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E.V.S. CONSULTANTS LTD.

~DV ~~1 _______________________ ~

EFFECTS OF SPECIES INTERACTIONS ON THE SURVIVAL AND RESPIRATION OF Limnodrilus hoffmeisteri AND

Tubifex tubifex (OLIGOCHAET A, TUBIFICIDAE) EXPOSED TO VARIOUS POllUTANTS AND

ENVIRONMENT Al FACTORS

Prepared For

Dr. R.O. Brinkhurst Chief, Ocean Ecology laboratory

Institute of Ocean Sciences P.O. Box 6000

9860 West Saanich Road Sidney, B.C.

V8l482

Prepared By

P.M. Chapman, M.A. Farrell and K. Teng E.V.S. Consultants Ltd. 195 Pemberton Avenue North Vancouver, B. C.

V7P 2R4

Project 657

August, 1981

~---E~S. consultants Itd .. ~-----.

August I, 1981

Dr. R.O. Brinkhurst Chief, Ocean Ecology laboratory Institute of Ocean Sciences P.O. Box 6000 9860 West Saanich Road Sidney, B~C. V8l4B2

Dear Dr. Brinkhurst:

Our File: 657

Re: lethal and Sublethal Tolerances of Aquatic Oligochaetes With Reference to Their Use as a Biotic Index of Pollution

We are pleased to present the final report on mixed species work with Limnodrilus hoffmeisteri and Tubifex tubifex. The title of this report is:

"Effects of Species Interactions on the Survival and Respiration of Limnodrilus hoffmeisteri and Tubifex tubifex (Oligochaeta, Tubificidae) Exposed to Various Pollutants and Environmental Factors."

Thank you for your review and comment on this report in draft format. We have enjoyed working with you on this most interesting project and will shortly be submitting this data to the journal Water Research. If you have any questions or concerns, please do not hesitate to call.

Sincerely,

E. V .S. CONSULTANTS lTD.

~ .. -r:t-~ U~ ~

Peter M. Chapman, h.D. Project Scientist/Manager

PMC:dcm

Encl.

195 Pemberton Avenue, North Vancouver, B.C. V7P 2R4 Telephone (604) 986-4331

Title Page

Letter of T ransm ittal

Table of Contents

List of Figures

List of Tables

List of Appendices

Acknowledgements

Abstract

Preface

1.0 INTRODUCTION

2.0

3.0

4.0

METHODS

RESULTS

DISCUSSION

iii

T ABLE OF CONTENTS

5.0 REFERENCES CITED

APPENDICES

ii

iii

iv

v

vi

vii

viii

ix

2

3

4

7

iv

LIST OF FIGURES

Figure

Respiration rate of mixed species at 10° and 20°e.

2 Respiration rate of mixed species exposed to various toxicants.

3 Respiration rate of mixed species exposed to various environmental conditions.

Table

2

v

LI ST OF TABLES

96h LCSO values for individual and mixed oligochaete species exposed to various pollutants and environmental factors

Relative tolerances to anoxia

3 Indexes of correlation and standard deviations for respiration data

i

r .!

Appendix

vi

LIST OF APPEND I CES

95% confidence limits for 96h LC50 values of mixed species exposed to various pollutants

2 Time (h) to 50% mortality (L T50) following exposure to different concentrations of pollutants and environmental factors

3 Survival times of mixtures of Limnodrilus hoffmeisteri and Tubifex tubifex exposed to various pollutants and environmental factors

4 Respiration data

- - . , , - - ~, ., >:: ~':"- ~.:. :.-:. ..... --:..-~.... . - ,

vii

ACKNOWLEDGEMENTS

We wish to thank Dr. R.O. Brinkhurst, Chief, Ocean Ecology Laboratory, lOS, for

his assistance and cooperation during the course of this study.

E. V.S. Consultants Ltd. would like to acknowledge the principal investigators Or.

Peter M. Chapman, Melody A. Farrell and Ken Teng, who undertook this

assignment, and the assistance of Dr. Gary A. Vigers, Ric Olmsted and Deedee

Kathman.

We would also like to thank Heather Larratt for assistance in computer

programming and statistical analyses and D. MacNicol for typing the manu

script. L. Borleske drafted the figures.

viii

PREFACE

In October, 1979, E.V.S. Consultants Ltd. were awarded DSS Contract

07SB.FP833-9-0965 through the UnsolicitedProposal Fund for UP-E-89 entitled:

"Lethal and Sublethal Tolerances of Aquatic Oligochaetes with Refer

ence to Their Use as a Biotic Index of Pollution."

The purpose of this project was to develop much needed toxicological informa

tion that would relate indicator organisms to the presence of particular

pollutants in the aquatic environment. The study consisted of four separate

Phases as follows: Phase I involved exposing eleven species of oligochaetes

(classified from ecological studies as tolerant, moderately tolerant and in

tolerant to organic pollution) to selected chemical pollutants and environmental

factors under defined bioassay conditions with and without sediments. Phase II

involved exposing four candidate species to pollutants in combination with a

range of abiotic factors and determining their tolerances under these combina

tions. Phase III involved measurements of respiratory stress imposed by exposure

of candidate species to sublethal concentrations of selected pollutants and

environmental factors. Phase IV involved measurements of respira.tory stress of

oligochaetes exposed to combinations of pollutants and environmental factors.

Following successful completion of Phases I and II and the production of

initial data for Phases III and IV, E.V.S. Consultants Ltd. were awarded a

contract extension to examine the importance of species interactions on relative

tolerances to pollutants and environmental factors and on respiration. The

present report is concerned with this supplemental work, and subject to approval

of the contract authorities it is intended that results will be published in a

refereed journal. Thus the present report is written in journal format with all

ancillary information in the form of Appendices. All data sheets and laboratory

notebooks related to this study are on file at E. V.S. Consultant~ Ltd and are

available on request. The projected authorship and title for publication of this

study are as follows:

Chapman, P.M., M.A. Farrell, K. Teng and R.O. Brinkhurst. Effects of

species interactions on the survival and respiration of Limnodrilus

hoffmeisteri and Tubifex tubifex (Oligochaeta, Tubificidae) exposed to

various pollutants and environmental factors.

ix

ABSTRACT

Laboratory tests were conducted to determine short-term (96h LCSO) and

physiological (respiration) effects of various pollutants (Cd, Hg, NaPCP) and

environmental factors (temperature, pH, salinity, anoxia) on mixtures of two

freshwater oligochaete species: Limnodrilus hoffmeisteri and Tubifex tubifex.

Comparisons with similar data for individual species indicated that mixed species

were significantly more tolerant to toxicants, similarly tolerant to environ

mental factors and less tolerant to anoxia. Although individually these species

regulated respiration rate, in mixed culture they were non-regulators and

respiratory changes in response to stress appeared to vary depending on test

conditions.

1.0 INTRODUCTION

Aquatic oligochaetes, in particular the eutrophic species Limnodrilus

hoffmeisteri and Tubifex tubifex, are commonly used as indicators of

organic pollution. Recent work has been done to characterize the

tolerance of these and other species to a variety of toxicants and

environmental factors (Chapman et al., 1981 a). Such tolerance tests

are commonly conducted with separate species and provide valuable

relative data, however the results may not be directly applicable to the

natural environment where monocultures are rare. L. hoffmeisteri and

!.. tubifex grow better and have a lower respiration rate in mixed as

opposed to pure culture (Brinkhurst et al., 1972; Chua and Brinkhurst,

1973; Brinkhurst and Austin, 1979). Thus it seems I ikely that mixed

species would have higher combined tolerances than individual species.

