Environmental Letters
ISSN: 0013-9300 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/lesa17
Field Studies on Ozone Inactivation of a Gymnodinium Breve Toxin W. J. Blogoslawski , F. P. Thurberg , M. A. Dawson & M. J. Beckage To cite this article: W. J. Blogoslawski , F. P. Thurberg , M. A. Dawson & M. J. Beckage (1975) Field Studies on Ozone Inactivation of a Gymnodinium Breve Toxin, Environmental Letters, 9:2, 209-215, DOI: 10.1080/00139307509435848 To link to this article: http://dx.doi.org/10.1080/00139307509435848
Published online: 02 Sep 2009.
Submit your article to this journal
Article views: 4
View related articles
Citing articles: 6 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=2esa20 Download by: [NUS National University of Singapore]
Date: 06 November 2015, At: 12:34
FIELD STUDIES ON OZONE INACTIVATION OF A GYMNODINIUM BREVE TOXIN
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
KEY WORDS:
Red t i d e , Gymnodinium breve, ozone
W. J. Blogoslawski, F. P. Thurberg, M. A. Dawson and M. J . Beckage
National Marine Fisheries Service Middle Atlantic Coastal Fisheries Center Milford Laboratory Milford, Connecticut 06460 ABSTRACT Water samples were collected from Boca Ciega Bay ( S t . Petersburg) during the April, 1974, red t i d e t h a t occurred on the Florida west coast.
The causative agent o f t h i s phenomenon
was the toxic dinoflagellate, Gymnodinium breve.
The toxic red
t i d e samples were treated with ozone gas and rendered nonlethal , as measured by mouse injection. INTRODUCTION Proliferation of the toxic dinoflagellate, Gymnodinium breve, has been implicated in massive f i s h kills and other animal mortalities i n the Gulf of Mexico1 y 2 y 4 ciated with the occurrence of toxic bivalves 3
and has been asso-
.
These red tides
have caused severe economic loss t o tourist industries and recently t o f a c i l i t i e s dealing i n commercially popular tropical fish4y5. 209 Copyright 0 1975 by Marcel Dekker, Inc. All Rights Reserved. Neither this work nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher.
210
'BLOGOSLAFISKI ET plt
Powdered residues o f
5.
breve from laboratory cultures
and from seawater containing natural blooms are lethal to
mice and f i s h upon i n j e ~ t i o n ~ ' ~Ozone . gas detoxifies similar extracts from laboratory cultures8.
The present study was
i n i t i a t e d t o evaluate ozone detoxification of f i e l d samples; Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
the 1974 red t i d e provided conditions o f high g. breve concent r a t i o n in natural seawater.
Successful ozone treatment of
red t i d e water would be useful t o f a c i l i t i e s t h a t depend on natural seawater for holding and maintaining marine organisms. Such removal of toxic metabolites f r m influent water would be particularly valuable t o those who depend e n t i r e l y on a seawater supply from an area i n which red tides frequently occur.
-
EXPERIMENTAL
On April 2 , 1974, about 80 l i t e r s of seawater containing '
22 x lo6 Gymnodinium breve c e l l s / l were collected from Boca
Ciega Bay, S t . Petersburg, Florida and sent to the Milford laboratory.
This water was shipped following an on-site v i s i t
t o the red t i d e area where preliminary ozone testing was conducted. Two-liter portions of the seawater were placed i n graduated cylinders and ozone gas was bubbled t h r o u g h the seawater for 5 min using aquarium airstones (Halvin 668). Gas flow rates of 25, 55, 110, and 220 ml/min were monitored continuously w i t h Gilmont rotameters, s i z e 1. A t o t a l of 9.5 1 of
OZONE INACTLVATION OF G. breve TOXIN
seawater was treated a t each ozone rate.
211
Ozone was produced
by a Pollution Control Industries generator (Model MWP-6) using oxygen as the feed gas f o r an effective ozone yield of 1 2 gm/hr (Figure 1 ) .
