Research Article

The Leptin Hormone Administration on the Estrus Intensity and the Appearance Time of Postpartum Estrus in Primiparous Bali Cattle

Desak Nyoman Dewi Indira Laksmi1*, I. Gusti Ngurah Bagus Trilaksana1, I. Wayan Sukernayasa1, Nyoman Oka Widiarta2, I. Wayan Nico Fajar Gunawan1, I Made Merdana3

1Department of Veterinary Clinic and Reproduction, Faculty of Veterinary Medicine, Udayana University, Jimbaran, Badung Regency, Bali, 80361, Indonesia; 2Bali Cattle Breeding Center, Department of Agriculture and Food Security, Badung Regency, 80351, Indonesia; 3Department of Veterinary Pharmacy, Faculty of Veterinary Medicine, Udayana University, Jimbaran, Badung Regency, 80351, Indonesia

Received | May 14, 2024; Accepted | June 22, 2024; Published | August 05, 2024

*Correspondence |Desak Nyoman Dewi Indira Laksmi, Department of Veterinary Clinic and Reproduction, Faculty of Veterinary Medicine, Udayana University, Jimbaran, Badung Regency, Bali, 80361, Indonesia; Email: dewiindira@unud.ac.id

Citation | Laksmi DNDI, Trilaksana IGNB, Sukernayasa IW, Widiarta NO, Gunawan IWNF, Merdana IM (2024). The leptin hormone administration on the estrus intensity and the appearance time of postpartum estrus in primiparous bali cattle. Adv. Anim. Vet. Sci. 12(9): 1700-1704.

DOI | https://dx.doi.org/10.17582/journal.aavs/2024/12.9.1700.1704

ISSN (Online) | 2307-8316; ISSN (Print) | 2309-3331

Copyright: 2024 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

The government’s efforts to increase the population of Bali cattle and provide meat nationally have been carried out continuously with several government programs. However, one barrier to success in its implementation is suboptimal reproductive function in Bali cattle. This condition will impact economic losses in the form of additional costs for treatment and breeding, long unproductive periods, an increase in abandoned livestock, and a population decline (Budiyanto et al., 2016). In addition to the low quality of seeds, the problems often encountered in developing Bali cattle (Chamdi, 2005) are also caused by less than optimal reproductive function (Suartini et al., 2013). The disruption of reproductive function will have an impact on low birth rates.

There are several things that can be used as indicators of suboptimal reproductive function in Bali cattle, one of which is the length of the calving interval due to the absence of estrus more than 3 months after giving birth (postpartum anestrus). According to (Guntoro, 2002), the calving interval for Bali cattle is very long, namely around 555 days. In fact, to give birth to one calf a year, the calving interval for cows should ideally be no more than 365 days or 12 months (Kamal et al., 2014). The return of the estrous cycle after delivery highly depends on the function of the hypothalamic-pituitary-ovarian axis. The hypothalamus secretes gonadotropin-releasing hormone (GnRH) at the right frequency and amount to stimulate the gonadotropin hormones, namely follicle-stimulating hormone (FSH) and luteinizing hormone (LH) from the anterior pituitary. An increase in GnRH frequency is the main factor in increasing FSH, LH, and follicular development circulation, where the follicles will synthesize and produce the hormone estrogen. Leptin is a metabolic signal regulating the hypothalamic pituitary ovarian axis (Zieba et al., 2003; Ahmadzadeh et al., 2011). (Kadokawa and Martin, 2006) stated that leptin concentration is correlated with the frequency and amplitude of the LH hormone. (Holtenius et al., 2002) reported that a minimum leptin concentration is required to induce an LH surge after delivery.