The present study was initiated to evaluate the importance of mixed

species interactions between !::.. hoffmeisteri and T. tubifex on toler

ance to phys ical and chem ical factors. A great deal of background data

are available for these species, which are commonly found together.

An experimental dose-response aproach was used to define acute lethal

tolerance, followed by measurements of respiration with exposure to

sublethal concentrations of toxicants and environmental factors. Mixed

species tolerances have not been previously determined while previous

mixed species respiration measurements have been conducted under

baseline conditions usually by taking single measurements of oxygen

consumption at relatively high ambient oxygen concentrations. In the

present study, respiration rates were measured in relation to a range of

oxygen values, allowing definition of comparative respiratory curves

under different test conditions described by Chapman et al. (1981 b, c).

2

2.0 METHODS

SedimeAts containing Limnodrilus hoffmeisteri and Tubifex tubifex

,were collected by Ponar grab from a site in the lower Fraser River,

B.C. (490 12.0'N, I 22oS4.S'W) and sieved (0.5 mm mesh) to remove fine

sediments. Worms were sorted under dissecting microscopes and

maintained in the dark at IO:0.S°C. 96h acute lethal bioassays, with

24h solution replacement, were conducted as described by Chapman et

al. (l98Ia) with three replicates per experimental concentration and 10

worms (5 of each species) per replicate. Tolerances were determined

without sediment in relation to pH, temperature, salinity, cadmium

(3CdS04.8H20), mercury (HgCI 2) and pentachlorophenol (NaPCP). All

tests excluding those" involving pH and temperature were run at 10:

O.SoC and pH 7.0:0.2. 96h LCSO's were calculated using the nomo

graphic procedures of Litchfield and Wilcoxon (1949) for toxicant

exposure and a linear graph of concentration vs percent mortality for

environmental factors.

Tolerance to anoxia was determined by allowing the worms to deplete

the oxygen in tightly capped 10 ml disposable syringes, and determining

time to mortality. Oxygen consumption was determined potentio

metrically with a Radiometer PHM73 Blood Gas Analyser and replicate

testing was conducted as detailed by Chapman et al. (1981 a).

Respiration rates (41 02/h/mg dry wt at STP) of equal numbers of mixed

species were measured over a range of external oxygen values under

baseline conditions (pH 7.0, lOoC, ° ppt salinity) and then during

exposure to sublethal concentrations of Hg, Cd and NaPCP and to

sublethal levels of salinity, pH and temperature regimes as detailed for

individual species by Chapman, et al. (l98Ib, c). Concentrations for

" 3.0

3

testing were chosen for comparison with previous work using non-lethal

96h LCSO test concentrations greater than 10% of the LCSO value.

Respiration rates were compared to external oxygen levels by compu

ter-assisted graphic curvilinear regression with data grouping (Ezekiel

and Fox, I 967).

RESULTS

Mixed species bioassay results are compared in Table I to similar data

for individual species, using a basic t-test comparison of 96h survivals

(Chapman et al., 198Ia). Significantly higher tolerances were noted for

mixed species exposed to toxicants; similar tolerances were noted in

the case of environmental factors. Mixed species were 3-4x less

tolerant to anoxia than individual species (Table 2).

Comparative respiration data are provided in Figures 1-3. Indexes of

correlation and standard deviations for mixed species data are given in

Table 3; the "former are all relatively high, indicating good fine fit and

low point scatter.

"At lDoC, h. hoffmeisteri and T. tubifex individually were respiratory

regulators, whereas together these species were non-regulators (Figure

I). However, mixed species respiration rate was generally lower than

that for individual species. At 200 C mixed species respiration was

highly elevated compared to that of L. hoffmeisteri but was similarly

indicative of partial regulation.

Toxicant-specific effects were observed on mixed species respiration

(Figure 2). With NaPCP, respiration was non-regulatory and elevated

over both baseline rate and that for L. hoffmeisteri. In contrast,

4

exposure to Hg resulted in respiratory regulation in a similar (but

elevated) pattern to that for b,. hoffmeisteri alone. Respiration rate

was similar to baseline with Cd exposure and was generally below that

for L. hoffmeisteri.

A non-regulatory respiration pattern was maintained with exposure to

sublethal levels of pH and salinity (Figure 3). At pH 4, respiration rate

was lower than both b,. hoffmeisteri alone andbaseline. At pH 10 and 5

ppt salinity, respiration was generally lower than L. hoffmeisteri and

differed from basel ine in rate.

4.0 DISCUSSION

The results of the present study support previous work with mixed

species. As suspected, b,. hoffmeisteri and I. tubifex were more

tolerant to toxicants in mixed than pure culture, which indicated that

mixed species interactions enhanced survival under stress. The impl ica

tions of this fact to future toxicant assessments both with oligochaetes

and other species will be discussed elsewhere.

Mixed species tolerances to NaPCP were significantly higher at lOoC

(96h LC50 = 0.58 mgt!) than determined by Whitley (1968) for mixed

species at 200 C (72h LC50 = 0.31 mgt!). Although this difference may

indicate a decreased tolerance with increasing temperature, it may also

be due to different test conditions as L. hoffmiesteri and T tubifex - -tested individually were significantly more tolerant to NaPCP at 200 C

than at IOoC (Chapman et al., 1981 d).

Tolerances to environmental factors were comparable to other study

results with individual and mixed L. hoffmeisteri and T. tubifex (i.e.

5

temperoture - Birtwell and Arthur, 1981; pH- Whitley, 1968; salinity -

Chapman and Brinkhurst, 1980; all factors - Chapman et al., 1981 a).

This lack of significant difference in tolerance of mixed and individual

species to environmental factors may be due to the fact that the

environmental test concentrations did not include intermediate con

centrations, e.g. concentrations were either lethal or non-lethal in all

tests.

However, mixed species were far less tolerant than individual species to

anoxia. This result was unexpected although consistent with the fact

that mixed species were less able to regulate respiration than individual

species. Previous anoxia testing indicated that the rank order of

tolerance reflected the status of species as indicators of trophy with

the exception of the oligotrophic species Rhyacodrilus montana and

Variechaeta pacifica which, due to lack of specimens, were tested

collectively and were exceptionally tolerant (Chapman et al., 1981 a).

Although the reason for this difference is not apparent, it may explain

why monocultures predominate under totally anoxic conditions.

The fact that mixed species did not regulate respiration was also

unexpected. While Provini and Marchetti (1976) found that respiration

of isolated tubificids was higher than in the presence of a substrate,

they also noted that combinations of 1:.. hoffmeisteri and I. tubifex in

natural sediments regulated respiration at 15.50 and 190 C. In a

previous respi ratory study without substrate, Chapman et al. (1981 b)

showed that mixtures of R. montana and V. pacifica were capable of

regulation; however, these species were not tested separately so dif

ferences between mixed and individual species could not be resolved.

6

Despite their lack of regulation, the respiration rate of mixed species

was generally below that for individual species, consistent with studies

by Chua and Brinkhurst (1973) which indicated lower respiration in

mixed as opposed to pure cultures. However, because these authors

used different techniques (e.g. I SoC vs lOoC; sediment vs no sediment;

IL vs Radiometer Blood Gas Analyser), respiration rates could not be

directly compared with present results.

Mixed species exposed to chemical and physical stresses maintained a

pattern of non-regulation with the exception of Hg exposure when a

change to regulation was noted. Changes from partial to complete

regulation have been noted for individual freshwater (Stylodrilus her

ingianus) and saltwater species (Monopylephorus -cuticulata) exposed to

Cd and to pH extremes (Chapman et al., 1981 c); however, a change

from non-regulation to regulation has not been previously reported.

