Insect Physiol., 1977, Vol. 23, pp. 1163 to 1167. Pergamon Press. Printed in Great Britain.

EXTRA-OVARIOLAR EGG RESORPTION IN A DUNG

BEETLE, EUONITICELLUS INTERMEDIUS

MARINA TYNDALE-BISCOE and J. A. L. WATSON

Division of Entomology, C.S.I.R.O., P.O. Box 1700, Canberra City, A.C.T. 2601, Australia

(Received 6 May 1977)

Abstract—Most oösorption in the dung beetle Euoniticellus intermedius takes place in the haemocoele, oöcytes being
extruded from the ovariole before the deposition of the chorion. Oösorption can be induced in the laboratory both by
prevention of oviposition and by starvation. For up to two days after the onset of starvation the terminal oöcyte appears
normal. After three days the prechorionic oöcyte may more through the ovariole wall; the yolk spheres are then disrupted.
On the fourth day little yolk remains in the extruded oöcyte, and most of the extruded cells are degenerating. We suggest
that extra-ovariolar egg resorption may be a mechanism for ensuring that the single ovariole is not occluded when
conditions are suitable for oviposition.

INTRODUCTION on the effects of starvation; these were carried out at 27º

OÖSORPTION is n common phenomenon in insects
(review by BELL and BOUM, 1975). It tends to occur
when food reserves are depleted, or conditions are
unsuitable for oviposition. Resorption can affect oöcytes
throughout their period of vitellogenesis, and need not
affect all oöcytes in a given ovary, there being some
relationship between the severity of nutritional stress and
the number of eggs completing vitellogenesis. It is thus a
mechanism of considerable importance for conserving
metabolites when conditions are unfavourable.
Most insects in which oösorption has been studied
resorb their eggs within the ovarioles. A very few
species—e.g. Encyrtus fuliginosus, a parasitic wasp, and
the cockroach Periplaneta americana—can extrude
resorbing chorionated eggs into the haemocoele
(FLANDERS, 1942; BELL, 1971). These examples of extra-
ovariolar resorption appear to be exceptions; Encyrtus
fuliginosus was the only species among several parasitic
encyrtid Hymenoptera studied by FLANDERS (1942) to
show such resorption, and in Periplaneta extrusion is a
facultative alternative to intraovariolar resorption (BELL,
1971). During recent studies on reproductive activity and
age determination of the scarabacine dung beetle
Euoniticellus intermedius, to be reported elsewhere
(TYNDALE-BISCOE, 1977), it became apparent that most
oösorption was occurring in the haemocoele, and that
oöcytes were being extruded before the deposition of the
chorion. In this paper we describe the process, examine
its rate of induction, and speculate on its possible
advantages to the beetle.
± 2ºC under a daily regime of 14 hr light/10 hr dark. The
first experiment provided material for a preliminary
histological study, while the major experiment was
concerned with the timetable of resorption induced by
starvation.
In the first experiment, 30 mated females, 12 days
post-emergence, were taken from normal culture
conditions and placed on moist peat in a container 120
cm in diameter; peat does not constitute an adequate diet.
Several females were dissected each day to check the
progress of resorption, and one ovary was fixed in
aqueous Bouin's fluid. The ovaries were embedded in
paraffin wax under vacuum, sectioned at 8 μm, and
stained with Ehrlich haematoxylin and eosin. Some
further ovaries were fixed and sectioned from females
that were found to have resorbed oöcytes under routine
culture conditions.
In the experiment on the timetable of resorption, two
sets of beetles, one 9 to 11 and the other 5 to 8 days after
emergence from the brood ball, and either breeding
normally or on the point of doing so, were transferred to
cages 30 x 30 x 20 cm containing moist peat 15 cm deep;
approximately 30 ♀ and 15 ♂ were placed in each
container. Similar numbers of controls were kept in
cages containing soil 15 cm deep, to which 500 ml of
fresh cow dung was added every three days. Each day
approximately 20 starved females and 20 controls were
dissected, and the incidence of resorption was recorded.
RESULTS
The structure of the ovary

The scarabaeine dung beetles are peculiar in having only

MATERIALS AND METHODS a single ovary, consisting of but one ovariole
(ENGELLMANN, 1970; RICHTER and BAKER, 1974). The
Beetles were reared, and dissected, as described in ovary is of a modified meroistic type, probably
TYNDALE-BISCOE (1977). Two experiments were run telotrophic in its early stages, although we have not seen

