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)

Inespecific system no antibody production

Cellular defense Humoral defense

• Phagocytosis• Respiratory burst• NO production

• Lysozyme• Lectins• Prophenoloxidase

SERUM SOLUBLE FACTORSHEMOCYTES

External barriers

Hemocytic infiltration

Immune system of bivalve molluscs

contaminants

depression in immune system

infection by pathogens

MORTALITYimportant economical losses in Galicia, one of the main Mediterranean mussel producers worldwide

To assess the influence of the Prestige fuel oil on the immune response of Mediterranean mussel.

Objective of this work

contaminants

depression in immune system

infection by pathogens

MORTALITYimportant economical losses in Galicia, one of the main Mediterranean mussel producers worldwide

To assess the influence of the Prestige fuel oil on the immune response of Mediterranean mussel.

Objective of this work

contaminants

depression in immune system

infection by pathogens

MORTALITYimportant economical losses in Galicia, one of the main Mediterranean mussel producers worldwide

To assess the influence of the Prestige fuel oil on the immune response of Mediterranean mussel.

Objective of this work

contaminants

depression in immune system

infection by pathogens

MORTALITYimportant economical losses in Galicia, one of the main Mediterranean mussel producers worldwide

To assess the influence of the Prestige fuel oil on the immune response of Mediterranean mussel.

Objective of this work

contaminants

depression in immune system

infection by pathogens

MORTALITYimportant economical losses in Galicia, one of the main Mediterranean mussel producers worldwide

To assess the influence of the Prestige fuel oil on the immune response of Mediterranean mussel.

Objective of this work

Methodology

+1 kg fuel +2 kg fuel control

Monthly sampling during four months

•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).

tank 1 tank 2 tank 3

Results

- No granulocytomes or neoplasia

- Mytilicola in 68% of sampled animals

1. HISTOLOGY

2. IMMUNE PARAMETRES

df Viab. Phagoc. CL NO Protein Lisoz.

Month (M) 3 0.008** 252.75 0.0003** 5273.16** 0.043 70.41**

Treatment (T) 2 0.002 266.59 0.0001* 536.08 0.081* 54.98**

M T 6 0.008** 143.37 0.0002** 2611.54* 0.010 24.92*

Error 36 0.002 146.12 0.0000 1015.16 0.020 8.65

2.1. Mean squares (34 factorial ANOVA):

•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 0.220a 11.080a 0.015a 52.770a 0.242a 7.507ab

1 kg 0.219a 13.890a 0.013a 50.120a 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

0,00

20,00

40,00

60,00

80,00

100,00

120,00

140,00

gill 1 kg gill 2 kg dig. gland1 kg

dig. gland2 kg

3 MP

2 MP

9 MP

1 MP

Prestige fuel oil

0

200400

600

8001000

1200

14001600

1800

Similar methylphenantrene profiles in mussel organs

and Prestige fuel oil real exposure to the fuel.

0,00

500,00

1000,00

1500,00

2000,00

2500,00

3000,00

3500,00

mes 1 mes 2 mes 3 mes 4

1 kg2 kgcontrol

0,00

5000,00

10000,00

15000,00

20000,00

25000,00

mes 1 mes 2 mes 3 mes 4

1 kg2 kgcontrol

TPAH gill

TPAH dig.gland

•Higher TPAH concentration (ng/ g) in digestive gland than in gill.

•Low TPAH conc. in control mussels.

•Excepting gill from the first month, organs treated with 2 kg of fuel show higher TPAH conc. than 1 kg- treated ones.

REGRESSION PROCEDURE:

• Model: [contam]=0 + 1 MONTH + 2 TREATMENT

• Number of PAHs depending on each independent variable:

^

only treatment

only month

treatment + month

no dependent

GLAND 9 4 7 5GILL 3 - 1 21WATER - - - 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.

Comparison between gland [PAH] and immune data (difficult due to

the experimental design).

Future analysis

Muchas gracias