The impact of ocean acidification,

temperature and a bacterial challenge on

the immune response of Mytilus edulis

Robert. P. Ellis*, H. Parry, S. Widdicombe,

T.H. Hutchinson, R. Pipe and J. I. Spicer
Ocean acidification – the other CO2 problem
Ocean acidification – the other CO2 problem

?
 Ocean acidification – the other CO2 problem

Physiology Behaviour

Morphology Life History

Ecosystem
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 Ocean acidification – the other CO2 problem

Physiology Behaviour

Morphology Immune Life History

System?

Ecosystem
?
 OA and the immune system – what do we know?

One published study to date – Bibby et

al. 2008

32 day study looking at several immune

parameters

OA suppressed phagocytosis in mussels

after 32 days, reducing immune capacity

Immune system
understudied with
respect to OA
 So why study the immune system?

Forms a major physiological mechanisms to ensure

host survival

Important to virtually all animals, an integral

component of the stress response (Snow 2007)

Offers an ideal system to study the ecological and

evolutionary impact of stressors

e.g. Temperature - (e.g. Monari 2007),

Salinity - (e.g. Gagniaire 2006),
Hypoxia - (e.g. Cheng et al 2004),
Anoxia – (e.g. Matozzo et al 2005)

(Wootton et al 2006)
 Experimental approach

The mussel, Mytilus edulis, maintained under

one of five pH treatments (8.05, 7.8, 7.6, 7.35 or 6.5)
and one of two temperatures (12ºC or 16ºC) for a total of 90 days

Immune response measured in surviving mussels after 83

days of exposure to low pH and increased temperature, (T0)

Remaining mussels in each aquaria inoculated with

pathogenic bacteria, Vibrio tubiashii
From Widdicombe and Needham 2007

Immune response measured again 24 hours (T1) and 7

days (T7) after bacterial challenge
 Experimental approach

Parameters measured

• Organism survival
•Total haemocyte counts
•Differential haemocyte counts
•Antibacterial activity of cell-free haemolymph

From Widdicombe and Needham 2007

Results
 Results – Impaired survival

12ºC 16ºC
100 100

80 80

60
% Survival

60

% Survival
40 40
8.05
7.8 8.05
7.6 7.8
7.35 7.6
20 20 7.35
6.5
6.5

0 0
1 21 42 56 77 91 1 21 42 56 77 91

Exposure duration (Days) Exposure duration (Days)

At low pH mussel survival significantly impaired

Increased temperature increases the sensitivity of mussels exposed to low pH

No additional mortality post bacterial inoculation

 Results – Immune system plasticity
Antibacterial activity of mussel haemolymph
1.2
T0 Post inoculation
1.0 * 0.4
*
0.3

Change in Growth Inhibition

0.8
Growth Inhibition

0.2
0.6

0.4 0.1

0.2 0.0

0.0 -0.1
8.00 7.80 7.60 7.35 6.50 8.00 7.80 7.60 7.35 6.50

pH pH

At low pH mussels have reduced ability to fight bacterial infection

Yet on exposure to bacteria mussels at low pH are able to

increase antibacterial activity
 Results – Gender differences
Antibacterial activity of mussel haemolymph

T0 Post inoculation
1.2
* 0.08

1.0 0.06

Change in Growth Inhibition

0.04
0.8
Growth Inhibition

0.02 *
0.6
0.00

-0.02
0.4

-0.04

0.2
-0.06

0.0 -0.08
Female Male
Female Male

Females have a higher antibacterial activity than males

Additionally females better able to up regulate antibacterial activity after

exposure to bacteria
 Results – Gender differences
Total cell count Differential cell count
Post inoculation Post inoculation
1000000 0.15

800000
* T1
T7

Change in proportion of granulocytes

* 0.10
Change in number of haemocytes

600000

400000
0.05
200000

0
0.00
-200000

-400000

-600000

-800000
-0.05
**
-0.10
-1000000

**
-1200000
Female Male -0.15
Female Male

No difference in the response of males or females at T0

Gender significantly affects change in total cell count, with females reducing
haemocyte numbers after bacterial inoculation and males increasing THC

Gender and stage post inoculation significantly interact to affect differential

cell counts
 Summary

• Exposure to low pH significantly reduces mussel survival, this is

compounded by an increase in temperature

• Mussels can trade-off immune response when exposed to stressful environmental

conditions, but critically maintain ability to increase it again when needed

• Significant differences noted in the immune response of males and females

exposed to stressful environmental conditions
 Future work
Histology

Metabolomics and Shell strength and

Lipid profiling composition

Antimicrobial peptide
From Grollman et al. (2007) PNAS, 104: 12129
expression
 Acknowledgements

Travel grant funding:

SEB - CoB travel grant
Marine Institute – Marine Institute Education Fund
Thank you for listening
Any questions?