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Comparative therapeutic effects of [PGF.sub.2[alpha]] via intramuscular and intra-vulvo submucosal route in crossbred cows.


The [PGF.sub.2[alpha]] and its synthetic analogues are capable of causing luteolysis when administered between 5th day and 16th day of oestrous cycle (Schams and Karg 1982). Both natural (Dinoprost Tromethamine, Tandle et al, 1997, Dutt and Kharche 2000) and synthetic analogues like Cloprostenol (Rowson et al., 1972), Fenprostaline (Martinez and Thibier 1984) and Luprostiol (Kharche et al., 1996) are luteolytic and are available commercially for clinical use. They have been administered through different routes like intramuscular (Horta et al., 1986, Ono et al., 1982), sub-cutaneous (Brogliatti et al., 2000), intra vulvo sub-mucosal (Horta et al., 1986, Chohan 1998), intravaginal (Ono et al., 1982), intra-uterine (Tervit et al, 1973, Louis et al, 1974), intra-luteal (Inskeep et al., 1975), intra-ovarian (Bermudez et al.,1999) and ischio rectal fossa (Colazo et al., 2002).

Administration of [PGF.sub.2[alpha]] causes luteolysis with a corresponding decrease in progesterone levels in circulation and decreasing progesterone concentrations facilitates final maturation of follicle and then expression of estrus followed by ovulation (Peterson et al., 1975). This series of events is indistinguishable from the normal events surrounding the endogenous release of [PGF.sub.2] a that occurs around 17th day of estrous cycle (Louis et al., 1975). Peterson et al (1975) recorded decline in blood progesterone before the pre estrus rise in estrogen and there was no pre estrus peak in progesterone levels and concluded that major activity of prostaglandin coincided with decline in progesterone and active stage of estrogen secretion.

Materials and Methods

The study was carried out by utilizing Holstein Friesian and Jersey crossbred cows belonging to farmers of 10 villages of Bengaluru north taluk, Karnataka. The animals selected for the study were 3-8 years of age (first to fifth lactation), apparently normal with good body condition and regular cyclicity. The animals had normal calving and had no history of retention of fetal membranes and uterine diseases like endometritis. All the animals were subjected for gynaecological examination and presence of corpus luteum was confirmed. As per the history collected and findings of rectal examination to confirm presence of corpus luteum, the selected animals were between 8 and 16 days of estrous cycle.

The [PGF.sub.2[alpha]] utilized in the study for the induction of estrus were Dinoprost tromethamine (Lutalyse (a), 5mg/ml) and Cloprostenol Sodium (Vetmate (b), 263 [micro]g/ml) which are natural and synthetic [PGF.sub.2[alpha]] respectively.

The study included one hundred crossbred dairy cows which were randomly assigned for ten treatment groups with ten animals in each group and received [PGF.sub.2[alpha]], during their luteal phase of the cycle as follows. The IVSM injection of [PGF.sub.2] a was ipsilateral to the ovary bearing corpus luteum (Table 1).

Collection of blood samples

Blood samples were collected by jugular vein puncturing with an 18 G disposable sterile needle from all the animals belonging to different experimental group just prior to administration of [PGF.sub.2[alpha]] and subsequently at 24, 48 and 72 hr. The blood samples were processed to yield serum and it was preserved at -20[degrees]C pending analysis for progesterone.

Estimation of Serum progesterone

Estimation of serum progesterone concentration by radio immunoassay method was done in RIA laboratory as per the protocol of Talwar (1983).

Results and Discussion Progesterone profile

The exogenous administration of natural [PGF.sub.2[alpha]] as well as its synthetic analogues induces luteolysis with concurrent decline in serum progesterone levels when injected between 5th and 16th day of the oestrous cycle (Hansel and Schechter 1972; Louis et al., 1975; Scams and Kharg 1982). The decreasing progesterone concentration after exogenous administration of [PGF.sub.2[alpha]] and luteolysis allow the final maturation of follicle, active estrogen secretion and expression of estrus followed by ovulation (Peterson et al., 1975; Louis et al., 1975). These events are indistinguishable from normal events with endogenous release of [PGF.sub.2[alpha]] which occurs around 17th day of oestrous cycle (Louis et al., 1975). If a mature corpus luteum was present at the time of exogenous administration of [PGF.sub.2] a the estrus was expressed in 2-5 days at which time animals could be inseminated (Williamson et al., 1972). The fertility at synchronized estrus with single or double injection schedule of [PGF.sub.2[alpha]] was normal presumably due to normal endocrine profile at [PGF.sub.2[alpha]] induced luteolysis (Laudedale et al., 1974; King and Robertson 1974; Hafs et al., 1975; Cooper and Rowson 1975).

