240
Y. Ruiz et al.
Purpose
The aim of this study is to describe the neoplastic disorders observed in the gonad
(mantle tissue) of M. galloprovincialis in the R´ ıa of Vigo, discussing their possible
relationship with the accumulation of persistent pollutants. This would allow one to
use these pathologies as indicators of the pollution risks on mussel farms as well as
an indicator of human food safety.
Material and Methods
Mussels (M. galloprovincialis Lmk.) were collected fortnightly between February
2001 and August 2002 from the floating rafts where are cultured in the farms of
the R´ ıa of Vigo (Galicia, NW Spain). For each sampling, 30 adult individuals from
6 to 8 cm in length were randomly collected at depths of 5 m. A small section
(0.5 1 cm) of the central part of the gonad (mantle tissue) was fixed in Bouin’s
solution, dehydrated and embedded in paraffin. Sections 5 m thick were stained
with Harris’ haematoxylin and eosin for histological analysis. The remaining tissues
were pooled, homogenized in a Virtis microprocessor of tissues, immediately frozen
and lyophilized for chemicals analysis.
For determination of the gametogenic stage the model proposed by Lubet (1959)
and Su´ arez et al. (2005) were followed. The comparative microscopical analysis of
histological slides allowed the observation of several histopathological alterations.
From lyophilized samples, PAHs, organochlorine compounds (PCBs and
OCPs) and trace metals were extracted and quantificated according to Vi˜ nas
Di´ eguez et al. (2002), De Boer (1988), Gonz´ alez-Quijano and Fumega (1996),
and Besada et al. (2002), respectively. A total of 13 PAHs were analyzed:
phenantrene, anthracene, pyrene, fluoranthene, benzo(a)anthracene, chrysene,
benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(e)pyrene, benzo(a)pyrene,
dibenzo(a,h)anthracene, benzo(g,h,i)perylene and indeno(1,2,3-cd)pyrene; 16
organochlorine compounds were measured: 10 PCBs (CB:28, 31, 52, 101, 105,
118, 138, 153, 156 and 180) and 6 OCPs (’HCH, ”HCH, p,p
0 -DDD, p,p
0 -DDE,
o,p
0 -DDT, p,p
0 -DDT); and Cd, Cu, Hg, As, Pb and Zn as trace metals.
Statistical analysis was performed using SPSS Package 15.0 (SPSS Inc., Microsoft Co.). The variation of each variable was assessed by one-way analysis of
variance (ANOVA). The level of significance was at p < 0.05. Correlations among
variables were computed from Pearson coefficient (r). Multivariate analyses of
principal component (PCA) and aggregation analyses (cluster) were carried out
in order to determine the relationship between the analyzed variables and their
distributions. Finally, an overlaying of the cluster analyses to PCA was performed
using PRIMER 6.0 program (Clarke and Gorley 2006).
Y. Ruiz et al.
Purpose
The aim of this study is to describe the neoplastic disorders observed in the gonad
(mantle tissue) of M. galloprovincialis in the R´ ıa of Vigo, discussing their possible
relationship with the accumulation of persistent pollutants. This would allow one to
use these pathologies as indicators of the pollution risks on mussel farms as well as
an indicator of human food safety.
Material and Methods
Mussels (M. galloprovincialis Lmk.) were collected fortnightly between February
2001 and August 2002 from the floating rafts where are cultured in the farms of
the R´ ıa of Vigo (Galicia, NW Spain). For each sampling, 30 adult individuals from
6 to 8 cm in length were randomly collected at depths of 5 m. A small section
(0.5 1 cm) of the central part of the gonad (mantle tissue) was fixed in Bouin’s
solution, dehydrated and embedded in paraffin. Sections 5 m thick were stained
with Harris’ haematoxylin and eosin for histological analysis. The remaining tissues
were pooled, homogenized in a Virtis microprocessor of tissues, immediately frozen
and lyophilized for chemicals analysis.
For determination of the gametogenic stage the model proposed by Lubet (1959)
and Su´ arez et al. (2005) were followed. The comparative microscopical analysis of
histological slides allowed the observation of several histopathological alterations.
From lyophilized samples, PAHs, organochlorine compounds (PCBs and
OCPs) and trace metals were extracted and quantificated according to Vi˜ nas
Di´ eguez et al. (2002), De Boer (1988), Gonz´ alez-Quijano and Fumega (1996),
and Besada et al. (2002), respectively. A total of 13 PAHs were analyzed:
phenantrene, anthracene, pyrene, fluoranthene, benzo(a)anthracene, chrysene,
benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(e)pyrene, benzo(a)pyrene,
dibenzo(a,h)anthracene, benzo(g,h,i)perylene and indeno(1,2,3-cd)pyrene; 16
organochlorine compounds were measured: 10 PCBs (CB:28, 31, 52, 101, 105,
118, 138, 153, 156 and 180) and 6 OCPs (’HCH, ”HCH, p,p
0 -DDD, p,p
0 -DDE,
o,p
0 -DDT, p,p
0 -DDT); and Cd, Cu, Hg, As, Pb and Zn as trace metals.
Statistical analysis was performed using SPSS Package 15.0 (SPSS Inc., Microsoft Co.). The variation of each variable was assessed by one-way analysis of
variance (ANOVA). The level of significance was at p < 0.05. Correlations among
variables were computed from Pearson coefficient (r). Multivariate analyses of
principal component (PCA) and aggregation analyses (cluster) were carried out
in order to determine the relationship between the analyzed variables and their
distributions. Finally, an overlaying of the cluster analyses to PCA was performed
using PRIMER 6.0 program (Clarke and Gorley 2006).