TitleResponse of a marine benthic invertebrate community and biotic indices to organic enrichment from

sewage disposal

Fiona E. Culhane*, Robert A. Briers, Paul Tett, Teresa F. Fernandes

*Corresponding author: Email: F.Culhane@liverpool.ac.uk Tel. +44 (0) 151 795 4646

Supporting Information

1

Table S1 Details of sample points at Ironotter PointStation Distance

from discharge

(m)

Depth (m)

Number of replicates in each year

1989 1992 1995 1998

A1 100 23 3 3 3 3A2 500 30 3 3 3 2A3 750 30 3 3 3 2A4 1000 - 0 0 3 0B1 100 23 2 3 3 0B2 500 - 0 0 3 0B3 750 28 3 3 3 0B4 1000 - 0 0 3 0C1 100 20 3 3 3 0C2 500 20 3 3 3 0C3 750 25 3 3 3 0D1 100 18 3 3 3 0D2 500 18 2 3 3 0D3 750 10 3 3 3 0E1 100 20 3 3 3 3E2 500 26 3 3 3 2E3 750 20 3 3 3 2E4 1000 - 0 3 0 0F1 100 20 3 3 3 0F2 500 28 3 3 3 0F3 750 31 3 3 3 0G1 100 22 3 3 3 0G2 500 28 3 3 3 0G3 750 30 3 3 3 0G4 1000 - 0 0 3 0H1 100 20 3 3 3 0H2 500 24 3 3 3 0H3 750 - 0 3 0 0

Notes on sampling

The team of taxonomists did not change over the course of the surveys and had a common leader throughout, therefore bias in species identification by different taxonomists is assumed not be a factor in this study.

The change in sampling regime in 1998 was due to a change in priorities and distribution of funding at this time, resulting in a minimal survey.

2

Testing for effects of autospatial correlation

Alternative models were fit to the data with and without an exponential spatial error term based on sample site coordinates using the ‘gls’ function within the ‘nlme’ package (Pinheiro et al. 2019) in R. A likelihood ratio test indicated that there was no significant difference between the fit of the two models (Likelihood ratio = 3.20 x10-8, p=0.9999). Alternative forms of the spatial error function (linear and spherical) did not affect the outcome.

A summary of the outcome of the two models is shown below.

Non-spatial model

Organic Carbon

df F p

Distance 8 2.56 0.023

Year 1 133.25 <0.0001

Transect 14 0.76 0.701

Distance*Year 1 12.83 0.0009

Distance*Transect 7 0.25 0.969

Year*Transect 7 0.45 0.862

Distance*Year*Transec

t

7 0.12 0.996

Exponential Spatial Error model

Organic Carbon

df Chi-squared p

Distance 8 20.49 <0.001

Year 1 133.25 <0.0001

Transect 14 10.67 0.712

Distance*Year 1 12.83 <0.0001

Distance*Transect 7 1.75 0.972

Year*Transect 7 3.16 0.869

Distance*Year*Transec

t

7 0.86 0.996

Reference:

3

Pinheiro J, Bates D, DebRoy S, Sarkar D, R Core Team (2019). _nlme: Linear and Nonlinear Mixed Effects Models. R package version 3.1-140, <URL: https://CRAN.R-project.org/package=nlme>.

4

Details of indices used in this assessment

Table S2 Assigned quality classifications according to index values for some commonly used indicesIndex Boundaries QualityAMBI (Borja, 2004) AMBI≤1.2

1.2<AMBI≤3.33.3<AMBI≤4.34.3<AMBI≤5.5AMBI>5.5

HighGoodModeratePoorBad

BOPA (Dauvin and Ruellet, 2007)

0.00000 ≤BOPA≤0.045760.04576<BOPA≤0.139660.13966<BOPA≤0.193820.19382<BOPA≤0.267610.26761<BOPA≤0.30103

HighGoodModeratePoorBad

ITI (Word, 1979) ≥80-100≥60-80≥30-600-30

Reference conditionsNormal conditionsChanged conditionsDegraded conditions

IQI(WFD-UKTAG, 2008)

≥0.750.64≤IQI<0.750.44≤IQI<0.640.24≤IQI<0.44IQI<0.24

HighGoodModeratePoorBad

BQI (Rosenberg et al, 2004; Culhane, 2012)

>17.28>12.96≤17.28>8.64≤12.96>4.32≤8.64≤4.32

HighGoodModeratePoorBad

5

Effect of distance on index quality

1989 1992 1995 19981020304050607080

Spec

ies R

ichne

ss, S

1989 1992 1995 19980

500

1000

1500

2000

2500

3000

Abun

danc

e, N

1989 1992 1995 19981

1.5

2

2.5

3

3.5

4

AMBI

1989 1992 1995 19980.6

0.620.640.660.68

0.70.720.740.760.78

IQI

1989 1992 1995 199815

25

35

45

55

65

ITI

1989 1992 1995 199889

1011121314151617

BQI

1989 1992 1995 199870

72

74

76

78

80

82

84

Taxo

nom

ic Di

stinc

tnes

s

1989 1992 1995 199805

1015202530354045

A/S

1989 1992 1995 19980

0.020.040.060.08

0.10.120.140.160.18

0.2

BOPA

1989 1992 1995 19980.4

0.450.5

0.550.6

0.650.7

0.750.8

0.850.9

Piel

ou's

Even

ness

, J'

Figure S1 Average values of indices with standard deviation. Blue line: shows all data from all years; Orange line: shows data only from transects A and E which were the only transects sampled in all years; Grey line: shows data only from sample points 100m and 500m from transects A and E (closest to the outfall where greatest impacts would be expected). 1989 n= 64, 1992 n= 66, 1995 n= 84, 1998 n=14)

