UNIVERSITI PUTRA MALAYSIA

SOME ASPECTS OF THE LIFE CYCLE OF FISH BLOOD FLUKE, Sanguinicola armata PLEHN, 1905 (DIGENEA: SANGUINICOLIDAE)

IN GRASS CARP (Ctenopharyngodon idellus CUVlER AND VALENCIENNES, 1884)

FINGERLINGS

KUA BENG CHU

FPSS 1995 4

SOME ASPECTS OF THE LIFE CYCLE OF FISH BLOOD FLUKE, Sanguinicola annata PLEHN, 1905 (DIGENEA: SANGUINICOLIDAE)

IN GRASS CARP

(Ctenopharyngodon idellus CUYlER AND VALENCIENNES, 1884) FINGERLINGS

By

KUA BENG CHU

Thesis submitted in fulfilment of the requirement for the degree of Master of Science in the Faculty of Fisheries and Marine Science,

Universiti Pertanian Malaysia

September 1995

ACKNOWLEDGEMENTS

I woul d l ike to express my deepest gra t itude to my

supervi sory committee , Dr . Fa i z ah Shaharom-Harr ison ,

Assoc i ate Professor Dr . Jambari Haj i Al i and Dr . Hassan

Mohd . Daud for the i r guidance , constant support a nd

advice . I woul d espec i a l ly l ike to thank Dr . Fa izah

Shaharom-Harr i son , the cha irman of the supervi sory

committee for her encouragement , t ime and end l ess

pat i ence throughout the proj ect .

spec i a l recogn i t ion i s a l so due to Professor J.W .

Lew i s of Roya l Ho l l oway , Un ivers ity of London who

con f i rmed the sporocyst generations in th i s study . Thanks

are a l so due to Professor Bj orn Berland of I nst itute o f

Zoology , Un ivers ity o f Bergen , Norway and Mr Maj eed o f

Faculty o f F i sher ies and Mar ine Sc ience ( Terengga nu

Branch ) for tran s l at ing some o f the l iterature in German

and Spanish . My s i ncere thanks a l so to B . W . Herbert of

Department of Pr imary I ndustr ies , Austra l i a for giving

some o f the l iterature needed f or revi ew .

I a l so w i sh to thank Mr Ng Sween Chu , owner of Ng ' s

Aquat i c Farm , Me l aka for supp lying the seed of l oca l ly

bred grass carp used in this study .

ii

I thank Mrs Karti n i Mohamad ( Terangganu Branch) who

helps w ith the scanning e lectron microscopy . Thanks a lso

to Mrs Jar i ah Sula iman and Mr Sabri Omar for helping me

with the photography and Mr . Mahmmud Jusoh who k indly l et

me use the fac i l ities in Microbr ian Laboratory .

My s i ncere gratitude go to IRPA ( Intens i f ication o f

Research for Prior ity Area ) No : 1 - 0 7 - 0 5 - 0 7 6 - 5 0 3 7 0 for

f inanc i a l ass i stance in this study . I am a l so grateful to

Professor Mohamed Shar i f f bin Mohamed D i n for h i s support

and help me in var i ous ways dur ing the pro j ect . I thank

Mrs Chan Soo Mum , Miss Prema la Arul and a l l fr iends who

supported me in every way throughout the proj ect .

F i na l ly , my s incere thanks to my dad , mum , s i sters

and brothers for the i r inva luable and endless

encouragement , sacr i f i ces , pat ience and l ove throughout

my study .

i i i

TABLE OF CONTENTS

Page

ACKNOWLEDGEMENTS . . . . . . . . . . . . . . . . . . • • . • • . . . . i i

LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . • . . . • vii

LIST OF FIGURES . . . . . . . . . . . . . . . . . . • . . . • . • . . • ix

LIST OF PLATES . . . . . . . . . . . . . . . . . . • • • . . . . . . . . x i i

LIST O F ABBREVIATIONS . . . . . . . . . . . . . . . . . . . . . . ABSTRACT • • • • • • • • • . • • • • • • • • • • • • • • • • • • • • • • • • • ABSTRAK • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

CHAPTER

I

I I

I NTRODUCTION . . . . . . . . . . . . . . . . . . . . . . Ba ckground . . . . . . . . . . . . . . . . . . . . . . . Sanguinico l iasis . . . . . . . . . . . . . . . . . Paras ito l og i ca l status of Sanguini col a arma ta i n Ma laysia . . Obj ect ives . . . . . . . . . . . . . . . . . . . . . . .