Variations in the rate of respiratory non-regulation were noted in

different tests. Respiration rate was elevated with exposure to NaPCP

and elevated temperature. Similar results were obtained for combina

tions of h. hoffmeisteri and I. tubifex in other studies involving NaPCP

exposure (Whitley and Sikora, 1970) and elevated temperature (Provini

and Marchetti, 1976).

That mixed species were at least as tolerant as individual species to

chemical and physical stress, despite their lack of respiratory regula

tion, confirms the contention of Chapman et al. (1981 b) that the abil ity

to regulate respiration is not directly related to tolerance. However,

although the present results do not conflict with previous work and

generally support it, they also serve to raise new questions concerning

the mechanisms governing short-term tolerance and sublethal (respira

tory) stress effects.

7

5.0 REFERENCES CITED

Birtwell, I.K. and D.R. Arthur. 1980. The ecology of tubificids in the Thames estuary with particular reference to Tubifex costatus (Claparede), p. 331-381. .!!!: R.O. Brihkhurst and D.G. Cook (eds.), Aquatic Oligochaete Biology. Plenum Press, N. Y.

Brinkhurst, R.O., K.E. Chua and N. Kaushik. 1972. Interspecific interactions and selective feeding by tubificid 01 igochaetes. Limnol. Oceanogr. 11: 122-133.

Brinkhurst, R.O. and M.J. Austin. 1979. Assimilation by aquatic oligochaetes. Int. Revue ges. Hydrobiol. 63: 863-868.

Chapman, P.M. & R.O. Brinkhurst. 1980. Salinity tolerance in some selected aquatic 01 igochaetes. Int. Revue ges. Hydrobiol. 65: 499-505.

Chapman, P.M., M.A. Farrell and R.O. Brinkhurst. 1981 a. Relative tolerances of selected aquatic oligochaetes to individual pollutants and environmental factors. Aquatic Toxicol. (in Press).

Chapman, P.M., M.A. Farrell, D. McCullough and R.O. Brinkhurst. 1981 b. The respiration rates of selected aquatic 01 igochaetes. Can. J. Zool (Submitted).

Chapman, P.M., M.A. Farrell, D. McCullough & R.O. Brinkhurst. 1981 c. Changes in the respiration rates of selected aquatic oligochaetes exposed to individual pollutants and environmental factors. Aquatic Toxicol. (Submitted).

Chapman, P.M., M.A. Farrell & R.O. Brinkhurst. 1981d. Relative tolerances of selected aquatic oligochaetes to combinations of pollutants and environmental factors. Aquat ic T oxicol. (in Press).

Chua, K.E. and R.O. Brinkhurst. 1973. Evidence of interspecific interactions in the respiration of tubificid oligochaetes. J. Fish. Res. Board Can. 30: 617-622.

Ezekiel, M. & K.A. Fox. 1967. Methods of Correlation and Regression Analysis, Linear and Curvilinear. John Wiley & Sons, Inc: London. 584 pp.

Litchfield, J.V. & F. Wilcoxon. 1949. A simplified method for evaluating dose effect experiments. J. Pharmac. expo Therap. 96: 99-113.

Provini, A. and R. Marchetti. 1976. Oxygen uptake of river sediments and benthic fauna. Boll. Zool. 43: 87-110.

Whitley, L.S. 1968. The resistance of tubificid worms to three common pollutants. Hydrobiologia 32: 193-205.

Whitley, L.S. and R.A. Sikora. 1970. The effect of three common pollutants on the respiration rate of tubificid worms. J. Water. Pollut. Control Fed. 42: R57-R66.

FIGURE I

RESPIRATION RATE OF MIXED SPECIES AT 10° AND 20°C.

Respiration rates for pure cultures of h,. hoffmeisteri and T.

tubifex (from Chapman et al., 1981 b, c) are shown as dashed

and dotted lines respectively where such information is avail

able. pH = 7.0

FIGURE 2

RESPIRATION RATE OF MIXED SPECIES EXPOSED TO VARI

OUS TOXICANTS. Respiration rates for h. hoffmeisteri under

the same test conditions (from Chapman et al., 1981 c) are

shown as a dashed line. Unless otherwise noted, pH = 7.0,

temperature = lOoC, salinity = a ppt. Figure 3 follows the same

pattern.

0.8

0.6

0.4

0.2

.... -c en E 0.8

"-'N o 0.6

~ = "'--" 0.4

NaPCP

Mercury

•• • •

• •

•

• • •

• ___ L.h. ----- ----.. •

• • • • • • . •• • • w ~ a::

0.2 ~-.-;-.... -.- -:- -,- - -.- - ~ - - - - - - - - - - -L.J.., • •• •

/.

:z o --c.. tJ) w a::

/

0.8

0.6

0.4

0.2

Cadmium

---~ . ,/

""- • ".

/ I

/

I

L." . •

•

• •

• •

• ..

15 30 45 60 75 90 105 120 135 150

X P02 (mm Hg)

~:

,

, ; . .' , i l ~-

f·

~. r r

FIGURE 3

RESPIRA TION RA TE OF MIXED SPECIES EXPOSED TO

VARIOUS ENVIRONMENTAL CONDITIONS

0.8 pH 4

0.6

0.4 ....... I..h • • .c •

T •

" --- •

".-- • 0.2 .; , • • ...... " • • • • • • •• •

3: / • • ( .. • •

7' • • • /

>- I

~

-C

en 0.8 pH 10 E "-ON 0.6 ...J ~ •

........ 0.4 \.. . h. ----W

0.2 I-< a: ./

" Z 0 5 ppt Salinity - 0.8 I-c::c c::: 0.6 -c.. tJ) W 0.4 •

L- ."' • c:: • -~-----.-• ---- -----• • • •

, -.-- .. -0.2 • :<-' • • • •

>. • • • .' ./ • / • • • /

, I

15 30 45 60 75 90 105 120 135 150 - P02 (mm Hg) X

Species

+Lirnnodrilus hoffmeisteri

Limnodrilus hoffmeisleri/ Tubifex tubifex

·T ubifex tubifex

n.s. not significant

• 0.05> p > 0.0 I

,... - p <0.001

TABLE I

96H LC50 VALUES FOR INDIVIDUAL AND MIXED OLiGOCHAETE SPECIES EXPOSED TO VARIOUS

POLLUT ANTS AND ENVIRONMENTAL FACTORS

NaPCP (mg/l) Hg (mg/l) Cd (mg/l) Law pH High pH Salinity (ppt) Temperature (oC)

0.33 0.18 0.17 3.5 10.5 10 34

*** • *** n.s. n.s. n.s. n.s.

0.58 0.23 0.58 3.5 10.5 8.8 34.5

**. .*. n.s. n.s. n.s. n.s.

0.38 0.14 0.32 3.6 10.5 9 35

doto froln Chapman et a!. (l98Ia)

'".", .-~- ·""'-~"",_C.-'~_-'-"R ... ........,~ •. ~, __ .• 'r-""··""""''''I'&'"·t ... -r'''--1''''-'''·_·'~·_~·'~ __ '._ .-< -.r·-.~··"--~~1"'-'·~- r'_" -'" •••• -."..,._~. "",~-,., .~.'r7"...,-,,,,,,,;:.,,,.!-.li_-~ :/~.'"~"-";'(~"=";:~.J,:>l"~

Species

L. hoffmeisteri/ -T. tubifex

+L. hoffmeisteri

+T. tubifex

TABLE 2

RELA TIVE TOLERANCES TO ANOXIA

Mean Survival (days)

5

23

16

+ data from Chapman et al. (I 981 a).

Number of Tests

6

3

3

TABLE 3

INDEXES OF CORRELATION AND STANDARD DEVIATIONS FOR RESPIRATION DATA Unless Otherwise Noted, pH = 7.0, temperature ii 10°C, salinity = 0 ppt.

