animals
Communication
Prediction of Parturition in Ferrets Using Ultrasonographic
Fetal Measurements
Giorgia Pettina 1 , Fabrizia Samiani 2 , Viola Zappone 1, * , Marco Quartuccio 1
Citation: Pettina, G.; Samiani, F.;
Zappone, V.; Quartuccio, M.; Pisu,
M.C. Prediction of Parturition in
Ferrets Using Ultrasonographic Fetal
Measurements. Animals 2023, 13,
3707. https://doi.org/10.3390/
ani13233707
Academic Editor: Jordi Miró Roig
Received: 14 October 2023
Revised: 10 November 2023
Accepted: 28 November 2023
Published: 29 November 2023
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
Animals 2023, 13, 3707. https://doi.org/10.3390/ani13233707
and Maria Carmela Pisu 2
1 Department of Veterinary Sciences, University of Messina, Viale Palatucci, 13, 98168 Messina, Italy;
giorgia.pettina@studenti.unime.it (G.P.); mquartuccio@unime.it (M.Q.)
2 VRC-Centro di Referenza Veterinario, 10138 Turin, Italy; fabriziasamiani@gmail.com (F.S.);
mariacarmela.pisu@vierreci.it (M.C.P.)
* Correspondence: viola.zappone@unime.it
Simple Summary: Gestation in ferrets is generally 42 days (38 to 42 days). Cases of maternal and
paternal cannibalism highlight the need to know the exact date of birth in order to separate the
parturient mother from the other ferrets in the herd. Unlike dogs and cats, there are no established
formulae for fetal ultrasound biometry and growth assessment in ferrets. The aim of this study was
to develop a formula to accurately date the birth of ferret litters to prevent cannibalism and ensure
proper management of the breed. The results showed a significant relationship between days to
parturition and biparietal diameter, which was therefore an accurate predictive ultrasound parameter.
Abstract: The gestation period in ferrets lasts 42 days (range 38–42). Numerous cases of maternal and
paternal cannibalism have been reported, so it is important for breeders to know the exact date of
parturition in order to separate the mother from the other ferrets in the herd. Fetal ultrasonographic
biometry and specific formulae for assessing fetal growth have not yet been developed in ferrets as
they have been in dogs and cats. The aim of this study was to develop a formula, similar to those
available for other domestic species, that could be used in ferrets to date the birth of a litter to within
one day. Among the different ultrasonographic parameters, the biparietal diameter (BP) gave a very
accurate prediction and showed a significant relationship with days before parturition. The formula
developed could allow better planning of care before, during and after parturition, thus helping to
reduce neonatal mortality.
Keywords: ferret gestation; parturition prediction formula; ultrasonographic parameters
1. Introduction
Gestation in ferrets lasts 38 to 42 days and consists of a pre-implantation period of
10 to 12 days and an active gestation period of 30 days [1,2]. Pre-implantation embryos
have an extended period in the oviduct at the end of which, from day 10 of gestation,
they can only be retrieved from the uterus. Implantation in the ferret is centralized,
with rapid invasion of the uterine epithelium by the trophoblast over a large area, which
eventually becomes a zonal band of endotheliochorial placenta [2]. The corpus luteum
begins to secrete progesterone immediately after ovulation, peaking between days 12
and 14 during the implantation period. Circulating progesterone levels decrease from day
24 until delivery on day 42, reaching basal concentrations about 7 days later [3]. The product
of conception does not influence the duration of the luteal phase, making pregnancy and
pseudopregnancy indistinguishable [4]. The gravid uterus can be visualized from day 12 of
gestation and pregnancy can be diagnosed through abdominal palpation from day 14–16.
Radiographic diagnosis through visualization of fetal skeletons can be performed from day
30 of pregnancy [5].
Fetal ultrasound biometry is commonly used to assess fetal growth during pregnancy
in many mammalian species and involves the measurement of various structures of the
https://www.mdpi.com/journal/animals
Animals 2023, 13, 3707 2 of 6

