1. Laboratorios Calier S.A.
PLUSET : MAIN FEATURES
Ramon Calonge
Laboratorios Calier S.A.
Barcelona - Spain
Presentation prepared by Calier and given at AETS Conference August 2004.

2. 2 vials of freeze-dried powder:
500 IU FSH IU LH each
1 vial of diluent:
Normal saline to 20 ml

3. Background
Developed by Serono (Italy)
First approval: 1991, in Italy. Later on in USA, Mexico, Argentina, Brazil, New Zealand.
1998: acquired by Laboratorios Calier, S.A. Production plant moved to Barcelona (Spain). Settlement of the industrial extraction and production process.
Approval in Colombia, Venezuela, Ecuador.
2003: Approved in 5 EU member states (UK, DE, NL, ES, PT)
2004: Approved in Australia

4. FSH LH Extraction Method Frozen Pituitary Glands Acetonic phase
(2 days)
Acetonic powder
Alcoholic phase
(3 days)
Alcoholic powder
Extraction phase
(4 days)
Crude FSH / LH
Separation phase
(4 days)
FSH LH LH
FSH

5. FSH IU FSH LH IU LH Final product Vial filling Lyophilisation
Biological analysis
(to confirm activity)
Biological analysis LH
FSH
IU LH
IU FSH

6. FSH: Steelman & Pohley method (ovarian weight)
Biological analysis
FSH: Steelman & Pohley method (ovarian weight)
LH: Parlow method (Ascorbic Acid depletion)
Described by the WHO
The only ones accepted by pharmacopoeial monographs to determine FSH and LH potency of therapeutic preparations
Measures biologically active hormones
No equivalence with other methods (RIA)

7. The same proportion of the hMG (human Menopausal Gonadotrophin)
LH content
The same proportion of the hMG
(human Menopausal Gonadotrophin)
LH content is known and constant
Proportion in weight is  25:1
FSH t ½: 2:30 h
LH t ½: 40 min

8. Recommended Administration Schedule
in Holstein-Friesian Dairy Cows I.U.
Day 1 AM PM
3 ml i.m.
Start treatment between 9th and 12th day
of the cycle, preferably on 11th day
Day 2 AM PM
2.5 ml i.m.
2 AI: Inseminate at 12h and 24h of onset of oestrus
3 AI: Inseminate at 12h, 24h and 36h of onset of oestrus
Optional: add a dose of GnRH or hCG at 1st AI to induce a contemporaneous ovulation
Flushing at 6-8 days (7 days) of the 1st AI
Day 3 AM PM
1.5 ml i.m.
PGF2
PGF2
Day 4 AM PM
1 ml i.m.
Onset of oestrus
* 1 ml Pluset: 50 I.U. FSH + 50 I.U. LH

9. Recommended Administration Schedule in Spain
in Beef cows I.U.
Day 1 AM PM
2.5 ml i.m.
Day 2 AM PM
2 ml i.m.
Beef cows: I.U.
Beef heifers: 500 I.U.
Dairy heifers: 500 I.U.
Bos Indicus (zebu): I.U.
Buffalo: 250 I.U.
Sheep: I.U.
Day 3 AM PM
1.5 ml i.m.
PGF2
PGF2
Day 4 AM PM
1 ml i.m. AI
Onset of oestrus
* 1 ml Pluset: 50 I.U. FSH + 50 I.U. LH

10. Dairy cows

11. Use of different commercial FSH preparations in the superovulation of high producing Holstein cows
Fuentes, S. and De la Fuente, J.
Proceedings, XXI World Buiatrics Congress.
Punta del Este, Uruguay, 2000
Total dose: 850 IU
Day 1: PRID insertion
Day 9: 175 IU IU
Day 10: 125 IU IU
Day 11: 75 IU + 75 IU
PRID removal
PG (AM and PM)
Day 12: 50 IU + 50 IU
Holstein Cows, 5-6 years old
3-4 calvings
Interval calving-SOV treatment: months
Mean milk prod: kg/day

12. Fuentes, S. and De la Fuente, J.
Study of different doses of Pluset for the lactating cow superovulation
Fuentes, S. and De la Fuente, J.
Unpublished data
Holstein Cows, 5 years old
2.5 calvings
Interval calving-SOV : 6 months
Day 1: PRID insertion
Day 11: PRID removal
PG (AM and PM)

13. Fuentes, S. and De la Fuente, J.
Study of the effect of a similar dose of PLUSET over dairy cows with different parity number
Fuentes, S. and De la Fuente, J.
Unpublished data
Total dose: 850 IU
Day 1: PRID insertion
Day 9: 175 IU IU
Day 10: 125 IU IU
Day 11: 75 IU + 75 IU
PRID removal
PG (AM and PM)
Day 12: 50 IU + 50 IU
Holstein Cows
Interval calving-SOV : months
Group A, First calving. n= 12.
Group B, Second calving. n= 14.
Group C, Third calving. n= 17.
Group D, Fourth calving. n= 14.