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
Untreated control water samples and the ozone-treated water samples were acidified with 12 fi HC1 t o a pH of 6.0 and extracted w i t h 200 ml of diethyl ether per l i t e r of seawater. Ether was evaporated from the samples, leaving the crude ext r a c t as a dry powder618.
.
The crude toxin was suspended in
d i s t i l l e d water and injected intraperitoneally into 19-21 gm mice (Carworth, CF-1 s t r a i n ) ,
Each mouse received 30 mg of
toxin i n 1 ml o f d i s t i l l e d water. RESULTS AND DISCUSSION Extracts from red tide water samples ozonized a t or above
55 mi/min were non-toxic, as measured by mouse assay (Table 1 ) . All mice injected with control extracts died i n 2.5-3.5 min, and showed characteristic symptoms of respiratory irregularity, violent twitching, and 'loss of coordination.
Neither a i r nor
oxygen bubbled through toxic solutions f o r 20 m i n a t 220 ml/min rendered the samples non-toxic.
The reddish color of
the red tide water gradually faded as ozone was bubbled through the solution, and the final dry powder was grayish i n color.
Ether extracts and the resulting dry powder from the
control solutions were green.
212
BLOGOSLAWSKI ET
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
7 +
+-2000
ROTAMETERS
ML
GRADUATED CYLINDERS
AIRSTONES
I f7 I
OXYGEN
eg
I
OZONE GENERATOR
FIG, 1, OZONE GENERATOR
AND
SPARGI NG APPARATUS
OZONE INACTIVATION OF G. breve TOXIN
213
TABLE 1
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
Loss of
5.
breve Toxicity w i t h Ozone Gas
m l 03/min
No. mice
Avg. death time (min)
No. a l i v e
220
10
-
10
110
10
-
10
55
9
-
9
25
6
7.0
0
0
25
2.8
0
(24 h r )
The toxicity of extracts from f i e l d samples of red t i d e water was lower t h a n extracts from laboratory cultures.
We
injected 30 mg o f crude toxic material into each mouse t o achieve death times below 10 min, whereas laboratory-grown ext r a c t s gave similar death times with a 6 mg dose8.
Alam
g
al.7 reported a similar discrepancy when purifying toxin from Gymnodinium breve cultures and from red tide water and a t t r i b uted t h i s difference t o an i n e r t contaminant. Ozone has been used t o s t e r i l i z e seawater contaminated by human pathogens a t l e a s t as early as 1929’, and i s presently used i n s h e l l f i s h depuration stations and i n marine laboratories dependent upon a non-toxic natural seawater supply1oy11. Toxic red t i d e metabolites present potential water quality problems to such f a c i l i t i e s . inactivation of
5.
The present study has demonstrated
breve toxin under f i e l d conditions during
BLOGOSLAWSKI ET
214
a natural bloom.
The results of t h i s study should be of i n -
t e r e s t t o those f a c i l i t i e s seeking a method o f obtaining non-toxic seawater f o r use i n r e a r i n g and maintaining marine organisms during red t i d e outbreaks. ACKNOWLEDGMENTS Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
The authors g r a t e f u l l y acknowledge t h e cooperation of Edwin Joyce, J r .
, Karen S t e i d i n g e r , Beverly Roberts, and Mary
Ann Burklew of t h e Marine Research Laboratory, Florida Department o f Natural Resources , S t . Petersburg, F l o r i d a , and Charles Bohara o f t h e Milford laboratory.
We a l s o thank
Rita Riccio f o r e d i t i n g and typing t h e f i n a l manuscript. REFERENCES 1.
Wilson, W. B. and S. M. Ray. 1956. The occurrence of Gymnodinium brevis i n %he western G u l f of Mexico. Ecology 37: 388.
2.
S i e v e r s , A. M. 1969. Comparative t o x i c i t y o f Gonyaulax monilata and Gymnodinium breve t o an'nelids, crustaceans , molluscs, and a f i s h . J . Protozool. 16:401-404.
3.
