University of
diversicolor during an 8-week trial fed with Huso huso sludge and Brown Valparaíso. Special thanks to Alice Turner and Guillermo Valenzuela for
et al. (2011) indicated a 2.1% day−1 in Nereis virens fed with H. hip- their help during the development of the experiments.
poglossus sludge during a 71-day trial. Lower growth rates were re-
ported by Suckow et al. (2010), where Nereis virens fed with D. labrax References
sludge obtained 0.9% day−1 in 6-week trial. As well, Fang et al. (2016)
reported in Perinereis aibuhitensis fed with P. olivaceus sludge a SGR of Akizuki, S., Toda, T., 2018. An anaerobic-aerobic sequential batch process with si-
0.64% day−1 (35-day trial) and Yearsley et al. (2011) indicated a SGR multaneous methanogenesis and short-cut denitrification for the treatment of marine
biofoulings. Waste Manag. 74, 168–176. https://doi.org/10.1016/j.wasman.2017.
of 0.39% day−1 in Arenicola loveni loveni implemented in a Haliotis 12.013.
midae aquaculture effluent. The differences in polychaete growth rates Alyakrinskaya, I.O., 2003. Some ecological features of the lugworm Arenicola marina L.
between studies could be due to the system design, density of organisms (Annelida, Polychaeta) and its morphological and biochemical adaptations to bur-
rowing. Biol. Bull. Russ. Acad. Sci. 30 (4), 411–418. https://doi.org/10.1023/
or the sludge quality (Bischoff, 2007; Palmer, 2010; Seekamp, 2017). A:1024878310385.
The highest growth rate of A. pusilla was obtained in C3 (test sludge) Andrade, C., Ríos, C., Gerdes, D., Brey, T., 2016. Trophic structure of shallow-water
and 60 and 75 org m−2 (test density). It could be expected that the benthic communities in the sub-Antarctic Strait of Magellan. Polar Biol. 39 (12),
2281–2297. https://doi.org/10.1007/s00300-016-1895-0.
highest growth rate would be obtained in the units with the highest APHA, 1998. Standard Methods for the Examination of Water and Wastewater, 20th ed.
TOM removal. However, other remediation studies reported similar American Public Health Association, American Water Works Association, Water
results, where the growth and sludge consumption by TOM analysis in Environment Federation, Washington, D.C.
Aruety, T., Brunner, T., Ronen, Z., Gross, A., Sowers, K., Zilberg, D., 2016. Decreasing
the sediment was not correlated (Bischoff, 2007; Bischoff et al., 2009;
levels of the fish pathogen Streptococcus iniae following inoculation into the sludge
Seekamp, 2017). The ammonia excretion rate by A. pusilla increases digester of a zero-discharge recirculating aquaculture system (RAS). Aquaculture
with the sludge proportion added to the system (Gómez et al., 2018). 450, 335–341. https://doi.org/10.1016/j.aquaculture.2015.08.002.
This could have affected the growth of the organisms in the units with Batista, F.M., e Costa, P.F., Matias, D., Joaquim, S., Massapina, C., Passos, A.M., Ferreira,
P.P., da Fonseca, L.C., 2003. Preliminary results on the growth and survival of the
10% of aquaculture sludge. Due to the implementation of the biofil- polychaete Nereis diversicolor (OF Muller, 1776), when fed with faeces from the carpet
tration system, only significant differences were observed in the con- shell clam Ruditapes decussatus (L., 1758). Bol. Inst. Esp. Oceanogr. 19 (1–4), 443(10.
centration of ammonia in the surface water until the day 7 of the ex- 1.1.556.4108&rep=rep1&type=pdf).
Bianchi, T.S., 2007. Biogeochemistry of Estuaries. Oxford University Press, New York
periments (test of sludge). However, it is possible that its concentration (688 pp).
was higher in the substrate (Bischoff, 2007). Also, the TOM removal Bischoff, A.A., 2007. Solid Waste Reduction of Closed Recirculated Aquaculture Systems
rate could be favored in the units with the highest density of poly- by Secondary Culture of Detritivorous Organisms. PhD-Thesis at the Leibniz-Institute
of Marine Sciences, Kiel, Germany, pp. 179.
chaetes by heterotrophic microorganisms consuming organic matter. Bischoff, A.A., Fink, P., Waller, U., 2009. The fatty acid composition of Nereis diversicolor
These are associated with aquaculture sludge and they can proliferate cultured in an integrated recirculated system: possible implications for aquaculture.
in remediation systems due to bioturbation processes (Batista et al., Aquaculture 296 (3–4), 271–276. https://doi.org/10.1016/j.aquaculture.2009.09.