LI TERATURE REVIEW . . . . . . . . . . . . . . . . . Genus Sanguini col a . . . . . . . . . . . . . . . L i f e Cyc l e . . . . . . . . . . . . . . . . . . . . . . .

Hosts . . . . . . . . . . . . . . . . . . . . . . . . . Transmi ss ion Patterns . . . . . . . . . .

Morphological Features . . . . . . . . . . . Adult sanguinicolid .......... . Eggs and Miracid ium . . . . . . . . . . . Sporocyst . . . . . . . . . . . . . . . . . . . . . Cercar iae . . . . . . . . . . . . . . . . . . . . .

Pathology . . . . . . . . . . . . . . . . . . . . . . . . Prevent ion and Contro l . . . . . . . . . . .

iv

xvi

xvi i

xx

1 1 3

5 7

8 9 1 3 1 3 1 6 2 1 21 22 23 24 26 2 9

III MORPHOLOGICAL FEATURES OF THE EGGS AND MIRACIDIA OF Sanguinicola armata ISOLATED FROM THE DEFINITIVE HOST , GRASS CARP (Ctenopharyngodon idellus) FINGERLINGS. • • • • • • • • • • • • • • • • • • • • • • • • • • 31 Mater i a l s and Methods . . . . . . . . . . . . . . . . 3 3

Grass Carp Fingerl ings . . . . . . . . . . . . 3 3 Eggs and Mirac idia . . . . . . . . . . . . . . . . 3 3

Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 4 Eggs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 4 Miracidia . . . . . . . . . . . . . . . . . . . . . . . . . 3 5

D iscussion . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3

IV SUITABILITY OF THE SNAIL Gyraulus convexiusculus ( GASTROPODA : PLANORBIDAE) AS AN INTERMEDIATE HOST OF THE FISH BLOOD FLUKE , Sanguinicola armata............. 4 8 Mater i a l s and Methods . . . . . . . . . . . . . . . . . 5 0

Ma intenance of Definit ive Host . . . . . 5 0 Mai ntenance of Intermediate Host . . . 5 0 Exper imenta l Des ign . . . . . . . . . . . . . . . . 5 3 Laboratory Exper imenta l Infections . 5 9 H i sto logy . . . . . . . . . . . . . . . . . . . . . . . . . . 6 0

Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 Pre-Tr i a l Exper iments . . . . . . . . . . . . . . 6 2 Laboratory Exper imenta l Infect i ons . 6 3 Sporocyst . . . . . . . . . . . . . . . . . . . . . . . . . . 6 4

D i scussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1

V MORPHOLOGICAL FEATURES OF Sanguinicola armata CERCARIAE . . . . . . . . . . 7 7 Mater i a l s and Methods . . . . . . . . . . . . . . . . . 7 8

Fresh Observation . . . . . . . . . . . . . . . . . . 7 8 Scanning Electron Microscopy . . . . . . . 7 9

Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 0 Observat ion of Cercariae Under Nomarski opt ics . . . . . . . . . . . . . . . . . . . . 8 0 Observation of Cercariae Under S canning E lectron Microscope . . . . . . . 8 2

D i s cuss ion . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 2

v

VI TRANSMISSION OF CERCARIAE OF Sanguinicola armata TO NAIVE GRASS CARP (Ctenopharyngodon idellus)

VII

FINGERLINGS . • • • • • • • . • • • . • • • • • • • • • • • • • • 99 Mater ials and Methods . . . . . . . . . . . . . . . . 1 0 0

Grass Carp Fingerl ings . . . . . . . . . . . . 1 0 0 Infected Sna i ls . . . . . . . . . . . . . . . . . . . 1 0 0 Laboratory Transmiss ion Exper iments . . . . . . . . . . . . . . . . . . . . . . . 1 0 1

Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 0 3 Observation o f Cercar iae Under The Dissecting Microscope . . . . . . . . . 1 0 3 Infected F ish . . . . . . . . . . . . . . . . . . . . . . 1 0 6 Adu lt F lukes . . . . . . . . . . . . . . . . . . . . . . 1 0 8 The Eggs and Mirac idia . . . . . . . . . . . . 1 1 0

D i scuss ion . . . . . . . . . . . . . . . . . . . . . • . . . . . 1 1 1

GENERAL DISCUSSION AND CONCLUSION . • • • General D i scuss ion . . . . . . . . . . . . . . . . . . . Conc lusion . . . . . . . . . . . . . . . . . . . . . . . . . . .

