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Post-Partum Involution of the Canine Uterus - Gross Anatomical and

Histological Features

Article  in  Reproduction in Domestic Animals · July 2009

DOI: 10.1111/j.1439-0531.2009.01388.x · Source: PubMed

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Reprod Dom Anim 44 (Suppl. 2), 152–155 (2009); doi: 10.1111/j.1439-0531.2009.01388.x
ISSN 0936-6768

Post-Partum Involution of the Canine Uterus – Gross Anatomical and Histological

Features
DC Orfanou1, HN Ververidis2, A Pourlis1, IA Fragkou1, AN Kokoli2, CM Boscos2, IA Taitzoglou2, A Tzora3, CM Nerou1,
L Athanasiou1 and GC Fthenakis1
1
Veterinary Faculty, University of Thessaly, Karditsa; 2School of Veterinary Medicine, Aristotle University of Thessaloniki, Thessaloniki; 3TEI
Epirus, Arta, Greece

Contents The genital tract was removed under sterile condi-

We aimed to study the normal puerperium in the bitch.
Ovariohysterectomy was performed in nine bitches, each at a
different day after normal whelping; their genital tract was
subject to gross anatomical examination, as well as to
histological examination and electron microscopy scanning.
Corpora albicans were evenly distributed in the left and right
ovaries and placental sites were evenly distributed among left
and right uterine horns. Placental sites were initially of dark
green to grey colour, later becoming dark brown; their length
and height progressively decreased. Height of the myometrium
and diameter of the uterine glands progressively decreased.
Trophoblast-like cells were consistently observed at the
placental sites and on the surface of the interplacental areas,
at all time points where hysterectomy had been performed. It is
suggested that involution of the canine genital tract can last up
to 3 months and is slow. Continuous (up to D84 post-partum)
presence of prominent placental sites should be considered a
normal feature of canine uterine post-partum involution.
Introduction
The puerperium is the period during which the genital
system prepares to return to cyclicity. In contrast to
other domestic mammalian species, literature on the
normal puerperium of the bitch is very limited. The aim
of this study was to describe the gross anatomical and
histological features of the post-partum involution of
the genital tract of the bitch, after normal whelping. It is
a part of a wider study into the canine puerperium. The
work was carried out under a licence for experimental
procedures issued by the Greek Ministry of Agriculture,
based on EU guidelines.
Materials and Methods
Nine primiparous Beagle bitches, aged 12–17 months,
which had whelped normally, were monitored for up to
3 months after parturition. Regular clinical examina-
tions of the animals were carried out during the study.
Furthermore, samples were collected after inserting a
sterile swab into the anterior part of the vagina, for
bacteriological and cytological examination by using
conventional techniques; details have been presented by
Orfanou et al. (2008).
Ovariohysterectomy was carried out on one experi-
mental animal at each of the following days (D) after
whelping: D7, D21, D28, D35, D42, D56, D70 and to
two animals on D84, for a detailed anatomical exam-
ination of the ovaries and the uterus. All animals
recovered uneventfully from surgery.
tions. A small incision was performed on the lateral
part of the uterine body and, by using an aseptic
technique, a sterile swab was used to sample the
content of the uterus. Subsequently, dimensions of the
ovaries were measured. Number and diameter of
corpora albicans were determined after performing
one longitudinal section on each ovary. The appear-
ance of the uterus was recorded. Length of uterine
horns (from the apex to the bifurcation) was measured.
Diameter of uterine horns was measured at placental
sites and interplacental areas. The uterus was then
dissected; the placental and interplacental sites were
assessed and their dimensions (length and height) were
measured.
Uterine swab samples were plated onto Columbia 5%
blood agar; the media were incubated aerobically at
37C for up to 72 h. Throughout this study, all bacteria
isolated were identified by using conventional tech-
niques (Euzeby 1997) and the ‘API SYSTEM’ quick
identification strips (BioMerieux S.A., Marcy-l’-Etoile,
France). Swabs were also rolled on glass slides and
stained by using the Giemsa technique. Finally, tissue
samples were collected for processing with haematoxy-
lin–eosin stain for histological examination and mor-
phometric analysis.
Tissue samples were also prepared for ultrastructural
examination by using scanning electron microscope. The
specimens were fixed in a sodium cacodylate (0.1 M)
buffered solution of 2% glutaraldehyde and 2% para-
formaldehyde. Consequently, they were washed in
several changes of sodium cacodylate buffer, transferred
for 1 h to 1% OsO4 and dehydrated in graded acetone.
Tissues were critically point dried in carbon dioxide,
mounted onto stubs and sputter coated with palladium
and gold in a Bal-Tec (Balzers, Liechtenstein) sputter
coater. They were observed in a JEOL (Tokyo, Japan),
JSM 840 scanning electron microscope.
Differences in findings between the various time
points, throughout the study were detected with the
Wilcoxon signed-rank test, as appropriate. Statistical
significance was set at p = 0.05.
Results
All animals whelped normally five to nine puppies
(median value: 7; total number of puppies born: 66) and
remained clinically healthy throughout the experimental
period. Results of bacteriological and cytological

 2009 The Authors. Journal compilation  2009 Blackwell Verlag GmbH

DC Orfanou, HN Ververidis, A Pourlis, IA Fragkou, AN Kokoli, CM Boscos, IA Taitzoglou, A Tzora, CM Nerou, L
Athanasiou and GC Fthenakis 153

examination of vaginal samples collected throughout the

study have been presented by Orfanou et al. (2008).

