pluset ® superovulation raised to maximum potency pluset : main features ramon calonge laboratorios...

plusetpluset®® Superovulation raised to maximum potency

PLUSET : MAIN FEATURES

Ramon CalongeLaboratorios Calier S.A.

Barcelona - Spain Presentation prepared by Calier and given at AETS Conference August 2004.


2 vials of freeze-dried powder:2 vials of freeze-dried powder:

• 500 IU FSH + 500 IU LH each500 IU FSH + 500 IU LH each

1 vial of diluent:1 vial of diluent:

• Normal saline to 20 mlNormal saline to 20 ml


BackgroundBackground

• Developed by Serono (Italy)Developed by Serono (Italy)

• First approval: 1991, in Italy. Later on in USA, Mexico, First approval: 1991, in Italy. Later on in USA, Mexico,

Argentina, Brazil, New Zealand.Argentina, Brazil, New Zealand.

• 1998: acquired 1998: acquired by Laboratorios Calier, S.A. Production plant by Laboratorios Calier, S.A. Production plant

moved to Barcelona (Spain). Settlement of the industrial extraction moved to Barcelona (Spain). Settlement of the industrial extraction

and production process.and production process.

• Approval in Colombia, Venezuela, Ecuador.Approval in Colombia, Venezuela, Ecuador.

• 2003: Approved in 5 EU member states (UK, DE, NL, ES, PT) 2003: Approved in 5 EU member states (UK, DE, NL, ES, PT)

• 2004: Approved in Australia2004: Approved in Australia


Frozen Pituitary GlandsFrozen Pituitary GlandsFrozen Pituitary GlandsFrozen Pituitary Glands

Acetonic Acetonic powderpowderAcetonic Acetonic powderpowder

Alcoholic Alcoholic powderpowderAlcoholic Alcoholic powderpowder

Crude FSH / LHCrude FSH / LHCrude FSH / LHCrude FSH / LH

SeparationSeparation phasephase (4 days)(4 days)

FSHFSH

LLHH

LLHH

FSFSHH

Acetonic phaseAcetonic phase (2 days)(2 days)

ExtractionExtraction phasephase (4 days)(4 days)

AlcoholicAlcoholic phase phase (3 (3 days)days)

Extraction MethodExtraction Method


FSHFSH

LLHH

LLHH

FSFSHH

Final productFinal product

Biological analysisBiological analysis

IU IU FSHFSH

IU LIU LHH

IU LIU LHH

IU IU FSHFSH

Vial fillingVial fillingLyophilisationLyophilisation

Biological analysisBiological analysis(to confirm activity)(to confirm activity)

Vial fillingVial fillingLyophilisationLyophilisation

Biological analysisBiological analysis(to confirm activity)(to confirm activity)


Biological analysisBiological analysis

FSH: Steelman & Pohley method FSH: Steelman & Pohley method (ovarian weight)(ovarian weight)

LH: Parlow method LH: Parlow method (Ascorbic Acid depletion)(Ascorbic Acid depletion)

• Described by the WHODescribed by the WHO

• The only ones accepted by pharmacopoeial The only ones accepted by pharmacopoeial

monographsmonographs to determine FSH and LH potency of to determine FSH and LH potency of

therapeutic preparationstherapeutic preparations

• Measures biologically active hormones Measures biologically active hormones

• No equivalence with other methods (RIA)No equivalence with other methods (RIA)


LH contentLH content

• The same proportion of the hMG The same proportion of the hMG

(human Menopausal Gonadotrophin)(human Menopausal Gonadotrophin)

• LH content is known and constantLH content is known and constant

• Proportion in weight is Proportion in weight is 25:1 25:1

• FSH t ½: 2:30 hFSH t ½: 2:30 h

• LH t ½: 40 minLH t ½: 40 min


Recommended Administration Schedule in Holstein-Friesian Dairy Cows - 800 I.U.

Day 1 AM

PM

Day 3 AM

PMDay 4 AM

PM

Day 2 AMPM

2.5 ml i.m.

2.5 ml i.m.

3 ml i.m.

3 ml i.m.

1 ml i.m.

1 ml i.m.

1.5 ml i.m.

1.5 ml i.m.

* 1 ml Pluset: 50 I.U. FSH + 50 I.U. LH

Start treatment between 9th and 12th dayof the cycle, preferably on 11th day

Onset of oestrus

PGF2

•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

PGF2


Day 1 AM

PM

Day 3 AM

PMDay 4 AM

PM

Day 2 AMPM

2 ml i.m.

2 ml i.m.

2.5 ml i.m.

2.5 ml i.m.

1 ml i.m.

1 ml i.m.

1.5 ml i.m.

1.5 ml i.m.

Onset of oestrus

PGF2

• Beef cows: 600-700 I.U. • Beef heifers: 500 I.U. • Dairy heifers: 500 I.U. • Bos Indicus (zebu): 200-300 I.U. • Buffalo: 250 I.U. • Sheep: 250-350 I.U.

PGF2

AI

* 1 ml Pluset: 50 I.U. FSH + 50 I.U. LH

Recommended Administration Schedule in Spain in Beef cows - 700 I.U.


