Polarographic Oxygen Sensors: Aquatic and Physiological Applications

377 Pages·1983·10.37 MB·English

by  J. M. Hale (auth.)

Polarographic
Oxygen Sensors
Aquatic and Physiological Applications
Edited by
E. Gnaiger and H. Forstner
With 142 Figures
Springer-Verlag
Berlin Heidelberg New York 1983
Dr. ERICH GNAIGER
Dr. HELLMUTH FORSTNER
Institut fUr Zoologie
Abteilung Zoophysiologie
Universitat Innsbruck
Peter-Mayr-StraBe 1 a
A-6020 Innsbruck, Austria
ISBN-13: 978-3-642-81865-3 e-ISBN-13: 978-3-642-81863-9
DOl: 10.1007/978-3-642-81863-9
Library of Congress Cataloging in Publication Data. Main entry under title: Polarographic
oxygen sensors. Bibliography: p. Includes index. I. Oxygen - Analysis. 2. Polarograph
and polarography. I. Gnaiger, E. (Erich), 1952-. II. Forstner, H. (Hellmuth), 1935--.
QP535.0IP58. 1983. 574.19'214. 82-19419.
This work is subject to copyright. All rights are reserved, whether the whole or part of the
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use a fee is payable to 'Verwertungsgesellschaft Wort', Munich.
© by Springer-Verlag Berlin Heidelberg 1983.
Softcover reprint of the hardcover lst edition 1983
The use of registered names, trademarks, etc. In this publication does not imply, even in the
absence of a specific statement, that such names are exempt from the relevant protective laws and
regulations and therefore free for general use.
2131/3130-543210
Preface: A Biologist's View
Es liegt ein tiefes und griindliches Gliick darin,
daf> die Wissenschaft Dinge ermittelt, die stand
halten und die immer wieder den Grund zu neuen
Ermittlungen abgeben: - es konnte ja anders sein!
Friedrich Nietzsche
Die frohliche Wissenschaft
Molecular oxygen comprises about 20% of our atmosphere, but less
than 5% of this amount is dissolved at equilibrium in water. As a con
sequence of this low solubility in seawater, in freshwater and in aqueous
body fluids, living cells are subjected to a universal problem of low
oxygen availability. Thus biologically relevant measurement of dis
solved oxygen extends from a cellular scale where sensors with diame
ters of less than 10-6 m are employed, to the oceanic scale where in
situ measurements are performed at depths beyond 104 m.
This book covers the wide spectrum of aquatic and physiological
applications of polarographic oxygen sensors. It is intended as a basic
introduction for the student, and as a readily available compilation of
detailed information for the specialist. Assessments of various methods
for monitoring aquatic environments and physiological processes aid
in overcoming practical problems frequently encountered in the labo
ratory and in the field. Concomitant with the provision of experi
mental guidelines, topics of bioenergetics are addressed on the basis of
respiratory oxygen exchange and related physiological mechanisms.
A respiratory physiologist would specify polarographic oxygen sen
sors as perfect oxygen conformers, their respiratory rate being linearly
dependent on Po, . He would find that their oxygen consumption is
proportional to the area of their oxygen-transducing membrane, with
a QIO of 1.4 and a constant of proportionality (about 2 nmol O2 h-1
mm-2) which might be typical of, for example, fish eggs. Although
the increasing unpredictability of their oxygen consumption with in
creasing age would be understandable, he might be surprised that, des
pite their vulnerability to desiccation, oxygen consumption remained
the same in air as in water. In these respects polarographic oxygen sen
sors are not different from organisms, except that microsensors con
sume oxygen proportional to their diameter instead of area. But do
we know that microorganisms do not behave Similarly?
These apparent phenomena are explained on the basis of physico
chemical principles in Part I. Why is this understanding so important?
Since oxygen concentration per unit partial pressure (co /PO =
solubility) is low in water, small concentration changes beco~e appar-
VI Preface: A Biologist's View
ent as large changes of POl' signalled by the sensor. This feature pre
destines polarographic oxygen sensors for aquatic application: The
sensor is 20 to 30 times more sensitive to respiratory rates in water
than in the same volume of air.
The operational principle of polarographic oxygen sensors is based
on oxygen diffusion to the polarized cathode, where oxygen reduc
tion (in micromoles O per second) generates the electrical signal (in
2
amperes). According to Faraday's law the ratio is calculated as
2.591 /lmol O2 S-1 A-I (:::: 9.328 nmol O2 h-1 /lA-I). Variations in
the construction of polarographic oxygen sensors are mainly related
to the problem of how oxygen diffuses to the cathode. In this respect
the actual design of every sensor necessarily involves a compromise,
since optimising particular functions, such as sensitivity and response
time, detracts from others, such as stability and stirring requirements.
The optimal design therefore depends upon the application.