Treatment Index of Correlation Standard Deviation

control 0.89 0.034

20°C 0.84 0.170

pH 4 0.69 0.043

pH 10 0.88 0.042

5 ppt salinity 0.58 0.052

0.10 mg/I Cd 0.75 0.046

0.10 mg/I Hg 0.77 0.047

0.32 mg/I NaPCP 0.79 0.051

APPENDIX I

95% CONFIDENCE LIMITS FOR 96h LC50 VALUES OF MIXED SPECIES

EXPOSED TO VARIOUS POLLUTANTS

96h LC50

95% confidence 1 imits

Hg (mg/l)

0.23

0.21-0.26

Cd (mg/l) NaPCP (mg/l)

0.58 0.58

0.54-0.62 0.54-0.62

Hg 1l!!9.ill

0.56

0.32

0.10

Cd

APPENDIX II

TIME (h) TO 50% MORTALITY (LT50) FOLLOWING EXPOSURE TO DIFFERENT

CONCENTRATIONS OF POLLUTANTS AND ENVIRONMENTAL FACTORS

NaPCP salinity low high L T50 lr!!9L!l LT50 ~ LT50 (ppt) LT50 Q.IL LT50 £tL

9.7 1.0 38 5.6 3.1 20 <1 3 1.6 11

14 0.56 >96 3.2 5.5 10 42 4 >96 10

>96 0.32 >96 1.0 54 5 >96 5 >96· 9

0.56 >96

0.32 >96

temperature LT50 (OC) LT50

1.4 40 2

>96 30 >96

>96 20 >96

APPEND I X II I

SURVIVAL TIMES FOR MIXTURES OF LimnodriZus hoffmeisteri &

Tubifex tubifex EXPOSED TO VARIOUS POLLUTANTS AND ENVIRON

MENTAL FACTORS

SURVIVAL (%) OF Limnodrilus hoffmeisteri/Tubifex tubifex

EXPOSED TO DIFFERENT CONCENTRATIONS OF NaPCP

HOURS TO PERCENT SURVIVAL

without sediment CONCENTRA TI or~ 1 2 4 8 18 24 48 72 96

5.6 mg/l 100 100 20 a a a a a 3.2 mg/l 100 100 60 7 7 a a a 1.0 mg/l 100 100 100 100 100 77 10 0

0.56 mg/l 100 100 100 100 100 100 100 60

0.32 mg/l 100 100 100 100 100 100 100 100

0.10 mg/l 100 100 100 100 100 100 100 100

control 100 100 100 100 100 100 100 100

SURVIVAL (%) OF LimnodriZus hoffmeisteri/Tubifex tubifex

_EXPOSED TO DIFFERENT CONCENTRATIONS OF MBRCURY


without sediment CONCENTRATION 1 2 4 8 18 24 48 72 96

0.56 mg/l 100 100 100 0 0 0 0

0.32 mg/l 100 100 100 27 7 7 7

0.10 mg/l 100 100 100 100 100 100 100

control 100 100 100 100 100 100 100

-.,~.-- ~"---- .. - ----:-. ~--"--'- .. -" ~-. _ ........ -•. ,.,.- .,- .... ~.~~~.,. .. ~ ... --~. ~~-...... ~~~~ .~". ~ ... -~,..-,. -, .... '-~··:l'l'-r"l"~~...,....,··'~~·· ..... ,--·· .. ·"-"'~'·,ft:.-..... --"r.<; ..... ='·~".;".:,;;;':~~':r-:"j.;,.q-,,<}"~";:;:"8>':::-':"'''l;;-;;i:;:;~~:

SURVIVAL (%) OF Limnod~iZus hoffmeiste~/Tubifex tubifex

EXPOSED TO DIFFERENT CONCENTRATIONS OF CADMIUM

HOURS TO PERCENT SURVI VAL

without sediment CONCENTRATION 1 2 '4 8 18 24 48 72 96

1.0 mg/l 100 100 100 100 27 17 0

0.56 mg/l 100 100 100 100 80 73 57

0.32 mg/l 100 100 100 100 100 100 100

0.10 mg/l 100 100 100 100 100 100 100

control 100 100 100 100 100 100 100

- - _._--_ .. _- - --...~--------------.-~-

SURVIVAL (%) OF LimnodriZus hoffmeisteri/Tubifex tubifex EXPOSED TO DIFFERENT pHis


without sediment CONCENTRATIor~ 1 2 4 8 18 24 48 72 96

3 100 a a a a a a a

4 100 100 100 100 100 100 100 100

5 100 100 100 100 100 100 100 100

7 100 100 100 100 100 100 100 100

9 100 100 100 100 100 100 100 100

10 100 100 100 100 100 100 100 100

11 100 30 3 a a a a a

"'-·"7~~-"'""'-·-'~.~ru'7:o:..=r,._=r.~,:c.·"· .. -_"r.~",--,.-.,~,r.""'=,"""","'~'<>J"""",,,,""='-"'-;-·'''''''''O-'''=-''''===--Z''''~'~'='''''C-_''-'''''_,-= ... .<.~ .... ."...~"-;:: •• - ··'''--'''''''~~'·'~·-~'''''-'J:''·'7;~.·~M~~~~",,,,,,~,,,,,-:-,·;:O''''~,,,,,,,-;r;r':':~·::0.--"-I:,~;;"""<"'.'·;;;'·,T:-t';'·"o:.·<"".r;'7'::;:I,;Jfo.Tl".:";;;':lL'l-"J;;':>5...~j;:;;;::::;';<;'"l;<T'{';i;$.!:.;I:;:~.:.'U~~~.'

SURVIVAL (%) OF Limnodrilus hoffmeisteri/Tubifex tubifex EXPOSED TO DIFFERENT SALINITIES


without sediment CONCENTRATIor~ 1 2 4 8 18 24 48 72 96

20 ppt a a a a a a a

10 ppt 100 100 100 97 40 40 30

5 ppt 100 100 100 100 100 100 100

a ppt 100 100 100 100 100 100 100

•

SURVIVAL (%) OF LimnodPiZus hoffmeisteri/Tubifex tubifex EXPOSED TO DIFFERENT TEMPERATURES


without sediment CONCENTRATION 1 2 4 8 18 24 48 72 96

400 C 100 50 27 a a a a

300 C 100 100 100 93 90 87 83

200 C 100 100 100 100 100 100 100

lODe 100 100 100 100 100 100 100

_v_ ,_ •• ,,_.,.,' ,'-.-~.'-:-.<" >','- --~o;~"";."'~~'~'.,"".,;,,, '"> .......... ~.~;~~.

