GIARDIA MURI S AN D GIARDI A DUODENALI S GROUPS :

ULTRASTRUCTURAL DIFFERENCE S BETWEE N T H E TROPHOZOITE S

M a r i a Inê s L . S O G A Y A R (1 ) & E l i s a A p a r e c i d a G R E G Ó R I O (2 )

SUMMARY

Trophozoites o f th e Giardi a muri s grou p fro m hamsters , domesti c rat s an d

mice an d o f th e Giardi a duodenali s grou p fro m hamster s an d domesti c rat s wer e
examined b y transmissio n electro n microscopy . Th e basi c ultrastructur e o f th e
trophozoites wa s similar. Difference s wer e show n in th e morpholog y o f th e ventro -
lateral flang e o f th e trophozoite s o f Giardi a muris an d Giardia duodenalis groups.
Marginal plate s are less developed in the species of th e Giardia duodenalis group. I n
this group , th e dista l extremit y o f th e latera l flang e i s shor t an d thic k an d th e
marginal plat e doe s no t penetrat e int o th e dista l extremit y o f th e flange . I n th e
Giardia muri s group , the ventro-latera l flang e is well developed and narrow an d the
marginal plate penetrate s th e distal extremity o f th e flange. The osmiophilic lamella ,
which accompanie s th e dorsa l surfac e o f th e margina l plat e i s see n onl y i n th e
Giardia muri s group .

K E Y W O R D S : Giardi a muris; Giardi a duodenalis; Trophozoite, Ultrastructure .

INTRODUCTION

Based o n th e media n bod y morpholog y MATERIAL AN D METHOD S

and th e trophozoite' s siz e an d shap e on e ca n
differentiate Giardi a muri s an d Giardi a
duodenalis b y ligh t microscopy . Th e dat a i n
the literatur e abou t suc h differentiatio n b y
electron microscop y ar e scarce. N E M A N I C et.
a l . wer e unabl e t o sho w difference s betwee n
12
trophozoites o f Giardi a muri s fro m mic e an d
those of Giardi a lamblia fro m man .
Taking i n accoun t thes e result s an d since
the morpholog y o f Giardi a from hamster s an d
domestic rat s ha d no t bee n describe d yet , w e
decided t o stud y comparativel y th e
ultrastructure o f Giardi a muri s an d Giardi a
duodenalis trophozoite s foun d i n hamsters ,
domestic rat s an d mice.
Giardia specie s fro m naturall y infecte d
4
animals wer e previousl y identifie d b y ligh t
microscopy. Smear s o f faece s or mucos a were
fixed wit h Schaudinn' s solutio n an d staine d
with Heidenhain' s haematoxylin . Th e species
of Giardi a wer e identifie d b y morphologica l
features an d trophozoit e lengt h an d widt h
measurements, at 1.00 0 x magnification .
Trophozoites o f th e Giardi a muri s grou p
from hamsters , domesti c rat s (Rattu s rattu s
rattus) an d mic e and Giardi a duodenalis grou p
from hamster s an d domesti c rat s wer e used .
Fragments o f th e host' s mucos a were fixe d i n
2.5% glutaraldehyd e buffere d a t p H 7. 3 wit h

(1) Departmen t o f P a r a s i t o l o g y , I B B , U N E S P , 1860 0 B o t u c a t u , S ã o P a u l o , B r a z i l .