uterus and fetal body [6–9]. In dogs and cats, the linear regression model is used to assess
correlations between measurements of these structures and the number of days to delivery.
In fact, for some anatomical structures, regression is significantly correlated with gestational
age, so it is possible to determine the expected date of delivery after performing ultrasound
measurements of the appropriate structures and applying mathematical formulae [10].
There are several ultrasonographic parameters that can be easily measured during preg-
nancy. These include, in the first half of pregnancy, internal chorionic cavity diameter (ICC),
external uterine diameter (OUD) and fetal crown–rump length (CRL). In the second half of
pregnancy, measurements of biparietal diameter (BP) and body diameter (BD) have been
shown to be predictive in dogs and cats [9].
In the ferret, to the authors’ knowledge, there are no specific formulae available
for calculating the day of parturition. The aim of this study was therefore to develop a
formula, similar to those already available for dogs and cats, to accurately predict the date
of parturition in ferret litters to within one day.

2. Materials and Methods

2.1. Study Design
The study was conducted from February to May 2023 and included 27 clinically healthy
pregnant female ferrets (primiparous and multiparous), privately owned, aged between 24
and 60 months (40.16 ± 12.22 months), weighing between 800 and 1300 g (1028.61 ± 145.21 g),
presented to the Veterinary Reference Centre of Turin for assessment of pregnancy status.
Written informed consent was obtained by all owners before enrolment. Clinical history
and physical examination data were recorded for each subject. All patients were in good
general and reproductive health. They were divided into two groups, the first consisting of
eighteen ferrets for formula design (group 1) and the second consisting of nine ferrets for
formula validation (group 2).

2.2. Ultrasound Examination

The B-mode examination for pregnancy assessment was performed by the same opera-
tor, who had 15 years of experience in this field, using a MyLab Sigma ultrasound machine
(MyLab Sigma, Esaote, Genoa, Italy) with a linear probe (7.5–15 MHz). Subjects were posi-
tioned dorsally or laterally and gently restrained without sedation, which was unnecessary
to improve animal compliance. The fur over the ventral part of the abdomen was carefully
wetted and moved. Alcohol and mating gel were applied to the skin. Standardized depth
settings were used as far as possible and overall gain, dynamic range, focal zone and time
gain compensation were optimized. A B-mode scan of the uterus was performed to assess
the pregnancy status and to assess various fetal biometric parameters.
In the first part of the study, the fetal biometric parameter BP was obtained through
a single ultrasound evaluation per animal between the 23rd and 27th day of gestation.
BP was measured by drawing a line in the same transversal scan as that for crown–rump
length, and the parietal bones of the skull have to be parallel in order to measure the correct
distance between them [11] (Figure 1). At the same time, the visualization of fetal organs
was noted. At least two BP measurements were recorded for each fetus and the mean
values were calculated. The date of delivery, as reported by the owner, was then recorded.
In the second part of the study, nine pregnant ferrets underwent the same ultra-
sound examination, and the expected date of parturition was calculated using the formula
developed in the first phase of the study.
 Animals 2023, 13, 3707
2023, 13, 3707 3 of 7 3 of 6

Figure 1. Ultrasonographic measurement of biparietal

Figure 1. Ultrasonographic diameter
measurement (BP) in ferrets
of biparietal around
diameter 21 days
(BP) post-around 21 days
in ferrets
fertilization: (a) ferrets of group 1 and (b) ferrets of group 2.
post-fertilization: (a) ferrets of group 1 and (b) ferrets of group 2.