002.
2003; Olive et al., 2009; Palmer, 2010; Palmer et al., 2018).
Brown, N., Eddy, S., Plaud, S., 2011. Utilization of waste from a marine recirculating fish
culture system as a feed source for the polychaete worm, Nereis virens. Aquaculture
5. Conclusion 322, 177–183. https://doi.org/10.1016/j.aquaculture.2011.09.017.
Bureau, D.P., Hua, K., 2010. Towards effective nutritional management of waste outputs
in aquaculture, with particular reference to salmonid aquaculture operations. Aquac.
The search for alternatives to reduce and treat the sludge generated Res. 41 (5), 777–792. https://doi.org/10.1111/j.1365-2109.2009.02431.x.
is a global priority to the sustainable development of aquaculture. The Callier, M.D., Rossi, F., Geoffroy, T., Hutting, S., Van Bruggen, V., Blancheton, J.P., 2014.
use of polychaetes for the removal of organic wastes contained in Bioremediation of fish farm waste by deposit-feeding polychaetes Capitella sp. and
Hediste diversicolor. In: Aquaculture Europe 2014, Meeting of the European
aquaculture sludge can increase the environmental sustainability of Aquaculture Society (EAS): Adding Value, 14–17 October 2014, Donostia-San
marine RAS and offers the possibility of recycling their valuable nu- Sebastián, Spain, . https://archimer.ifremer.fr/doc/00228/33945/.
trients. Chen, S., Coffin, D.E., Malone, R.F., 1997. Sludge production and management for re-
circulating aquacultural systems. J. World Aquacult. Soc. 28 (4), 303–315. https://
In this study, the remediation performance of a species of the genus doi.org/10.1111/j.1749-7345.1997.tb00278.x.
Abarenicola was evaluated for the first time. The results obtained de- Crane, R.L., Merz, R.A., 2012. Sediment properties and burrowing of Abarenicola pacifica
monstrated that A. pusilla is a good candidate for its implementation in and A. claparedi vagabunda in False Bay. In: Blinks/BEACON/NSF Research
Experience for Undergraduates, pp. 21. https://digital.lib.washington.edu/
remediation systems associated with marine aquaculture systems on

382
S. Gómez, et al. Aquaculture 507 (2019) 377–384

researchworks/handle/1773/27068. 002.
Cubillo, A.M., Ferreira, J.G., Robinson, S.M., Pearce, C.M., Corner, R.A., Johansen, J., Mason, W.T., Lewis, P.A., Weber, C.I., 1983. An evaluation of benthic macroinvertebrate
2016. Role of deposit feeders in integrated multi-trophic aquaculture: a model ana- biomass methodology. Environ. Monit. Assess. 3 (1), 29–44. https://doi.org/10.
lysis. Aquaculture 453, 54–66. https://doi.org/10.1016/j.aquaculture.2015.11.031. 1007/BF00394030.
Drennan II, D.G., Hosler, K.C., Francis, M., Weaver, D., Aneshansley, E.D., Beckman, G., Mayhead, E., Silkina, A., Llewellyn, C.A., Fuentes-Grünewald, C., 2018. Comparing nu-
Johnson, C.H., Cristina, C.M., 2006. Standardized evaluation and rating of biofilters. trient removal from membrane filtered and unfiltered domestic wastewater using
II. Manufacturer's and user's perspective. Aquac. Eng. 34 (3), 403–416. https://doi. Chlorella vulgaris. Biology 7 (1), 12. https://doi.org/10.3390/biology7010012.
org/10.1016/j.aquaeng.2005.07.001. Mongirdas, V., Žibienė, G., Žibas, A., 2017. Waste and its characterization in closed re-
Ebeling, J.M., Timmons, M.B., 2012. Recirculating aquaculture systems. In: Tidwell, J.H. circulating aquaculture systems: a review. J. Water Secur. 3. https://doi.org/10.