6

Table S3 Quality classification based on average index value at each sample point in each year for Ironotter Point according to five indices

YearSample IQI BQI BOPA AMBI ITI

Year

1989 A100 Good Good Good Good Changed Similar 19921989 A500 Good Good Good Good Changed Similar 19921989 A750 Good Good Good Good Changed Similar 19921989 B100 High Good Good Good Changed Disagree 19921989 B750 Good Good Good Good Normal Agree 19921989 C100 Good Good Good Good Changed Similar 19921989 C500 Good Mod Good Good Normal Similar 19921989 C750 Good Good Good Good Normal Agree 19921989 D100 High Mod Good Good Changed Disagree 19921989 D500 High Mod Good Good Normal Disagree 19921989 D750 Good Mod Good Good Changed Similar 19921989 E100 Good Good Good Good Normal Agree 19921989 E500 High Mod Good Good Changed Disagree 19921989 E750 High Mod High Good Changed Disagree 19921989 F100 Good Good Good Good Changed Similar 19921989 F500 High Mod Good Good Changed Disagree 19921989 F750 High Poor High Good Changed Disagree 19921989 G100 Good Good Good Good Changed Similar 19921989 G500 Good Good High Good Changed Disagree 19921989 G750 Good Good Good Good Normal Agree 19921989 H100 High Good High Good Changed Disagree 1992

Agreement Level

YearSample IQI BQI BOPA AMBI ITI

Year

1989 A100 Good Good Good Good Changed Similar 19921989 A500 Good Good Good Good Changed Similar 19921989 A750 Good Good Good Good Changed Similar 19921989 B100 High Good Good Good Changed Disagree 19921989 B750 Good Good Good Good Normal Agree 19921989 C100 Good Good Good Good Changed Similar 19921989 C500 Good Mod Good Good Normal Similar 19921989 C750 Good Good Good Good Normal Agree 19921989 D100 High Mod Good Good Changed Disagree 19921989 D500 High Mod Good Good Normal Disagree 19921989 D750 Good Mod Good Good Changed Similar 19921989 E100 Good Good Good Good Normal Agree 19921989 E500 High Mod Good Good Changed Disagree 19921989 E750 High Mod High Good Changed Disagree 19921989 F100 Good Good Good Good Changed Similar 19921989 F500 High Mod Good Good Changed Disagree 19921989 F750 High Poor High Good Changed Disagree 19921989 G100 Good Good Good Good Changed Similar 19921989 G500 Good Good High Good Changed Disagree 19921989 G750 Good Good Good Good Normal Agree 19921989 H100 High Good High Good Changed Disagree 1992

Agreement Level

7

Table S3 continued

ITIYear

Sample IQI BQI BOPA AMBI ITIChanged Similar 1992 A100 Good Good Good Good ChangedChanged Similar 1992 A500 Good Good Good Good ChangedChanged Similar 1992 A750 Good Good Good Good ChangedChanged Disagree 1992 B100 Good Good Good Good ChangedNormal Agree 1992 B750 High Good Good Good NormalChanged Similar 1992 C100 High Good Good Good NormalNormal Similar 1992 C500 High Good Good Good ChangedNormal Agree 1992 C750 Good Good Good Good NormalChanged Disagree 1992 D100 High Good Good Good ChangedNormal Disagree 1992 D500 Good Mod Good Good ChangedChanged Similar 1992 D750 Good Good Good Good NormalNormal Agree 1992 E100 Good Good Good Good ChangedChanged Disagree 1992 E500 Good Mod Mod Good NormalChanged Disagree 1992 E750 Good Mod Good Good ChangedChanged Similar 1992 F100 Good Good Mod Mod ChangedChanged Disagree 1992 F500 Good Good Good Good NormalChanged Disagree 1992 F750 Good Mod Good Good ChangedChanged Similar 1992 G100 Good Good Good Good ChangedChanged Disagree 1992 G500 High Good Good Good NormalNormal Agree 1992 G750 High Good Good Good ChangedChanged Disagree 1992 H100 Good Good Good Good NormalNormal Similar 1992 H500 High Good Good Good Normal

Agreement Level

8

Heavy metal contamination data were only available for 1995. The mean, standard deviation, max

and min are shown below.

Table S4 Trace metals concentration µg/g dry weight sediment for the <500µm fraction from 1995

Cd Cr Cu Mn Ni Pb ZnMean 0.12 112.96 61.62 676.32 24.66 94.04 185.54Standard Deviation 0.09 43.99 126.38 502.02 6.95 35.52 62.49Max 0.40 196.00 703.00 2399.00 46.00 178.00 315.00Min 0.00 46.00 15.50 235.00 14.20 50.00 90.00

9

Supporting Information References

Borja, A. 2004. The biotic indices and the Water Framework Directive: the required consensus in the new benthic monitoring tools. Marine Pollution Bulletin, 48, 405-408.

Culhane, 2012. The use of benthic communities in environmental health assessment. PhD Thesis, Edinburgh Napier University, UK.

Dauvin, J. C. & Ruellet, T. 2007. Polychaete/amphipod ratio revisited. Marine Pollution Bulletin, 55, 215-224.

Rosenberg, R., Blomqvist, M., Nilsson, H. C., Cederwall, H. & Dimming, A. 2004. Marine quality assessment by use of benthic species-abundance distributions: a proposed new protocol within the European Union Water Framework Directive. Marine Pollution Bulletin, 49, 728-739.

WFD-UKTAG, 2008. UKTAG coastal water assessment method benthic invertebrate fauna. Invertebrates in soft sediments (Infaunal quality Index (IQI)). Edinburgh, Scotland.

Word, J. Q. 1979. The Infaunal Trophic Index. In: BASCOM, W. (ed.) Southern California Coastal Water Research Project, Annual Report 1978. El Segundo, California.

10