Gross anatomical findings

No significant differences (p > 0.05) were seen in the
dimensions of the ovaries from D7 to D84. Corpora
albicans were evenly distributed in the left and right
ovaries of the animals: in total 32 and 30, respectively;
median diameter was 4 mm on D7, 3.5 mm on D35 and
<2 mm on D84 (p > 0.05).
Externally, the uterus was pink, with longitudinal folds
and mild vascularization. The two horns were symmet-
rical between them and laterally oblate; macroscopically,
the placental sites could be appreciated even up to D84.
No significant differences in the median length of the
uterine horns were evident among the various time points
(on D7: 9.0 cm, on D35: 7.1 cm, on D84: 8.3 cm;
p > 0.5). Median diameters of the uterine horns were
different across the various time points, albeit only at the
p < 0.1 level: 2.60 cm (same for placental sites and
interplacental areas) on D7 vs 1.70 ⁄ 1.20 cm (placental
sites ⁄ interplacental areas) on D35 vs 0.95 ⁄ 0.70 cm (pla-
cental sites ⁄ interplacental areas) on D84.
There was a small amount of viscous, dark-brownish
fluid inside the uterus of four bitches (D21, D28, D35
and D42). Placental sites were evenly distributed in the
left and right uterine horn of the animals: in total 35 and
33, respectively. They were initially of dark green (D7)
to grey colour (D21–D28), whereas later they were dark
brown (D35 and thereafter). These were easily recog-
nized, thickened and with foamy surface. All placental
sites of each uterus (i.e. of the same animal) were
macroscopically similar. Their length was 8–10 mm up
to D35 and 3–5 mm on D84; their height was 3–4 mm
up to D35 and 1–2 mm on D84. The interplacental areas
were white to pink colour with mild longitudinal
wrinkles thereon. Their length varied from 8 to 10 mm
in all tissue samples; their height (i.e. uterine wall
thickness) was 6 mm on D7, 2.5 mm on D35 and
1.5 mm on D84 (p < 0.1).

Bacteriological findings
The bacteria (Arcanobacterium pyogenes or Escherichia
coli) isolated from the uterine content, as well as from
the anterior vagina (Orfanou et al. 2008) on each day of
the study, are shown in Table 1. In one of the two
instances, where bacteria were isolated from uterine
Table 1. Bacteria isolated from uterine content after ovariohysterec-
tomy and dissection of the uterus and from swabs from anterior vagina Fig. 1. Uterus of a bitch, 7 days post-partum (a) prominent placental
sites (pl) on uterus lumen; (b) uterine glands (ug) with single-layer,
Day after Bacteria from uterine Bacteria from
cuboidal epithelium (·100, H&E); (c) Trophoblast-like cells (tl) at a
whelping content anterior vagina
placental site (·250, H&E)
D7 Arcanobacterium pyogenes –
D21 A. pyogenes A. pyogenes
D28 – – swab, there was fluid in the uterus; in the other, there
D35 – Escherichia coli was not.
D42 – –
D56 – –
D70 – – Histological findings
D84 – –
A simple columnar epithelium was evident in the
N-dash denotes no bacteria isolated. interplacental endometrium of all experimental animals.