The many new commercial oxygen sensors that have come onto
the market during the past few years include, to some extent, original
ideas, or are merely copies of existing sensors. For instance, the in
formation that a polarographic oxygen sensor for marine applications
(InterOcean Systems, San Diego) is virtually the same sensor at a price
six times that of the original (Ingold, CH) probably comes too late for
some purchasers.
Proficiency in solving respirometric problems facilitates research
into the physiological mechanisms and functional interpretations of
gas exchange. With this in mind, respirometric and in situ monitoring
methods, of which the polarographic oxygen sensor forms an integral
part, are outlined in the detail that is of practical importance but
usually neglected (Parts II and III). Methods related to studies of
oxygen exchange are also dealt with. Responses to environmental
variables and to toxicological or pharmacological agents, metabolic
patterns and biological rhythms can be resolved by automatic long
term monitoring of oxygen consumption with polarographic oxygen
sensors. In combination with simultaneous measurements of, e.g.,
locomotory activity, ventilatory rates, chloroplast migration, biomass
production and notably heat dissipation such investigations can be
most fruitful.
It is noteworthy that bioenergetics and the recognition of oxygen
have a common root in Lavoisier's ingeneous concept of combustion
and his classical experiments on direct and indirect calorimetry. Keep
ing in line with the extension of the direct and indirect calorimetric
approach to the bioenergetics of aquatic animals, the thermodynamic
interpretation of oxygen consumption in aquatic organisms is revised
in an Appendix and more closely aligned to the needs of ecological
energetics. Several case studies illustrate the practical application of
the methods and point out concurrent conceptual advances.
Preface: A Biologist's View VII
The respirometers employed in these studies are no longer simply
closed or open systems. Further differentiation has given rise to
specific types such as the "rubber mask", "twin-flow" or "slurp
gun" respirometers. Various types have invaded lakes, coastal regions
and even the deep sea, yet respirometers are still most common in the
shallow, constant temperature water baths on the bench. Restocking
with new variations and their association with other instumental
groups seems rewarding, especially in some laboratories where respiro
meters have become fossilized or extinct.
Previous summaries of the applications of polarographic oxygen
sensors in medicine and cellular physiology can be found in Kessler M.
et al., eds. (Oxygen Supply. Urban Schwarzenberg, Mtinchen-Berlin
Wien, 1973) and Fatt I. (Polarographic Oxygen Sensors. CRC Press,
Cleveland, 1975). For an extended theoretical discussion the reader
is referred to Hitchman L.M. (Measurement of dissolved oxygen.
Wiley, New York and Orbisphere Laboratories, Geneva, 1978).
We hope that the joy and benefits of interdisciplinary communica
tion as experienced by the editors of this book may be shared by our
readers. Ideas and technological advances in one field may unex
pectedly provide the key to solving problems in other apparently un
related disciplines (compare e.g. Chaps. 1.4 and III.2, or p. 117 and
247). Thus perhaps even those readers whose special knowledge or
special needs are neglected or inadequately dealt with may find some
inspiration.
The plan for this book originated during a workshop held at the
Institut ftic Zoophysiologie der Universitat Innsbruck, Austria, in
October 1978. Several methodological developments and the editorial
work were supported by the Fonds zur F6rderung der wissenschaft
lichen Forschung in Osterreich, projects no. 2919, 2939, 3307 and
3917 (Univ. Prof. Dr. W. Wieser, Univ. Innsbruck, principal investiga
tor), by the Forschungsfdrderungsbeitrag der Vorarlberger Landesre
gierung and by a British Council Scholarship at the Institute for
Marine Environmental Research, Plymouth, England. I thank all con
tributors for their encouraging cooperation. I want to express my
special gratitude to Mrs. J. Wieser for her experienced help in improv
ing the English style of several manuscripts, and I thank my colleagues
and friends for their support and fruitful comments.