(2) Departmen t o f M o r p h o l o g y , I B B , U N E S P , 1860 0 B o t u c a t u , S ã o P a u l o , B r a z i l .
0.1M phosphat e buffer, post-fixe d i n 1 % osmic
tetroxide i n th e sam e buffer an d embedde d i n
Araldite. Th e ultrathi n section s wer e staine d
with urany l acetat e an d lea d citrat e an d
examined wit h a n E M 30 1 Philip s electro n
microscope.
RESULTS
The basi c ultrastructur e o f th e tropho -
zoites o f Giardi a uiuri s grou p fro m hamsters ,
domestic rat N an d mic e an d Giardia duodenalis
group fro m hamster s and domestic rat s is simi-
lar. Th e trophozoite s hav e a microtubuia r ad -
hesive disk surrounded b y an adjacent cytoplas -
mic flang e know n a s th e v entro-lateral flang e
and fou r pair s o f flageil a (Fig , 1 ) whose basa l
bodies ar e locate d betwee n th e tw o nuclei . Os-
miophilic materia l i s seen nexi t o th e axonemes
of th e flageil a an d vacuole s ar e presen t i n th e
dorsal regio n o f th e trophozoite . Scattere d i n
the cytoplasm , w e hav e notice d cytoplasmi c
clefts withou t bein g envelope d b y membranes ,
with a littl e dens e homogeneou s conten t (Fig .
1), glycoge n an d fre e ribosomes . Tw o paralle l
rows o f microtubule s (funis ) accompan y the in -
tracyioplasmic trac k o f th e caudal flageila. The
median bod y i s forme d b y microtubule s wi -
thout apparen t relationshi p t o othe r structures .
Mitochondria, endoplasmi c reticulu m an d Gol-
gi apparatu s ar e no t noticed .
We have not observe d differences betwee n
Giardia duodenali s grou p fro m hamster s an d
domestic rat s o r amon g Giardi a muri s grou p
from hamsters , domesti c rat s an d mice .
Differences were , however , evidence d i n th e
morphology o f th e ventrolatera l flang e o f
trophozoites o f Giardi a muri s an d Giardi a
duodenalis groups .
In th e tw o group s studie d th e flang e i s
arranged peripherall y t o th e striate d dis k
presenting a cytoplasmi c li p supporte d b y
marginal plate. * whic h ar e associate d with on e
of th e axoneme s of th e anterolatera l flageila .
In the Giardia duodenalis group bot h from
hamsters (Fig . 2) and fro m domesti c rat s (Fig.
3), th e cytoplasmic lip that makes up the ventro -
lateral flang e i s shor t an d thick , an d doe s no t
show electro n dens e materia l i n it s dista l
extremity. Th e plate s tha t suppor t th e flang e
are striated , littl e develope d an d ar e jus t
associated wit h th e plasmi c membran e o f th e
ventral surfac e o f th e ventrolatera l flange . I n
this group , th e osmiophili c lamell a i s no t
observed.
 In Giardi a muri s grou p fro m hamster s
(Fig. 4) , domestic rat s (Fig . 5) and mice (Fig. 6)
the dista l extremit y o f th e cytoplasmi c li p i s
narrow an d developed , an d i t i s darkene d b y
dense cytoplasm . Th e margina l plate s see m t o
occupy the whole flang e extension. They extend
from th e dorso-latera l surfac e o f th e striate d
disk int o th e ventro-latera l flang e bein g
associated wit h th e flang e plasmi c membran e
either dorsall y o r ventrally . Thes e margina l
plates sho w striation s an d penetrat e throug h
the flang e and become lost i n the dense material
of thei r dista l extremity . A dens e lamell a o f
osmiophilic materia l tha t accompanie s th e
dorsal surface of th e plat e i s noticed associated
with th e margina l plates .
DISCUSSION
Giardia muri s an d Giardi a duodenali s
groups fro m hamster s an d domesti c rats ,
described for th e first time in this paper, show a
basic ultrastructura l similaritie s t o th e othe r
species already describe d in th e literature , such
as Giardi a muri s fro m mice ; Giardi a
256 7 1 4
lamblia fro m m a n ; Giardi a duodenali s
1 1 0 1 5
from rabbits an d Giardia canis from dogs .
3 13
 Our result s show that Giardia belonging t o
the sam e grou p (duodenali s o r muri s groups )
do no t presen t ultrastructura l difference s whe n
located in different animal s (hamsters-domestic
rats an d hamsters-domesti c rats-mice ,
respectively).