2.3. Statistical Analysis

2.3. Statistical Analysis
Statistical
Statistical analysis analysis was
was performed performed
using GraphPad using GraphPad
Prism Prism 10(GraphPad
10 for Windows for Windows (GraphPad
Software, San Diego, CA, USA). The relationship between
Software, San Diego, CA, USA). The relationship between BP growth and days before par-BP growth and days before
parturition was analyzed using a linear regression model.
turition was analyzed using a linear regression model. The growth equations were de- The growth equations were
rived as y = a + bx (y = days before parturition, a = intercept coefficient, b = regression b = regression
derived as y = a + bx (y = days before parturition, a = intercept coefficient,
coefficient and xcoefficient and x =inmeasurement
= measurement mm BP), andin mm
the BP), and coefficients
correlation the correlation
werecoefficients
analyzed were analyzed
using Student’s t test (p < 0.001).
using Student’s t test (p < 0.001).
3. Results
3. Results
Fetal organs were visualized during the ultrasound evaluation. In particular, the
Fetal organs were visualized
difference during the
in echogenicity ultrasound
between evaluation.
the liver In particular,
and lungs the dif-
was assessed, and the kidneys,
ference in echogenicity between the liver and lungs was assessed, and the kidneys,
where the outline of the cortical part could be seen, and the filling and whereemptying of the
the outline of the corticaland
stomach part could be
bladder seen,
were and the filling and emptying of the stomach
observed.
and bladder were observed.
The results of the ultrasonographic measurements of BP in the eighteen ferrets of
The resultsgroup
of the ultrasonographic
1 are measurements
presented in Table 1. of BP in the eighteen ferrets of
group 1 are presented in Table 1.
Table 1. The table shows the mean of the biparietal diameter measurements of the eighteen ferrets
Table 1. The table(group
shows1)the mean
used of the biparietal
to calculate diameter measurements of the eighteen ferrets
the formula.
(group 1) used to calculate the formula.
Biparietal Diameter Day before
Group 1 Group
Biparietal 1
Diameter (mm) Min–Max (mm) Min–Max (mm)
Day before ParturitionParturition
(mm)
Subject 1 4.81
Subject 4.84.5–4.9 4.5–4.9 16 16
Subject 2 5.12
Subject 5.15.0–5.4 5.0–5.4 16 16
Subject 3 Subject
5.23 5.25.2–5.4 5.2–5.4 16 16
Subject 4 Subject
6.34 6.36.0–6.4 6.0–6.4 14 14
Subject 5 6.1 5.9–6.4 12
Subject 5 6.16
Subject 5.35.9–6.4 5.3–5.4 12 12
Subject 6 5.37
Subject 5.95.3–5.4 5.8–6.3 12 11
Subject 7 Subject
5.98 6.35.8–6.3 6.3–6.8 11 11
Subject 8 Subject
6.39 6.46.3–6.8 6.2–6.6 11 11
Subject 10 6.5 6.2–6.8 9
Subject 9 6.411
Subject 6.66.2–6.6 6.5–6.7 11 9
Subject 10 6.512
Subject 6.76.2–6.8 6.5–7.3 9 9
Subject 11 Subject
6.613 6.76.5–6.7 6.7–6.8 9 9
Subject 12 Subject
6.714 7.36.5–7.3 7.2–7.5 9 9
Subject 15 7.4 7.1–7.7 9
Subject 13 6.7
Subject 16 7.1
6.7–6.8 6.6–7.3
9 8
Subject 14 7.317
Subject 7.57.2–7.5 7.1–7.6 9 8
Subject 15 7.418
Subject 7.87.1–7.7 7.6–7.9 9 8
Subject 16 7.1 6.6–7.3 8
Subject 17 7.5 7.1–7.6 8
Subject 18 7.8 7.6–7.9 8
Animals 2023, 13, 3707 4 of 7