14. Superovulation with different doses of PLUSET in relation to parturition number in high production dairy cows
Fuentes, S. and De la Fuente, J.
IX Jornadas sobre Producción Animal, ITEA, Zaragoza, abril’01
Day 1: PRID insertion
Day 11: PRID removal
PG (AM and PM)
Group A, 800 IU, one calving. n= 15
Group B, 850 IU, two calving. n= 15
Group C, 1000 IU, three calving. n= 14
Interval calving-SOV :
6 months (3-7 m)

15. Superovulation of Holstein-Friesian cows with reduced dose of FSH-p
Bényei B, Barros CWC
Arq. Fac. Vet. UFRGS, Porto Alegre, V. 27, n. 1, 1999 (supl.): p. 203
Control: 1000 IU FSH/4 days (n=99)
Experiment: 600 IU FSH/4 days (n=83)
PG at the 3rd day; AI at 45 and 60h

16. Arq Bras Med vet Zootec Vol 52, n 4, Belo Horizonte Aug 2000
Effect of superovulation on performance of bovine embryo donors imported from temperate zone to tropical climate during the first two years of adaptation
Bényei B, Barros CCW
Arq Bras Med vet Zootec Vol 52, n 4, Belo Horizonte Aug 2000
Holstein-Friesian heifers imported pregnant from Hungary to Brazil
In the 2nd year were superovulated with Folltropin or Pluset (600 IU / 4 days)
FSH in decreasing doses for 4 days, PG at 3rd day, AI at 60-72h

17. Heifers

18. Superovulation in cattle: effect of FSH type and methods of administration on follicular growth, ovulatory response and endocrine patterns
Kelly P., Duffy P., Roche J.F. and Boland M.P. Animal Reproduction Science 46 (1997): 1-14
Group SF: (n=23) Single Folltropin. 270 mg NIH-FSH-P1 in a single s.c injection.
Group MF: (n=23) Multiple Folltropin, 270 mg in 4 days
Group SP: (n=22) Single Pluset, IU in a single s.c injection
Group MP: (n=22) Multiple Pluset, IU in 5 days

19. Effect of timing of oestradiol benzoate injection relative to gonadotropin treatment on superovulatory response, and on embryo yield and quality in beef heifers
Yaakub H, Duffy P, O’Callaghan D, Boland MP
Animal Reproduction Science (1998) 52:
Group I: only CIDR for 9 days
Group II: 5 mg ODB at CIDR insertion
Group III: 5 mg ODB 1.5 days after CIDR insertion
Group IV: 5 mg ODB 3 days after CIDR insertion
Sov started 6.5d after CIDR insertion
3 doses: 400, 600, 800 IU Pluset
in 8 equal i.m injections each 12h
RESULTS:
No interaction between treatment and dose of Pluset
Combination of ODB and CIDR do not improve embryo yield
best results achieved with CIDR only

20. Mantovani R et al - 12nd Meeting AETE - Lyon, 1996: 168
Dose effect of porcine gonadotropin extract containing FSH and LH on superovulatory response and fertility of Friesian heifers
Mantovani R et al - 12nd Meeting AETE - Lyon, 1996: 168
60 cycling heifers, months
Dose in 9 i.m doses, each 12h (4 1/2 days), giving 35%, 25%, 20%, 15% and 5% of the total dose
Results: Optimal response with 500 IU FSH, with no effects on subsequent reprod. Performance
higher dose affects embryo yield and quality, but not fertility

21. Beef cows

22. Effects of prepartum lipid supplementation on FSH superstimulation and embryo quality in multiparous beef cows
Bader JF et al
Theriogenology, v 57, n 1, 2002: 760
40 mature cows, prepartum lipid supplement 40 days prior to calving
WSB: whole soybean supplement
GnRH-7 days- GnRH- 7 days- PG: 28 cows responded
Superovulation started days 8-10 cycle (60d postpartum)
600 IU at decreasing doses for 4 days, PG: 96h and 108h after start treatment
27 cows showed oestrus: AI 0, 12 and 24h after onset oestrus

23. Evaluation of an estrus synchronization protocol prior to superovulation in beef cows
Kojima FN et al
Theriogenology, v 57, n 1, 2002: 766
24 crossbred non-lactating beef cows
Protocol 7-11 synch (n=11) or control (n=13)
7-11 Synch: MGA for 7 days + PG 7th day + GnRH 11th day
Control synchronised with 2 PG 14 days apart
2 days after GnRH: 600 IU FSH, decreasing doses twice daily, for 4 days
PG: 2 adm. on the last day FSH - 5th Day: PG, AI at 48h and 60h

24. Zebu (Bos indicus)

25. Total Doses Recommended in Brazil
Breed
Age
Minimum
Maximum
Nelore
Heifer
220 UI
260 UI
Cow
250 UI
350 UI
Simmental
280 UI
320 UI
400 UI
Holstein
420 UI
500 UI
Crossbred
Zebu/European
230 UI

26. Efficiency of two different hormonal products for superovulation of
Nelore cows
Diniz EG et al
Rev. Bras. Reprod. Anim., v.23, n.3, 1999:
Comparison of 250 IU Pluset vs. 20 mg NIH Folltropin in Nelore Cows (Bos indicus)
FSH in decreasing doses, in 4 days. PG: 2 doses (AM-PM) of the 3rd day
AI: h post-oestrus

27. Rev. Bras. Reprod. Anim., v.26, n.3, 2002: 246 - 248
Infuence of age on the superovulatory response of Nelore donors: preliminary results
Oliveira MAL et al
Rev. Bras. Reprod. Anim., v.26, n.3, 2002:
Comparison of superovulatory response of 100 Nelore donors allocated by age:
GI: 2-5 years; GII: 6-8 years; GIII: 9-12 years; GIV: >12 years
400 IU FSH in decreasing doses, in 4 days. PG: on the 3rd day.
AI: h post-oestrus
Group
N cows
Total
structures
Viable
embryos
Degenerate
Ova
2-5 y 25
15.8 ± 10.9
9.8 ± 8.4
2.3 ± 2.2
2.9 ± 3.7
6-8 y
18.5 ± 12.3
11.5 ± 10.0
3.3 ± 3.7
4.0 ± 4.8
9-12 y
13.6 ± 13.6
6.5 ± 5.1
3.1 ± 2.9
4.3 ± 5.5
> 12y
9.2 ± 5.0
4.0 ± 2.6
1.6 ± 2.0
3.6 ± 4.3

28. Now available in Australia!
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