APPENDIX IV

• RESPIRATION DATA

OLIGOCHAETE RESPIRATION DATA

Species: LimnodriZus hoffmeisteril Trea tment: none Tubifex tubifex

Temperature (oC): 10 pH ~ 7.0 Salinity (ppt) . 0 Barometric pressure: (mm Hg): 762

Worm weights (mg dry weight) Calibration value (mm Hg) : 157.6

Replicate A: 21.4 Replicate .B: 20.6 Replicate C: 20.4

Page 1 of 3

Aliquot readings (rrun 02) Syringe X P02

Respiration volume (ml) Rate

Replicate Time 1 2 3 X initial final (mm Hg) (ul/mg/h)

A 1020 117.9 B 1025 113.7 C 1030 114.7

A 1155 92.0 89.6 89.5 90.4 10 9 B 1210 90.5 91.1 91. 3 91.0 10 9 C 1225 89.9 89.5 90.0 89.8 10 9

A 1340 61.2 59.0 58.1 59.4 9 8 74.9 calibra-B l350 61.3 60.4 60.3 60.7 9 8 75.9 ti on pro-C 1400 66.0 66.9 67.3 66.7 9 8 78.3 blems

A 1505 42.6 39.3 39.9 39.5 8 7 49.5 .25 B 1515 42.4 42.0 41.5 42.0 8 7 51.4 .25 C 1525 48.0 48.2 47.9 48.0 ·8 7 57.4 .25

A 1625 21.8 19.6 18.9 19.3 7 5.6 29.4 .24 B 1635 23.5 23.1 22.9 23.2 7 5.6 32.6 .23 C 1645 31.1 31.1 30.9 31.0 7 5.6 39.5 .21

A 1745 8.6 7.1 6.3 7.3 5.6 4 l3.3 .11 B 1755 8.5 8.4 7.4 8.1 5.6 4 15.7 .15 C 1805 11. 7 11.9 11. 7 11.8 5.6 4 21.4 .19


Species: LimnodriZus hoffmeisteri/ Trea tment: none Tubifex Subifex

Temperature ( C): 10 pH: 7.0

Salinity (ppt) 0 Barometric pressure (mm Hg): 765



Page 2 of 3

Aliquot readings (mrn 02) Syringe X P02


Replicate Time 1 2 3 X . initial final (mm Hg) (ul/mg/h)

A 0940 125.5 B 0942 123.8 C 0945 122.2

A 1010 109.9 108.9 108.5 109.1 10 9 B 1020 114.4 114.7 114.6 114.6 10 9 C 1030 112.8 112.4 112.3 112.5 10 9

A 1125 89.7 88.9 89.3 9 8 99.2 .34 B 1135 102.7 103.1 103.0 102.9 9 8 108.8 .31 C 1145 97.3 97.1 97.0 97.1 9 8 104.8 .33

A 1240 72.6 69.2 68.5 68.9 8 7 79.1 .31 B 1250 89.6 89.7 89.8 89.7 8 7 96.3 .31 C 1300 84.2 84.4 84.9 84.5 ·8 7 90.8 .24

A 1405 50.9 51.0 50.9 7 6 59.9 .21 B 1425 73.3 73.4 72.8 73.2 7 6 81. 5 .27 C 1435 67.3 66.7 66.6 66.9 7 6 75.7 .23

A 1555 31.1 29.7 29.7 6 5 40.3 .17 B 1605 51.3 52.1 51.7 51.7 6 5 62.5 .28 C 1615 47.7 47.2 47.2 47.4 6 5 57.2 .21

~ ~ '" ~~

OLIGOCHAETE RESPIRATIDN DATA ~ f~

Species: Limnodrilus hoffmeisteri/ Treatment: none ~ Tubifex bubifex

Temperature ( C): 10 pH: 7.0 Sal inity (ppt) 0 Barometric pressure~ (mm Hg): 760 ! Worm weights (mg dry weight) Calibration value (mm Hg) : 157

Replicate A: 4.8 Replicate B: 5.8 Replicate C: 6.1

Page 3 of 3

Aliquot readings (mm 02) Syri nge X P02


Replicate Time 1 2 ~ X initial final (mm Hg) (ul/mg/h)

A 0935 128.8 B 0945 129.8 C 0955 128.5

A 1035 124.3 124.2 124.3 8.8 7.8 B 1045 122.9 122.8 122.8 122.8 9 7.4 C 1055 122.6 122.9 122.8 122.8 9 8

A 1135 119.0 118.7 119.9 7.8 6.8 122.1 .36 B 1145 117.5 116.9 116.8 117.1 7.4 6.4 120.0 .35 C 1155 117.0 117.2 117.2 117.1 8 6.8 120.0 .36

A 1240 113.3 112.9 112.5 112.9 6.8 5.6 116.4 .46 B 1250 110.1 110.1 109.5 109.9 6.4 5.2 113.5 .36 C 1300 110.2 110.5 110.4 110.4 6.8 5.8 113.8 .34

A 1355 106.1 105.6 105.9 5.6 4 109.4 .33 B 1405 101.1 100.9 100.4 100.8 5.2 4 105.4 .32 C 1415 102.3 102.0 102.0 102.2 5.8 4 106.3 .30


Species: LimnodriZus hoffmeisteri/ Tubifex tubifex

Temperature (oC): 20

Salinity (ppt) 0

Worm weights (mg dry weight)

Treatment: elevated temperature

pH: 7.0 Barometric pressure; (mm Hg): 763 Calibration value (mm Hg) : 155.9

Replicate A: 5.6 Replicate ,B: 6.4 Replicate C: 8.2

Page 1 of 1

Replicate

A B C

A B C

A B C

A B C

A B C

A B C

B

Time

1045 1046 1050

1115 1125 1135

1240 1250 1300

1350 1400 1410

1450 1500 1510

1550 1605 1615

1640

Aliquot readings (mm 02) 1 2 3 X

144.6 142.3 141. 3

128.8 128.4 127.8 126.7 126.1 126.3 120.7 120.3 120.5

93.5 92.6 92.0 98.5 99.5 99.9 89.1 88.2 86.8

60.7 58.8 57.9 71.8 71.7

57.2 56.9 57.4

32.8 30.9 48.7 48.9 47.2 33.2 32.2 28.9

5.4 3.1 17.5 17.0 15.4 10.2 6.6 5.7

9.1 6.5 4.0

128.3 126.4 120.5

92.7 99.3 88.0

59.1 71.8 57.2

31. 9 48.3 31.4

4.3 16.6 7.5

6.5

Syringe volume (ml)

initial final

10 10 10

8 9 9

6.6 8 8

5.4 7 7

4 5.8 6

4.4

8 9 9

6.6 8 8

5.4 7 7

4 5.8 6

3 4.4 4

3

X P02 (mm Hg)

110.5 112.9 104.3

75.9 85.6 72.6

45.5 60.1 44.3

18.1 32.5 19.5

11. 6

!

Respiration Rate . (ul/mg/h)

1.42 1. 07 1.00

1.34 1.17 1.02

1.04 1.02 0.87

0.78 1.07 0.64

0.48


Species: Limnodrilus hoffmeisteri/ Tubifex tubifex

Temperature (oC): 10 Salinity (ppt) . 0


Treatment: 0.1 mg/l Cd

pH: 7.0 Barometric pressure'~ (mm Hg): 761

Calibration value (mm Hg) 157.2


Page 1 of 2

Replicate

A B C

A B C

A B C

A B C

A B C

B

Time

0930 0932 0935

0955 1005 1015

1105 1115 1125

1235 1250 1300

1355 1405 1415

1510

Aliquot readings (mm 02) 1 2 3 X

58.0 58.0 51.6

41.8 40.6 48.3 49.6 48.5 41.1 41.0 40.5

35.6 31.6 30.5 39.5 38.9 38.2 30.4 30.0 29.2

17.8 19.1 18.4 29.6 29.7 29.0 19.2 18.1 17.6

6.5 5.6 4.6 17.8 18.9 17.8 9.0 8.0 7.8

9.7 10.7 10.8

41.2 48.8 40.9

31.1 38.9 29.9

18.4 29.4 18.3

5.6 18.2 8.3

10.4

Syringe volume (ml)

initial final

10 10 10

9 9 9

8 8

·8

6.2 6.6 6

5.4

9 9 9

8 8 8

6.2 6.6 6

5 5.4 5

4

X P02 (mm Hg)

36.2 43.9 35.4

24.8 34.2 24.1

12 23.8 13.3

14.3

Respiration Rate (ul/mg/h)

.28

.26

.28

.24

.18

.20

.22

.21

.16

.14



Temperature (oC): 10 Salinity (ppt) . 0


Treatment: 0.1 mg/l Cd

pH: 7.0 .

