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Table 5.

Summary of the split-split plot four-way ANOVAs on the effect of overwintering temperature, bivalve species, tissue and sampling date on the phospholipid to sterol ratio, unsaturation index, total PUFA and major individual PUFA of polar lipids

PL/ST
Unsaturation
PUFA
22:6n-3
20:5n-3
20:4n-6
22:2 NMI
20:2 NMI
Source of variationd.f.FPFPFPFPFPFPFPFP
Main plot analysis                  
   T°C 1.4 0.373 1.1 0.446 0.4 0.677 2.6 0.225 2.1 0.264 1.6 0.343 1.4 0.372 2.6 0.222 
   Error A                 
Subplot analysis                  
   Sp. 73.2 0.003 238.8 0.001 625.4 0.001 6.0 0.090 161.6 0.001 0.4 0.588 470.7 <0.001 1522.1 <0.001 
   T°C×Sp. 0.0 0.967 0.5 0.644 0.5 0.652 0.6 0.621 1.5 0.356 0.8 0.517 0.8 0.517 2.0 0.280 
   Error B                 
Sub-subplot analysis                  
Tissue 103.2 <0.001 7.6 0.008 151.7 <0.001 24.6 <0.001 165.4 <0.001 221.9 <0.001 162.0 <0.001 150.5 <0.001 
   T°C×Tissue 1.2 0.313 0.3 0.710 0.3 0.723 0.4 0.673 1.0 0.376 0.1 0.934 0.4 0.668 0.9 0.432 
   Species×Tissue 0.1 0.733 6.2 0.016 0.0 0.973 1.0 0.311 73.3 <0.001 2.0 0.161 2.2 0.140 224.0 <0.001 
   T°C×Species×Tissue 0.6 0.537 0.2 0.807 0.4 0.652 0.1 0.932 1.0 0.375 0.3 0.758 0.5 0.585 0.6 0.573 
Date 10.0 <0.001 60.6 <0.001 31.4 <0.001 55.6 <0.001 122.4 <0.001 88.4 <0.001 4.1 0.006 1.2 0.313 
   T°C×Date 0.3 0.965 1.1 0.379 0.5 0.845 1.1 0.356 2.1 0.049 0.6 0.778 1.0 0.464 1.0 0.460 
   Species×Data 1.1 0.348 0.8 0.550 0.1 0.973 2.1 0.095 7.7 <0.001 26.6 <0.001 5.1 0.002 1.8 0.136 
   T°C×Species×Date 0.4 0.921 0.7 0.715 0.5 0.823 1.1 0.383 1.4 0.209 2.6 0.017 1.9 0.083 1.4 0.234 
Tissue×Date 1.0 0.402 4.5 0.003 6.4 0.001 2.1 0.100 3.0 0.029 2.5 0.053 2.3 0.075 1.0 0.431 
   T°C×Tissue×Date 1.8 0.107 1.0 0.448 1.1 0.437 1.0 0.479 0.7 0.681 1.2 0.348 1.6 0.139 1.1 0.358 
   Species×Tissue×Date 1.0 0.412 0.9 0.492 0.6 0.639 0.4 0.803 1.0 0.413 1.3 0.295 0.3 0.869 2.4 0.064 
   T°C×Species×Tissue×Date 1.6 0.157 0.7 0.662 0.8 0.576 0.7 0.721 1.3 0.246 1.3 0.268 1.0 0.433 0.2 1.340 
Error C 54                  
Error A: Tank (T°C)                  
Error B: Species×Tank (T°C)                  
Error C: Tissue×Tank (T°C), d.f.=3; Species×Tissue×Tank(T°C), d.f.=3; Date×Tank (T°C), d.f.=12;Species×Date×Tank (T°C), d.f.=12; Tissue×Date×Tank(T°C), d.f.=12; Species×Tissue×Date×Tank (T°C),d.f.=12                  
PL/ST
Unsaturation
PUFA
22:6n-3
20:5n-3
20:4n-6