(Ed.), Aquaculture production systems. Wiley-Blackwell, Hoboken, pp. 245–277. 15544/jws.2017.002.
https://doi.org/10.1002/9781118250105.ch11. Moreno, R.A., Rozbaczylo, N., Sepúlveda, R.D., Marinkovic, M.I., Arntz, W.E., Thatje, S.,
Edwards, P., 2015. Aquaculture environment interactions: past, present and likely future 2007. On some ecological aspects of the lugworm Abarenicola affinis chiliensis Wells,
trends. Aquaculture 447, 2–14. https://doi.org/10.1016/j.aquaculture.2015.02.001. 1963 (Polychaeta: Scolecida: Arenicolidae) from shallow soft bottoms of northern
Fang, J., Zhang, J., Jiang, Z., Du, M., Liu, Y., Mao, Y., Gao, Y., Fang, J., 2016. Chile. Helgol. Mar. Res. 61 (1), 1–7. https://doi.org/10.1007/s10152-006-0047-2.
Environmental remediation potential of Perinereis aibuhitensis (Polychaeta) based on Nelson, E.J., MacDonald, B.A., Robinson, S.M.C., 2012. The absorption efficiency of the
the effects of temperature and feed types on its carbon and nitrogen budgets. Mar. suspension-feeding sea cucumber, Cucumaria frondosa, and its potential as an ex-
Biol. Res. 12 (6), 583–594. https://doi.org/10.1080/17451000.2016.1177653. tractive integrated multi-trophic aquaculture (IMTA) species. Aquaculture 370,
Gómez, S., Hurtado, C.F., Orellana, J., Valenzuela-Olea, G., Turner, A., 2018. Abarenicola 19–25. https://doi.org/10.1016/j.aquaculture.2012.09.029.
pusilla (Quatrefages, 1866): a novel species for fish waste bioremediation from Nimptsch, J., Woelfl, S., Osorio, S., Valenzuela, J., Ebersbach, P., von Tuempling, W.,
marine recirculating aquaculture systems. Aquac. Res. 49 (3), 1363–1367. https:// Palma, R., Encina, F., Figueroa, D., Kamjunke, N., Graeber, D., 2015. Tracing dis-
doi.org/10.1111/are.13562. solved organic matter (DOM) from land-based aquaculture systems in north
Görlitz, S., 2012. The Lugworm Abarenicola affinis (Arenicolidae, Polychaeta) in Tidal Patagonian streams. Sci. Total Environ. 537, 129–138. https://doi.org/10.1016/j.
Flats of Otago, Southern New Zealand. PhD-Thesis at University of Otago, New scitotenv.2015.07.160.
Zealand, pp. 188. http://hdl.handle.net/10523/2330. Olive, P. J. W., Craig, S., Cowin, P. B. D., 2006. Washington, DC: U.S. Patent and
Guerdat, T.C., Losordo, T.M., DeLong, D.P., Jones, R.D., 2013. An evaluation of solid Trademark Office. U.S. Patent No. 7,004,109.
waste capture from recirculating aquaculture systems using a geotextile bag system Olive, P.J., Duangchinda, T., Ashforth, E., Craig, S., Ward, A.C., Davies, S.J., 2009. Net
with a flocculant-aid. Aquac. Eng. 54, 1–8. https://doi.org/10.1016/j.aquaeng.2012. gain of long-chain polyunsaturated fatty acids (PUFA) in a lugworm Arenicola marina
10.001. bioturbated mesocosm. Mar. Ecol. Prog. Ser. 387, 223–239. https://doi.org/10.3354/
Guerrero, S., Cremades, J., 2012. Integrated Multi-Trophic Aquaculture (IMTA): A meps08088.
Sustainable, Pioneering Alternative for Marine Cultures in Galicia. Regional Pajand, Z.O., Soltani, M., Bahmani, M., Kamali, A., 2017. The role of polychaete Nereis
Government of Galicia, Spain, pp. 111. https://hal.archives-ouvertes.fr/hal- diversicolor in bioremediation of wastewater and its growth performance and fatty
00743395/. acid composition in an integrated culture system with Huso huso (Linnaeus, 1758).