In responded cows

The serum progesterone concentration of crossbred cows which responded with overt estrus symptoms following administration of different doses of Cloprostenol sodium and Dinoprost tromethamine by IM and IVSM route is presented in Table 2. The serum progesterone concentration of cows treated with 500 [micro]g of Cloprostenol sodium by IM route and 250 and 125 [micro]g of Cloprostenol sodium either by IM or IVSM routes showed significant decline to reach basal levels by 72 hours after treatment. Similar trend in serum progesterone concentration was observed after administration of Dinoprost tromethamine at the dose of 25mg by IM route and 12.5 and 6.25mg either by IM or IVSM route to reach below the basal levels by 72 hours.

The significant (P [greater than or equal to] 0.05) stage to stage decline in mean serum progesterone concentration followed a similar trend after administration of both Cloprostenol sodium and Dinoprost trome thamine at different doses by IM or IVSM route. Similar observation with no difference in the pattern of fall in blood progesterone levels between cows administered with synthetic (Cloprostenol sodium 500 [micro]g IM) and natural (Dinoprost tromethamine 25mg IM) [PGF.sub.2[alpha]] was reported by Baishya et al., (1994). Further, similar to the present findings the fall in progesterone concentration below 1ng/ ml by 72 hours following administration of 500 [micro]g of Cloprostenol Sodium by IM route was reported by Godfrey et al. (1989). The decline in progesterone levels to less than 1 ng/ml by 72 hours after administration of 250 [micro]g Cloprostenol Sodium by IVSM route was reported by Holy (1984). Chauhan et al. (1986) recorded decline of blood progesterone below the basal level by 72 hours after administration of Cloprostenol Sodium at 500 [micro]g by IM route and 125 [micro]g by ivsm route but the progesterone decline to basal level was not seen with administration of 62.5 [micro]g Cloprostenol Sodium by IVSM route. They opined that clinical symptoms of estrus following [PGF.sub.2[alpha]] administration were related to fall in blood progesterone level.

In the present study, the decline of progesterone in responded cows followed a similar trend to reach basal levels by 72 hours following administration of different doses of Cloprostenol sodium and Dinoprost tromethamine either by IM and IVSM route. Considering the percentage of cows responded with overt signs of estrus; the Cloprostenol sodium at 125 [micro]g by IVSM route and 250 [micro]g by IM route induced luteolysis and estrus. Similarly, Dinoprost tromethamine at 6.25 mg by IVSM route and at 12.5 mg by IM route was capable of inducing luteolysis and estrus.

In non responded cows

The serum progesterone in non responded cows following [PGF.sub.2[alpha]] treatment (Cloprostenol sodium and Dinoprost tromethamine) showed significant decline from stage to stage but it did not reach basal level by 72 hours after treatment. However, the mean serum progesterone concentration of two cows which received 125 [micro]g of Cloprostenol sodium by IVSM route declined below the basal level but there were no overt signs of estrus which could be identified by the farmer. Further, the gynaecological examination of these two cows revealed the presence of a well developed follicle, partially regressed corpus luteum and tonicity of uterus. As the present study was mainly indented to induce estrus which could be identified by farmer, these two cows were considered as non responders. However in organized dairy farms with constant Veterinary supervision these two cows could have been considered as responded. The data of non responded cows pertaining to 25mg and 12.5mg of Dinoprostromethamine by IM route was not subjected to statistical analysis as there was only one nonresponded cow in each group.

The significant stage to stage decline of serum progesterone levels in non responded cows without reaching the basal levels by 72 hours after [PGF.sub.2[alpha]] administration was indication of partial luteolysis. So complete luteolysis as indicated by decline of progesterone to basal levels is the primary requirement for estrus induction with overt symptoms. The clinical symptoms of estrus were directly related to fall in blood progesterone levels (Chauhan et al., 1986).


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(a)--Brand of Upjohn, USA

(b)--Brand of Cargill Animal Health, Bengaluru

Narasimha Murthy (1), M. Devaraj, A. Krishnaswamy and T.G. Honnappa

Department of Veterinary Gynaecology and Obstetrics Veterinary College Karnataka Veterinary, Animal and Fisheries Sciences University (KVAFSU) Hebbal Bengaluru--560024 (Karnataka)

* Part of Post Graduate Thesis submitted by first author to KVAFSU, Bidar, Karnataka.