1 19 1 1 9 1 2 6

BIBLIOGRAPHY • • • • • • • . . . • • . • . . . • • • . • • • . . . • . . • • . • • • 130

APPENDICES . • . . • • . . . . • . . . . . . . . . . . . . • • • . • . • • • . • . . . 2 0 7 A - ADDITIONAL TABLES . . . . . . . . . . . . . . 140-150 B - ADDITIONAL FIGURES . . . . . . . . . . . . . 151-1� C - TAXONOMI CAL KEY . . . . . . . . . . . . . . . . 1 5 5

BIOGRAPHICAL SKETCH . • . • • . • . . . . . . . . • • • • • • • • . • • 1 5 6

vi

LIST OF TABLES

Table Page

1 Observation of the f ish b lood f luke of the genus Sanguini col a in the f ina l hosts and the i r wor ld d istri but ion . ( Adapted from Ong , 1 9 9 4 ) . . . . . . . . 1 1

2 Intermedi ate and definit ive hosts of the f i sh blood f luke of the genus Sanguini col a . ( Adapted f rom Ong , 1 9 9 4 ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 4

3 Morphol ogy of the eggs of sangu i n i co l id a s reported by var ious authors . . . . . . . . . . . . . . . . 2 2

4 Measurements of the eggs of S . arma ta i s o l ated f rom var ious organ of thirty f inger l ings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 5

5 Measurements of S . arma ta eggs by var i ous authors . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 6

6 Percentage of sna i l inf ect ion rate at d i fferent exposure t imes . . . . . . . . . . . . . . . . . . . . . . 6 2

7 Percentage of sna i l infect i on rate in laboratory infection experiments . . . . . . . . . . . . . . 63

8 Mean measurements of S . arma ta cercariae under l ight microscope . . . . . . : . . . . . . 8 1

9 Measurements o f mature cercari a e of S . arma ta under the scanning e lectron m i croscope . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 3

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10 Observation of the adults , eggs and miracidia of S . armata in i n fected gras s carp f i nger l ings . . . . . . . . . . . . . . . . . . . . . . . 10 7

11 Measurements of the eggs of S . armata i n vari ous organs . ( Raw data - i n m icrometre ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 0

1 2 Measurements of the mirac i d i a o f S . armat a . ( Raw data - in m icrometr e ) . . . . . . . . 1 4 2

13 Measurements of S . armata cerca r i a e under l ight microscope . ( Raw data - in m icrometre ) . . . . . . . . . . . . . . . . . . . 14 3

1 4 Measurements of S . arma ta cerca r i ae under scann ing electron m i croscope ( Raw data - in m icrometre ) . . . . . . . . . . . . . . . . . . . 1 4 4

15 Measurements of the adu lt f lukes o f S . arma ta . ( Raw data - in m i crometre ) . . . . . . . . 1 4 5

1 6 water qua l ity mea surements i n pre-tr i a l exper iments ( I , I I and I I I ) and the l aboratory in fect i on exper iment ( A ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 4 6

1 7 Two- sampl e ana lys i s results for exper iment I ( F ish and sna i l s left together throughout the exper iment ) and experiment I I ( 2 4 hours exposure ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 4 9

1 8 Two-samp l e analys is resu lts for exper iment A ( Observat ion of two sna i l each day ) and B ( Observat i on o f a l l the post-exposed sna i l s ) . . . . . . . . . . . . . . 14 9

1 9 Water qua l ity measurements i n transmiss i on exper imenta l tanks . . . . . . . . . . . . . . 1 5 2

