A post graduate credit seminar

on

Tocolysis in Veterinary
Reproduction
Speaker: DHIREN B.BHOI
Reg.No.-04-00303-07
VOBG-900

Major Guide: Minor Guide:

Dr. V. K. Sharma Dr. J. N. Mistry
Tocolysis is a Greek derivative;
tokos = childbirth or labor
lysis = dissolution

DEFINITION

Reduced uterine contractions following administration of

myometrial relaxing agents: Tocolytic drugs.
 TOCOLYTIC DRUGS: GENERAL CONCEPT

These drugs emerged for human use and gradually linked with
animal medicine.
Smooth muscles innervated by the sympathetic nervous system:
α and β receptors
α receptors:- Muscular contractions
β receptors:- Relaxation

β1:confined to heart and small intestine

β2:vascular smooth muscle, myometrium, bronchial tree

Innes and Nicherson, 1975; Putnam et al., 1985

Stimulation of β2 receptor cause bronchodilation, relaxation of
the uterus and vasodilation in most species.

Zerobin and Kondig,1980

So, drugs that produce this effect are termed uterine myorelaxants,
tocolytic drugs or simply tocolytics.
 DRUGS USED AS TOCOLYTICS

 β-sympathomimetics –
e.g. Isoxsuprine, Ritodrine, Terbutaline,
salbutamol, Orciprenaline, Fenoterol,
Clenbuterol, Meluadrine Tartrate, KUR-1246.
Calcium antagonists –
e.g. Nifedipine, Verapamil, Nicardipine,
Nitrendipine.
Oxytocin antagonists –
e.g. Atosiban and all Beta-sympathomimetics.
Prostaglandin antagonists –
e.g. Indomethacin
Other agents –
e.g. glyceryl trinitrate, Extracts of Bryophyllum
pinnatum, omega-3 long chain
Polyunsaturates and
Magnesium sulfate.

Norwitz et al., 1999

 β-SYMPATHOMIMETICS
Tocolytic agents
Isoxsuprine lactate was 1st agents to be used for treatment of labor.

Kauppila et al., 1978, Schenken et al., 1980

Subsequently new agents invented that stimulate β2 receptors and

used as tocolytics without side effects,
Ritodrine
For examples Salbutamol,
Terbutaline,
Clenbuterol,
Orciprenaline.
Vamerzani et al., 1996, Garg et al., 2004
 COMMERCIALLY AVAILABLE TOCOLYTIC DRUGS

Sr. no. Proprietary name Ingredient Pharmaceutical Company

1 A. Duphaspasmin Isoxsuprine lactate Philips-Duphar (Holland)
B. Duvadilan Duphar Interfran, Bombay (India)
2 Yutopar Ritodrine chloride Philips-Duphar (UK)
3 Brethine Terbutaline Novartis Pharmaceuticals
Sulfate (USA)
4 Ventolin Salbutamol Allen & Hanbury's Pharma.
hemisulfate (Canada)
5 Alupent Orciprenaline Boehringer-lngelheim (Canada)
sulfate
6 Berotec Fenoterol Boehringer-lngelheim
bromide (Canada)
7 Ventipulmin, Planipart Clenbuterol Boehringer-lngelheim
hydrochloride (Canada, Germany)

Major side effects: Cardiovascular complications

 MECHANISM OF ACTION
Drug administration Drug transportation through
carrier protein or diffused
Adenyle cyclase activation osmosis; within 10-15 min.

ATP cAMP

Protein kinese stimulation

Protein phosphorilation

Intracellular calcium sequesterization

Deminished contractile protein activity

Uterine myometrial relaxation

(www.Drug.com)
 PHARMACOKINETICS
Unknown in ruminants
Absorption Entire intestine in monogastric animal
PH affects the absorption
Higher/lower PH:- Reduce absorption
Neutral PH:- Increase absorption (Smith, 1998)

Peak plasma level occurs within 1-3 hr

Distribution after oral dose. (Morgan, 1990)

Metabolism Phase: I; Oxidation, Reduction, Hydrolysis

Phase: II; Conjugation

Excretion Milk, Urine and feaces

Urine:-Clenbuterol in bovine.
(Smith and Paulson, 1997)
Clenbuterol: Dave et al, 1998
Terbutaline: Melanie et al, 2007
CLINICAL USE OF TOCOLYTIC DRUGS

Threatened abortion / Preterm labour

Controlled calving / Nocturnal delivery

To reduce neonatal morbidity and mortality

in dystocia.
To aid obstetrical operations; cesarean section
and fetotomy

Treatment of uterine prolapse

Embryo biotechnology
PRETERM LABOUR / THRETENED ABORTION

Incidence of premature labor in animals is around 5–7 %.