However, Giardi a belongin g t o differen t
groups (muri s an d duodenalis ) fro m th e sam e
host (hamste r o r domesti c rat ) sho w marke d
differences i n th e ventro-latera l flang e
ultrastructure regarding thei r size , arrangemen t
of th e margina l plate s an d presenc e o f
osmiophilic lamella .
 According t o H O L B E R T O N , i t seem s
7
that th e for m adopte d b y th e ventro-latera l
flange ma y ac t o n th e attachmen t o r
detachment o f th e hos t t o th e substrat e a s the
magnitude o f th e suctio n forc e cause d by th e
striated dis k depend s o n th e calibe r o f th e
lateral channel s betwee n th e ventro-latera l
flange an d th e substrate . Therefore , th e
differences foun d betwee n muri s an d
duodenalis group s i n relatio n t o th e
ventro-lateral flang e sugges t tha t ther e i s some
difference i n th e attachmen t o f thes e species t o
the substrate . Th e fac t tha t th e flang e i s les s
developed i n Giardi a duodenali s lead s u s t o
believe tha t i t i s no t probabl e t o hav e th e
formation o f latera l channel s i n thi s group .
This woul d lea d th e specie s o f G i a r d i a
duodenalis grou p t o attac h weakl y t o th e
substrate. Suc h a fact coul d explai n the lac k o f
any mirror-imag e attachmen t o f th e striate d
disk t o th e intestin e i n case s o f huma n
biopsies .11
Previous publication s di d no t provid e
evidence o f ultrastructura l difference s betwee n
G i a r d i a muri s an d G i a r d i u lambli a
t r o p h o z o i t e s , althoug h K U L D
12
A&
N O R Y N K O V Á * i n a revie w abou t Giardi a
ultrastructure, hav e suggested that th e marginal
plates i n Giardi a intestinalis ar e les s develope d
than i n Giardi a muris.
It i s wel l know n tha t th e taxonom y o f th e
Giardia specie s i s still somewha t controversial .
Three morphologica l differen t group s hav e
been recognized among the species described by
light microscope : Giardia agilis group whic h i s
found i n amphibians , Giardi a muri s an d
Giardia duodenali s groups, which ar e foun d i n
mammals, eac h havin g a numbe r o f species . 9
F I L I C E define d thes e group s a s specie s bu t
4
according t o K U L D A & N O R Y N K O V Á * the y
apparently represen t highe r tax .
The ultrastructura l difference s betwee n
Giardia muri s an d Giardi a duodenali s group s
and th e similaritie s withi n eac h grou p o f
Giardia, show n i n thi s paper , confir m thi s
general tendenc y tha t poin t t o th e recognitio n
of tw o morphologica l group s o f Giardi a i n
mammals. Althoug h w e do no t kno w i f thes e
ultrastructural difference s hav e som e
significance a t th e leve l o f species , w e ca n use
the morphologica l feature o f th e ventro-latera l
flange as an auxiliaire morphologica l characte r
to hel p a differentia l diagnosi s among group s
of Giardi a in favorabl e sections examined with
electron microscope , even if th e host is infecte d
by more tha n on e type o f Giardia .
The axeni c cultur e o f Giardi a duodenali s
group obtaine d i n laborator y hav e permitte d
the us e o f othe r method s beside s thos e
morphological feature s t o stud y difference s o r
similarities amon g isolate s o f Giardi a fro m
different anima l species. However , these results
have no t ye t bee n use d to systematiz e Giardia
species .
9
 RESUMO
Giardia muri s e Giardia duodenalis :
diferenças ultraestruturai s entr e trofozoito s
Neste trabalh o fo i analisada , atravé s d e
Microscopia Eletrônic a d e Transmissão, a mor -
fologia d e trofozoitos d e Giardia muri s obtido s
de hamsters , rato s peridomiciliare s e camun -
dongos, be m com o d e trofozoito s d e Giardi a
duodenalis obtido s d e hamster s e ratos perido -
miciliares. Basicament e a ultraestrutur a do s
trofozoitos estudado s fo i semelhante . Fora m
observadas, n o entanto, diferenças n a morfolo-
gia d a franj a ventro-latera l do s trofozoito s d e
Giardia muri s quand o comparado s co m o s d e
Giardia duodenalis , diferença s esta s relaciona -