(1.) Assistant Professor and Corresponding author. E-mail:
Table 1: Therapeutic regime followed for each group of animals

Group (N=10)      [PGF.sub.2[alpha]]         Dose

Group I          Cloprostenol sodium     500 [micro]g
Group II         Cloprostenol sodium     250 [micro]g
Group III        Cloprostenol sodium     125 [micro]g
Group IV         Cloprostenol sodium     250 [micro]g
Group V          Cloprostenol sodium     125 [micro]g
Group VI        Dinoprost tromethamine      25 mg
Group VII       Dinoprost tromethamine     12.5 mg
Group VIII      Dinoprost tromethamine     6.25 mg
Group IX        Dinoprost tromethamine     12.5 mg
Group X         Dinoprost tromethamine     6.25 mg

Group (N=10)            Route

Group I             Intramuscular
Group II            Intramuscular
Group III           Intramuscular
Group IV        Intravulvo-submucosal
Group V         Intravulvo-submucosal
Group VI            Intramuscular
Group VII           Intramuscular
Group VIII          Intramuscular
Group IX        Intravulvo-submucosal
Group X         Intravulvo-submucosal

Table 2: Serum progesterone (ng-ml) profile following
[PGF.sub.2[alpha]] administration (mean [+ or -] SE)

[PGF.sub.2[alpha]]   Route       Dose              0 hr (BT)

Cloprostenol         IM      500mg (N=10)    4.62 [+ or -] 0.29 (a)
sodium                       250mg (N=10)    4.79 [+ or -] 0.27 (a)
                             125mg (N=05)    4.45 [+ or -] 0.32 (a)
                     IVSM    250mg (N=10)    4.29 [+ or -] 0.30 (a)
                             125mg (N=08)    4.83 [+ or -] 0.35 (a)

Dinoprost            IM       25mg (N=09)    5.13 [+ or -] 0.52 (a)
tromethamine                 12.5mg (N=09)   5.02 [+ or -] 0.35 (a)
                             6.25mg (N=06)   3.75 [+ or -] 0.15 (a)
                     IVSM    12.5mg (N=07)   4.06 [+ or -] 0.38 (a)
                             6.25mg (N=07)   5.40 [+ or -] 0.42 (a)

[PGF.sub.2[alpha]]   Route       Dose                24 hr

Cloprostenol         IM      500mg (N=10)    2.58 [+ or -] 0.16 (b)
sodium                       250mg (N=10)    2.74 [+ or -] 0.20 (b)
                             125mg (N=05)    2.88 [+ or -] 0.29 (b)
                     IVSM    250mg (N=10)    2.75 [+ or -] 0.18 (b)
                             125mg (N=08)    2.76 [+ or -] 0.32 (b)

Dinoprost            IM       25mg (N=09)    2.87 [+ or -] 0.13 (b)
tromethamine                 12.5mg (N=09)   3.55 [+ or -] 0.23 (b)
                             6.25mg (N=06)   2.48 [+ or -] 0.12 (b)
                     IVSM    12.5mg (N=07)   2.92 [+ or -] 0.26 (b)
                             6.25mg (N=07)   2.84 [+ or -] 0.21 (b)

[PGF.sub.2[alpha]]   Route       Dose                48 hr

Cloprostenol         IM      500mg (N=10)    1.24 [+ or -] 0.14 (c)
sodium                       250mg (N=10)    1.30 [+ or -] 0.16 (c)
                             125mg (N=05)    1.65 [+ or -] 0.26 (c)
                     IVSM    250mg (N=10)    1.33 [+ or -] 0.13 (c)
                             125mg (N=08)    1.59 [+ or -] 0.34 (c)

Dinoprost            IM       25mg (N=09)    1.64 [+ or -] 0.18 (c)
tromethamine                 12.5mg (N=09)   1.47 [+ or -] 0.17 (c)
                             6.25mg (N=06)   1.54 [+ or -] 0.34 (c)
                     IVSM    12.5mg (N=07)   1.70 [+ or -] 0.17 (c)
                             6.25mg (N=07)   1.79 [+ or -] 0.16 (c)

[PGF.sub.2[alpha]]   Route       Dose                72 hr

Cloprostenol         IM      500mg (N=10)    0.68 [+ or -] 0.18 (d)
sodium                       250mg (N=10)    0.96 [+ or -] 0.15 (d)
                             125mg (N=05)    0.73 [+ or -] 0.13 (d)
                     IVSM    250mg (N=10)    0.80 [+ or -] 0.09 (d)
                             125mg (N=08)    0.99 [+ or -] 0.22 (d)

Dinoprost            IM       25mg (N=09)    0.92 [+ or -] 0.11 (d)
tromethamine                 12.5mg (N=09)   0.85 [+ or -] 0.11 (d)
                             6.25mg (N=06)   0.70 [+ or -] 0.26 (d)
                     IVSM    12.5mg (N=07)   0.96 [+ or -] 0.13 (d)
                             6.25mg (N=07)   0.95 [+ or -] 0.15 (d)

Note: BT-before treatment Means bearing different superscripts in
rows vary significantly with each other (P [greater than or equal
to] 0.05)
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Title Annotation:Research Article
Author:Murthy, Narasimha; Devaraj, M.; Krishnaswamy, A.; Honnappa, T.G.
Publication:Intas Polivet
Geographic Code:9INDI
Date:Jan 1, 2016
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