Gaspar et al.., 2005; King et al., 2008

Statistics indicate that preterm birth is the leading factor

causing neonatal morbidity and mortality in animals.

Haram et al., 2003, Tucker and McGuire 2004

Progesterone therapy is generally advocated for the treatment
of premature labour, but it has some adverse effects:

When labour pain has reached to its maximum amplitudes,

the drug does not work or ineffective.
It prolongs gestation length, when administered during
advanced pregnancy, as a result, chances of dystocia
increases due to additional weight gain of the fetus.

Clinical trial

300 μg, Clenbuterol, i/m, injected in a cow for the treatment

of premature labour. The treatment induced inhibition of
uterine contractions without any side effects.
Albeck, 1981
 Tocolytic effects of KUR-1246 & Ritodrine hydrochloride on
Oxytocin-Induced Uterine Contraction in Pregnant Sheep.
Ritodrine hydrochloride KUR-1246
(Oxytocin dose 0.77–0.94 IU/kg/min) (Oxytocin dose 1.00 IU/kg/min)
Dose (μg/kg/min) uterine relaxation Dose (μg/kg/min) uterine relaxation
rate (%) rate (%)
0.1 -14.3 0.001 31.1

0.3 14.9 0.003 63.1

1.0 35.3 0.010 69.6
3.0 68.8 0.030 91.3
10.0 75.5 0.100 105.9
30.0 79.4 0.300 96.3
50% inhibition dose (μg/kg/min):-- 1.5 50% inhibition dose (μg/kg/min):-0.0024
Efficacy is adjudged on: Cardiovascular parameters of dam.
: General metabolism of dam and fetus.
Kiguchi et al., 2002
Experimental trial
Effect of Meluadrine tartrate and ritodrine hydrochloride on maternal
systolic, diastolic and mean blood pressure in pregnant goats.
Control Oxytocin 30 min 60 min 90 min 120 min
Meluadrine tartrate treated goats (0.03, 0.1, 0.3, 1 μg/kg/min)
Systolic 114 ± 10 116 ± 11 115 ± 11 119 ± 11 115 ± 11 111 ± 10
(mmHg)
Diastolic 69 ± 7 72 ± 7 67 ± 6 67 ± 4 62 ± 4 60 ± 5
(mmHg)
Mean 84 ± 8 87 ± 8 83 ± 7 84 ± 6 80 ± 6 77 ± 6
(mmHg)
Ritodrine hydrochloride treated goats (1, 3, 10, 30 μg/kg/min)
Systolic 120 ± 8 120 ± 8 115 ± 8 118 ± 10 118 ± 11 116 ± 9
(mmHg)
Diastolic 72 ± 5 73 ± 4 67 ± 6 66 ± 5 61 ± 7 60 ± 6
(mmHg)
Mean 88 ± 6 89 ± 4 83 ± 6 83 ± 6 80 ± 7 78 ± 7
(mmHg)

Matsuda et al., 2002

Effect of Clenbuterol and Nifedipine on xylazine-induced alterations
in the frequency of uterine contractions, 5 min-1 in adult goats.