 2009 The Authors. Journal compilation  2009 Blackwell Verlag GmbH

154
Fig. 2. Uterus of a bitch, 84 days post-partum (a) presence of
involuted placental sites (pl) on uterus lumen; (b) uterine glands (ug)
with single-layer, cuboidal epithelium (·100, H&E); (c) trophoblast-
like cells (tl) at a placental site (·250, H&E)
In general, the diameter of uterine blood vessels was
smaller at later time points; the height of myometrium
was 460 lm on D7 and 200 lm on D84.
The median diameter of the uterine glands’ transverse
sections was 79 lm on D7, 53 lm on D56 and 52 lm on
D84 (p = 0.042). However, no significant differences
were seen in the median height of glandular epithelial
 2009 The Authors. Journal compilation  2009 Blackwell Verlag GmbH
Post-Partum Features of Canine Uterus
Fig. 3. (a) Profiles of uterine glands in the wall of the uterus of a bitch,
70 days post-partum (·110, SEM). (b) Higher magnification of
squared area in (a) (·1500, SEM). (c) Profile of a placental site in
the uterus of a bitch, 84 days post-partum (·1300, SEM)
cells: 6.6 lm on D7, 7.6 lm on D56, 7.2 lm on D84
(p = 0.55).
Soon after whelping, there was no inflammatory
cellular infiltration in the inner (circular) layer of the
myometrium. On D28, there was some cellular infiltra-
tion (macrophages, eosinophils and plasma cells),
 DC Orfanou, HN Ververidis, A Pourlis, IA Fragkou, AN Kokoli, CM Boscos, IA Taitzoglou, A Tzora, CM Nerou, L
Athanasiou and GC Fthenakis
whereas at later time points, this became extensive
throughout this layer.
Trophoblast-like cells were consistently observed at
the placental sites and on the surface of the interplac-
ental areas, at all time points where hysterectomy had
been performed (Figs 1–3) .
Discussion
In general, there is a paucity of detailed descriptions of
the normal post-partum involution of the genital tract of
the bitch. The findings, coupled with clinical, bacterio-
logical and cytological results (Orfanou et al. 2008),
provide gross and microscopic anatomical as well as
microbiological data, which can be used as reference
information for further studies into pathologic condi-
tions of the canine post-partum uterus.
Our findings suggest that the normal involution of the
genital tract in the bitch is slow and can last up to
3 months. The slowness of this process is indicated by the
lack of significance in size change of uterine glands and
epithelial cells from D7 to D84. Thus, one may propose
that involution might continue even subsequently to that
date. This would be in contrast to Al-Bassam et al.
(1981a), who mentioned that uterine involution in
bitches is completed 84 days after whelping.
Another interesting finding was the presence of
trophoblast-like cells, observed throughout the study,
even at 84 days post-partum. This lends further support
to the above hypothesis. These cells were always
observed on the surface of the uterine epithelium, which
is consistent with a normal involution process (Al-Bas-
sam et al. 1981b). Finally, another supporting finding is
the late invasion (D28) of blood cellular elements into
the myometrium.
Trophoblast-like cells seen in the vaginal discharge at
late post-partum stages are believed to indicate subin-
volution of placental sites, especially in primiparous
bitches (Reberg et al. 1992), as were the animals used in
the present study. However, in the present work, these
cells were consistently detected on the uterine epithelium
and at the placental tissues, as well as in smears of
vaginal discharge obtained from the bitches (Orfanou
et al. 2008). We, thus, propose that their significance be
re-assessed, and, perhaps, their presence possibly be
 2009 The Authors. Journal compilation  2009 Blackwell Verlag GmbH
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155
considered as normal. Within that context, however, the
quantity of such cells, as well as other signs that might
indicate subinvolution, should be considered.
Finally, the continuous presence of prominent pla-
cental sites, even at 3 months after whelping, should be
considered a normal feature of canine uterine post-
partum involution.
Author contributions
Work described in present paper is part of PhD studies undertaken by
DCO. The studies are supervised by GCF and co-supervised by CMB
and IAT. Further to that, other authors had equivalent contributions.
Conflicts of interest
The authors have declared no conflicts of interest.
References
Al-Bassam MA, Thomson BG, O’Donnell L, 1981a: Normal
postpartum involution of the uterus in the dog. Can J Comp
Med 45, 217–232.
Al-Bassam MA, Thomson BG, O’Donnell L, 1981b: Involu-
tion abnormalities in the postpartum uterus of the bitch. Vet
Pathol 18, 208–218.
Euzeby JP, 1997: List of bacterial names with standing in
nomenclature: a folder available on the internet. Int J Syst
Bacteriol 47, 590–592. (List of Prokaryotic Names with
Standing in Nomenclature. Last full update 7 September
2006. URL: http: ⁄ ⁄ www.bacterio.net).
Orfanou DC, Ververidis HN, Pourlis A, Fragkou IA, Kokoli
AN, Boscos CM, Taitzoglou IA, Tzora A, Nerou CM,
Athanasiou L, Fthenakis GC, 2008: Experimental Study of
the Post-partum Involuting Genital Tract of the Bitch. Part
I: Clinical, Bacteriological and Cytological Features. Pro-
ceedings of the 6th International Symposium of Canine
and Feline Reproduction, Vienna, 9–11 July 2008, pp. 175–
176.
Reberg SR, Peter AT, Blevins WE, 1992: Subinvolution of
placental sites in dogs. Compend Contin Educ Pract Vet 14,
789–796.
Submitted: 30 June 2008
Author’s address (for correspondence): George Fthenakis, Veterinary
Faculty, University of Thessaly, PO Box 199, 43100 Karditsa, Greece.
E-mail: gcf@vet.uth.gr