Animals 2023, 13, 3707 4 of 6

The data obtained in the first phase of the study allowed us to develop an equation
with two
The unknowns,
data obtainedfinding the phase
in the first intercept (b)study
of the and the slopeus(a),
allowed Y= (−b ×an
to develop X equation
mm) + a, which
resulted
with twoinunknowns,
the following
findingformula:
the intercept (b) and the slope (a), Y = (−b × X mm) + a, which
resulted in the following formula:
Y = (−2.887 × X) + 29.39 (R2 = 0.7887) (1)
2
Y = (−2.887 × X) + 29.39 (R = 0.7887) (1)
where Y indicates the days before parturition (which will always be a negative number),
and X indicates
where Y indicatesthe
themean of theparturition
days before biparietal (which
diameters (mm). be
will always R2 arepresents coefficient of
negative number),
determination. The results of regression analysis showed 2 a significant relationship
and X indicates the mean of the biparietal diameters (mm). R represents coefficient of be-
tween days before
determination. The parturition and biparietal
results of regression analysis diameter (p < 0.001)relationship
showed a significant (Figure 2).between
The accuracy
ofdays
the before
formulaparturition
was 94%and biparietal
at +/− diameter
1 day and 100%(pat< 0.001) (Figure 2). The accuracy of the
+/− 2 days.
formula was 94% at +/− 1 day and 100% at +/− 2 days.

Figure 2. Relationship between biparietal diameter (BP) and days before parturition in ferrets.
Figure 2. Relationship between biparietal diameter (BP) and days before parturition in ferrets.
In the second phase of the study, the formula was applied to nine pregnant ferrets.
TheIn the second
expected date phase of the
calculated study,
using the formula
the formula was applied
was compared withtothe
nine pregnant
actual date offerrets.
The expected
parturition date calculated
provided using
by the owners, the formula
resulting at ±compared
in 77.77%was withwith
1 day and 100% the aactual
margindate of
of ± 2 days.
parturition provided by the owners, resulting in 77.77% at ±1 day and 100% with a margin
of ±2 days.
4. Discussion
Neonatal mortality on ferret farms has been significantly reduced by modern breeding
4. Discussion
practices, but, despite this, it can still occur at alarming rates that can cause severe economic
Neonatal
losses [12]. mortality on ferret farms has been significantly reduced by modern breed-
ing practices,
Accurately but, despite athis,
predicting birthit would
can still occur
allow at monitoring
close alarming rates
of it, that
which can cause
can help severe
prevent these serious
economic losses [12]. consequences of an otherwise normal birth. Over the past few
decades, several studies have proposed different methods to estimate the
Accurately predicting a birth would allow close monitoring of it, which can help pre- date before
parturition in dogs consequences
vent these serious and cats [7,9–17].ofMore recently, ultrasound
an otherwise (US)-based
normal birth. Over the parameters
past fewand decades,
formulae have been developed, which have shown variable accuracy, mainly related to the
several studies have proposed different methods to estimate the date before parturition in
stage of pregnancy and litter size [11,18–23].
dogs and cats [7,9–17]. More recently, ultrasound (US)-based parameters and formulae
Ultrasound assessment of fetal and extra-fetal characteristics is essential for pregnancy
have been
staging and developed,
parturitionwhich have shown
date estimation whenvariable
matingaccuracy, mainly
or ovulation related
data are to the stage of
not available.
pregnancy and litter size
The most commonly [11,18–23].
studied extra-embryonic and extra-fetal parameters include inner
Ultrasound
chorionic assessment
cavity (ICC), of fetal
outer uterine and extra-fetal
diameter (OUD) andcharacteristics is essential
placental thickness. for preg-
In addition,
embryonic
nancy andand
staging fetal parturition
parameters such
dateasestimation
crown–rump length
when (CRL),or
mating body diameterdata
ovulation (BD),are not
biparietal The
available. diameter
most(BP), deep partstudied
commonly of the diencephalo-telencephalic
extra-embryonic and extra-fetalvesicle (DPTV) and
parameters in-
kidney length are considered. The data obtained from measuring these structures
clude inner chorionic cavity (ICC), outer uterine diameter (OUD) and placental thickness. have
been used to develop formulae and tables for determining gestational age or days before
In addition, embryonic and fetal parameters such as crown–rump length (CRL), body di-
birth (DBP) [24].
ameter (BD), biparietal diameter (BP), deep part of the diencephalo-telencephalic vesicle
In dogs and cats, BP is the most accurate fetal parameter in late pregnancy [8] and can
(DPTV) and kidney
be detected as early length areofconsidered.
as day 30 pregnancy [6]. TheBPdata
hasobtained from measuring
good accuracy these struc-
at 5 and 6 weeks’
tures have(95.2%
gestation been used to develop
and 88.4%, formulae
respectively, and ±tables
within 2 days),foralthough
determining gestationala age or
it is considered
days before
reliable birth (DBP)
parameter up to 8[24].
weeks’ gestation (85.3%, ±2 days) [22].
In dogs and cats, BP is the most accurate fetal parameter in late pregnancy [8] and
can be detected as early as day 30 of pregnancy [6]. BP has good accuracy at 5 and 6 weeks’
gestation (95.2% and 88.4%, respectively, within ±2 days), although it is considered a reli-
able parameter up to 8 weeks’ gestation (85.3%, ±2 days) [22].
Animals 2023, 13, 3707 5 of 6

