PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY
182
Survival and Development of Leucochloridiomorpha
constantiae (Trematoda) in the Chick Coelom1
BERNARD FRIED, STEVEN A. CANTNER, AND DAVID L. TEMPEST
Biology Department, Lafayette College, Easton, Pennsylvania 18042
ABSTRACT: To determine the adaptability of Leucochloridiomorpha constantiae to a nonintestinal site,
metacercariae and adults were surgically implanted into the chick coelom. Sixteen (44.4%) of 36 implanted adults and 42 (24.0%) of 175 implanted metacercariae were recovered live on various surfaces
of the viscera from 18 hr to 15 days postimplantation. Crural contents varied and consisted of transparent
or refractile material in worms recovered from the peritoneum or adipose tissue and dark-brown material
in those obtained from the liver. Metacercariae became ovigerous in the coelom within 4 days and by 10
days contained amber eggs with fully developed pear-shaped miracidia.
Although the small intestine of the raccoon,
Procyon I. lotor, has been experimentally infected with Leucochloridiomorpha constantiae
(Mueller, 1935), in naturally or experimentally
infected birds this parasite appears to live
exclusively in the bursa of Fabricius (Gower,
1938; Allison, 1943). To determine the adaptability of L. constantiae to nonintestinal sites,
adults and metacercariae were implanted into
the chick coelom and results are reported
herein.
Materials and Methods
Adults, 14 to 21 days old, obtained from
the bursa of Fabricius of chicks previously infected with metacercariae by the "cloacal
drop" method of Allison (1943), and metacercariae dissected from the uteri of Campeloma
decisum snails (Fried and Harris, 1971) were
washed in three changes of Locke's (Paul,
1960) solution prior to coelomic implantation
into uninfected 7-day-old white Leghorn chicks.
Recipient chicks were starved overnight prior to
surgery, anesthetized with Equi-Thesin (Fried
and Berry, 1961), and a 1-cm incision was
made through the ventral body wall behind
the sternum. Each chick received either six
adults or 25 metacercariae pipetted into the
coelom in a drop of Locke's. The incision was
sutured with cotton thread and chicks were
necropsied from 18 hr to 15 days postimplantation. Most flukes recovered from the coelom
were examined live under light coverslip pres1 Supported in part by a research grant from the
Lafayette College Committee on Advanced Study and
Research.
sure (Figs. 2, 4). An in situ preparation of
a 2-week-old coelomic fluke attached to the
serosa of the liver was prepared as paraffin
sections fixed in AFA and stained with Heidenhain's iron alum hematoxylin (Figs. 5. 6).
Results
Sixteen (44.4%) of 36 implanted adults
were recovered live from the coelom between
18 hr and 15 days postimplantation. Worms
were attached by their acetabula to the liver
serosa, peritoneum, pericardium, adipose tissue, and intestinal and air sac surfaces. Four
days after implantation adults still contained
sperms in the seminal receptacle and eggs with
fully developed miracidia. The fact that adult
implants were live and looked essentially like
bursal worms led to studies with metacercariae.
Forty-two (24.0%) of 175 implanted metacercariae were recovered live on various surfaces of the viscera from 3 to 14 days postimplantaion. Egg development was first seen
in 4-day-old worms, sperms were present in
the seminal receptacle, and except for a day
lag in development these worms appeared essentially as those observed in the bursa (Allison,
1943; Fried and Harris, 1971). Seven-day-old
worms recovered from the surface of the liver
contained hepatic cells in the crura and eggs
with well-developed embryos (Figs. 2, 4 ) .
Ten-day-old worms contained amber eggs with
fully developed pear-shaped miracidia. Crural
contents varied and consisted of transparent or
refractile material in worms recovered from the
peritoneum or adipose tissue, and dark-brown
material in those obtained from the liver.
Copyright © 2011, The Helminthological Society of Washington
OF WASHINGTON, VOLUME 39, NUMBER 2, JULY 1972
183
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' • :;'« :• • , .'
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^ '•'" •'-.. »
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Figures 1-6. Development of L. constantiae in the chick coelom. Abbreviations: (a) acetabulum, (b)
blood clot, (c) crura, (ci) cirrus, (e) eggs, (er) erythrocytes, (h) hepatic cells, (1) liver, (o) ovary, (p) pharynx, (s) serosa of the liver, (t) testis, (v) vitellaria, (vr) vitelline reservoir. 1. Anterior aspect of live metacercaria stained intravitally with neutral red. 2. Anterior aspect of an adult 7 days after metacercarial
inoculation into the coelom. Note liver tissue in the pharynx and crura of this fluke which was recovered
from the liver surface. 3. Posterior aspect of inetacercaria seen in Figure 1. 4. Posterior aspect of fluke seen
in Figure 2. 5. In situ histologic preparation of metacercaria cultivated for 2 weeks in the coelom and recovered on the liver surface. Note blood clot associated with the fluke. 6. Enlarged view of bracketed area
in Figure 5.
Copyright © 2011, The Helminthological Society of Washington
PROCEEDINGS OF THE HELMINTHOLOGICAL SOCIETY
184
Sections of a 2-week-old coelomic worm attached to the liver revealed the presence of
a clot containing erythrocytes and some entrapped hepatic cells (Figs. 5, 6).
Discussion
L. constantiae metacercariae contain genital
primordia (Mueller, 1935; Fig. 3), and Allison
(1943) reported lipoidal material in the crura.
Unpublished studies on cryostat sections of
metacercariae stained with Oil Red O (Lillie,
1944) have confirmed Allison's findings. Although further studies are needed to elucidate
differences in the ingesta of metacercariae
and adults this fluke appears nonspecific in
its feeding habits, ingesting blood, columnar
epithelium, and lymphoidal follicles in the
bursa (Fried and Lang, 1971) and at least
hepatic and peritoneal tissue in the coelom.
Gross observations on metacercariae cultivated for 10 days in the chick coelom indicate
that such worms appear morphologically similar to 7-day-old bursal worms obtained from
double-worm infections in chicks (Fried and
Harris, 1971). Further studies are needed to
determine why the chick coelom is suitable
for the cultivation of this fluke. Other coelomic implantation studies on hermaphroditic
digenes revealed non survival of gorgoderids
in the frog coelom (Goodchild, 1954, 1955)
and philophthalmids in the chick coelom
(Fried, 1964), survival of adult Fasciola hepatica and adults of Paragonimus westermani
and P. ohirai in the rat coelom (Lienert, 1959;
Omura, 1960), suboptimal development of
Echinostoma revolutum in the chick coelom
(Fried and Fink, 1968), and sexual maturation
of Clinostomum complanatum in the mouse
coelom (Dowsett and Lubinsky, 1966).
Acknowledgments
The authors are grateful to Mr. Kevin R.
Harris, Institute for Pathobiology, Lehigh
University, Bethlehem, Pa., for the use of
unpublished material on the histochemistry of
Leucochloridiomorpha constantiae.
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