Superovulation

Superovulation, also called superstimulation, is a treatment intended

to increase the ovulation rate and thus the number of available oocytes
in the donor animal without disrupting the physiological and
endocrinological processes associated with oocyte maturation,
ovulation, and fertilisation, as well as subsequent embryonic and fetal
development.
If exogenous FSH is supplied, pure or associated with LH these follicles can
be " recuperated " and thereby avoid atresia. It is the principle of
stimulatory treatments of ovarian functions (hMG or pure FSH) which lead to
multiple ovulations.
Principle of superovulation

To induce more ovulations than normal rate by giving a gonadotrophin

stimulus (at critical moments of follicular development), followed by
control of luteolysis, synchronous ovulation, high fertilisation and early
embryonic development rates.
The most favorable and optimal time for superovulation treatments is
between the 8th and 14th day of the cycle.

Superovulation treatment will result in release of multiple eggs

at a single estrus.

Cows or heifers that are properly treated can release 10 or more

viable eggs.
Nearly 85 % of the donors respond to superovulation treatment protocols.
An ideal response of 5 to 12 good quality embryos is obtained from about
one third of the donors
Hormones used in treatment
• Equine chorionic gonadotrophin (eCG) derived from the serum of
pregnant mares (also called pregnant mare’s serum gonadotrophin,
PMSG);
• Extracts of domestic animal pituitaries, particularly those of the pig, of
various degrees of purity and FSH to LH ratios;
• Recombinant FSH; and
• Gonadotrophins of pituitary origin extracted from human post
menopausal urine (human menopausal gonadotrophin, hMG)
PMSG
The pregnant mare’s serum gonadotrophin (PMSG) is a glycoprotein that
produces both FSH and LH biological effects. Contains a high content of
carbohydrate side chains and sialic acid.

Half life: A very long half-life of about 5 days .

Dose: A single dose of 1500 to 3000 IU of PMSG.
Time: During the mid-luteal phase of the estrous cycle.
A luteolytic dose of prostaglandin F2 D or an analogue is administered 2 to 3
days later.
The result is an interval of about 4 days between starting PMSG
treatment and the onset of oestrus.
Adverse effect of PMSG
The long half-life of PMSG and its LH activity might produce adverse
effects on superovulatory response, particularly on the quality of
embryos.
The long half-life of PMSG often induces extrafollicular growth during
the first follicular wave postovulation, with a resultant increased
estradiol secretion that persists during the postovulatory period. This
may have a deleterious effect on early embryonic development.
To minimise the adverse effect of PMSG on fertilisation and embryonic
development, the injection of an antiserum against PMSG has been
proposed. The PMSG-antiPMSG treatment resulted in better
superovulatory responses, including higher ovulation rate, decreased
number of unruptured follicles and decreased number of cysts.

Neutralisation of PMSG at any time before the LH surge, worthy

interferes with the normal stimulation of follicles. The administration
of the anti-PMSG serum shortly (6 to 8 hours) after the preovulatory
LH surge, decreases the excessive follicular development subsequent to
PMSG injection, and therefore results in an increase in the ovulation rate
and the number of normal embryos recovered.
FSH
The most widely used procedures consists of administrating two daily
injections of FSH at approximately 12-hours intervals over a 4-day period,
more frequently in decreasing doses.
The half-life of FSH is believed to be approximately 5 hours, which
justifies the multiple injection protocols.

Treatment: The most common protocol for superovulation with FSH was
5, 5, 4, 4, 3, 3, 2 and 2 mg with prostaglandin F2alpha or an analogue.
The prostaglandin F2alpha is given simultaneously to the 5th or 6th
injection of FSH to induce the lysis of the donor's corpus luteum.
FSH-LH ratio
One of the main factors influencing the effectiveness of superovulation is
the quality of the gonadotrophin preparation used.
Especially the proportion of FSH/LH plays an important role, since it
controls the growth of the ovarian follicles.
The activity ratio of LH to FSH preparation used affects the response, with
the higher LH content of the gonadotrophin preparation generally resulting
in a lower superovulatory response.
The LH must be present in preparations used for superovulation with a
FHS:LH ratio of approximately 5: 1
Recombinant bovine FSH (rbFSH)
The recombinant bovine FSH (rbFSH) has been shown to posses high
biological activity.
The embryo production with rbFSH appears to be comparable with data
for other superovulatory compounds, but embryo quality seems to be
increased using rbFSH when compared with the result from
superovulation with pituitary FSH.
Although reducing the number of injections of FSH results in decreased
superovulatory responses, superstimulation with a single subcutaneous
injection of FSH has been reported with encouraging results.
Probably the FSH dissolved in polyvinylpyrrolidone is capable of
achieving a similar profile to that obtained with conventional multiple-
injection procedure.
Other hormones
The availability of recombinant bovine somatotrophin (rbST) led to its
use in combination with FSH treatment, being a co-gonadotrophin. The co-
treatment with rbST enhances the superovulatory response and embryo
yield. The hMG is a protein purified from urine collected from menopausal
women; at presents both FSH and LH activities. 600 IU of hMG
administered twice daily over a 3-day period produced a recovery rate of
high quality embryos higher than that obtained following treatment with 20
mg of FSH. More recently, it was described that superovulation can be
adequately induced using only one injection of 450 to 600 IU of HMG in
polyvinylpyrrolidone.
Treatment with a horse anterior pituitary extract (HAP) was found to be
an acceptable alternative to FSH, but there is no reduction in the variability
of the superovulatory response.