Immune parametres of mussels
treated with Prestige fuel oil
M. Camino Ordás, Joan Albaiges, Josep M. Bayona,
Amando Ordás and Antonio Figueras
Instituto de Investigaciones Marinas (Vigo)
Centro de Investigación y Desarrollo (Barcelona)
Misión Biológica de Galicia (Pontevedra)
Immune system of bivalve molluscs
Inespecific system  no antibody production
 External barriers
 Hemocytic infiltration
Cellular defense
HEMOCYTES
•
•
•
Phagocytosis
Respiratory burst
NO production
Humoral defense
SERUM SOLUBLE FACTORS
•
•
•
Lysozyme
Lectins
Prophenoloxidase
contaminants
depression in immune system
infection by pathogens
MORTALITY
important economical losses in Galicia, one of the
main Mediterranean mussel producers worldwide
Objective of this work
To assess the influence of the Prestige fuel oil on the
immune response of Mediterranean mussel.
contaminants
depression in immune system
infection by pathogens
MORTALITY
important economical losses in Galicia, one of the
main Mediterranean mussel producers worldwide
Objective of this work
To assess the influence of the Prestige fuel oil on the
immune response of Mediterranean mussel.
contaminants
depression in immune system
infection by pathogens
MORTALITY
important economical losses in Galicia, one of the
main Mediterranean mussel producers worldwide
Objective of this work
To assess the influence of the Prestige fuel oil on the
immune response of Mediterranean mussel.
contaminants
depression in immune system
infection by pathogens
MORTALITY
important economical losses in Galicia, one of the
main Mediterranean mussel producers worldwide
Objective of this work
To assess the influence of the Prestige fuel oil on the
immune response of Mediterranean mussel.
contaminants
depression in immune system
infection by pathogens
MORTALITY
important economical losses in Galicia, one of the
main Mediterranean mussel producers worldwide
Objective of this work
To assess the influence of the Prestige fuel oil on the
immune response of Mediterranean mussel.
tank 1
tank 2
tank 3
Methodology
Monthly sampling
during four months
+1 kg fuel +2 kg fuel
control
•Histology.
•Immune parametres:
 Cellular: hemocyte viability, phagocytosis, NO production,
chemiluminescence (CL).
 Humoral: protein concentration and lysozime activity.
•PAH concentration in water and mussel tissue (gills
and digestive gland).
Results
1. HISTOLOGY
- No granulocytomes or neoplasia
- Mytilicola in 68% of sampled animals
2. IMMUNE PARAMETRES
2.1. Mean squares (34 factorial ANOVA):
Month (M)
Treatment (T)
MT
Error
df
Viab.
3
2
6
36
0.008**
0.002
0.008**
0.002
Phagoc.
252.75
266.59
143.37
146.12
CL
0.0003**
0.0001*
0.0002**
0.0000
NO
5273.16**
536.08
2611.54*
1015.16
Protein Lisoz.
0.043
0.081*
0.010
0.020
70.41**
54.98**
24.92*
8.65
•No significant differences in phagocytic activity and significant
differences in protein concentration.
•Remaining parametres: interaction between month and treatment.
2.2. One way ANOVA (without taking into account the
sampling month): significant differences in protein
concentration and lysozyme activity.
2.3. Least square means:
Treatment
Viab.
Phagoc.
CL
NO
Protein
Lysoz.
2 kg
1 kg
0.220a
0.219a
11.080a
13.890a
0.015a
0.013a
52.770a
50.120a
0.242a
7.507ab
0.283ab 9.636a
0 kg
0.236a
19.108a
0.010a
61.200a
0.381b
5.943b
Means followed by the same letter in a column are not statistically different according
to the Waller-Duncan k- ratio t Test.
3. PAH CONCENTRATION
Prestige fuel oil
Similar methylphenantrene profiles in mussel organs
and Prestige fuel oil  real exposure to the fuel.
TPAH gill
•Higher TPAH
concentration (ng/ g)
in digestive gland than
in gill.
•Low TPAH conc. in
control mussels.
TPAH dig.
gland
•Excepting gill from
the first month, organs
treated with 2 kg of
fuel show higher TPAH
conc. than 1 kgtreated ones.
REGRESSION PROCEDURE:
^
• Model: [contam]=
0 + 1  MONTH + 2  TREATMENT
• Number of PAHs depending on each independent variable:
GLAND
GILL
WATER
only
treatment
9
3
-
only
month
4
-
treatment
no
+ month dependent
7
5
1
21
25
For comparisons between [PAH] and immune parametres, it
would be more appropriate to use data from digestive gland,
since [PAH] in gill is independent of our experimental design.
Future analysis
Comparison between gland [PAH] and immune data (difficult due
to the experimental design).
Muchas gracias