1163
1164 MARINA TYNDALE-BISCOE and J. A. L. WATSON
nutritive cords (BRYAN, 1954; TINDALE-BISCOE, 1977). It
is effectively panoistic in the later stages of
vitellogenesis, commencing with the massive deposition
of proteinaceous yolk, during which the basal nutritive
cells are inconspicuous, but the follicle cells are enlarged
and appear active, suggesting direct transfer of yolk
proteins from the haemolymph through the follicle to the
developing oöcyte.
The oöcytes develop sequentially, the terminal
oöcyte being the most fully developed (TINDALE-BISCOE,
1977); however, deposition of yolk is extensive in the
subterminal and following oöcyte when the terminal
oöcyte appears to be mature i.e. is on the point of being
laid. Yolk is laid down as discrete spheres, apparently
sequestered in the oöplasm.
At the lower end of the ovariole, above the cuticular
lining of the oviduct, there is a short region of which the
outermost layer appears to consist of widely spaced,
narrow bundles of muscle fibres, enclosing loose lobes of
large, vacuolated cells.
The extrusion and resorption of the oöcyte
Shortly after a reproductively active female E.
intermedius is transferred into conditions unfavourable
for egg laying, which can include lack of sites, crowding
and starvation, her terminal oöcyte breaches the wall of
the lower ovariole, sometimes at several places, just
above its junction with the oviduct, and enters the
haemocoele. We have observed only pre-chorionic
oöcytes in the haemocoele, lacking the thin chorion that
the egg has when laid. The oöcyte remains connected
with a small intra-ovariolar residue (Fig. 1). The extra-
ovariolar mass of egg is gradually resorbed, and after 48
to 60 hr little or no material remains (Fig. 2). The intra-
ovariolar residue also degenerates, leaving a yellow or
brown body (corpus luteum) inside the neck of the
ovariole, similar to that formed after repealed oviposition
(TINDALE-BISCOE, 1977). Beetles in the process of
resorbing eggs have fat bodies that are noticeably more
granular and more opaque than those of beetles that are
laying normally.
Female beetles that had emerged five weeks
previously, and had been prevented from laying since
emergence by absence of oviposition sites, all showed
signs of extra-ovariolar resorption, and all had corpora
lutea, of varying sizes. When these beetles were returned
for a week to conditions favourable for oviposition, each
laid from 2 to 5 eggs; the ovaries then showed far more
yellow body than would have been expected from the
number of eggs that had been laid, and there were no
degenerating eggs outside the ovariole.
The histological material gave some further
information on the processes of extrusion and resorption.
Pre-chorionic terminal oöcytes of females from normal
culture, and from cultures after one and two days
of starvation, appeared similar; the yolk was dispersed
evenly, in spheres, and the superficial oöplasm was free
of yolk, with the large, irregularly shaped nucleus
embedded in it on one side of the oöcyte.
The columnar, hexagonal follicle cells had large nuclei
(Figs. 3 and 4). In the ovariole immediately below the
follicle were spaces filled with amorphous material,
presumably fluid in nature, that extended between the
loose clusters of cells out to the investing musculature
(Fig. 3).
Sections of an ovary taken on the third day of
starvation, after an egg had been extruded, suggested that
a pre-chorionic oöcyte had moved through the spaces at
the lower end of the ovariole, accompanied by most of
the follicle cells, leaving behind some yolk and cellular
material (Fig. 4); presumably this detritus would later
have formed a corpus luteum. The extruded oöcyte
lacked a chorion. The yolk spheres were substantially
disrupted, forming large, irregular masses, and numerous
cells had invaded the peripheral parts of the yolk. The
origin of these cells is not known.
An ovary from a female ex stock culture provided an
extruded oöcyte at a late stage of resorption, with the
relic enclosed in a membrane, much of the yolk
dissipated, cells occupying a major part of the resorption
body, and some degenerating cells, marked by pycnotic
nuclei (Fig. 5). No chorion was discernible.
In a female starved for four days, one oöcyte had
been extruded, and resorbed to the point at which the
yolk had almost disappeared, and most of the cells
appeared to be degenerating. lo addition, the terminal
oöcyte was resorbing within the ovariole; the nucleus
was not recognisable, the yolk spheres had fused, and the
follicle was substantially disrupted, although there was
little cellular invasion of the yolk. The terminal oöcyte
lacked chorion. The subterminal oöcyte was also
abnormal, in that vitellogenesis had not commenced.
The onset of resorption
In the experiment on the onset of resorption
following starvation, the frequencies of resorption were
closely similar in the two sets of beetles, despite the
differences in their ages; there were no significant
differences between the two sets of controls, or between
the two starved batches. The data were therefore pooled;
the resulting percentages are shown in Fig. 6.
Analyses showed that the actual frequencies of
resorption were homogeneous in the controls throughout
the five days, but the frequencies in starved beetles were
significantly heterogeneous (χ2 = 69.40 with 3 d.f.; P <
0.001). There was no significant difference in the
frequencies shows by starved beetles and controls after
one or two days of starvation, but after three and four
days, the frequencies of resorption were significantly
higher in the starved beetles than in the controls (χ2 =
19.80 and 35.13 respectively, with 1 d.f.; P < 0.001). The
changes in frequency of resorption in starved beetles
between the second and third days, and between the third
and fourth days, were also significant (χ2 = 20.21 and
3.75 respectively with 1 d.f.; P < 0.001 and ~0.05).
 1165