Group Basal 5 min 15 min 30 min

Xylazine, (0.1mg/kg, i/v)
7.6 ± 1.6 25.8 ± 2.0 17.2 ± 2.0 14.8 ± 2.0

Xylazine, (0.1mg/kg,)
+Clenbuterol (4μg/kg) i/v 7.4 ± 1.7 4.2 ± 1.5 8.4 ± 2.1 7.4 ± 1.4

Xylazine, (0.1mg/kg,)
+Nifedipine (80 μg/kg) i/v 7.6 ± 1.7 2.2 ± 1.6 7.8 ± 1.1 7.0 ± 1.2

Perez et al., 1997

CONTROLLED CALVING / NOCTURNAL DELIVERY

Using PGF2α / Corticosteroids:-

More incidence of retained placenta (Lewing, 1985)
Tocolysis:- Prediction of calving time

Bovine , Ovine & Porcine

Prediction of calving time

Physical signs:-slackening of sacrosciatic ligament, relaxation

of perineum and vulva, distension of udder, vaginal mucous
discharge are difficult to quantify and their development in
relation to calving varied considerably.
Body temperature :-Too variable to be used in prediction of
parturition time (either side of 38.9ْ C).

Ewbank, 1963
 Probability (%) of cow calving within a specified period
in relation to plasma progesterone concentration.

Progesterone Concentration Period before calving (Hours)

(ng/ml)
0-6 0-12 0-15 0-18 0-24

0 (Undetectable) 42 80 88 93 97

0.4 18 57 70 79 90

1.0 3 20 30 41 59

>1.5 0 0 0 1 3

Parker et al., 1988

 Clenbuterol was employed to regulate the calving time (i.e. to
avert night calving) by suppressing the uterine contractions. A
dose of 300μg, i.m, arrested uterine contractions for about 5 hr,
in 95 cows without obvious harmful effect.
Ballarini, 1978
300 μg, i/m, Clenbuterol suppressed uterine contractions and delay
parturition in cattle on an average 5 - 8 hrs without any ill effect on
cow or calf, expulsion of placenta and fertility of cow.
Ballarini et al., 1978

Terbutaline,5mg/kg, i/m, has potential as a tocolytic drug to delay

parturition and for various bovine obstetrical maneuver.

Melanie et al., 2007

FACTORS AFFECTING TOCOLYSIS

Stage of labour

Parity of animal

Cervical dilatation & fetal position

Pelvic area of dam

 18 cross bred cows used in a trial and found that duration of
delaying calving in the first stage of labour was much more
compare with the second stage of labour.

Group Particular Delay in

Parturition
I Control (No treatment) ------
II Clenbuterol 300 μg in first stage of labour 7-10 hours
III Clenbuterol 300 μg in second stage of labour 2-3 hours

Vamerzani et al., 1996

Once cervix is fully dilated or fetal feet are passing into cervical area
Clenbuterol will only delay labour for maximum of a few hours.
Arbeiter and Holler, 1980
300 μg, Clenbuterol Hydrochloride, i/v, was used to postponed
parturition in cows.

Clinical observations of Clenbuterol on nocturnal delivery

Cervix Duration of
Animal (No.) Degree of dilation Minutes to Duration of parturition Retension
at treatment (cm.) Tocolysis Tocolysis after of
resumption of Placenta
2-4 cm 4 cm (Minutes) (Hours)
labour (Hours) (No.)
Mean 17.18 Mean 4.9 0.74
Heifers - 25 19 6 Min 10.00 Min 3.5 0.5 2
Max 25.00 Max 7.5 1.5
Mean 21.44 Mean 4.21 0.49
Cows - 7 1 6 Min 15.00 Min 2. 50 0.25 0
Max 35.00 Max 6.00 0.75

Zerobin and Kundig, 1980

Clenbuterol 300μg, i/m, was used in heifers for postponement of
parturition.

Influence of Clenbuterol on the length (minutes) of stage-I of

parturition in heifers with large and small pelvic areas.

Control Clenbuterol
Large Pelvic areas
Heifers (No.) 10 13
Average 119 minute 468 minute
Range 30 - 230 minute 75 – 1350 minute
Small Pelvic areas
Heifers (No.) 10 13
Average 130 minute 381 minute
Range 30 - 330 minute 30 – 900 minute

Putnam et al., 1985

Influence of Clenbuterol on the length (Minutes) of stage-II of
Parturition in heifers with large and small pelvic areas.