das ao tamanho, arranj o da s lâminas marginai s
e presença de lamel a osmiofílica .
ACKNOWLEDGMENTS
This wor k wa s supported b y a Grant fro m
the Conselh o Naciona l d e Desenvolviment o
Científico e Tecnológic o ( C N P q , Proc . n ?
403177/82) an d Fundaçã o d e Ampar o à
Pesquisa do Estad o d e Sã o Paul o ( F A P E S P ,
Proc. n ? 83/1428-7) . Th e author s ar e indebte d
to Prof . Fernand o Mott a d e Azeved o Corrê a
for revisio n o f th e manuscrip t an d t o Mari a
Aparecida M. Ramo s and Nivalde A . Bass o fo r
technical assistance.
REFERENCES
1. B A R B I E R I , D . ; D E B R I T O , T . ; H O S H I D O , O .
N A S C I M E N T O F? , O.B. ; M A R T I N S CAMPOS
J . V . ; Q U A R E N T E I , G . & M A R C O N D E S , E . — Giar
diasis i n c h i l d h o o d . A b s o r p t i o n test s a n d biochemistry
histochemistry, ligh t a n d electro n m i c r o s c o p y o f j e j u
n a l m u c o s a . A r c h . D i s . C h i l d h . , 45:466-472 , 1970 .
2. B R U G E R O L L E , G . — C o n t r i b u t i o n a l'etud e cytolo -
gique e t phylétiqu e de s diplozoaire s ( Z o o m a s t i g o p h o -
rea, D i p l o z o a D a n g e a r d , 1910) . V . Nouvell e interpreta -
tion d e l'organisatio n cellulair e d e G i a r d i a . Protistolo-
g i c a , 11 :99-109, 1975 .
3. C H E I S S I N , E . M . — Ultrastructur e o f L a m b l i a duode-
nalis. I . B o d y surface , suckin g dis c a n d media n bodies .
J . P r o t o z o o l . , 11 :91-98, 1965 .
4. F I L I C E , F . P . — Studie s o n th e cytolog y a n d lif e his -
tory o f a G i a r d i a f r o m th e laborator y rat . U n i v . C a l i f .
P u b l . Z o o l . , 57 :53-146, 1952 .
5. F R I E N D , D . S . — T h e fin e structur e o f G i a r d i a muris.
J . C e l l B i o l . , 29 :317-331, 1966 .
6. H O L B E R T O N , D . V . — F i n e structur e o f th e ventra l
disk apparatu s a n d th e mechanis m o f attachmen t i n th e
flagellate G i a r d i a m u r i s . J . C e ll B i o l . , 13 .11-41, 1973 .
7. H O L B E R T O N , D . V . — A t t a c h m e n t o f G i a r d i a , a h i -
d r o d y n a m i c mode l base d o n flagella r activity . J . C e l l.
S c i . , 60 :207-221, 1974 .
8. K U L D A , J . & N O R Y N K O V Á , E . — G i a r d i a a nd G i a r -
diasis. I n : K R E I E R , J . P . , ed . — Parasiti c P r o t o z o a .
New Y o r k , A c a d e m i c P r e s s , 1978 . v o l . 2 , p.69-138 .
9. M E Y E R , E . A . — T h e epidemiolog y o f giardiasis . P a -
rasit. today , 1 :101-105, 1985 .
•10. M O R E C K I , R . & P A R K E R , J . C . — Ultrastructura l
studies o f th e h u m a n G i a r d i a lamblia an d subjacen t j e -
j u n a l m u c o s a i n a subject wit h steatorrhea . Gastroente -
r o l o g y , 52 :151-164, 1967 .
11. M U E L L E R , J . C ; J O N E S , A . L . & B R A N D B O R G ,
L . L . — S E M observation s i n h u m a n giardiasis . I n :
J O H A R I , O . , e d . — Scannin g electro n microscopy .
C h i c a g o , I I T Researc h Institute, 1973 . p.557-564 .
12. N E M A N I C , P . C . ; O W E N, R . L . ; S T E V E N S , D . P . &
M U E L L E R , J . C . — Ultrastructur e observation s o n
giardiasis i n a mouse m o d e l . I I . E n d o s y m b i o s i s a n d or -
ganelle distributio n i n G i a r d i a muri s a n d G i a r d i a l a m -
blia. J . infect. D i s . , 140 :222-228, 1979 .
13. S O G A Y A R , M . I . T . L . & G R E G Ó R I O , E . A . — U l t r a s -
tructure o f G i a r d i a canis trophozoites . Z b l . V e t . - M e d .
B . , 31 :107-114, 1984 .
14. S O L O V I E V , M . M . & C H E N T S O V , J . S . — E l e c t r o n
microscope stud y o f L a m b l i a muris with regar d t o pe -
culiarities o f th e ecolog y o f th e parasite . M e d . P a r a z i t .
( M o s k . ) , 35 :667-672, 1966 .
; 15. T A K A N O , J . & Y A R D L E Y , J . H . — J e j u n a l lesion s i n
, patients wit h giardiasi s a n d m a l a b s o r p t i o n . A n electro n
- m i c r o s c o p y study . B u l l . J o h n s H o p k . H o s p . , 116 :413-
, 429, 1965 .
-
Recebido par a publicaçã o e m 17 /2/1989.