Barometri c pressure. (mm Hg): 762 Calibration value (mm Hg) 157.4


Page 2 of 2

Replicate

A B C

A B C

A B C

A B C

A B C

Time

0950 0953 1000

1030 1040 1050

1150 1200 1210

1325 1335 1345

1500 1515 1525

Aliquot readings (mm 02)

1 2 3 X

137.6 123.2 128.4

126.5 124.9 124.9 124.9 110.3 109.9 109.7 110.0 120.3 120.4 120.5 120.4

112.5 112.0 111.4 112.0 94.6 93.6 93.5 93.9 106.~ 107.1 106.9 106.9

96.8 74.5 93.7

82.0 55.9 81. 7

95.5 73.1 94.2

79.9 56.9 82.0

95.2 72.5 94.4

79.0 54.5 82.0

95.8 73.4 94.1

79.5 55.7 81.9

Syringe volume (ml)

initial final

10 10 10

9 9 9

8 8

·8

7 7 7

9 9 9

8 8 8

7 7 7

6 6 6

X P02 (mm Hg)

118.5 102.0 113.7

103.8 83.7

100.5

87.7 64.6 88.0


.30

.38

.40

.28

.36

.28

.25

.26

.22


Species: Limnodrilus hoffmeisteril Tuhifex tuhifex


Sal inity (ppt) 0


Treatment: 0.10 mg/l Hg

pH: 7.0

Barometri c pressure'~ (mm Hg): 765



Page 1 of 3

Replicate

A B C

A B C

A B C

A B C

A B C

A B C

A B C

Time

0945 0947 0950

1020 1030 1040

1145 1155 1205

1315 1325 1340

1445 1455 1505

1625 1635 1645

1750 1800 1810


1 2 3 X

139.8 136.1 134.0

126.2 125.8 125.4 121.4 121.1 121.2 123.0 122.8 123.1

107.3 106.7 106.1 103.5 103.1 102.8 105.9 106.3 106.2

83.6 82.9 83.6 83.7 83.7 85.5 85.5 85.0

62.8 62.2 62.0 61.7 61.4 67.8 68.0 67.4

36.2 34.9 42.7 43.4 43.9 45.1 45.3 45.8

12.9 12.2 24.5 25.9 24.6 14.5 14.2 13.2

125.8 121.2 123;0

106.7 103.1 106.1

83.3 83.7 85.3

62.5 61.7 67.7

35.6 43.3 45.4

12.5 25.0 14.0

Syringe volume (ml)

initial final

10 10 10

9 9 9

7.6 8

·8

6.6 7 7

5.6 6 6

4.6 5 3.6

9 9 9

7.6 8 8

6.6 7 7

5.6 6 6

4.6 5 3.6

3 4 2

X P02 (mm Hg)

116.3 112.2 114.6

95.0 93.4 95.7

72.9 72.7 76.5

49.1 52.5 56.6

24.1 34.2 29.7


.37

.33

.30

.:37

.30

.30

.28

.30

.25

.28

.19

.23

.23

.19

.31


Species: LimnodriZus hoffmeisteril Tuhifex tuhifex


Sa 1 ini ty (ppt) : 0


Treatment: 0.10 mg/l Hg

pH: 7.0 Barometri c pressure; (mm. Hg): 764 Cal ibration value (mm Hg) : 157.8


Page 2 of 3

Aliquot readings (mm 02) Syringe X P02

Respiratio volume (ml) Rate

Re~licate Time 1 2 3 X initial final (mm Hg} - (ul/m /h

A 0938 125.9 B 0940 122.0 C 0945 118.0

A 1230 109.2 108.9 108.9 109.0 10 9 B 1240 95.9 96.2 95.7 95.9 10 9 C 1250 93.5 93.8 92.9 93.4 10 9

A -1430 97.7 96.4 96.5 96.9 9 8 103.0 .33 B 1440 79.6 77.7 77 .8 78.4 9 8 87.2 .31 C 1450 76.0 76.3 76.2 76.2 9 8 84.8 .28

A 1640 81.6 80.4 79.8 80.6 8 7 88.8 .35 B 1650 56.9 55.0 54.7 55.5 8 7 67.0 .33 C 1700 59.2 59.7 59.2 59.4 8 7 67.8 .22


Species: Limnodrilus hoffmeisteri/ . Treatment: 0.10 mg/l Hg Tuhifex tubifex

Temperature (oC): 10 pH: 7.0

Salinity (ppt) 0 Barometric pressure·~ (mm Hg): 765

Worm weights (mg dry wei ght) . Calibration value .(mm Hg) : 158

Replicate A: 12.4 ReplicateB: 11.1 Replicate C: 13.9

Page 3 of 3

Aliquot readings (mm O2) Syringe X P02 Respiration

volume (ml) Rate Replicate Time 1 2 3 X initial final (mm Hg) (ul/mg/h)

A 1108 44.5 B 1110 48.0 C 1113 45.6

A 1210 37.0 37.3 37.1 37.1 10 9 B 1220 43.6 45.5 45.2 45.4 10 9 C 1230 34.4 35.7 33.8 34.6 10 9

A 1325 26.7 27.1 27.7 27.2 9 7 32.2 .28 B 1335 34.7 34.7 34.5 34.6 9 7 40.0 .33 C 1345 23.3 22.7 22.2 22.7 9 7 28.7 .30

A 1445 18.2 18.9 18.1 18.4 7 5 22.8 .18 B 1455 25.5 26.1 26.6 26.1 7 5 30.4 .19 C 1505 13.7 13.4 12.8 13.3 7 5 18.0 .17

A 1605 7.6 6.7 7.5 7.3 5 3 12.9 .16 B 1625 14.8 14.3 14.6 5· 3 20.4 .17 C 1635 6.7 6.3 4.9 6.0 5 3 9.7 .08


Species: LimnodriZus hoffmeisteril Treatment: 320 ppb NaPCP Tubifex tubifex


Salinity (ppt) 0 Barometric pressure~ (mm Hg): 761

Worm weights (mg dry weight) Calibration value (mm Hg) : 157.2 i

. Replicate A: 15.0 Replicate B: 13.6 Replicate C: 13.6 I

Page 1 of 2 r I I I I

Syringe Aliquot readings (mm 02) X P02 Respiratiorl

volume (ml) Rate I I

Replicate Time 1 2 3 X initial final (mm Hg) (ul/mg/h) I !