Fig. 1. Ovariole (OR) and oviduct (OD) with extruded terminal oöcyte (EX).
Fig. 2. Ovariole (OR) and oviduct (OD) with remnants of resorbing oöcyte (EX) adjacent to neck
of ovariole.
 1166

Fig. 3. Longitudinal section through lower ovariole showing unextruded terminal oöcyte (TO) and
spaces (S) below follicle wall (F). (Scale = 0.05 mm)
Fig. 4. Similar field after extrusion of terminal oöcyte, with yolk and cellular material (Y) in neck
of ovariole and (EX) in haemocoele; new terminal oöcyte (TO) on left, and oviduct (OD) right.
(Scale = 0.10 mm)
Fig. 5. Section of remnant of resorbing oöcyte, with residual yolk (Y) and peripheral degenerating
cells (C). (Scale = 0.10 mm)
 Extra-ovariolar egg resorption in a dung beetle
Fig. 6. Incidence of extra-ovariolar resorption in starved
Euoniticellus intermedius.
Thus starvation for more than two days abruptly
induced extrusion of the terminal oöcyte into the
haemocoele, leading to extra-ovariolar resorption.
DISCUSSION
Although we have studied details of extra-ovariolar egg
resorption only in Euoniticellus intermedius, it is known to
occur in at least one other species of Euoniticellus, and in
several species of Onthophagus, both African and
Australian (TINDALE-BISCOE, unpublished data). It thus
appears to be a far more general phenomenon in the
Scarabaeinae than it is in other major groups of insects.
We interpret extra-ovariolar resorption as a mechanism
which enables insects to combine the capacity to lay eggs at
short notice with a capacity to recycle reserves. As
FLANDERS (1942) has pointed out, intraovariolar resorption
could prevent oviposition, presumably because the resorbing
oöcytes could occlude the ovarioles. Insects have evolved at
least two alternative ways to overcome this problem. Certain
insects, such as some species of Musca, have developed an
oöstatic system, enabling mature eggs to be held for long
periods before laying (ADAM el al., 1968; TINDALE-BISCOE
and HUGHES, 1969), Continuous oögenesis combined with
extra-ovariolar resorption is the alternative.
It is difficult to interpret the situation in Periplaneta;
BELL (1971) has indicated that extra-ovariolar resorption
occurs if the cockroaches are starved, but little more. The
situation is clearer in Encyrtus fuliginosus (FLANDERS,
1942). The adults do not feed, and
1167
oögenesis proceeds continuously. Extra-ovariolar resorption
thus permits E. fuliginosus to lay eggs at short notice, an
important characteristic for a parasitic insect.
Extra-ovariolar resorption in the Scarabaeinae appears
to have a similar rationale, correlated with the possession of
a single ovariole. Euoniticellus intermedius is to some
degree an opportunist; it feeds on, and breeds in, a scattered,
intermittent resource, Dung generally remains in a ‘usable’
condition for only a short time after dropping. It is thus
advantageous for the beetle to have an egg available for
laying whenever the opportunity occurs, with further eggs
maturing at short intervals, equivalent to the time taken to
produce the dung balls in which the eggs are laid. Under
laboratory conditions, this interval is approximately 4 day-
degrees at temperatures above 19.5°C (TINDALE-BISCOE,
1977). To have the single ovariole occluded by a resorbing
oöcyte could be highly disadvantageous under such
circumstances; continued oögenesis and extra-ovariolar
resorption, rapidly induce by the failure to find a suitable
site for oviposition, provides the solution.
Acknowledgement—We thank NOEL CALL, of the
Department of Zoology, Australian National University, for
help with some of the histological work.
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