Cumins, J. M . , A. C. Jones and A. A. Stevens. 1971. Occurrence of t o x i c bivalve molluscs during a Gymnodinium breve "red t i d e . " Trans. Am. F i s h . SOC. 100: 1 12- 116-
4.
S t e i d i n g e r , K. A. and E. A. Joyce, Jr. 1973. Florida r e d t i d e s . Marine Research Lab. Educational S e r i e s Report No. 17, S t . Petersburg, Florida.
5. Moe , M. 1974. Personal communication. s e a r c h , S t . Petersburg, Florida. 6.
Aqua1 i f e Re-
Sasner, J. J . , J r . , M. Ikawa, F. Thurberg and M. A l a m . 1972. Physiological and chemical s t u d i e s on Gymnodinium breve Davis toxin. Toxicon lo: 163-172.
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
OZONE INACTIVATION OF G. breve TOXIN
2 15
7.
Alam, M., J. J . Sasner, J r . and M. Ikawa. 1973. Isolation of Gymnodinium breve toxin from Florida red t i d e water. Toxicon 11:201-202.
8.
Blogoslawski, W. J . , F. P. Thurberg and M. A. Dawson. 1973. Ozone inactivation of a Gymnodinium breve toxin. Water Research 7: 1701-1703.
9.
Voille, H. 1929. S t e r i l i z a t i o n of sea water w i t h ozone: application of this method t o the purification of contaminated s h e l l f i s h . Rev. Hyg. 1: 1-5.
10.
Blogoslawski, W. J., C. Brown, E. W. Rhodes and M. BroadIn Press. Ozone disinfection o f a seawater supply system. Proc. 1 s t Int. Symp. on Ozone f o r \.later and Wastewater Treatment. Wash. , D. C.
hurst.
11.
Anonymous. shellfish.
1972. Use of ozone i n sea water for cleansing Effluent and Water Treatment J . E: 260-262.
The use of t r a d e names is merely t o f a c i l i t a t e description and does not imply endorsement by -the National Marine Fisheries Service, NOAA.
ISSN: 0013-9300 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/lesa17
Field Studies on Ozone Inactivation of a Gymnodinium Breve Toxin W. J. Blogoslawski , F. P. Thurberg , M. A. Dawson & M. J. Beckage To cite this article: W. J. Blogoslawski , F. P. Thurberg , M. A. Dawson & M. J. Beckage (1975) Field Studies on Ozone Inactivation of a Gymnodinium Breve Toxin, Environmental Letters, 9:2, 209-215, DOI: 10.1080/00139307509435848 To link to this article: http://dx.doi.org/10.1080/00139307509435848
Published online: 02 Sep 2009.
Submit your article to this journal
Article views: 4
View related articles
Citing articles: 6 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=2esa20 Download by: [NUS National University of Singapore]
Date: 06 November 2015, At: 12:34
FIELD STUDIES ON OZONE INACTIVATION OF A GYMNODINIUM BREVE TOXIN
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
KEY WORDS:
Red t i d e , Gymnodinium breve, ozone
W. J. Blogoslawski, F. P. Thurberg, M. A. Dawson and M. J . Beckage
National Marine Fisheries Service Middle Atlantic Coastal Fisheries Center Milford Laboratory Milford, Connecticut 06460 ABSTRACT Water samples were collected from Boca Ciega Bay ( S t . Petersburg) during the April, 1974, red t i d e t h a t occurred on the Florida west coast.
The causative agent o f t h i s phenomenon
was the toxic dinoflagellate, Gymnodinium breve.
The toxic red
t i d e samples were treated with ozone gas and rendered nonlethal , as measured by mouse injection. INTRODUCTION Proliferation of the toxic dinoflagellate, Gymnodinium breve, has been implicated in massive f i s h kills and other animal mortalities i n the Gulf of Mexico1 y 2 y 4 ciated with the occurrence of toxic bivalves 3
and has been asso-
.
These red tides
have caused severe economic loss t o tourist industries and recently t o f a c i l i t i e s dealing i n commercially popular tropical fish4y5. 209 Copyright 0 1975 by Marcel Dekker, Inc. All Rights Reserved. Neither this work nor any part may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, microfilming, and recording, or by any information storage and retrieval system, without permission in writing from the publisher.