A 0937 109.5 I I f

B 0940 124.5 , I

C 0942 110.8 I A 1025 104.1 103.7 103.2 103.7 10 9 ! B 1035 113.8 114.2 113.9 114.0 10 9 C 1045 91.8 91.4 91.5 91.6 10 9

A 1135 81.9 81. 7 81.8 9 8 92.8 .55 B 1145 94.4 94.3 94.3 94.3 9 8 104.2 .54 C 1155 74.9 73.6 73.2 73.9 9 8 82.8 .49

·A 1310 61.8 62.5 62.2 62.2 8 7 72.0 .32 B 1320 74.6 74.5 73.7 74.3 8 7 84.3 .36 C 1330 55.6 55.0 53.8 54.8 8 7 64.4 .35

A 1425 41.5 39.0 37.8 39.4 7 6 50.8 .41 B 1435 52.9 53.5 53.2 53.2 7 6 63.8 .42 C 1445 36.2 36.8 36.5 7 6 45.7 .37

A 1545 20.9 19.7 20.3 6 5 29.9 .28 B 1555 32.2 33.4 33.5 33.0 6 5 43.1 .33 C 1605 18.6 18.3 17.6 18.2 6 5 27.4 .29

A 1645 8.8 8.7 8.8 5 4 14.6 .19 B 1655 16.8 16.4 16.6 5 4 24.8 .29 C 1705 6.3 5.2 4.9 5.5 5 4 11.9 .23

dLIGOCHAETE RESPIRATION DATA

Species: LimnodriZus hoffmeisteri/ Tubi fex tubi fex


Salinity (ppt) 0


Treatment: 320 ppb

pH: 7.0 Ba-rometric pressure'~ (mm Hg): 762



Page 2 of 2

Replicate

A B C

A B C

A B C

A B C

A B C

A B C

Time

1005 1007 1015

1100 1110 1120

1220 1230 1240

1355 1405 1415

1540 1550 1600

1710 1720 1730


1 2 3 X

115.2 113.6 115.4

102.4 101.3 101.2 95.8 95.9 96.6

100.7 100.6 100.2

83.0 75.6 82.6

61.0 54.0 63.9

40.8 28.5 42.5

8.0 19.4

83.4 74.9 82.6

61.0 52.7 63.2

39.0 27.7 43.0

18.0 7.7

19.9

83.1 74.6 82.3

60.4 52.8 63.2

39.2 27.9 44.0

16.7 7.2

19.6

Syringe volume (ml)

initial final

10 9 10 9 10 9

9 9 9

8 8

·8

7 7 7

6 6 6

8 8 8

7 7 7

6 6 6

5 5 5

X P02 (mm Hg)

92.4 85.6 91.5

72.0 64.1 73.0

50.3 40.6 53.3

28.6 17.8 31.4


.38

.35

.32

.34

.27

.26

.26

.25

.21

.27

.20

.25

-OLIGOCHAETE RESPIRATION DATA

Species: Limnodrilus hoffmeisteri/ Treatment: salinity Tuhi fex tuhi fex

Temperature (oC): 10 pH: 7.0 Sal inity (ppt) 5 Barometric pressure (mm Hg):' 765

. Worm weights (mg dry weight) Calibration value (mm.Hg) 158.0

Replicate A: 15.4 Replicate .B: 19.8 Replicate C: 19.7 I

Page 1 of 4 i I I


Respi rat; 01 volume (ml) Rate I

Reelicate Time 1 2 3 X initial final (mm Hg) (ul/mg/h} !

I

A 0952 133.4 I B 0955 136.0 I C 0958 131.8 I

A 1050 119.7 119.7 119.1 119.5 10 9 I !

B 1100 118.4 118.2 118.1 118.2 10 9 C 1110 117.4 117.2 117.3 117.3 10 9

A 1215 104.3 104.0 103.3 103.9 9 8 111. 7 .30 B 1225 100.7 100.4 100.3 100.5 9 8 109.4 .27 C 1235 99.7 99.5 98.8 99.3 9 8 108.3 .27

A 1350 84.6 85.1 84.9 84.9 8 7 94.4 .29 B 1400 76.4 76.4 76.1 76.3 8 7 88.4 .29 C 1410 81.1 81.1 80.8 81.0 8 6.8 90.2 .22

A 1515 68.3 67.4 67.8 67.8 7 6 76.4 .26 B 1525 56.4 56.1 56.3 56.3 7 6 66.3 .23 C 1535 67.0 66.4 66.2 66.5 6.8 5.8 73.8 .17

A 1655 45.7 45.9 45.9 45.8 6 5 56.8 .24 B 1705 34.0 33.5 33.8 33.8 6 5 45.1 .19 C 1715 46.7 47.1 47.1 47.0 5.8 4.8 56.8 .16

A 1820 27.4 28.4 28.2 28.0 5 4 36.9 .19 B 1830 11.6 11. 2 10.8 11.2 5 4 22.5 .19 C 1840 26.8 27.6 27.6 27.3 . 4.8 3 37.2 .16


Species: Limnodrilus hoffmeisteri/ Treatment: salinity Twifex tubifex

Temperature (oC): 10 pH: 7.0 Salinity (ppt) 5 Barometric pressure (mm Hg): 765

Worm weights (mg dry weight) Calibration value (mm Hg) : 158


Page 2 of 4




A 1100 67.0 B 1102 55.0 C 1105 52.7

A 1135 49.7 48.4 49.1 10 9 B 1145 48.5 47.4 47.8 47.9 10 9 C 1200 42.3 41.8 42.7 42.3 10 9

A 1255 43.8 42.7 42.7 9 7.8 45.9 .20 B 1305 40.0 41.1 39.8 40.3 9 8 44.1 .31 C 1315 34.3 31. 7 29.5 31.,8 9 7 37.1 .34

A 1415 36.1 35.8 36.0 7.8 6 39.4 .18 B 1425 33.5 32.9 32.3 32.9 8 6 36.6 .27 C 1435 19.8 19.2 19.6 19.5 7 5 25.7 .29

A 1545 28.5 27.3 27.9- 6 4 32.0 .15 B 1555 25.0 24.6 24.1 24.6 6 4 28.8 .20 C 1615 7.3 7.0 6.7 7.0 5 3 13.4 .17

A 1645 16.4 16.3 16.5 16.4 4 3 22.2 .21 B 1655 15.6 15.5 15.4 15.5 4 3 20.1 .22




Salinity (ppt) 5


Treatment: salinity

pH: 7.0

Barometric pressure (mm Hg): 763 Calibration value (mm Hg) : 157.6


Page 3 of 4 i

Aliquot readings (mm O2)

Replicate Time 1 2 3 X

A 0930 136.4 B 0932 134.7 C 0935 135.1

A 1200 106.1 105.4 105.8 . B 1210 111.0 111.4 111. 3 111.2 C 1220 120.0 120.6 120.5 120.4

A 1400 91.7 91.2 91.5 B 1410 100.6 101. 3 100.5 101. 5 C 1420 112.3 112.3 112.3 112.3

A 1605 72.9 71.5 72.2 B 1620 83.8 84.5 84.2 82.8 C 1630 100.4 100.4 100.5 100.4

Syringe volume (ml)

initial final

10 9 10 9 10 9

9 8 9 8 9 8

8 7 8 7

'8 7

·X PO 2 (mm Hg)

98.7 106.4 116.4

81.9 92.2

106.4

i ResPiratid Rate . I

I .27 .23 .33

.30

.36

.39

I




Salinity (ppt) : 5


Treatment: salinity

pH: 7.0

Barometric pressure (mm Hg): 761 Calibration value (mm Hg) : 157.2

Replicate A: 16.3 Replicate.8: 6.8 Replicate C: 12.2

Page 4

Aliquot readings (mm O2) Syringe X P02 volume (ml) Replicate Time 1 2 3 X initial final (mm Hg)

A 1535 46.7 B 1537 47.4 C 1540 35.3

A 1605 37.0 38.0 37.4 37.5 10 8 B 1625 35.2 34.8 35.9 35.3 10 7.8 C 1635 27.8 27.2 28.6 27.9 10 8

A 1710· 27.4 28.0 26.8 27.4 8 7 32.5 B 1720 27.5 28.8 27.8 28.1 7.8 6.8 31. 7 C 1730 20.9 20.5 19.9 20.4 8 6 24.2

of 4


.22

.22

.21


Species: LimnodriZus hoffmeisteri/ Treatment: high pH Tubi fex tubi fex


Salinity (ppt) 0 Barometric pressure; (mm Hg): 764


Replicate A: 15.1 Replicate .B: 16.9 Replicate C: 16.3 I Page 1 of 2 I

. i 1

Syringe R . . I Aliquot readings (mm 02) X P02

espl ratlonl volume (ml) Rate

Replicate Time 1 2 3 X initial final (mm Hg) (ul/mg/h) i A 0937 83.5

t B 0940 84.0 C 0943 84.9

A 1045 69.9 69.6 69.3 69.6 10 9 B 1055 75.6 75.3 74.4 75.1 10 9 C 1105 74.3 73.6 73.8 73.8 10 9

A 1210 59.8 59.4 59.1 59.4 9 7 64.7 .21 B 1220 62.9 63.5 63.3 63.2 9 7 69.2 .22 C 1230 61.9 62.0 62.1 62.0 9 7 67.9 .22

A 1335 48.7 50.2 47.7 48.9 7 5.8 54.2 .17 B 1345 54.0 53.5 54.0 53.8 7 6 58.5 .13 C 1355 53.5 52.3 51.0 52.3 7 6 57.2 .14