The aim of this study was to develop a formula, similar to those that exist for dogs
and cats, that could be used in ferrets to date the parturition of a litter with only one
day’s discard, thus avoiding the possibility of cannibalism, avoiding issues with males or
other ferrets in the breeding group and having the best attention from the breeder for the
management of the birth. The assessment of BP in the second half of the pregnancy, around
the 21st day after insemination, on the other hand, provided reliable measurements. In fact,
our data show that BP is a reliable indicator of gestational age, as evidenced by a coefficient
of determination (R2 ) greater than 0.7. Based on the results of the measurements in the first
phase of the study, it was therefore possible to develop a formula for predicting the date of
delivery.
The formula predicted the date of birth with an accuracy of 94% at ±1 day and 100%
at ±2 days. The accuracy at ±2 days was higher than that at ±1 day. This result was to be
expected, as increasing the time from ±1 day to ±2 days increases the likelihood that the
actual and predicted date will fall within the same range.
Using the formula in clinical practice, 77.77% of ferrets giving birth on the expected
date within one day is not a low prediction rate, not least because in reality, the percentage
at +/− 1 day should be 88.88% (8/9), as one subject gave birth a few minutes after midnight.
The reason for this result is that there are a number of intrinsic and extrinsic factors which
can have an influence on the time of birth.
To the authors’ knowledge, this is the first formula developed for ferret birth dating.
Neonatal mortality in pets can have significant emotional and economic consequences for
owners, so accurate prediction of birth date could be of great benefit to both breeders and
veterinarians. The formula developed could allow better planning of care before, during
and after birth, helping to reduce neonatal mortality.

5. Conclusions
The development of a formula for predicting ferret parturition dates represents a
significant advance in the understanding of reproductive biology and may have practical
applications in breeding programs and veterinary care.
In our study, assessment of BP in the second half of pregnancy provided reliable
measurements. In fact, our data show that BP is an accurate indicator of gestational age, as
evidenced by a coefficient of determination (R2 ) greater than 0.7.

Author Contributions: Conceptualization, M.C.P.; methodology, G.P., F.S. and V.Z.; software, M.Q.
and V.Z.; validation, M.C.P. and G.P.; formal analysis, G.P. and F.S.; data curation, M.C.P. and F.S.;
writing—original draft preparation, M.C.P., G.P. and F.S.; writing—review and editing, G.P., F.S. and
V.Z.; supervision, M.C.P. and M.Q. All authors have read and agreed to the published version of the
manuscript.
Funding: This research received no external funding.
Institutional Review Board Statement: All treatment, housing and care of the animals were in
accordance with Directive 2010/63/EU on the protection of animals used for scientific purposes.
The study was conducted during a normal visit to assess pregnancy status and each animal enrolled
in the study underwent an ultrasound examination at the free choice of its owners, who gave their
informed consent. Therefore, according to national legislation, no ethics committee approval was
required for the study.
Informed Consent Statement: These were client-owned animals, and written informed consent was
obtained from the owner of the animals (or an authorized agent for the owner).
Data Availability Statement: The data presented in this study are available on justified request from
the corresponding author.
Acknowledgments: The authors acknowledge support from the University of Messina through the
APC initiative.
Conflicts of Interest: The authors declare no conflict of interest.
Animals 2023, 13, 3707 6 of 6

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