DISTRIBUTION AND FEEDING PREFERENCES OF FREE-LIVING NEMATODES ASSOCIATED WITH THE KELP, Nacrocystis integrifolia (LAMINARIALE~) David Bruce Trotter B.Sc., McGill University, 1977 A THESIS SUBMITTED IN PARTIAL FULFILLIIENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in the Department of Biological Sciences @ DAVID TROTTER 1981 SIMON FRASER UNIVERSITY April 1981 All rights reserved. This thesis may not be reproduced in whole or in part, by photocopy or other means, without permission of the author. Approval I Name : David B . T r o t t e r Degree: Master of Sc ience T i t l e o f Thes i s : D i s t r i b u t i o n and f e e d i n g p r e f e r e n c e s o f f r e e - l i v i n g nematodes a s s o c i a t e d w i t h t h e k e l p , M?crocyst- is t n t o g r i f o l i a !T,aminariales) Examining Commit tee: Cha irman : D r . Rober t C . Brooke 'Dr. J . M. Webster , S e n i o r S u p e r v i s o r D r . L. D. Druehl - -- D r . Glen H. Geen, P r o f e s s o r B i o l o g i c a l S c i e n c e s Simon F r a s e r U n i v e r s i t y Pub1i c Examiner \ - Date approved '- 3 !%d//j'p\ PART l AL COPYR IG tiT L l CENSE I hereby g r a n t t o Simon Fraser U n i v e r s i t y t h e r i g h t t o lend my t h e s i s , p r o j e c t o r extended essay ( t h e t i t l e o f which i s shown below) t o users o f t h e Simon Fraser U n i v e r s i t y L i b r a r y , and t o make p a r t i a l o r s i n g l e cop ies o n l y f o r such users o r i n response t o a reqbes t f rom t h e l i b r a r y o f any o t h e r u n i v e r s i t y , o r o t h e r educa t iona l i n s t i t u t i o n , on i t s own beha l f o r f o r one o f i t s users. I f u r t h e r agree t h a t permiss ion f o r m u l t i p l e copy ing o f t h i s work f o r s c h o l a r l y purposes may be granted by me o r t h e Dean o f Graduate Stud ies. I t i s understood t h a t copy ing o r p u b l i c a t i o n o f t h i s work f o r f i n a n c i a l g a i n s h a l l n o t be a l lowed w i t h o u t my w r i t t e n permiss ion. T i t l e o f Thes is /Pro ject /Extended Essay D i s t r i b u t i o n and f eed ing p re f e r ences of f r e e - l i v i n g nematodes a s s o c i a t e d w i t h t h e ke lp , Macrocyst is i n t e g r i f o l i a (Laminar ia les) Author : ( s i g n a t u r e ) David B. T r o t t e r ABSTRACT , Free-living nematodes are a major component of the fauna of marine interstitial environments and are frequently associated with intertidal and subtidal algae. A study was done to categorize the nematodes of a kelp, Macrocystis integrifolia Bory, in the Bamfield region of Barkley Sound, British Columbia, Canada. The feeding preferences of the pre- dominant free-living nematode species found on the M-. integrifolia blades were evaluated. Nine species (belonging to six families) of nematodes were found on the blades and three of them comprised 91-99% of the nematode fauna. Two monhysterids (Monhystera disjuncta and M_. refringens) and a chromadorid Prochromadorella neapolitana) occurred in all monthly samples. About 50% of the nematodes on the blades from July to October was M. refringens, which was at its peak population level - during these months. During the fall, the M. refringens - population declined to low levels prior to a progressive increase in population commencing in February. M-. disjuncts was relatively abundant throughout the year and, in particular, during the late winter and early spring. P. neapolitana - occurred in relatively large numbers in July and subsequently declined to a very low population in the winter and spring. Nematode distribution on M_. inteqrifolia appeared to be related to blade age and the associated food sources on the blade. Both monhysterids occurred in greatest abundance on the bottom blades of Macrocystis plants and on the middle b l a d e s o f p l a n t s i n deep w a t b r . Low numbers o f P . n e a p o l i t a n a o c c u r r e d t h r o u g h o u t t h e e n t i r e d e p t h g r a d i e n t on b o t h bot tom and middle b l a d e s . Few o f t h e s e t h r e e s p e c i e s o c c u r r e d on the t o p b l a d e s . Nematode s p e c i e s o t h e r t h a n t h e s e t h r e e con- t r i b u t e d l i t t l e t o t h e abundance and d i s t r i b u t i o n p a t t e r n s cbse rved on t h e M a c r o c y s t i s b l a d e s . A l l t h r e e predominant nematode s p e c i e s showed s p e c i f i c r e s p o n s e s t o a v a i l a b l e b a c t e r i a l and d ia tom food s o u r c e s i n f e e d i n g e x p e r i m e n t s . -P. n e a p o l i t a n a e x h i b i t e d a s i g n i f i c a n t p r e f e r e n c e f o r d i a t o m s , p a r t i c u l a r l y Cocconeis s c u t e l l u m and Gramatophora m a r i n a , o v e r t h e b a c t e r i a s p e c i e s under b o t h s i m u l a t e d summer and w i n t e r c o n d i t i o n s . M. r e f r i n g e n s showed - no p r e f e r e n c e towards b a c t e r i a o r d ia toms b u t e x h i b i t e d some p r e f e r e n c e f o r p a r t i c u l a r d i a t o m o r b a c t e r i a s p e c i e s . Under b o t h e n v i r o n m e n t a l s i m u l a t i o n s M . disjuncts showed a marked p r e f e r e n c e f o r rod-shaped b a c t e r i a . The p r o b a b i l i t y t h a t t h e s e a s o n a l abundance and d i s t r i - b u t i o n o f t h e s e mar ine nematodes i s c o r r e l a t e d w i t h t h e s e a s o n a l i t y o f t h e i r food s o u r c e s , b a c t e r i a and d i a t o m s , i s s u g g e s t e d . ACKNOWLEDGEMENTS I wish to sincerely thank my senior supervisor, Dr. John Webster, for his valuable advice, inspiration, support and patience in all aspects of this study. I would like to extend special thanks to Dr. Louis Druehl for his interest and advice concerning the seaweed aspects of this study, Mr. Bruce Hopper of the Canada Department of Agriculture, Ottawa, for his valuable confirmation of the nematode species, Dr. ~ o ~ cLekwi n, Department of Botany, University of Washington, for identification of the diatom species, Dr. Glen Geen and the staff at the Bamfield Marine Station. Special thanks go to M. Shivji and A. Bergey for their help in the field work and for their s'ense of humour. In addition, thanks to F. Heijs, J. Brouns, W. Roland, J. Shzrma and many more friends for their encouragement and support. Finally, my special thanks and appreciation to M. Jacques and M, Amat for just listening. .v i . TABLE'OF CONTENTS Page Title Page ........................................ i Approval Page ..................................... ii Abstract .......................................... iii ~cknowledgements. ................................. v Table of Contents ................................. ' vi List of Tables .................................... viii List of Figures ................................... xi List of Appendices ................................ xiv INTRODUCTION ...................................... 1 DESCRIPTION OF STUDY SITE AND ASSOCIATED KELP ..... 8 MATERIALS AND METHODS ............................. 12 I . Field Survey ............................. 12 a) Sampling procedure ................... 12 b) Treatment of Samples ................. 13 c) Statistical Treatment ................ 14 d) Environmental Parameters ............. 14 e) Taxonomic notes ...................... 14 I1 . Feeding Preference Experiments ........... 16 a) Culture of Nematodes ................. 16 b) Culture of ~ o o dS ources .............. 17 c) Experimental Design .................. 19 d) Statistical Treatment ................ 21 .v ii . Page RESULTS I ........................................ 22 a) 0bservatior.s on Physical Parameters ..... 22 b) Nematode Species Composition ............ 22 c) Seasonal Species Changes ................. 22 d) Species nistrihutior? on Uelp ............. 34 a) Prochromadorella neapolitana ............. 63 b) Monhystera refringens .................... 68 d) Life Cycle Observations on Monhystera disiuncta ................................ 78 DISCUSSION I1 ..................................... 80 GENERAL DISCUSSION ................................ 84 APPENDICES ........................................ 89 REFERENCES ........................................ 113 - viii - L I S T OF TABLES Table Page 1 Diatom (Dl-D8) and bacteria (BI-B~)s pecies isolated from Macrocystis integrifolia blades and used in the feeding preference experiments. 2 Checklist of free-iiving nematodes found in blade samples of Macrocystis integrifolia. 3 Total number/0.5 m2 and per cent abundance of males, females and juveniles of Prochromadorella neapolitana over sampling period. 4 Total number/0.5 m2 and per cent abundance of males, females and juveniles of Monhystera - refringens over sampling period. 5 Total number/0.5 m2 and per cent abundance of males, females and juveniles of Monhystara disjuncta over sampling period. 6 Distribution of Proch.romadorella neapolitana with reference to Macrocystis integrifolia blade position and depth F values for a grouped two-level ANOVA calculated on the number of nematodes/0.5 m2 for each sample month. 7 Distribution qf Monhystera refringens with reference to Macrocystis integrifolia blade position and depth F values for a grouped two-level ANOVA claculated on the number of nematodes/0.5 m2 for each sample month. 8 Distribution of Monhystera disjuncta with reference to Macrocystis integrifolia blade position and depth F values for a grouped two-level ANOVA calculated on the number of nematodes/0.5 m2 for each sample month. 9 Distribution of the three predominant nematode species with reference to Macrocystis integrifolia blade position and depth F values for an unreplicated three-level ANOVA calculated on the number of nematodes/0.5 m2 for each sample month. Page Total male and female mean accumulation of 6 4 Prochromadorella neapolitana to diatom and bacteria species (see able 1) under simulated summer conditions. Analysis of attractiveness and preference of Prochrornadorella neapolitana to diatom and bacteria species (see Table 1) using Chi-square two-level ANOVA and Student- Newrnan-Ruels testlr,g under simulated suimer and water conditions. Total male and female mean acchulation of . Prochromadorella neapolitana to diatom and bacteria spe~ies( see Table 1) under simulated winter conditions. Total male and female mean accumulation of Monhystera refringens to diatom and bacteria species (see Table 1) under simulated summer conditions. Analysis of attractiveness and preference of ~onhysterar efringens to diatom and bacteria species (see Table 1) using Chi-square, two- level ANOVA and Student-Newman-Kuels testing under simulated summer and winter conditions. Total male 2nd female mean accumulation of Monhystera refringens to diatom and bacteria species (see Table 1) under simulated winter conditions. Total male and female mean accumulation of Monhystera disjuncta to diatom and bacteria species (see Table 1) under simuiated. summer conditions. Analysis of attractiveness and preference of Monhystera disjuncta to diatom and bacteria species (see Table 1) usinq Chi-square, two- level ANOVA and Student-Newrnan-Kuels testing under simulated summer and winter conditions. Total male and female mean accumulation of Monhystera disjuncta to rod and coccoid bacteria species (see Table 1) under simulated summer conditions.