A 1545 27.3 27.1 27.2 5.8 4.8 38.1 .19 B 1555 32.3 32.6 33.2 32.7 6 5 43.3 .17 C 1605 30.0 29.7 29.9 6 5 41.1 .18

A 1710 15.7 15.8 13.7 15.1 4.8 3.8 21.2 .13 B 1720 20.6 20.6 21.1 20.8 5 3.8 26.8 .12 C 1730 17.9 17.7 17.7 17.8 5 4 23.9 .13

A 1815 6.3 4.9 4.1 5.1 3.8 2 10.1 .11 B 1825 6.7 6.6 6.5 6.6 3.8 2 13.7 .14 C 1835 6.6 5.5 3.9 5.3 4 2 11.6 .14


Species: Limnodrilus hoffmeisteri/ Treatment: high pH Tubifex tubifex


Salinity (ppt) 0 Barometri c pressure'~ (mm Hg): 764



Page 2 of 2




A 0940 137.0 B 0943 137.0 C 0946 133.9

A 1040 120.6 120.1 119.8 120.2 10 9 B 1100 120.4 120.5 120.2 120.4 10 9 C 1110 110.6 110.3 110.2 110.4 10 9

A 1210 100.7 100.2 100.2 100.4 9 8 110.3 .32 B 1220 102.9 102.9 102.2 102.7 9 8 111.6 .45 C 1230 91.8 91.2 91.0 91. 3 9 8 100.9 .34

A 1330 81.5 81.4 81. 5 8 7 91.0 .31 B 1340 86.6 87.7 87.2 87.2 8 7 95.0 .35 C 1350 77.3 75.7 75.4 76.1 8 6.6 83.7 .24

A 1450 60.7 61.3 60.1 60.7 7 6 71.1 .29 B 1500 71.8 72.2 71.6 71.9 7 6 79.6 .30 C 1510 57.1 58.1 56.6 57.3 6.6 5 66.7 .25

A 1610 42.4 41. 7 41.4 41.8 6 5 51.3 .23 B 1620 56.5 56.6 56.5 56.5 6 5 64.2 .26 C 1630 36.5 35.2 36.4 36.0 5 4 46.7 .21


Species: LimnodriZus hoffmeisteri/ Treatment: low pH Tubifex -Subifex

pH: 4.0 Temperature ( C): 10 Salinity (ppt) 0 Barometric pressure·; (mm Hg): 764


Replicate A: 14.8 Replicate B: 14.2 Repl icate C: 14.1

Page 1 of 3 i I

! I


Respiratio~ volume (ml) Rate


A 0930 88.7 B 0932 92.0 C 0934 94.3

A 1015 82.7 80.8 81.8 10 9 B 1025 86.4 87.1 87.3 86.9 10 9 C 1035 83.2 83.9 82.8 83.3 10 9

A 1140 73.7 72.8 73.3 9 8 77 .6 .18 B 1150 77.7 77.9 77.7 77.7 9 7 82.3 .20 C 1200 72.9 72.5 72.7 72.7 9 7 78.0 .23

A 1305 61.2 61.2 61.2 8 6 67.3 .22 B 1315 68.8 68.9 69.5 69.1 7 6 73.4 .14 C 1325 65.8 65.1 64.2 65.0 7 6 71. 4 .13

A 1515 42.5 42.0 42.3 6 4.8 51.8 .17 B 1525 50.3 50.5 50.2 50.3 6 4.8 59.7 .18 C 1535 44.5 44.2 43.8 44.2 6 5 54.6 .20

A 1640 24.2 23.2 23.7 4.8 3.6 33.0 .21 B 1650 38.7 39.0 39.0 38.9 4.8 3.8 44.6 .13 C 1700 31.3 31.6 31.6 31.5 5 4 37.9 .15


Species: Limnodrilus hoffmeisteri/ Treatment: low pH Tubifex tubifex

Temperature (oC): 10 pH: 4.0 Salinity (ppt) . 0 Barometric pressure~ (mm Hg): 764

Worm weights (mg dry weight) Calibration value (mm Hg) 157.8


Page 2 of 3




A 0932 140.3 . B 0935 137.5 C 0937 137.6

A 1020 123.8 123.1 122.8 123.2 10 9 B 1030 120.6 120.5 120.3 120.5 10 9 C 1050 117.6 117.1 117.3 117.3 10 9

A 1140 104.6 104.0 104.3 9 8 113.8 .27 B 1150 101. 7 102.8 101.3 101. 9 9 8 111.2 .36 C 1200 100.7 101.1 100.8 100.9 9 8 109.1 .28

A 1300 86.1 85.7 85.9 8 7 95.1 .24 B 1310 84.0 82.9 82.2 83.0 8 7 92.5 .32 C 1320 85.7 85.0 84.2 85.0 8 7 93.0 .21

A 1420 66.9 66.1 66.5 7 6 76.2 .22 B 1430 64.2 63.5 63.5 63.7 7 6 73.4 .29 C 1440 67.2 66.2 66.6 66.7 7 6 75.9 .22

A 1540 46.8 45.8 46.4 6 4.8 56.5 .19 B 1550 43.0 42.3 42.3 42.5 6 4.8 53.1 .27 C 1600 48.4 49.8 49.5 49.2 6 5 58.0 .18

OLiGOCHAETE RESPIRATION DATA

Species: LimnodriZus hoffmeisteri/ Treatment: low pH Tuhifex Suhifex

Temperature ( C): 10 pH: 4.0 Salinity (ppt) 0 Barometric pressure~ (mm Hg): 765

Worm weights (mg dry weight) Calibration value (mm Hg) : 158


Page 3 of 3 I I I

I

Syringe R . . I Aliquot readings (mm 02) X P02

espwatl o~ volume (ml) Rate I

Re~licate Time 1 2 3 X initial final (mm Hg} (ul/mg/h} l A 1155 . 48.5 t B 1158 40.0 I

C 1202 46.6 l A 1205 33.8 34.0 34.8 34.2 10 8.8 I

I

B 1215 34.9 34.3 34.0 34.4 10 8.8 I C 1225 43.9 45.3 44.3 44.5 10 8.8 I

I I

A 1320 25.7 25.3 25.5 8.8 7.4 29.9 .17 I B 1330 27.0 26.1 25.4 26.2 8.8 7.6 30.3 .14 C 1340 37.6 38.6 38.8 38.3 8.8 7.6 41.4 .14

A 1425 17.9 15.4 16.7 7.4 6.2 21.1 .17 B 1435 19.9 18.5 18.5 19.0 7.6 6.2 22.6 .12 C 1445 27.8 28.5 28.3 28.2 ·7.6 6 33.3 .22

A 1530 8.2 7.3 7.8 6.2 5 12.3 .14 B 1540 9.5 10.3 8.9 9.6 6.2 5 14.3 .13 C 1550 19.2 19.1 19.1 19.1 6 5 23.7 .16

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