Haryana Vet.
(March, 2022) 61(SI), 91-93 Short Communication
ARTIFICIAL INSEMINATION IN SHE DOGS WITH LIQUID SEMEN
MALATI NAIK*, SANTOSH SARANGAMATH1, D. JAHANGIRBASHA 2
and D.S. SUPRITH
1
Department of Veterinary Gynaecology and Obstetrics, Department of Veterinary Medicine,
2
Department of Veterinary Surgery and Radiology, Veterinary College, Gadag–582 101, India
Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar
Received: 04.05.2021; Accepted: 16.06.2021
SUMMARY
Semen collection, semen evaluation and artificial insemination are essential skills for small animal practitioners. Semen collection was
performed using digital manipulation technique among all the different breeds of dogs and several tests including macroscopic (semen volume,
semen colour, hydrogen ion concentration) and microscopic tests (mass motility, progressive motility, sperm concentration, live and dead sperms by
eosin nigrosin staining) were performed to estimate the fertility of the male. Even exfoliative vaginal cytology (EVC) was also performed to detect
the proper phase of oestrous cycle. She dogs which had more than 90% cornification index, were selected for AI and evaluation of conception was
done. Finally concluded that digital manipulation of semen collection is a cost-effective method and AI should be conducted for those bitches which
had issues with natural mating.
Keywords: Artificial insemination, Canines, EVC, Semen collection, Semen evaluation
How to cite: Naik, M., Sarangamath, S., Jahangirbasha, D. and Suprith, D.S. (2022). Artificial insemination in she dogs with liquid
semen. Haryana Vet. 61(SI): 91-93.
Canine breeding is a rapidly growing industry and previous two to three oestrous cycles. On clinical
there is influx of exotic breeds of dogs into India for examination, all physiological parameters were found to
breeding purposes (Singh et al., 2019). Among different be normal. It was observed that two females were too
assisted reproductive techniques, artificial insemination aggressive and three were inexperienced. In one female, the
(AI) is one that involves collection of semen from a stud height was relatively shorter. On per-vaginal examination,
male and introducing it into genital passage of a female or it was observed that two female dogs had a small hard mass
female reproductive tract, so that fertilization can occur in of 3 cm diameter which was not occupied fully on the
the absence of natural mating (Mason, 2018). In animals vaginal floor and able to pass finger. Vaginal smear was
and humans, AI is one of the earliest techniques employed collected by cotton swab technique and study of vaginal
for assisted reproduction. It took longer period to be cells (EVC) was done as recommended by Antonov
implemented and standardised in pet dogs due to species (2017). Those females having more than 90%
specific particularities. Abbe Spallanzani performed first cornification index (more of anuclear keratinized and
successful AI in dogs in 1780. By 1950’s, the use of stored superficial cells) upon EVC were selected for AI (Fig. 3).
dog semen for AI was practiced. Later by 1990’s, the EVC was performed using cotton swab technique as
technology of frozen semen was developed for breeding recommended by Aydin et al. (2011). The smears were
practice in dogs (Foote, 2002; England and Millar, 2008). stained using Giemsa staining and observed under 100x.
Currently, there has been a huge development in Semen collection: The most commonly employed
reproductive biology and biotechnology. However, initial methods of AI are artificial vagina, manual collection and
efforts to improve AI failed, mainly due to the two electrical stimulation (Payan-Carreira et al., 2011). In the
limitations i.e., unexplained reproductive physiology of present study, the semen collection was performed using
dog and unresponsiveness of dog sperm to freezing as manual collection. For semen collection, male was placed
opined by Hermansson and Forsberg (2006). The most in quite, non-stressful environment and adequately spaced,
common reasons attributed for AI in dogs are that the male non slippery surface for footing. Semen collection was
or female may be shy, inexperienced or too aggressive. performed using digital pressure and massage technique as
There may be presence of obstructions in the genital recommended by Jahangirbasha et al. (2018). In those
passage such as vaginal band, small hard mass at vaginal dogs which did not show sexual interest, semen collection
floor. Other possible reasons may be that both male and using digital manipulation was followed using the she dog
female would have grown together and know each other to in estrus being positioned infront of male dog and aiding it
aid in controlling the risk of sexually transmitted diseases to mount the female. When the male dog exhibited sexual
on either gender (Johnston et al., 2001). interest, using a gloved hand, the preputial skin was pushed
Case presentation and observation: Seven female dogs caudally, exposing the glans penis. The base of the penis
were presented to Veterinary Clinical Complex, Veterinary behind the bulbous glandis was grasped through prepuce
College, Gadag with a history of vaginal bleeding since six and firm and constant pressure was applied by the palm
to ten days and unwillingness to mate naturally. The details and fingers. Pressure was continuously applied with
are given in Table 1. Further, history revealed that natural backward and forward movements until erection was
mating was not possible in four out of seven dogs during brought (Fig. 1). The ejaculated semen was collected in a
*Corresponding author: [email protected] sterile test tube for further evaluation.
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Fig. 1. Semen Collection Fig. 2. Artificial Insemination Fig. 3. Anuclear keratinized and superficial
cells under EVC
Table 1
Age and parity (Mean± SE) et al. (2016). Macroscopic and microscopic tests were
carried out to evaluate the effectiveness of semen for AI.