According to (Kadokawa et al., 2000), in the early postpartum, there is a decrease in leptin concentration until it reaches the nadir concentration and then increases again to stimulate the hypothalamus to secrete GnRH. In addition, the longer the interval from giving birth until leptin reaches the nadir concentration will prolong the appearance of estrus and the first ovulation after giving birth. Research by (Laksmi, 2018) said that peripartum leptin levels were lower than leptin levels during postpartum estrus. Likewise, research conducted by (Liefers, 2004) in Holstein dairy cows found that a leptin concentration of 4.2 ng/ml achieved after giving birth would cause the appearance of postpartum estrus. Leptin injection into infertile mice can increase FSH and LH concentrations in serum and increase ovary weight, testicular weight, and sperm count. Administration of leptin can restore fertility, growth, and reproductive organ function in mice that lack the leptin gene (Zieba et al., 2005)

MATERIALS AND METHODS

This research was approved by the Animal Ethics Commission, Faculty of Veterinary Medicine, Udayana University, Indonesia, with approval Number B/283/UN14.2.9/PT.01.04/2023. It is a true experiment using a Randomized Pre Post Group Control Design. For three months, leptin administration and observing the estrus appearance in Bali cattle was conducted at SIPADU and the Breeding Center in Sobangan Village, Mengwi District, Badung Regency. The sample for this study was 28 cattle based on Federer’s formula (1963): the number of treatments (n) = 2, and the repetition was carried out 14 times. This cattle gave birth once and did not show postpartum estrus for over three months.

Recombinant hormone Ovine Leptin (Brand Cedarlane) at a dose of 200 µg/ml dissolved in 2 ml of sterile aquadest, 3 CC syringes, cotton, and alcohol were provided. The recombinant leptin at a 200 µg/ml dose was administered twice subcutaneously, with an interval of 12 hours. (Laksmi, 2016 ). The estrus was observed twice daily, every morning (06.00-09.00 AM) and afternoon (06.00-09.00 PM). Furthermore, the signs observed were restlessness, swollen and red vulva covered by transparent mucus, increased blood flow to the vaginal mucosa and mucus fluid accumulated, urination, tail lifted, and ready to receive the male (Hafez and Hafez, 2000).

A score from 1 to 3 is used to compare the level of estrus intensity, with 1, 2, and 3 indicating less clear, moderate intensity, and clear intensity estrus (Yusuf, 1990). The value of 1 was assigned to cattle showing symptoms of less mucus discharge (++), unclear vulva condition (swelling, wet and red) (+), appetite does not appear to decrease (+), less restless as well as no visible symptoms of climbing and silent when climbed by fellow female cattle (-). Meanwhile, a score of 2 was assigned to livestock that showed all of the above estrus symptoms with the symbol ++, including the signs of riding other female livestock and silence when climbed by other females with an intensity of moderate. The estrus intensity with a score of 3 (clear) was awarded to female cattle clearly showing all estrus symptoms (+++).

This research data were evaluated descriptively, and the Shapiro-Wilk test was employed to determine the sample data’s mean distribution status. An inferential test was conducted to examine the difference in mean significance in all treatment groups.

RESULTS AND DISCUSSION

The appearance time and intensity of estrus in Bali cattle were observed by administering leptin until the symptoms developed. The signs of estrus were characterized by the discharge of clear transparent mucus from the vagina, the vaginal mucosa appearing reddish, and the presence of vulvar swelling. Furthermore, the intensity level of estrus was compared by a score of 1 to 3; the ratings of 1, 2, and 3 indicate less clear, moderate, and clear intensity estrus.

The body condition score (BCS) of Bali cattle with and without leptin in several SIPADU in Sobangan village was 3. The average appearance time and intensity of estrus in treated and untreated cattle are shown in Table 1. When leptin was administered, the average appearance time to estrus was faster at 31.81 days. The results of statistical testing with Mann-Whitney showed a difference (P<0.05) between the leptin-treated and the non-leptin-treated groups.

 

Table 1: Average (x ± SD) Appearance Time and Intensity of Estrus in Bali Cattle Treated with Concentrate Treatment.