Control Clenbuterol
Large Pelvic areas
Heifers (No.) 10 13
Average 86 minute 107 minute
Range 20 – 150 minute 15 - 145 minute
Small Pelvic areas
Heifers (No.) 10 13
Average 120 minute 54 minute
Range 45 - 270 minute 5 - 240 minute

Putnam et al., 1985

During Clenbuterol treatment in cows, the duration of tocolysis
depends on the position of the fetus at the time of treatment, being
longest at the beginning of the labour process (Stage-I).
Greene, 1981
Clenbuterol 300 μg, i/m is useful to postpone the parturition at
different stages in cows.
Group Cows Average interval from
injection to calving
Group-I (2 fingure cervical dilation) 32 23.4 hours

Group-II (4 fingure cervical dilation) 40 14.2 hours

Group-III (full hand Cervical dilation) 19 9.9 hours

Group-IV (Fetal parts in cervix) 9 5.2 hours

Greene, 1981
 OVINE

Clenbuterol, 240 μg, i/m, abolished uterine motoricity for 8-10 hrs
suspending overnight lambing in 91% of ewes subjected to trial.
Delatour and Roizard, 1979
Clenbuterol treatment in ewes effectively resulted in to delayed
parturition for at least 10 hr. in most ewes.
Plant and Bowler, 1988
Nifedipine 80 μg/kg, i/v given to sheep and found that it delayed
the parturition for 6-7 hours.
Parez et al., 1997

Clenbuterol, 6 ml, i/m given to ewes to delay lambing for 7 hours.

Hirst et al., 2005

PORCINE
Interruption of parturition
Nocturnal delivery

Clenbuterol had been used in 13 pigs for postponement of

parturition at dose rate of 150 μg, i/v (Planipart).

Suitable for all the phases of farrowing,

Interrupts labour for several hours,

Results into unhindered farrowing without affecting piglet vitality.

Zerobin and Kundig, 1980

Interruption of parturition
Gilts (No) No. of piglets Onset of Duration of Duration of No. of
already born Action Tocolysis parturition after piglets born
(Hours) resumption of (Stillborn)
(min.)
labour (Hours)
7 13 Mean 8.28 2.83 2.14 7 – 14 (0-3)
Min 5.00 2.5 1.5
Max 12.00 3.25 2.5
Postponement of parturition
Gilts (No.) Duration of Duration of parturition Number of piglets born
postponement (hrs) (Hours) Alive Stillborn
Mean 14.89 3.0
5 Min 12.5 2.5 7 - 12 0–1
Max 17.0 -

Zerobin and Kundig, 1980

 DYSTOCIA
Tocolytic drug Dose & Tocolytic Duration of
Species effect within tocolysis Reference
Isoxsuprine 5-10 ml: Cow &
lactate Mare Horvath and
1-2 ml: Ewes Bacsfay, 1981
1-4 ml: Saw 10-15 min. 1-1.5 hours Erkert and
0.2-1 ml: Bitch Macallister,
2002
0.1 ml: Cat
Clenbuterol 300 μg: Cow 15-30 min 1.5-2 hours Menard, 1994
hydrochloride 300 μg: Cow 15-30 min 1.5-2 hours Jonker et al.,
1991
100 μg: Sheep 10-15 min. 1-1.5 hours Zerobin and
Kundig, 1980
300 μg: Mare 10-15 min. 1-1.5 hours Riepe, 1981
ADVANTAGES OF TOCOLYSIS IN OBSTETRICAL MANEUVER

Less requirement of epidural anaesthesia

Easy and correct diagnosis of obstetrical defects

Facilitated obstetrical maneuvers

Repositioning of head and neck deviations.
Corrections of malpresentations and malpostures
Repulsion and rotation of fetus
Correction of forelimb retension, hock flexion,
and breech presentation

Low incidence of retained placenta (Menard, 1994)

Low incidence of genital prolapse (Albeck, 1981)

 REQUIREMENT OF EPIDURAL ANAESTHESIA

300 μg, i/m, Clenbuterol given to cows in dystocia cases

Number of animals Epidural anaesthesia

Treatment 219 80 (37%)
Control 456 319 (70%)