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LIST OF FIGURES

F igure

1 Transm i s s ion patterns o f sanguinico l id

Page

b lood f l uke . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . 17

2 D i agrams o f S . armata eggs show ing vari ou s stages o f deve lopment . . . . . . . . . . . . . . . . . 3 8

3 A c ompos ite drawing of the ful ly formed mirac id ium o f S . arma ta with in the egg . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 0

4 D iagrams showing a sequence of m i ra c i d i a l movement w ithi n the egg shel l and l i beration of miracidium from the egg . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2

5 Laboratory-bred sna i l s G. convexi us cul us exposed to g i l l t i s sue infected with S . arma t a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 4

6 Pre-tri a l experiment I and I I to con f i rm G . convexi usculus a s the intermediate host of S . arma ta . Abbreviat ions : I F ( Infected Fish) , UF ( Uninfected F i s h ) ,

us ( Un i n f ected Sna i l s ) . . . . . . . . . . . . . . . . . . . . . . . . 5 7

7 Pre-tr i a l exper iment I I I to conf irm G . convexi uscu l us a s the intermediate host o f S . armat a . Abbreviat i ons : GT ( G i l l T i s su e ) , UGT ( Uninfected G i l l Ti s su e ) , I F ( In fected F i s h ) , UF ( Uninfected F i s h ) a nd U S ( Uninfected Sna i ls ) . . . . . . . . . . . . . . . . . . . . 5 8

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8 Two sets of exper iments carr i ed out to infect the sna i l s G . convexiu s cu l us with S . arma ta miracidium . . . . . . . . . . . . . . . . . . . . . 61

9 Diagrams showing the immature stages o f S . armata . Scale bar = 15 �m . . . . . . . . . . . . . . 6 6

10 A compos ite draw ing of mature cerca r i a of S . armata . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 5

11 Laboratory transmiss ion exper iments of S . arma ta cercariae to na i ve gras s carp f inger l ings . . . . . . . . . . . . . . . . . . . . . . . . 10 2

12 The a ct i ve and pass ive pos i t i on s of cercaria of S. arma ta in the water . . . . . . . . . . . . 10 4

13 D iagrams showing the penetration process ( A , B , C , D and E ) of S. armata cercaria entering the abdominal area of the f i sh host , grass carp (C. idel l us ) f ingerl ings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 5

14 Morpho logical f eatures o f immature worm (A) found on day 12 and the mature worm (B) on day 18 . . . . . . . . . . . . . . . . . . . . . 10 9

15 The l i fe cyc le of sanguinicol a armata . . . . . . . . . 12 7

16 Map of Peninsul a Ma lays ia showing the locat i on of gras s carp farm . • sa lak South Baru , S e l angor

( I nfected f inger l ings ) �KPg . Pandan , Me laka ( Uninfected f inger l ings ) . . . . . . . . . . . . . . . . . 14 0

17 Procedures f or d a i ly fresh observation on the eggs , miracidium and sporocyst of S . arma t a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 1

x

18 Procedures for fresh observation on cercariae of S . arma ta . . . . . . . . . . . . . . . . . . . . . 1 4 2

19 Experiments to i nfect the sna i l s G. convexiusculus with S . armata m i r a c id ium for h istological stud i e s . . . . . . . . . . . 143

xi

LI ST OF PLATES

P late Page

1 Var i ous stages o f S . arma ta eggs . . . . . . . . . . . . . . 37

2 Mature egg of S . armata showing the miracidia stage . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

3. The mature mira c id ium (MI) about to hatch w ith the granu les ( G ) be ing pushed to one s i de . ( Nomarski opt ics ) S c a l e bar = 1 0 j.Lm • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 4 1

4 . Emergence o f the mirac i di um from egg shel l . Scale bar = 2 j.Lm • • • • • • • • • • • • • • • • • • • • • • • 4 1

5 . The defin it ive host , grass carp (C. i del l u s ) f inger l ings used in the study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

6. The i ntermedi ate host , sna i l G . convexi uscul u s used i n the study. Sca l e bar = 1 mm . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . 5 2

7. Sporocyst stages i n the sna i l G . convexi uscul u s . Sca l e bar = 2 6 J.Lm . . . . . . . . . . 6 5

8 . Var ious stages o f daughter sporocyst conta i n ing immature cercar i a in a thin membrane . Scale bars : A , B , C and D ( 2 0 j.Lm ) • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 67