Innsbruck-Plymouth Erich Gnaiger
Contributors
You will find the addresses at the beginning of the respective contributions
Baldwin, R.J. 298 Hitchman,M.L. 18,31
Bals, I. 102 Johansen, K. 127
Baumgard, H. 37 Knapp, W. 195
Bridges, C.R. 219 Lomholt, J.P. 127
Broda, H. 190 Lilbbers, D.W. 37
Bucher, R. 66 Mickel, TJ. 81,184
Bilhler, H. 76 Newrkla, P. 274
Childress, J.J. 81 Ott, J. 285
Dalla Via, G.J. 202 Pamatmat, M.M. 167
Fatt, I. 234 Quetin,L.B.81,176,184
Forstner, H. 90, 111,321 Revsbech, N.P. 265
Gnaiger, E. 31,134,245,321, Schweiger, G. 190
334,337,352 Schweiger, H.G. 190
Golterman, H.L. 346 Smith, K.L., Jr. 298
Grabner, W. 86 Svoboda, A. 285
Hale, J.M. 3,73,102 Wolff, D. 190
Contents
Part I Principles of Sensor Design and Operation
Chapter I.l Factors Influencing the Stability of Polarographic
Oxygen Sensors
J.M. Hale (With 2 Figures) ................. 3
Chapter 1.2 Calibration and Accuracy of Polarographic
Oxygen Sensors
M.L. Hitchman (With 2 Figures) ............. 18
Chapter 1.3 A Thermodynamic Consideration of Permeability
Coefficients of Membranes
M.L. Hitchman and E. Gnaiger (With 1 Figure) ... 31
Chapter 1.4 Microcoaxial Needle Sensor for Polarographic
Measurement of Local O Pressure in the Cellular
2
Range of Living Tissue. Its Construction and
Properties
H. Baumgartl and D.W. LUbbers (With 13 Figures) 37
Chapter 1.5 Electrolytes
R. Bucher ........................... 66
Chapter 1.6 The Action of Hydrogen Sulfide on Polarographic
Oxygen Sensors
J.M. Hale ............................ 73
Chapter 1.7 A Double-Membrane Sterilizable Oxygen Sensor
H. BUhler (With 1 Figure) ................. 76
Chapter 1.8 Construction of a Polarographic Oxygen Sensor
in the Laboratory
T.J. Mickel, L.B. Quetin, and J.J. Childress
(With 2 Figures) ....................... 81
X Contents
Chapter 1.9 A Polarographic Oxygen Sensor Designed for
Sewage Work and Field Application
W. Grabner (With 1 Figure). . . . . . . . . . . . . . .. 86
Chapter 1.10 Electronic Circuits for Polarographic Oxygen
Sensors
H. Forstner (With 8 Figures) . . . . . . . . . . . . . .. 90
Chapter 1.11 The Application of a Microprocessor to Dissolved
Oxygen Measurement Instrumentation
I. Bals and J.M. Hale (With 3 Figures) ......... 102
Part II Laboratory Applications of Polarographic Oxygen Sensors
Chapter ILl An Automated Multiple-Chamber Intermittent-Flow
Respirometer
H. Forstner (With 7 Figures) ............... 111
Chapter 11.2 The Application of Polarographic Oxygen Sensors
for Continuous Assessment of Gas Exchange in
Aquatic Animals
J.P. Lomholt and K. Johansen (With 4 Figures) ... 127
Chapter 11.3 The Twin-Flow Microrespirometer and Simultaneous
Calorimetry
E. Gnaiger (With 20 Figures) ............... 134
Chapter 11.4 Simultaneous Direct and Indirect Calorimetry
M.M. Pamatmat (With 3 Figures) ............ 167
Chapter 11.5 An Automated, Intermittent Flow Respirometer for
Monitoring Oxygen Consumption and Long-Term
Activity of Pelagic Crustaceans
L.B. Quetin (With 5 Figures) ............... 176
Chapter 11.6 Sealed Respirometers for Small Invertebrates
L.B. Quetin and T.J. Mickel (With 3 Figures) .... 184
Chapter 11.7 A Method for the Simultaneous Long-Term
Recording of Oxygen Evolution and Chloroplast
Migration in an Individual Cell of Acetabularia
H.G. Schweiger, H. Broda, D. Wolff, and
G. Schweiger (With 3 Figures) .............. 190
Contents XI
Chapter 11.8 A Respirometer for Monitoring Homogenate and
Mitochondrial Respiration
W. Knapp (With 2 Figures) ................ 195
Chapter 11.9 Bacterial Growth and Antibiotics in Animal
Respirometry
GJ. Dalla Via (With 3 Figures) .............. 202
Chapter 11.1 0 Po and Oxygen Content Measurement in Blood
Sa~ples Using Polarographic Oxygen Sensors
C.R. Bridges (With 4 Figures) ............... 219
Chapter 11.11 Determination of the In Vivo Oxygen Flux into
the Eye
I. Fatt (With 8 Figures) ................... 234
Part III Field Applications of Polarographic Oxygen Sensors
Chapter 111.1 In Situ Measurement of Oxygen Profiles in Lakes:
Microstratifications, Oscillations, and the Limits of
Comparison with Chemical Methods
E. Gnaiger (With 11 Figures) ............... 245
Chapter III.2 In Situ Measurement of Oxygen Profiles of
Sediments by Use of Oxygen Microelectrodes
N.P. Revsbech (With 5 Figures) ............. 265
Chapter III.3 Methods for Measuring Benthic Community
Respiration Rates
P. Newrkla (With 7 Figures) ................ 274
Chapter I1I.4 In Situ Measurement of Community Metabolism
in Littoral Marine Systems
A. Svoboda and J. Ott (With 11 Figures) ....... 285
Chapter I1I.5 Deap-Sea Respirometry: In Situ Techniques
K.L. Smith, Jr. and R.J. Baldwin (With 10 Figures) 298
Appendix A Calculation of Equilibrium Oxygen Concentration
H. Forstner and E. Gnaiger ................ 321
AppendixB Calculation of Po in Water Equilibrated with a
Mixture of Room 2A ir and Nitrogen
E. Gnaiger (With 1 Figure) ................ 334