Gunning, D., Maguire, J., Burnell, G., 2016. The development of sustainable saltwater- Aquac. Res. 48 (10), 5271–5279. https://doi.org/10.1111/are.13340.
based food production systems: a review of established and novel concepts. Water 8 Palmer, P.J., 2010. Polychaete-assisted sand filters. Aquaculture 306 (1–4), 369–377.
(12), 598. https://doi.org/10.3390/w8120598. https://doi.org/10.1016/j.aquaculture.2010.06.011.
Herath, S.S., Satoh, S., 2015. Environmental impact of phosphorus and nitrogen from Palmer, P.J., Wang, S., Houlihan, A., Brock, I., 2014. Nutritional status of a nereidid
aquaculture. In: Feed and Feeding Practices in Aquaculture, pp. 369–386. https://doi. polychaete cultured in sand filters of mariculture wastewater. Aquac. Nutr. 20 (6),
org/10.1016/B978-0-08-100506-4.00015-5. 675–691. https://doi.org/10.1111/anu.12129.
Hiebert, T.C., 2014. Abarenicola pacifica. In: Hiebert, T.C., Butler, B.A., Shanks, A.L. Palmer, P.J., Wang, S., Mann, D., Nash, W.J., 2018. Polybridge season 3: Ecosystem ef-
(Eds.), Oregon Estuarine Invertebrates: Rudys' Illustrated Guide to Common Species, fects of polychaete-assisted sand filters. In: Technical Report. Queensland
3 ed. University of Oregon Libraries and Oregon Institute of Marine Biology, Government, Brisbane, Australia, pp. 44. http://era.daf.qld.gov.au/id/eprint/6150/.
Charleston, Oregon. https://library.uoregon.edu/scilib/oimb/OEI. Pechenik, J.A., Cerulli, T.R., 1991. Influence of delayed metamorphosis on survival,
Honda, H., Kikuchi, K., 2002. Nitrogen budget of polychaete Perinereis nuntia vallata fed growth, and reproduction of the marine polychaete Capitella sp. I. J. Exp. Mar. Biol.
on the feces of Japanese flounder. Fish. Sci. 68 (6), 1304–1308. https://doi.org/10. Ecol. 151 (1), 17–27. https://doi.org/10.1016/0022-0981(91)90012-L.
1046/j.1444-2906.2002.00568.x. Piedrahita, R.H., 2003. Reducing the potential environmental impact of tank aquaculture
Hopkins, K.D., 1992. Reporting fish growth: a review of the basics. J. World Aquacult. effluents through intensification and recirculation. Aquaculture 226 (1–4), 35–44.
Soc. 23 (3), 173–179. https://doi.org/10.1111/j.1749-7345.1992.tb00766.x. https://doi.org/10.1016/S0044-8486(03)00465-4.
Hylleberg, J., 1975. Selective feeding by Abarenicola pacifica with notes on Abarenicola Pombo, A., Baptista, T., Granada, L., Ferreira, S.M., Gonçalves, S.C., Anjos, C., Sá, E.,
vagabunda and a concept of gardening in lugworms. Ophelia 14 (1–2), 113–137. Chainho, P., da Fonseca, L.C., e Costa, P.F., Costa, J.L., 2018. Insight into aqua-
https://doi.org/10.1080/00785236.1975.10421972. culture's potential of marine annelid worms and ecological concerns: a review. Rev.
Ito, M., Ito, K., Ohta, K., Hano, T., Onduka, T., Mochida, K., Fujii, K., 2016. Evaluation of Aquac. https://doi.org/10.1111/raq.12307.
bioremediation potential of three benthic annelids in organically polluted marine Retraubun, A.S.W., Dawson, M., Evans, S.M., 1996. Spatial and temporal factors affecting
sediment. Chemosphere 163, 392–399. https://doi.org/10.1016/j.chemosphere. sediment turnover by the lugworm Arenicola marina (L.). J. Exp. Mar. Biol. Ecol. 201
2016.08.046. (1–2), 23–35. https://doi.org/10.1016/0022-0981(96)00016-0.