9. Hor i z onta l secti on of d igestive t i ssue ( TB) of uninfected sna i l on day 1 4 . Th i ckness ( 5 J.Lm ) H & E. (lOOx) . . . . . . . . 69

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1 0 . Hor i z onta l section o f d igest ive t i s sue (TB) of s na i l with heavy infect i on of immature stages o f s. armata (Arrowed) on day 1 4 . Thi ckness ( 5 �m) H & E . ( 1 0 0 x ) . . . . . . . . . . . . . . . . 6 9

1 1 . Close-up o f hor i z onta l secti on o f d i gestive t i s sue of sna i l showi ng the immature stages of s. armata (Arrowed) in between the tubul e system (TB) , Thi ckness ( 5 �m) H & E . Sca l e bar = 45 fJ.m . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 0

1 2 . A mature cercar i a of S . arma ta w i th body (B) consisting of dorsa l f i n ( DF ) , the sensory ha i r (SH) at the ta i l (T) a nd forked part of the ta i l (FT) . ( Nomarski optics ) Scale bar = 2 6 �m . . . . . . . . . . . 8 4

1 3 . The body of cercar ia with dorsa l f i n ( DF ) and c ircu lar musc les ( eM ) a t the anter i or of cerca r i a l ta i l . ( Nomarsk i opt ics/vital sta i n ) Scale bar = 1 0 � rn • • • • • • • • • • • • • • • • . • • • • • • . • • • • • • • . . • • • • . . • • • • • 8 6

1 4 . The l ong and tubular ta i l of cercar i a w i th the numerous sensory ha i r ( SH) . ( Nomarski opt ics ) Sca l e bar = 1 6 �m . . . . . . . . . . . 8 6

1 5 . The f orked part of cercar i a l ta i l A . The forked ta i l was surrounded by the membrane ( M ) . B . Only one sensory h a i r ( SH) present on both sides of forked ta i l . ( Nomarsk i opt i cs ) Sca le bars : A a nd B ( 6 �m ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 7

1 6 . The body (b) o f S . arma ta cercar i a w ith the constr i ct ion of dorsal f infold ( df) and body spines (bs) on the tegument . Scale bar = 2 0 �m . . . . . . . . . . . . 8 8

x i i i

1 7 . C lose-up of cepha l ic organ whi ch i s encircled by f ive rows of barbed-sp ines (Arrowed) at the anterior part of cepha l ic organ . The pap i l lae (pa) arise i n the midd l e of cerca r i a l mouth . Sca l e bar = 5 ,urn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 8

1 8 . The body surface (Arrowed) and the two pores of excretory system (fc) in the middle of the cerca r i a l body . Scale bar = 5 ,um • • • • • • • • • • • • • 8 9

1 9 . The body (b) and the ta i l (t ) was separated by a deep constr iction (Arrowed) . Scale bar = 5 ,um • • • • • • • • • • • • • • • • • • 89

2 0 . Numerous ta i l spines (Arrowed) and sensory hair (sh ) at the cercar i a l ta i l . Scale bar = 1 0 ,urn • • • • • • • • • • • • • • • • • • • • • • 9 0

2 1 . C l ose-up o f a pa i r of sensory ha i r (sh ) and the ta i l spines (Arrowed) at the cercar i a l ta i l . S c a l e bar = 5 ,urn • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 9 0

2 2 . C los e-up of the spines ( Arrowed ) at the forked ta i l of the cerca r i a . Scale bar = 5 ,urn • • • • • • • • • • • • • • • • • • • • • • • • • • • • • 9 1

2 3 . C lose-up of the dorso-ventr a l f in f o l d (Arrowed) and pore (p ) a t the end of furc a l ta i l reg i on o f cercari a . S c a l e bar = 0 . 1 ,um • • • • • • • • • • • • • • • • • 9 1

2 4 . Abundance of immature eggs of S . arma ta (Arrowed ) on the g i l l f i l aments o f dead f i sh on day 2 4 . ( Nomarski opt i cs ) ( 1 0 0 x ) . . . . . . . . . . . . . . . . . . . . . 1 0 6

xiv

2 4 . A mature worm of s. armata sta i ned w ith acridine orange on day 1 8 . S c a l e bar = 4 8 �m . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 0 9