210
'BLOGOSLAFISKI ET plt
Powdered residues o f
5.
breve from laboratory cultures
and from seawater containing natural blooms are lethal to
mice and f i s h upon i n j e ~ t i o n ~ ' ~Ozone . gas detoxifies similar extracts from laboratory cultures8.
The present study was
i n i t i a t e d t o evaluate ozone detoxification of f i e l d samples; Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
the 1974 red t i d e provided conditions o f high g. breve concent r a t i o n in natural seawater.
Successful ozone treatment of
red t i d e water would be useful t o f a c i l i t i e s t h a t depend on natural seawater for holding and maintaining marine organisms. Such removal of toxic metabolites f r m influent water would be particularly valuable t o those who depend e n t i r e l y on a seawater supply from an area i n which red tides frequently occur.
-
EXPERIMENTAL
On April 2 , 1974, about 80 l i t e r s of seawater containing '
22 x lo6 Gymnodinium breve c e l l s / l were collected from Boca
Ciega Bay, S t . Petersburg, Florida and sent to the Milford laboratory.
This water was shipped following an on-site v i s i t
t o the red t i d e area where preliminary ozone testing was conducted. Two-liter portions of the seawater were placed i n graduated cylinders and ozone gas was bubbled t h r o u g h the seawater for 5 min using aquarium airstones (Halvin 668). Gas flow rates of 25, 55, 110, and 220 ml/min were monitored continuously w i t h Gilmont rotameters, s i z e 1. A t o t a l of 9.5 1 of
OZONE INACTLVATION OF G. breve TOXIN
seawater was treated a t each ozone rate.
211
Ozone was produced
by a Pollution Control Industries generator (Model MWP-6) using oxygen as the feed gas f o r an effective ozone yield of 1 2 gm/hr (Figure 1 ) .
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
Untreated control water samples and the ozone-treated water samples were acidified with 12 fi HC1 t o a pH of 6.0 and extracted w i t h 200 ml of diethyl ether per l i t e r of seawater. Ether was evaporated from the samples, leaving the crude ext r a c t as a dry powder618.
.
The crude toxin was suspended in
d i s t i l l e d water and injected intraperitoneally into 19-21 gm mice (Carworth, CF-1 s t r a i n ) ,
Each mouse received 30 mg of
toxin i n 1 ml o f d i s t i l l e d water. RESULTS AND DISCUSSION Extracts from red tide water samples ozonized a t or above
55 mi/min were non-toxic, as measured by mouse assay (Table 1 ) . All mice injected with control extracts died i n 2.5-3.5 min, and showed characteristic symptoms of respiratory irregularity, violent twitching, and 'loss of coordination.
Neither a i r nor
oxygen bubbled through toxic solutions f o r 20 m i n a t 220 ml/min rendered the samples non-toxic.
The reddish color of
the red tide water gradually faded as ozone was bubbled through the solution, and the final dry powder was grayish i n color.
Ether extracts and the resulting dry powder from the
control solutions were green.
212
BLOGOSLAWSKI ET
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
7 +
+-2000
ROTAMETERS
ML
GRADUATED CYLINDERS
AIRSTONES
I f7 I
OXYGEN
eg
I
OZONE GENERATOR
FIG, 1, OZONE GENERATOR
AND
SPARGI NG APPARATUS
OZONE INACTIVATION OF G. breve TOXIN
213
TABLE 1
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
Loss of
5.
breve Toxicity w i t h Ozone Gas
m l 03/min
No. mice
Avg. death time (min)
No. a l i v e
220
10
-
10
110
10
-
10
55
9
-
9
25
6
7.0
0
0
25
2.8
0
(24 h r )
The toxicity of extracts from f i e l d samples of red t i d e water was lower t h a n extracts from laboratory cultures.
We
injected 30 mg o f crude toxic material into each mouse t o achieve death times below 10 min, whereas laboratory-grown ext r a c t s gave similar death times with a 6 mg dose8.