Jørgensen, E.H., Christiansen, J.S., Jobling, M., 1993. Effects of stocking density on food Robinson, G., Caldwell, G.S., Jones, C.J.W., Slater, M.J., Stead, S.M., 2015. Redox stra-
intake, growth performance and oxygen consumption in Arctic charr (Salvelinus al- tification drives enhanced growth in a commercially important deposit-feeding in-
pinus). Aquaculture 110 (2), 191–204. https://doi.org/10.1016/0044-8486(93) vertebrate: implications for bioremediation and integrated aquaculture. Aquac.
90272-Z. Environ. Interact. 8, 1–13. https://doi.org/10.3354/aei00158.
Lamprianidou, F., 2015. Development of a Model for Evaluating and Optimizing the Seekamp, I., 2017. Utilization of Excess Nutrients from Land-Based Aquaculture Facilities
Performance of Integrated Multitrophic Aquaculture (IMTA) Systems. PhD-Thesis at by Hediste diversicolor (OF Müller, 1776). Production of Polychaete Biomass and its
University of Stirling, United Kingdom, pp. 226. https://dspace.stir.ac.uk/handle/ Potential Use in Fish Feed. Master-Thesis at the Norwegian University of Science and
1893/22916. Technology, Trondheim, Norway, pp. 96. http://hdl.handle.net/11250/2451314.
LeDuc, D., Probert, P.K., Frew, R.D., Hurd, C.L., 2006. Macroinvertebrate diet in intertidal Silva, N., Vargas, C.A., Prego, R., 2011. Land–ocean distribution of allochthonous organic
seagrass and sandflat communities: a study using C, N, and S stable isotopes. N. Z. J. matter in surface sediments of the Chiloé and Aysén interior seas (Chilean northern
Mar. Fresh. Res. 40 (4), 615–629. https://doi.org/10.1080/00288330.2006. Patagonia). Cont. Shelf Res. 31 (3–4), 330–339. https://doi.org/10.1016/j.csr.2010.
9517450. 09.009.
Luo, G.Z., Ma, N., Li, P., Tan, H.X., Liu, W., 2015. Enhancement of anaerobic digestion to Srithongouthai, S., Tada, K., 2017. Impacts of organic waste from a yellowtail cage farm
treat saline sludge from recirculating aquaculture systems. Sci. World J. 2015, 1–5. on surface sediment and bottom water in Shido Bay (the Seto Inland Sea, Japan).
https://doi.org/10.1155/2015/479101. Aquaculture 471, 140–145. https://doi.org/10.1016/j.aquaculture.2017.01.021.
MacDonald, C.L., Stead, S.M., Slater, M.J., 2013. Consumption and remediation of Suckow, B., Bischoff, A., Buck, B.H., Simon, M., 2010. Growth and biochemical compo-
European seabass (Dicentrarchus labrax) waste by the sea cucumber Holothuria for- sition of Nereis virens (Sars, 1835) applied as a bioconverter of solid waste from land-
skali. Aquac. Int. 21 (6), 1279–1290. https://doi.org/10.1007/s10499-013-9629-6. based fish culture. In: Seafarming Tomorrow, Aquaculture Europe 2010 - the Annual
Madariaga, S.T., Marín, S.L., 2017. Sanitary and environmental conditions of aquaculture Meeting of the European Aquaculture Society, October 5–8, Porto, Portugal, .
sludge. Aquac. Res. 48 (4), 1744–1750. https://doi.org/10.1111/are.13011. https://epic.awi.de/id/eprint/33982/.
Marques, B., Calado, R., Lillebø, A.I., 2017. New species for the biomitigation of a super- Tal, Y., Schreier, H.J., Sowers, K.R., Stubblefield, J.D., Place, A.R., Zohar, Y., 2009.
intensive marine fish farm effluent: combined use of polychaete-assisted sand filters Environmentally sustainable land-based marine aquaculture. Aquaculture 286 (1–2),
and halophyte aquaponics. Sci. Total Environ. 599, 1922–1928. https://doi.org/10. 28–35. https://doi.org/10.1016/j.aquaculture.2008.08.043.
