MAIN


©1994-2021. All Rights Reserved. Online Journal of Veterinary Research. You may not store these pages in any form except for your own personal use. All other usage or distribution is illegal under international copyright treaties. Permission to use any of these pages in any other way besides the before mentioned must be gained in writing from the publisher. This article is exclusively copyrighted in its entirety to OJVR publications. This article may be copied once but may not be, reproduced or re-transmitted without the express permission of the editors


OJVRTM

 

Online Journal of Veterinary Research©

 

Volume 23 (7):628-645, 2019.


Evaluation of a muco-adhesive nano liposome low dose progesterone-PMSG

 intra-vaginal tablet on estrus, ovulation and lambing in ewes.

 

Iman Rasool Al-Shati and Najlaa Sami Ibrahim.

 

1Department of Surgery and Obstetric; Faculty of College of Veterinary Medicine University of Baghdad, Iraq.

 

SUMMARY

 

Al-Shati IR, Ibrahim NS., Evaluation of a muco-adhesive nano liposome low dose progesterone-PMSG intra-vaginal tablet on estrus, ovulation and lambing in ewes, Onl J Vet Res., 23 (7):628-645, 2019. We evaluated an intravaginal adhesive muco-nano liposomes tablet with low dose 5mg progesterone (P4) and 50IU pregnant mare serum gonadotropin PMSG (MAG) on onset and duration of estrus, ovulation, pregnancy, lambing, litter, twins and serum P4 in groups on 10 ewes each. The formulations were retained in a 50% hydrogel 0.5cm radius 0.1cm thick tablet placed on the dorsal wall of the vagina. Results were compared with standard higher dose 50mg P4 and 200IU PMSG intra-vaginal sponges (MAPS) in 10 ewes. Ten control ewes were inserted empty tablet and another 10 were not treated.  Onset, duration of estrus, ovulation were recorded and synchronization and pregnancy confirmed by blood P4 and ultrasound. Liposomes retained ~81% P4 and ~88% PMSG. Mean induction of estrus was 90% with MAG and 80% with MAPS compared with only 40-50% in controls (P < 0.05). In ewes inserted MAG, mean estrus was evident at ~34h, MAPS ~55h and controls 71-82h (P < 0.05) which then lasted 37h, 29h and 18-19h, respectively (P < 0.05). Mean pregnancy and lambing with MAG or MAPs was 90% and controls 40-50% (P < 0.05). Mean litter size was 1.33 with MAG, 1.25 with MAPS, and 1 in controls. At day 17 after insertion, serum progesterone reached 4.87±0.15ng/ml in ewes inserted MAPS and 2.93±0.99ng/ml in ewes inserted MAGS but only 1.02±0.01 ng/ml in controls inserted empty tablet and 1.64±0.03 ng/ml in those not treated. Progesterone levels in those give MAPS were higher than MAG. Lambing and twinning were correlated with higher progesterone. We found no difference in control lamb genders but more females (~66%) in ewes inserted MAG or MAPS. Birth weight in MAPS ranged 3.73-4.41kg MAG 3.47-4.01 and controls 4.40-4.72kg. We find that the low dose P4 and PMSG tablet yielded similar or better reproductive outcomes in ewes compared with standard high dose sponges.

 

Key words: liposome, PMSG, ewe, MAG Lipo P4, Lipo PMSG, Iraq, Estrous synchronization, superovulation. Raw data Provided.


INTRODUCTION

 

Exogenous hormones for estrus synchronization and superovulation are used regularly in sheep to control mating and insemination (González et al., 2001). However reduced fertility and spermatozoa possibly due to failure in cervical reservoirs, trauma, infection and immunity have been reported (Martinez-Ros et al., 2018;Armstrong and Evans 1983;Fornazari et al., 2018). WHat are the problem with sponges?????? Discuss hereNano-liposomes allow delivery of water-insoluble agents and sustain release during a prolonged period of time (Underwood and Van Eps, 2012). Authors report effect of muco-adhesive nano-liposomal progesterone PMSG on estrus and ovulation in ewes.

 

MATERIALS AND METHODS

 

This experiment was done at The Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Iraq following guidelines for the care and use of laboratory animals in research as stipulated by Al-Bayati and Khamas, (2015). During Spring to Summer (NH) 2017, forty 2 to 4 year old ~45Kg I(mean????) standard error???) primiparous and/or multiparous ewes were fed natural pastures ad libitum supplemented with 400g concentrate (daily???). For testing, ewes were grouped into straw-bedded cages.

 

No detailed descriptions of formulations here This more suited for your other formulation article Only short description of tablet and standard sponge required for this study!!! Groups of 10 ewes were inserted intravaginal adhesive muco-nano liposomes tablet with low dose 5mg progesterone (P4) ~10 days and then with 50IU pregnant mare serum gonadotropin PMSG (MAG) as described (unpublished data, 2019). Another 10 ewes were inserted standard higher dose 20mg P4 and 200IU PMSG vaginal sponges (MAPS) as dsecibed??? Refrences????. Ten control ewes were inserted empty tablet and another 10 were not.

 

Cycle status was determined by vaginal smear and blood progesterone (in blood your ,easured?????. as described by??? The perineum was washed with 1% potassium permanganate and saline; tablet??? were inserted into vagina via sterile applicator with MAG as depicted in Figure 1 below. Standard progesterone-PMSG sponges were inserted as described by ???? reference??

 

 

 

 

 

 

 

 

 

 

B

D

E

C

F

B

A

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


Figure 1. Insertion of muco-adhesive liposome (MAG) tablet impregnated with 5mg progesterone and 200IU PMSG. A) applicator, B) flat disc C) close view applicator, D) Vagina opened with speculum, E) insertion on dorsal vagina and F) adhering to dorsal vagina.