S.No. Age (yrs) Parity
Macroscopic tests:
Male Female Female 1. Semen volume: The volume of ejaculate was
1 2 2 0 assessed using graduated or calibrated test tube. The
2 3 2 0 mean ± SE of volume was 11.07 ± 0.51 ml (Table 2).
3 3 3 1 2. Semen colour: Semen colour was milky-white.
4 2 3 2
5 4 2 0 3. Hydrogen ion concentration (pH): the mean pH of
6 2 4 1 dog semen was 6.24 ± 003 (Table 2).
7 3 3 0 Microscopic tests:
Mean± SE 2.71±0.28 2.714 ±0.28 0.57±0.29 1. Progressive motility: Normal progressive motility
Procedure of Artificial Insemination: Female was placed test was performed immediately after semen
in quite, non-stressful environment and adequately spaced collection in order to assess the forward motion and
non-slippery footing surface. In order to facilitate easy rapid progression of the motile spermatozoa to
insemination, the she dog was positioned as per the method correlate with plasma membrane integrity as
described by Jahangirbasha et al. (2018). The hind quarters recommended by Roberts et al. (2016). A drop of
of the she dog was elevated to an angle 45 to 60 degree semen was diluted with normal saline and placed in
from the surface of the examination table with the help of a grease-free pre-warmed glass slide and was
an assistant (Fig. 2). Perineal area of the vulval lips was covered with coverslip (Roberts et al. (2016). The
cleaned with antiseptic solution and vaginal speculum was mean progressively motile spermatozoa was 86.42 ±
inserted aseptically. Sterile AI sheath was then introduced 0.92 percent (Table 2).
as much as possible to the level of external os of the cervix. 2. Sperm concentration: Sperm concentration was
Another assistant was directed to connect the free end of estimated following the gold standard technique using
the AI sheath with the syringe pre-loaded with the semen. haemocytometer as described by Payan-Carreira et al.
The semen was then injected slowly to the level of external (2011). The mean sperm concentration in the present
os. The she dog was maintained in the required position for study was 1037 ± 149.99 million/ml (Table 2).
10 minutes to prevent the back flow of the semen. The 3. Live and dead staining: Live and dead spermatozoa
procedure was performed once in all the cases. were estimated using Eosin-nigrosin staining. The
Semen Evaluation: The evaluation of collected semen mean viable spermatozoa in current study was 85.42
was performed as per the procedures described by Roberts ± 0.53 percent (Table 2).
Table 2
Microscopic and macroscopic evaluation of canine semen (Mean±SE)
S.NO. Parameters
Semen Volume Sperm Concentration Hydrogen ion Progressive Motility Live sperms
(ml) (million/ml) concentration (percent) (percent)
1 10.5 1500 6.3 85 85.0
2 9.5 1680 6.2 90 86.5
3 11.0 850 6.4 85 85.0
4 12.5 700 6.2 85 85.0
5 11.5 835 6.3 90 88.0
6 13.0 680 6.2 85 83.5
7 9.5 1020 6.1 85 85.0
Mean± SE 11.07 ± 0.51 1037±149.99 6.24±0.03 86.42±0.92 85.42 ± 0.53
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Pregnancy diagnosis: On 35-40 days post insemination, were confirmed pregnant with the presence of foetal parts
pregnancy diagnosis was carried out ultrasonographically. in foetal sacs. Normal whelping was recorded among six
In the present study, the AI was carried out in seven she dogs at 62 to 65 days of gestation. However, in one she
she dogs using the fresh semen collected from male dog, the puppies had to be delivered with professional
counterpart using manual manipulation technique where assistance on 68th day of gestation.
constant application of pressure behind the bulbous The present study showed that digital manipulation
glandis was prime important in maintaining a proper is cost-effective technique for semen collection in dogs.
pressure continuously in a pulsating manner for collecting Semen evaluation and artificial insemination is a valuable
sufficient quantity of semen. These findings were similar provision that can readily be performed in females and/or
to those observed by Shukla (2011) and Jahangirbasha et male dogs having issues with natural mating. The success
al. (2018). of AI depends on proper timing of the insemination, proper
The findings of current study varied in normal range. skill and expertise with regard to evaluation of EVC in
The mean volume of semen was 11.07 ± 0.51 ml which was order to identify the stage of oestrus. Good quality semen
similar to observation made by Payan-Carreira et al. collected from healthy sire with a mean progressive
(2011). The volume of semen depends upon age, breed of motility 86.42±0.92 percent, mean live spermatozoa 85.42
dog, the frequency of semen collection, size of prostate ± 0.53 percent and mean volume of 11.07 ± 0.51 ml semen
gland, size of the dog and third fraction of the ejaculate and along with hygienic collection and handling are very
is not a factor for semen quality assessment in dogs as essential. In the present study single insemination was
opined by Payan-Carreira et al. (2011). They opined that sufficient for successful conception.
the decrease in volume of ejaculate could be due to ACKNOWLEDGMENTS
prostatic cysts, benign prostatic hyperplasia, inflammatory The authors would like to thank the Dean, VCG and
lesions on prostate and epididymis. all the teaching staffs of Veterinary Clinical Complex,
Normal colour of dog semen is greyish-white. The Gadag for their provision to conduct clinical research and
volume of the third fraction of the ejaculate is reflective of aided the development of this paper.
the colour of semen. The appearance of semen colour in REFERENCES
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