1016/j.scitotenv.2017.05.121. Toulmond, A., 1975. Blood oxygen transport and metabolism of the confined lugworm
Marques, B., Lillebø, A.I., Ricardo, F., Nunes, C., Coimbra, M.A., Calado, R., 2018. Adding Arenicola marina (L.). J. Exp. Biol. 63 (3), 647–660.
value to ragworms (Hediste diversicolor) through the bioremediation of a super-in- van Rijn, J., 2013. Waste treatment in recirculating aquaculture systems. Aquac. Eng. 53,
tensive marine fish farm. Aquac. Environ. Interact. 10, 79–88. https://doi.org/10. 49–56. https://doi.org/10.1016/j.aquaeng.2012.11.010.
3354/aei00255. Vergara, J.M., Lopez-Calero, G., Robaina, L., Caballero, M.J., Montero, D., Izquierdo,
Martins, C.I.M., Eding, E.H., Verdegem, M.C., Heinsbroek, L.T., Schneider, O., M.S., Aksnes, A., 1999. Growth, feed utilization and body lipid content of gilthead
Blancheton, J.P., Roque d'Orbcastel, E., Verreth, J.A.J., 2010. New developments in seabream (Sparus aurata) fed increasing lipid levels and fish meals of different
recirculating aquaculture systems in Europe: a perspective on environmental sus- quality. Aquaculture 179 (1–4), 35–44. https://doi.org/10.1016/S0044-8486(99)
tainability. Aquac. Eng. 43 (3), 83–93. https://doi.org/10.1016/j.aquaeng.2010.09. 00150-7.

383
S. Gómez, et al. Aquaculture 507 (2019) 377–384

Wang, H., Seekamp, I., Malzahn, A., Hagemann, A., Carvajal, A.K., Slizyte, R., Standal, Animals. Academy Press, London (175 pp).
I.B., Handå, A., Reitan, K.I., 2019. Growth and nutritional composition of the poly- Yearsley, R.D., Jones, C.L., Britz, P.J., Vine, N.G., 2011. Integrated culture of silver kob
chaete Hediste diversicolor (OF Müller, 1776) cultivated on waste from land-based Argyrosomus inodorus and bloodworm Arenicola loveni loveni in abalone farm effluent.
salmon smolt aquaculture. Aquaculture 502, 232–241. https://doi.org/10.1016/j. Afr. J. Mar. Sci. 33 (2), 223–228. https://doi.org/10.2989/1814232X.2011.600290.
aquaculture.2018.12.047. Yogev, U., Sowers, K.R., Mozes, N., Gross, A., 2017. Nitrogen and carbon balance in a
Wells, G.P., 1966. The lugworm (Arenicola)—a study in adaptation. Neth. J. Sea Res. 3 novel near-zero water exchange saline recirculating aquaculture system. Aquaculture
(2), 294–313. https://doi.org/10.1016/0077-7579(66)90016-0. 467, 118–126. https://doi.org/10.1016/j.aquaculture.2016.04.029.
Weston, D.P., Penry, D.L., Gulmann, L.K., 2000. The role of ingestion as a route of con- Yokoyama, H., 2013. Growth and food source of the sea cucumber Apostichopus japonicus
taminant bioaccumulation in a deposit-feeding polychaete. Arch. Environ. Contam. cultured below fish cages — potential for integrated multi-trophic aquaculture.
Toxicol. 38 (4), 446–454. https://doi.org/10.1007/s002449910059. Aquaculture 372, 28–38. https://doi.org/10.1016/j.aquaculture.2012.10.022.
White, C.A., Woodcock, S.H., Bannister, R.J., Nichols, P.D., 2017. Terrestrial fatty acids as Zhang, X., Spanjers, H., van Lier, J.B., 2013. Potentials and limitations of biomethane and
tracers of finfish aquaculture waste in the marine environment. Rev. Aquac. https:// phosphorus recovery from sludges of brackish/marine aquaculture recirculation
doi.org/10.1111/raq.12230. systems: a review. J. Environ. Manag. 131, 44–54. https://doi.org/10.1016/j.
Winberg, G.G., Duncan, A., 1971. Methods for the Estimation of Production of Aquatic jenvman.2013.09.016.

384