Alam
g
al.7 reported a similar discrepancy when purifying toxin from Gymnodinium breve cultures and from red tide water and a t t r i b uted t h i s difference t o an i n e r t contaminant. Ozone has been used t o s t e r i l i z e seawater contaminated by human pathogens a t l e a s t as early as 1929’, and i s presently used i n s h e l l f i s h depuration stations and i n marine laboratories dependent upon a non-toxic natural seawater supply1oy11. Toxic red t i d e metabolites present potential water quality problems to such f a c i l i t i e s . inactivation of
5.
The present study has demonstrated
breve toxin under f i e l d conditions during
BLOGOSLAWSKI ET
214
a natural bloom.
The results of t h i s study should be of i n -
t e r e s t t o those f a c i l i t i e s seeking a method o f obtaining non-toxic seawater f o r use i n r e a r i n g and maintaining marine organisms during red t i d e outbreaks. ACKNOWLEDGMENTS Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
The authors g r a t e f u l l y acknowledge t h e cooperation of Edwin Joyce, J r .
, Karen S t e i d i n g e r , Beverly Roberts, and Mary
Ann Burklew of t h e Marine Research Laboratory, Florida Department o f Natural Resources , S t . Petersburg, F l o r i d a , and Charles Bohara o f t h e Milford laboratory.
We a l s o thank
Rita Riccio f o r e d i t i n g and typing t h e f i n a l manuscript. REFERENCES 1.
Wilson, W. B. and S. M. Ray. 1956. The occurrence of Gymnodinium brevis i n %he western G u l f of Mexico. Ecology 37: 388.
2.
S i e v e r s , A. M. 1969. Comparative t o x i c i t y o f Gonyaulax monilata and Gymnodinium breve t o an'nelids, crustaceans , molluscs, and a f i s h . J . Protozool. 16:401-404.
3.
Cumins, J. M . , A. C. Jones and A. A. Stevens. 1971. Occurrence of t o x i c bivalve molluscs during a Gymnodinium breve "red t i d e . " Trans. Am. F i s h . SOC. 100: 1 12- 116-
4.
S t e i d i n g e r , K. A. and E. A. Joyce, Jr. 1973. Florida r e d t i d e s . Marine Research Lab. Educational S e r i e s Report No. 17, S t . Petersburg, Florida.
5. Moe , M. 1974. Personal communication. s e a r c h , S t . Petersburg, Florida. 6.
Aqua1 i f e Re-
Sasner, J. J . , J r . , M. Ikawa, F. Thurberg and M. A l a m . 1972. Physiological and chemical s t u d i e s on Gymnodinium breve Davis toxin. Toxicon lo: 163-172.
Downloaded by [NUS National University of Singapore] at 12:34 06 November 2015
OZONE INACTIVATION OF G. breve TOXIN
2 15
7.
Alam, M., J. J . Sasner, J r . and M. Ikawa. 1973. Isolation of Gymnodinium breve toxin from Florida red t i d e water. Toxicon 11:201-202.
8.
Blogoslawski, W. J . , F. P. Thurberg and M. A. Dawson. 1973. Ozone inactivation of a Gymnodinium breve toxin. Water Research 7: 1701-1703.
9.
Voille, H. 1929. S t e r i l i z a t i o n of sea water w i t h ozone: application of this method t o the purification of contaminated s h e l l f i s h . Rev. Hyg. 1: 1-5.
10.
Blogoslawski, W. J., C. Brown, E. W. Rhodes and M. BroadIn Press. Ozone disinfection o f a seawater supply system. Proc. 1 s t Int. Symp. on Ozone f o r \.later and Wastewater Treatment. Wash. , D. C.
hurst.
11.
Anonymous. shellfish.
1972. Use of ozone i n sea water for cleansing Effluent and Water Treatment J . E: 260-262.
The use of t r a d e names is merely t o f a c i l i t a t e description and does not imply endorsement by -the National Marine Fisheries Service, NOAA.