Dairy cows


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 insertionDay 9: 175 IU + 175 IUDay 10: 125 IU + 125 IUDay 11: 75 IU + 75 IU

PRID removalPG (AM and PM)

Day 12: 50 IU + 50 IU

Holstein Cows, 5-6 years old3-4 calvingsInterval calving-SOV treatment: 5.8-6.5 monthsMean milk prod: 27-30 kg/day

n Totalembryos

Unfertilisedembryos

Transferableembryos

Pluset 66 9.4 ± 4.7 2.3 ± 2.6 6.2 ± 4.0a

Ovagen 35 7.3 ± 4.8 2.1 ± 2.8 3.9 ± 3.4b

FSH-p 33 7.8 ± 5.2 3.7 ± 4.3 3.2 ± 2.9c

a vs b: P< 0.05. a vs c: P< 0.001


Study of different doses of Pluset for the lactating cow superovulation

Fuentes, S. and De la Fuente, J.Unpublished data

Holstein Cows, 5 years old2.5 calvingsInterval calving-SOV : 6 months

800 IU 850 IU 1000 IUAM PM AM PM AM PM

Day 9 150 IU 150 IU 175 IU 175 IU 175 IU 175 IUDay 10 125 IU 125 IU 125 IU 125 IU 150 IU 150 IUDay 11 75 IU 75 IU 75 IU 75 IU 100 IU 100 IUDay 12 50 IU 50 IU 50 IU 50 IU 75 IU 75 IU

Day 1: PRID insertionDay 11: PRID removal

PG (AM and PM)

Dose N Total embryos Transferableembryos

Unfertilised ova

800 30 10,9 5,3a 6,9 4,1a 2,8 2,5850 75 9,5 5,1ab 5,9 3,9ab 3,1 3,1

1000 30 7,2 4,2b 4,2 3,0b 2,7 3,2a vs b: P< 0.01


Study of the effect of a similar dose of PLUSET over dairy cows with different parity number


Unpublished dataHolstein CowsInterval calving-SOV : 4.5-6 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.

Total dose: 850 IU

Day 1: PRID insertionDay 9: 175 IU + 175 IUDay 10: 125 IU + 125 IUDay 11: 75 IU + 75 IU

PRID removalPG (AM and PM)

Day 12: 50 IU + 50 IU

Calving N E.TOT E.VIA NF1 12 9,2 ± 5,9 7,1 ± 3,9 a 1,2 ± 1,3 a

2 14 9,8 ± 4,1 7,8 ± 3,1 a 0,9 ± 0,9 a

3 17 10,7 ± 4,7 7,1 ± 4,5 a 2,9 ± 2,6 a

4 14 9,0 ± 2,3 5,7 ± 2,5 a 2,7 ± 2,2 a

ETOT: Total embryos mean. EVIA: Viable embryos mean.NF: Non-fecundated ova.

a vs. b: P<0.05


Superovulation with different doses of PLUSET in relation to parturition number in high production dairy cows


IX Jornadas sobre Producción Animal, ITEA, Zaragoza, abril’01

800 IU 850 IU 1000 IUAM PM AM PM AM PM

Day 9 150 IU 150 IU 175 IU 175 IU 175 IU 175 IUDay 10 125 IU 125 IU 125 IU 125 IU 150 IU 150 IUDay 11 75 IU 75 IU 75 IU 75 IU 100 IU 100 IUDay 12 50 IU 50 IU 50 IU 50 IU 75 IU 75 IU

Day 1: PRID insertionDay 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)

Calving Dose N ETOT EVIA NF1 800 15 11,1 4,1 8,5 4,0 1,7 1,02 850 15 9,5 4,6 7,7 3,0 1,1 1,43 1000 14 9,6 4,0 8,3 2,8 1,0 2,4

ETOT: Total embryos EVIA: Viable embryos NF: Non-fecundated ova


Superovulation of Holstein-Friesian cows with reduced dose of FSH-pBényei 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

600 IU 1000 IUNo. Ovulations 13.6 6.0 12.3 6.9Good quality embryos 5.3 3.9 4.8 3.7Discarted ova 4.8 3.9 4.1 3.6Cystic ovaries 3.6% 20.2%


Effect of superovulation on performance of bovine embryo donors imported from temperate zone to tropical climate during the first two

years of adaptationBényei 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

Folligon Pluset Folltropin

Cows flushed 6 40 15Total structures 7.4 0.9a 11.7 6.9b 7.7 5.4a

Degen. / unfertil. 3.4 0.6a 5.6 4.8a 4.8 4.9a

Viable embryos 4.0 0.8a 6.0 4.4b 2.9 3.3c


Heifers


Superovulation in cattle: effect of FSH type and methods of administration on follicular growth, ovulatory response and endocrine

patternsKelly 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, 1.000 IU in a single s.c injection