 

Onset of estrus was detected twice daily for 1h with rams and wether markings recorded as described by Evans and Maxwell (1987). Duration of estrus was determined as the interval between onset of estrus and refusal to mount (Godfrey et al., 2001) with days between onset of 2 consecutive estruses (Chemineau et al., 1992a). Estrus was calculated as ewes in heat per number of synchronized ewes x 100 as described by Cinar et al., (2017) below:

 

 

Ultrasounds were performed to determine ovulations 48 h after onset of estrus. The probe was inserted into rectum directed ventrally but pushed cranially to visualize the bladder as guide the turned slightly cranial-laterally directed by uterine horn to ovaries (refrences??). Superovulation and counting of follicles; >4 mm at onset of estrus was done as described by Kühholzer et al., (2007) and Rickard et al., (2017).

 

Jugular blood was taken prior to insertion of MAG Lipo P4 and control sponges 4 days after then every 2 days to day 8 when progesterone levels confirmed estrus synchronization (references???). Blood was kept at 25 Cº for serum centrifuged 3000 rpm for 5 min stored at -18Cº as described by (Sanjay et al., 1997).

 

Pregnancy was diagnosed by serum progesterone at day 17 after withdrawal of MAG tablet or MAPS confirmed by ultrasound after mating (Ishwar, 1995). Ewes were scanned on days 30, 60 and 120 days post mating using ultrasound scanner with a 5 MHz trans-rectal transducer probe (manufacturer???. The lubricated probe was inserted into cleaned rectum to detect urinary bladder then rotated clockwise 90º and anti-clockwise 180º for reproductive organs (Buckrell, 1988). Pregnancy was calculated as pregnant/synchronized ewes with estrus and mated (Amer and Hazzaa, 2009 and Fornazari et al., 2018).

 

 

Lambing was determined as number of ewes birthed per number of pregnant ewes as defined by Schoenian & Burfening, (1990) and Manvi, (2014).

 

 

Kidding type and twins were determined as described by Marco, (1988) and Manvi, (2014).

 

 

 

Litter size was determined as number of total lambs per number of lambing ewes (Schoenian and Burfening, 1990).

 

Gender was determined at birth time in single and twins (Fornazari et al., 2018). Lamb weights were determined at birth by electronic balance (manufacturer??) (Marco, 1988). Two-way ANOVA was used to determine differences between means (p˂0.05) as described by Lopes et al., (2014).

RESULTS

 

Results are shown in Figures 2-8 and Tables 1-10 below

 

 

 

Figure 2: Mean + SE (where is SE????) serum progesterone in ewes inserted lose dose progesterone tablet (MAG) or standard sponge (MAPS) over 9 days.

 

Table 1. Mean + SE (%) (where is SE???) estrus induced in ewes inserted low dose progesterone tablet (MAG) and standard sponge (MAPS) over 9 days.

 

Not treated

Tablet control

MAG

MAPS

Ewes

4

5

9*

8*

Estrus (%)

40

50

90*

80*

 

 

 

*(P < 0.05)

 

Table 2. Mean + SE time to estrus (h) in ewes (n=10 each) inserted low dose progesterone-PMSG tablet (MAG) or standard dose intravaginal sponges (MAPS)

 

Not Treated

Tablet control

MAG Tablet

MAPS Sponge

Time hours

82.40±8.05

71.59±6.12 *

33.81± 4.10 **

55.72± 8.44 ***

*(P < 0.05)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Figure 3.  Mean + SE time for estrus (h) in ewes (n = 10 each) inserted low dose progesterone tablet (MAG) or standard sponge (MAPS)

Table 3.  Mean + SE (h) duration of estrus in ewes ( n= 10 each) inserted low dose progesterone (MAG) or standard sponge (MAPS).

 

Not Treated

Tablet controls

MAG

MAPS

Hours

18.49±2.82

19.64±3.61*

36.78±4.52**

29.1±6.81**

*(P < 0.05) **(P < 0.01) ** (P < 0.001)????.

A

C

B

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


Figure 4. Ultra scan of ovaries showing superovulation (stars) in ewes inserted low dose progesterone tablet (A) or standard sponge (B) C. and Tablet control????

 

Table 4. Mean + SE where is SE???? pregnancy or lambing rates (%) in groups of 10 ewes treated with progesterone tablet (MAG) or standard sponge (MAPS).

 

Not treated

Tablet controls

MAG

MAPS

Pregnancy or lambing ( %)

40

50

90*

100*

The data presented as mean ± SE Whre is SE????, *(p<0.05

A

B

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


Figure 5. Ultrasound of fetus (yellow) or single (red) A. Treated ewes B. Controls.

Table 6. Mean + SE litters in ewes (n= 10) inserted low dose progesterone-PMSG tablet (MAG) or standard sponge (MAPS.

 

Not treated

 Tablet control

MAG

MAPS

Litter size

1.00±1.00

1.00±1.00

1.33±0.22*

1.25±0.05**

* (p<0.05)

 

 Table 7. Mean + SE serum progesterone (ng/ml) at day 17 of intravaginal insertion in groups of 10 ewes each with low dose progesterone-PMSG tablet (MAG) or standard sponge (MAPS).

 

Not Treated

Tablet control

MAG

MAPS

Progesterone (ng/ml)

1.02±0.01

1.64±0.03*

4.87±0.15**

2.93±0.99***

*(p<0.05)

YOU should make this figure 2 or just delete figure 2 as same data repeated!!!.

Table 8: Serum progesterone (Mean + SE), pregnancy, lambing and twins in groups of 10 ewes each inserted low dose progesterone tablet (MAG) or standard sponges (MAPS)

 

Not treated

Tablet controls

MAG

MAPS

Pregnant

4

5

9

8