22:2 NMI
20:2 NMI
Source of variationd.f.FPFPFPFPFPFPFPFP
Main plot analysis                  
   T°C 1.4 0.373 1.1 0.446 0.4 0.677 2.6 0.225 2.1 0.264 1.6 0.343 1.4 0.372 2.6 0.222 
   Error A                 
Subplot analysis                  
   Sp. 73.2 0.003 238.8 0.001 625.4 0.001 6.0 0.090 161.6 0.001 0.4 0.588 470.7 <0.001 1522.1 <0.001 
   T°C×Sp. 0.0 0.967 0.5 0.644 0.5 0.652 0.6 0.621 1.5 0.356 0.8 0.517 0.8 0.517 2.0 0.280 
   Error B                 
Sub-subplot analysis                  
Tissue 103.2 <0.001 7.6 0.008 151.7 <0.001 24.6 <0.001 165.4 <0.001 221.9 <0.001 162.0 <0.001 150.5 <0.001 
   T°C×Tissue 1.2 0.313 0.3 0.710 0.3 0.723 0.4 0.673 1.0 0.376 0.1 0.934 0.4 0.668 0.9 0.432 
   Species×Tissue 0.1 0.733 6.2 0.016 0.0 0.973 1.0 0.311 73.3 <0.001 2.0 0.161 2.2 0.140 224.0 <0.001 
   T°C×Species×Tissue 0.6 0.537 0.2 0.807 0.4 0.652 0.1 0.932 1.0 0.375 0.3 0.758 0.5 0.585 0.6 0.573 
Date 10.0 <0.001 60.6 <0.001 31.4 <0.001 55.6 <0.001 122.4 <0.001 88.4 <0.001 4.1 0.006 1.2 0.313 
   T°C×Date 0.3 0.965 1.1 0.379 0.5 0.845 1.1 0.356 2.1 0.049 0.6 0.778 1.0 0.464 1.0 0.460 
   Species×Data 1.1 0.348 0.8 0.550 0.1 0.973 2.1 0.095 7.7 <0.001 26.6 <0.001 5.1 0.002 1.8 0.136 
   T°C×Species×Date 0.4 0.921 0.7 0.715 0.5 0.823 1.1 0.383 1.4 0.209 2.6 0.017 1.9 0.083 1.4 0.234 
Tissue×Date 1.0 0.402 4.5 0.003 6.4 0.001 2.1 0.100 3.0 0.029 2.5 0.053 2.3 0.075 1.0 0.431 
   T°C×Tissue×Date 1.8 0.107 1.0 0.448 1.1 0.437 1.0 0.479 0.7 0.681 1.2 0.348 1.6 0.139 1.1 0.358 
   Species×Tissue×Date 1.0 0.412 0.9 0.492 0.6 0.639 0.4 0.803 1.0 0.413 1.3 0.295 0.3 0.869 2.4 0.064 
   T°C×Species×Tissue×Date 1.6 0.157 0.7 0.662 0.8 0.576 0.7 0.721 1.3 0.246 1.3 0.268 1.0 0.433 0.2 1.340 
Error C 54                  
Error A: Tank (T°C)                  
Error B: Species×Tank (T°C)                  
Error C: Tissue×Tank (T°C), d.f.=3; Species×Tissue×Tank(T°C), d.f.=3; Date×Tank (T°C), d.f.=12;Species×Date×Tank (T°C), d.f.=12; Tissue×Date×Tank(T°C), d.f.=12; Species×Tissue×Date×Tank (T°C),d.f.=12                  

PL/ST, phospholipid to sterol ratio

Independent variables were overwintering temperature (T°C; 0°C,4°C and 9°C), bivalve species (mussel and oyster), tissue (gills and digestive glands) and sampling date (31 January, 14 February, 12 April, 4 May and 8 June)

Significant probabilities are in bold. Data of PL/ST, 20:2 and 22:2 NMI were log+1 transformed

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