Group MP: (n=22) Multiple Pluset, 1.000 IU in 5 daysDay 1

(AM/PM)Day 2

(AM/PM)Day 3

(AM/PM)Day 4

(AM/PM)Day 5

(AM/PM)MF (mg NIH) 50/50 40/40 28/28 17/17 --MP (UI) 150/150 125/125 100/100 75/75 50/50

Treatment SF SP MF MP No. CL 10.5 2.6a

22.6 3.5 b 17.7 2.1c

23.3 2.3 b

No. Embryos Rec. 5.7 1.1 a 13.6 2.2 b

12.2 1.7 b 16.4 1.4 c

No. Gr. 1+2 2.2 0.5 a 2.6 0.8 a

5.5 1.2 c 3.8 1.0 b

No. Gr. 3 1.0 0.3 a 1.3 0.4 a

2.7 0.9 b 2.7 0.9 b

No. Gr. 4+5 0.7 0.2 a 2.5 0.8 b

1.6 0.4 c 1.5 0.4 c

No. Unfert/Deg. 1.9 0.6 a 7.2 1.9 b

2.4 0.9 a 8.4 1.8 b

a-c Means with the same superscript within rows do not differ (p>0.05)


Effect of timing of oestradiol benzoate injection relative to gonadotropin treatment on superovulatory response, and on embryo yield and quality

in beef heifersYaakub H, Duffy P, O’Callaghan D, Boland MP

Animal Reproduction Science (1998) 52: 191-204

Group I: only CIDR for 9 daysGroup II: 5 mg ODB at CIDR insertionGroup III: 5 mg ODB 1.5 days after CIDR insertionGroup IV: 5 mg ODB 3 days after CIDR insertion

Sov started 6.5d after CIDR insertion3 doses: 400, 600, 800 IU Plusetin 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

Dose of FSH (IU) 400 IU 600 IU 800 IUNo. heifers 31 29 31No. CL 11.7 1.0 a 15.8 1.4 a,b 18.0 1.9 b

No. ova/Emb. recovered 5.8 0.9 a 7.0 0.8 a,b 9.1 1.0 b

No. transferable emb. 2.8 0.7 a 4.2 0.6 b 4.1 0.7 a,b

No. degen. ova/emb. 1.0 0.5 a 2.8 0.5 a 5.0 0.9 b


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, 13-16 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 fertilityDose of Pluset

100 IU 300 IU 500 IU 700 IUNo. Heifers 15 16 14 15Recovered Embryos 1.3 4.2 8.0 3.4Unfertilised ova 0.2 2.5 0.8 2.6Transferable embryos 1.2 3.0 6.5 2.5Grade 1+2 embryos 1.2 2.6 5.9 2.2Service period (days) 42 41 60 49No. Services 2.2 2.7 2.5 2.4Pregnancy rate (%):

At 1st AI 46.1 38.8 47.1 27.8After 3 months 86.1 76.8 75.7 84.9


Beef cows


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

Total Embryos

Transferable Embryos

Degenerate Embryos

Unfertilised Ova

WSB 14.73 3.54 10.33 2.49 3.33 1.13 1.07 0.47 Control 17.5 2.95 13.58 2.61 1.58 1.68 2.33 1.23


Evaluation of an estrus synchronization protocol prior to superovulation in beef cowsKojima FN et al

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

Theriogenology, v 57, n 1, 2002: 766

Treatment Total

structures Total

embryos Transferable

embryos Freezable embryos

Degenerated embryos

Unfertilized ova

7-11 Synch 20.9 3.2 21.0 1.8 9.9 1.6 8.3 1.4 2.1 0.7 8.9 2.7 Control 20.8 3.1 17.7 2.9 15.3 2.6 13.1 2.3 2.3 0.8 3.1 1.1


Zebu (Bos indicus)


Breed Age Minimum Maximum

Nelore Heifer 220 UI 260 UICow 250 UI 350 UI

Simmental Heifer 280 UI 350 UICow 320 UI 400 UI

Holstein Heifer 350 UI 420 UICow 400 UI 500 UI

Crossbred

Zebu/European

Heifer 230 UI 280 UICow 260 UI 350 UI

Total Doses Recommended in BrazilTotal Doses Recommended in Brazil


Efficiency of two different hormonal products for superovulation of Nelore cowsDiniz EG et al

Rev. Bras. Reprod. Anim., v.23, n.3, 1999: 319-321

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: 12 + 24h post-oestrus

NViable

embryosDegenerated

Non-fecundated

Totalstructures

Folltropin 200 5.9 5.1 5.8 8.8 1.1 3.7 10.7 8.6Pluset 33 6.8 7.3 5.1 5.9 0.7 1.8 12.6 8.4


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: 246 - 248

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: 12 + 24h post-oestrus

Group N cows Totalstructures

Viableembryos

Degenerateembryos

Ova

2-5 y 25 15.8 ± 10.9 9.8 ± 8.4 2.3 ± 2.2 2.9 ± 3.7

6-8 y 25 18.5 ± 12.3 11.5 ± 10.0 3.3 ± 3.7 4.0 ± 4.8

9-12 y 25 13.6 ± 13.6 6.5 ± 5.1 3.1 ± 2.9 4.3 ± 5.5

> 12y 25 9.2 ± 5.0 4.0 ± 2.6 1.6 ± 2.0 3.6 ± 4.3


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pluset ® superovulation raised to maximum potency pluset : main features ramon calonge laboratorios...

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