Menard, 1994
Results of Clenbuterol, i/v, 0.6-0.8 μg/kg body wt. before 15-20 min
of obstetrical corrections in cow.
Dystocia Total Tocolysis
Fetal
Oversize 6
Sacro-pubic position 35
Transverse presentation 2 Very good
Forelimb Retension 25
Head Deviation 41
Breech presentation 41
Hock Flexion 5
Maternal
Uterine Torsion 70 Very good
Delayed cervical dilation 7
Total 232

Menard, 1994
 Influence of Clenbuterol treatment on Retension of placenta

Dystocia Retension of Placenta Total

Yes No
Treated 48 (20.68%) 184 (79.32%) 232
Control 150 (32.89%) 306 (67.11%) 456
Total 198 490 688

Incidence of retension of placenta was lower in dystocia cases

treated with Clenbuterol compared to non-treated cows.

Menard, 1994
 CAESAREAN SECTION
Advantages
Facilitate veterinarians’ work.
Easy extraperitoneal lifting of the uterus.
(Hassett and Sloss, 1984)
No risk of anaesthetic induced recumbancy.
Impermeable closure of uterine muscles.
(Horvath and Bacsfay, 1981)
Easy suturing of uterus and no incidence of ROP.
(DeNooij, 1984)
Prevent post operative adhesions.

Reduce mortality of dam and calf.

(Menard and Diaz, 1987)
Cows treated with Clenbuterol 0.6 μg/kg body wt. 15-30 min
prior to caesarean, then good proportion of exteriorization was
noticed than in the non-treated animals.

Uterine Exteriorization
Caesareans Total
Yes No
Treated (n=63) 54/63 (85.7%) 9/63 (14.3%) 63

Non-treated (n=90) 27/90 (30%) 63/90 (70%) 90

Total 81 72 153

Menard and Diaz, 1987

 The incidence of retension of placenta and loss of cows and calf
also noticed very less in caesarean performed in cows using
Clenbuterol, 0.6 μg/kg body wt.

Treated Group (n=63) Non-treated Group (n=90)

Retension Uterine Deaths Retension Uterine Deaths
of placenta Adhesions Cow Calf of placenta Adhesions Cow Calf

(8/63) (11/63) (5/63) (4/42) (38/90) (44/90) (14/90) (8/57)

12.7% 17.46% 7.94% 9.52% 42.2% 48.88% 15.56 14.04
% %

Menard and Diaz, 1987

Good plane of uterine relaxation observed in cows affected with
dystocia treated with 5 mg/kg, i/v, terbutaline.
Melanie et al., 2007
 Uterine Exteriorization

66 66
59

40
Total animals

90% 26 Exteriorization
60% achieved
7
40% No response
10%
Treatment Group Control Group
10 ml Isoxsuprine, i/m, given in cows during caesarean section

Ahlers and Anderson, 1967

 Sheep
Very good uterine relaxation, improved ease of manipulation
and exteriorization of uterus was noticed during Caesarean
performed in Sheep (n=3) using Clenbuterol, 0.8 μg/kg b. wt.
Menard and Diaz, 1987

Pig
A cesarean section performed in a Berkshire saw treated with
6 ml Isoxsuprine, i/m, a good uterine relaxation was noticed
and uterus could be easily exteriorized.
Narasimhan and Thangaraj, 1969
 FETOTOMY
Conditions Drugs Animals Doses & Uterine Reference
route Relaxation
Fetal Isoxsuprine Cows 10 ml, i/m Very good Horvath and
Postural Bacsfay, 1981
defects Clenbuterol Cows 300μg, i/m Very good DeNooij, 1984

Clenbuterol equine 75μg, i/m Very good DeNooij, 1984

Clenbuterol Cows 300μg, i/m Very good Menard and Diaz,

1987

Fetal Isoxsuprine Cows 10 ml, i/m Very good Horvath and

Emphysema Bacsfay, 1981

Isoxsuprine Cows 10 ml, i/m Very good Narasimhan and

Thangaraj, 1969
 UTERINE PROLAPSE
Clinical traits of uterine
Animals Drug Doses Onset prolapse Reference
& of
action Reposition Reoccurrence
route
Cows Clenbuterol 3 mg, i/m 15 min. Achieved No Murling, 1983

Isoxsuprine 10 ml, i/m 15 min. Achieved No Narasimhan and

Thangaraj, 1969

Clenbuterol 0.3 mg, 15 min. Achieved No Albeck, 1981

i/m

Clenbuterol 0.3 mg, 10-20 Achieved No DeNooij, 1984

i/m min

Buffaloes Isoxsuprine 10 ml, i/m 15 min Achieved No Narasimhan and

Thangaraj, 1969

Sheep Clenbuterol 0.001mg/ 15 min Achieved No Zennetti, 1983

kg,i/m
Seven cases of bovine uterine prolapse were treated with uterine
relaxant Isoxsuprine with varying doses from 30-100 μg i/m,
according to size of the animal and good results were achieved.