2 5 . G i l l f i l ament with two immature eggs of S . armata (Arrowed) on day 77 . ( Nomarski opt ic s ) ( 1 0 0 x ) . . . . . . . . . . . . . . . . . . . . . 1 1 0

xv

B & b bs CM cm DF E ego FC FT G GC GT H & E I C I F I GT L M m l mm pa p PG ppm R SA SC SD S EM SH & sh T & t TB TEM UG UGT US VC Z J.1.m

LIST OF ABBREVIATIONS

Body Body spines C ircular Muscles cent imetre Dorsal F i n Eyespot For Examp le Flame Ce l l Forked Ta i l Granu l e Germ Ce l l G i l l T i s sues Haematoxy l in and Eos i n Immature Cercar ia Infected Fish Infected G i l l T i ssues Litre Membrane Mi l l i l itre M i l l imetre Pap i l la Pores Penetrat i on Gl and Part Per-Mi l l i on Rod let Sac Segmented Ce l l Standard Deviation Scanning Electron M icroscopy Sensory Ha ir Ta i l Tubule Transmission Electron Microscopy Unknown Gland Un infected G i l l T i ssues Un infected Sna i l vite l l ine Ce l l Zygote Micrometre

xvi

Abstract of thes i s submitted to the Senate o f Univer s i t i Pertani an Ma l ays ia i n ful f i lment o f the

r equirements for the degree of Master o f Sc i enc e .

SOME ASPECTS OF THE LIFE CYCLE OF FISH BLOOD FLUKE, sanguinicola armata PLEHN, 1905 (DIGENEA:

SANGUINICOLIDAE) IN GRASS CARP (Ctenopharyngodon idellus CUVIER AND VALENCIENNES 1884) FINGERLINGS

By

KUA BENG CHU

September 1 9 9 5

Cha i rman Fa i z ah Shaharom-Harr i son, Ph . D

Facu lty Fi sheries and Mar i ne S c i e nce

The l i fe-cycle of the f i sh blood f luke , Sanguinicol a

arma t a P lehn, 1 9 05 wa s stud i ed i n the l a boratory . The

sna i l Gyraulus convexiuscul us Hutton , 1 8 4 9 wa s ident i f ied

a s the i ntermed i ate host of S . arma ta from grass carp

(ctenopharyngodon idellus ) f i nger l i ngs .

Two types of exper iment ( pre-tr i a l a nd laboratory

exper imenta l i nfect ions ) were car r i ed out in order to

i nfect the l a boratory bred sna i l s . The pre -tr i a l study was

d iv i ded i nto three exper iments . In the f irst exper iment ,

twenty i nfected f i sh were exposed to the thirty uni nfected

sna i l s a nd was l eft for the duration of the exper iment .

For the second exper iment , s imi lar number of f i sh a nd

xvi i

sna i ls were l eft together for 2 4 hour s . I n the third

experiment, sna i ls were exposed to g i l l t i s sues conta i n i ng

m i r a c i d i a for 12 and 2 4 hours . Only sna i l s which were

exposed to l ive i nfected f ish w ith S . armata for 2 4 hours

exposure a nd left together throughout the experiment

became i nfected . The range of the percentage of sna i l

i nf ecti on rate i n the f irst and second exper iment were 0 -

2 5 % and 1 0 - 4 6% respect i vely . For l aboratory exper imenta l

i nfections , s im i lar procedures were performed as i n the

pre-tri a l study except that ten i nfected f i sh were used .

The range of the percentage of sna i l i nfect i on rate i n

l aboratory experiment a l i nfections were 3 2 - 4 8 % .

cercar iae of S . armata produced from the laboratory

exper iment a l i nfect ions were used to i nfect the uninfected

grass carp f i nger l i ngs . Three hundred cercar i ae were

exposed to 1 0 2 un i nfected grass carp f inger l ings for 2 4

hour s . Two f i sh were exam i ned da i ly for over a per i od of

1 0 5 days . A h i gh rate o f i nfecti on ( 74 % ) with a low rate

o f morta l ity ( 4 % ) was obta i ned from the study .

