230
A.A. Martinez et al.
and growth alterations (Santodonato 1997; Morton 1988; Bostr¨ om et al. 2002;
Pufulete et al. 2004; Ono et al. 2008; Veeramachaneni 2008; Sanders et al. 2009).
In marine invertebrates, histopathological and endocrine alterations have also been
related to exposure by PAHs and others chemicals (Alonso et al. 2001; Aarab et al.
2011; Au 2004; Lavado et al. 2006; Ortiz-Zarragoitia and Cajaraville 2006; JingJing et al. 2009; Sch¨ afer and K¨ ohler 2009; Ruiz et al. 2011). However, few studies
have been undertaken to demonstrate these alterations in the laboratory.
Purpose
Taking into account the concentrations of accumulated hydrocarbons measured in
mussels cultured in the R´ ıa of Vigo by Ruiz et al. (2011), we conducted a study,
under laboratory controlled conditions, to determine the gonadal histopathologies
induced by a tar mixture, commonly used in cleaning mussel culture rafts.
Method
Analytical Composition of Tar
The chemical analysis of the tar mixture was contracted to the Spanish Institute of Oceanography (IEO, Center of Vigo), where analytical quality control is
supported by the international intercalibration exercises from Quality Assurance
of Information for Marine Environmental Monitoring in Europe (QUASIMEME
2003) or International Atomic Energy Agency (IAEA) (Villeneuve et al. 2004). The
separation and quantification of PAHs was carried out by high-performance liquid
chromatography (HPLC) with fluorimetric detection according to Vi˜ nas Di´ eguez
(2002). A total of 13 PAHs were quantified: phenanthrene (Phe), anthracene
(An), fluoranthene (Flt), pyrene (Py), benzo(a)anthracene (BaA), chrysene (Chry),
benzo(e)pyrene (BeP), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF),
benzo(a)pyrene (BaP), dibenzo(a,h)anthracene (DBA), benzo(g,h,i)perylene (Bper)
and indeno(1,2,3,-c,d)pyrene (IP). Results were expressed as mg/kg of mussel
tissue.
Experimental Design
A total of 200 mussels (Mytilus galloprovincialis), between 6 and 8 cm in valve
length were randomly collected from floating rafts in the R´ ıa of Vigo (NW Spain)
in April of 2009. In the laboratory, mussels were placed in five 30 L polyethylene
tanks in an open system of 20 m filtered seawater, with a flow rate of 2 mL/min.
A.A. Martinez et al.
and growth alterations (Santodonato 1997; Morton 1988; Bostr¨ om et al. 2002;
Pufulete et al. 2004; Ono et al. 2008; Veeramachaneni 2008; Sanders et al. 2009).
In marine invertebrates, histopathological and endocrine alterations have also been
related to exposure by PAHs and others chemicals (Alonso et al. 2001; Aarab et al.
2011; Au 2004; Lavado et al. 2006; Ortiz-Zarragoitia and Cajaraville 2006; JingJing et al. 2009; Sch¨ afer and K¨ ohler 2009; Ruiz et al. 2011). However, few studies
have been undertaken to demonstrate these alterations in the laboratory.
Purpose
Taking into account the concentrations of accumulated hydrocarbons measured in
mussels cultured in the R´ ıa of Vigo by Ruiz et al. (2011), we conducted a study,
under laboratory controlled conditions, to determine the gonadal histopathologies
induced by a tar mixture, commonly used in cleaning mussel culture rafts.
Method
Analytical Composition of Tar
The chemical analysis of the tar mixture was contracted to the Spanish Institute of Oceanography (IEO, Center of Vigo), where analytical quality control is
supported by the international intercalibration exercises from Quality Assurance
of Information for Marine Environmental Monitoring in Europe (QUASIMEME
2003) or International Atomic Energy Agency (IAEA) (Villeneuve et al. 2004). The
separation and quantification of PAHs was carried out by high-performance liquid
chromatography (HPLC) with fluorimetric detection according to Vi˜ nas Di´ eguez
(2002). A total of 13 PAHs were quantified: phenanthrene (Phe), anthracene
(An), fluoranthene (Flt), pyrene (Py), benzo(a)anthracene (BaA), chrysene (Chry),
benzo(e)pyrene (BeP), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF),
benzo(a)pyrene (BaP), dibenzo(a,h)anthracene (DBA), benzo(g,h,i)perylene (Bper)
and indeno(1,2,3,-c,d)pyrene (IP). Results were expressed as mg/kg of mussel
tissue.
Experimental Design
A total of 200 mussels (Mytilus galloprovincialis), between 6 and 8 cm in valve
length were randomly collected from floating rafts in the R´ ıa of Vigo (NW Spain)
in April of 2009. In the laboratory, mussels were placed in five 30 L polyethylene
tanks in an open system of 20 m filtered seawater, with a flow rate of 2 mL/min.