Kind of animal Doses Onset of action

Non-descript cow-2 30 mg 25 minutes

Non-descript cow-1 60 mg 20 minutes

Non-descript cow-1 100 mg 15 minutes

Cross-bred jersey cow-1 100 mg 25 minutes

She buffalo-1 50 mg 20 minutes

She buffalo-1 100 mg 15 minutes

Rajasekaran et al., 1980

 UTERINE TORSION

Doses & Onset of Uterine

Animal Drug route action relaxation Detorsion Reference

Isoxsuprine 10 ml, i/m 15 min Good Achieved Horvath and

Bacsfay, 1981

Clenbuterol 0.3 mg, i/m 20 min Good Achieved Albeck, 1981

Cow

Clenbuterol 0.6μg/kg, i/v 15 min Good Achieved Menard, 1994

 EMBRYO TRANSFER
Non-surgical method

Recipients received 300μg Clenbuterol, i/m, 30-90 min. prior to

Embryo transfer.
Pregnancy rates varied from 19-56 % in Clenbuterol treated cases.
The results indicate no improvement in pregnancy rates. But
treatment resulted into a relaxant effect on uterine myometrium.
Wenkoff, 1986
Surgical method

Clenbuterol used to relax non-pregnant uterus & increased pregnancy

rates in cattle recipients undergoing surgical embryo transfer.
Coulthard, 1982
In a field trial, Clenbuterol, 364 μg (10 ml) was given by i/m,
immediately following the embryo transfer in cattle,

Recipient Pregnancy Rate

No Transfer Clenbuterol Control Total

1 Surgical 30/49 62.5 % 24/47 51.0 % 54/95 56.8 %

2 Non-surgical 57/97 58.7 % 58/98 59.2 % 115/195 58.9 %

3 Non-surgical 47/92 47.1 % 41/87 51.1 % 88/179 49.2 %

4 Surgical 31/68 45.58 % 28/71 39.43 % 59/139 42.4 %

Total 163/305 53.4 % 153/303 50.5 % 316/608 51.9 %

Barnes and First, 1985

The effects of Planipart (Clenbuterol), Duvadilan (Isoxsuprine)
and Aerolin (Salbutamol) were tested on pregnancy rates of cows
receiving the embryos transferred by non - surgical method.

Drug Dosage Transferred Embryos Pregnancy

Control ------ 38 N=15 39 %

Clenbuterol (Planipart) 0.3mg 34 N=13 38 %

Isoxsuprine (Duvadilan) 20 mg 77 N=37 48 %

Salbutamol (Aerolin) 4.8mg 24 N=11 46 %

Gregory et al., 1986

 Total 300 μg Clenbuterol given i/m to cattle recipients at the time
of ovarian palpation, 30 - 180 min. prior to embryo transfer had
effectively improved pregnancy rate after surgical embryo transfer.
Group Number of Pregnancy Rate
animals
Surgical Control 344 45%
Embryo Transfer
Treatment 264 55%

Group Number of Pregnancy Rate

animals
Non-surgical
Control 213 51%
Embryo Transfer
Treatment 307 49%

Maplotoft et al., 1986

 CONCLUSIONS

Among the β-sympathomimetic drugs used in reproduction, only

Clenbuterol and Isoxsuprine have been used widely in clinical
management of obstetrical disorders apart from embryo
biotechnology with encouraging post therapeutic results.

The efficacy of these drugs is mostly assessed clinically and

pharmacokinetic of each needs to be studied in detail to ensure wide
use with awareness of adverse effects of drug metabolites, if any.