A m inimum t ime of 4 0 to 4 3 days was needed to

complete the l i fe-cyc l e of S . armata . Newly l a id eggs

conta i ned several v i te l l i ne c e l l s and embryos . within 6 to

xvi i i

8 days , the eggs became mature a nd possessed a movi ng

c i l i ated miracidia . The eggs whi ch were found i n the

k idney , heart , l i ver and spleen were e ncapsul ated but the

eggs found in the g i l l t issues were not . Miracidia only

hatched in the gi l l tissues a nd swam freely before

penetra t i ng the sna i l , G . convexi u s cu l u s .

Upon enter ing the sna i l , the m i racid ium formed a

mother sporocyst, wh ich then produced a daughter sporocyst

by asexu a l reproduct ion. The shape o f the sporocyst was

var iabl e , th in-wall ed , nonmot i le and unbranched . A max imum

of two to three immature cercar iae were found ins ide the

th in membrane . Forked-ta i l c ercar iae deve l oped from two

sporocyst generat i ons within 14 to 1 5 days in the sna i l .

They were releas ed from the sna i l s and swam i n the water

towards the abdomina l region of grass carp finger l ings .

The ta i l of the cercar iae were shed when penetrat ion of

the cerca r i a l body was completed . cerca r i a took 1 8 days to

undergo the process from penetrat ion to migrat ion into the

b lood vesse l , matured

tri angu lar-shaped eggs .

into an adult and r e l eased

Adu lt f luke wh ich inhabited the

bu lbus arter iosus was ident i f i ed from its l anceo late shape

with margi na l sp ines on both s ides of its body , butter f ly­

shaped ovary and 10 pairs of testes .

x i x

Abstrak t e s i s yang d ikemukakan kepada Senat Univers i t i Pertania n Ma lays i a , sebaga i keperluan untuk mendapat

I j a z ah Master S a i ns .

BEBERAPA ASPER RITARAN HIDUP FLUR DARAH IRAN, Sanguinicola armata PLEHN, 1905 (DIGENEA: SANGUINICOLIDAE) DARI FRI IRAN RAP RUMPUT

(Ctenopharyngodon idellus CUVIER AND VALENCIENNES 1884)

O leh

RUA BENG CHU

September 1995

Pengerus i Fa i z ah Shaharom-Harr ison, Ph . D

Faku l t i Per ikana n d a n S a i ns S amudera

Ki taran hidup f luk darah ikan, Sanguinicol a armata

P l ehn, 1905 telah d ikaji d i d a l am makma l . Siput Gyraulus

convexiuscul us Hutton , 1849 telah d ikena lpasti sebaga i

perumah perantaraan untuk S. armata dar i f r i ikan kap

rumput (Ctenopharyngodon i del l us ) .

Dua eksper imen ( Pra-percubaan dan eksper imen

j a ngk itan makma l ) t e l ah d i jalankan untuk menja ngk i t i s iput

yang d ib i ak da l am makma l . Ka j i an pra-percubaan d ibahagikan

kepada t iga eksper imen . Dalam eksper imen pertam a , dua

puluh ikan j a ngk itan d idedahkan kepada t iga pu luh s iput

yang t idak d i j a ngk i t i dan d i b iarkan sepanjang masa

eksper imen . Untuk eksper imen kedua , j um l ah ikan dan s i put

xx

yang sarna telah digunakan dan d ibiarkan se lama 2 4 j am .

Bagi eksperimen ketiga , s iput d i dedahkan kepada t i su

i nsang yang mengandung i mira s i d i a untuk tempoh 12 dan 2 4

j am . Hanya s iput yang d idedahkan dengan ikan h idup bagi 2 4

j am dan d ib iarkan

ber j angkit . Ju lat

sepanj ang masa eksper imen d idapati

peratus s iput yang d i j a ngk i t i untuk

eksper imen pertama dan kedua ada lah 0 - 2 5 % dan 10 - 4 6 %

mas i ng-mas i ng . Untuk j angk itan eksper imen makma l , kaedah

sarna seperti da lam kaj ian pra -percubaan kecua l i hanya 10

ikan j a ngk itan d igunakan . Ju lat peratus s iput yang

d i j angk iti da lam j a ngk itan eksper imen makma l ada lah 3 2 -

4 8 % .

Serkaria s. armata yang d ihas i lkan dar i j angk itan

eksper imen makma l d igunakan untuk menj a ngk i t i fri ikan kap

rumput yang be lum d i j angk it i . T iga ratus serkar ia te lah

d idedahkan kepada seratus dua ekor ikan kap rumput untuk

2 4 j am dedahan . Dua ekor ikan diperiksa set i ap hari untuk

j angkamasa 1 0 5 har i . Kadar j a ngkitan yang t i nggi ( 7 4 % ) dan

kadar yang k ematian rendah ( 4 % ) t e l ah d ipero lehi dar i

ka j ia n .

Tempoh masa mi nimum selama 4 0 h i ngga 4 3 har i

d iperlukan untuk melengkapkan k i taran h idup s. arma ta .

xxi

Te lur yang baru d ike luarkan mengandung i beberapa sel

v i t e l ine dan embr io. Dalam j angkamasa 6 h i ngga 8 h ar i ,

te lur menj a d i matang dan mempunya i mirasidia bers i l ia yang

bergerak akt i f . Telur yang d i j umpa i pada organ g i nj a l ,

j antung , hati dan l impa ada l ah berkapsu l manaka la t e lur

pada organ i nsang t idak sedemik i a n . Mira s i d i a menetas

hanya pada t i su insang dan berenang bebas di d a l am a ir

sebe lum menj a ngk i t i s iput , G . convexiu scul us .

Apabi l a mema suk i perumah perantara a n , m i r a s i d i a

membentuk i bu sporos ista d i mana kemud ian i a mengh a s i lkan

per i ngkat a nak spor os i sta me l a lu i pemb i akan a s eksua l .

Sporo s i sta mempunya i pelbaga i bentuk sepert i membran yang

nip is , t idak bergerak dan tidak bercabang . Maks imum dua

h i ngga t i ga serka r i a t idak matang boleh d i j umpa i d a l am

membra n nip i s sporo sista . Serkar i a berekor dwi cabang

berkembang dar ipada dua gener a s i sporos i st a d a l am

j a ngkamasa 1 4 h i ngga 15 hari di dalam s iput . Mereka akan

ke luar dari s i put dan berenang ke arah baha g i a n abdomen

f r i ikan kap rumput . Ekor serkar i a tangga l apabila badan

serka r i a telah menembusi abdomen ikan sepenuhnya . Serkar i a

mengambi l 1 8 hari untuk melalu i proses j angk itan k e d a l am

sa luran darah dan menj adi dewasa serta mengeluarkan te lur

berbentuk segit iga . Fluk dewasa yang mend i am i bu lbus

xx i i

arteriosus d ikena lpasti dari bentuknya seperti daun dengan

margi na l

berbentuk

testes .

spi na d i

s epert i

kedua bahagian

kupu-kupu dan

xx i i i

s i s i bada n,

mempunya i 10

ova r i

pasang

CHAPTER 1

INTRODUCTION

Background

The grass carp , ctenopharyngodon i del l u s ( Fami ly :

Cypr inida e ) or locally known a s 'chow hu' among

Ma l ays ians is a very popular food f i sh wh ich fetches a

h igh pr ice in the market ( Sh i r eman and Smith , 1 9 83) .

Grass carp i s one of the ma in Ch inese Carp spe c i es whi ch

was i ntroduced extens ively into Ma l ays i a i n the 1 8 0 0 ' s

from China ( Welcomm i e , 1 9 8 1 ) . I n i t i a lly grass carp was

i ntroduced pr imar i ly for controlling submerged

vegetat ion . Due to i t s fast growth rate , it soon became

an i ntegral part of compos i te f i sh culture . It is the

most cult ivated species in the Southeast As ian reg i on

( P i llay , 1 9 7 6 ) .

The product ion o f grass carp i s Ma lays i a i s however ,

limited because o f the inadequate supp ly o f s eed ma i n ly

due to var ious problems encountered in its breed i ng

techni ques in the la st decade . The increase in demand has

thus f orced some local farmers to import grass carp seed

from other countr ies such as Ta i wan and Hong Kong s ince

early 1 9 7 4 ( LOW , 1 9 7 4 ) . I nevitably , the importation of

1