Group	Appearance Time of Estrus (day)	Estrus Intensity (Score)
Without Leptin	31.81 ± 2.56a	2.69 ± 0.48a
Leptin	7.56 ± 0.81b	2.81 ± 0.40a

 

Description: The superscripts with different letters (lowercase) towards the column show a significant difference (P<0.05).

 

Bali cattle leptin hormone levels during peripartum will decline to 2.60 ng/mL and increase slowly to 5.42 ng/mL. The higher level of peripartum leptin, the faster of postpartum estrus appearance times.(Laksmi et al 2018). Administration of o-leptin 200 µg/ml in postpartum anestrus of Bali cattle increases FSH 13.47 + 1.05 mIU/ml and the average levels of FSH before treatment 11.4 + 0.28 mIU/ml. The ovarian follicle diameters were 8.45 + 0.38 mm and Ovarian follicle diameter before treatment were 5.00 + 0.37 mm. (Laksmi et al, 2016). (Leifer, 2004), that leptin acts directly on the anterior pituitary to stimulate the release of FSH and LH. (Peter et al., 2009) stated that an increase in FSH concentration causes one of the follicles to be selected to grow into the dominant follicle. And after the peak FSH concentration is reached, there will be a decrease in the FSH concentration but follicle growth continues to occur with growth from 4.0 to 8.5 mm.

Metabolic hormones such as leptin, growth hormone (GH), insulin, and insulin-like Growth Factor-1 (IGF-I) are mediators of food intake and energy balance in the fertility level of cattle, which plays a role in the development of ovarian follicles (Diskin et al., 2003). The metabolic status of animals due to negative energy balance affects the circulation of various hormones such as leptin, insulin, GH, IGF-1, cortisol, or thyroxine, which affect the proliferation of bovine follicular cells and steroidogenesis.

Leptin is one of the central mediators indicating body fat status and acute changes in feed intake in cattle. The decrease in leptin, accompanied by an acute reduction in the feed, was usually followed by a decrease in insulin and IGF-1 (Mihm and Bleach, 2003). The condition is related to leptin, insulin, and insulin-like growth factor-I (IGF-I) (Mihm and Bleach, 2003), which contribute to increased feed intake and protein binding to benefit follicular growth and maturation of the dominant follicle. Therefore, it increases estrogen, which will cause a second postpartum estrus and LH for ovulation. Leptin stimulates estrogen production activity by increasing P450arom mRNA, P450arom protein expression, and aromatase activity by directly acting on granulosa cells. Leptin then increases estrogen production, which is stimulated by FSH and/or IGF-1, which is an aromatase stimulator. (Leifer, 2004) stated that leptin has a local influence on the ovaries by stimulating estrogen through leptin receptors in granulosa cells. animals with high leptin concentrations had a better expression of estrus.

(Garcia et al., 2000) stated that the estrus cycle occurs in beef cattle that have matured when the leptin concentration reaches 15-20 ng/ml. The time of appearance of estrus in this study was 7.56±0.81 days faster than that without leptin, which was 31.81 days. Meanwhile (Suartini et al., 2013) repeated administration of buserelin (GnRH agonist) at 24 hour intervals to Bali cattle experiencing post partum anestrus resulted in an average of 5.22 ± 0.833 days of estrus. The difference in the appearance of estrus is due to differences in target organs. When administering leptin, the target organ is the hypothalamus to produce GnRH which will stimulate pituitary activity. Meanwhile, buserelin is a synthetic GnRH preparation that stimulates pituitary activity. When leptin is administered, the GnRH produced is endogenous GnRH so that the anterior pituitary is more responsive to producing gonadotropins (FSH, LH) compared to synthetic GnRH (buserelin). In addition, leptin also works directly on granulosa cells to stimulate estrogen production by increasing mRNA for P450 aromatase.

The intensity is the estrus quality displayed by cattle broodstock. Many symptoms are shown during the estrus, including restlessness, clear discharge from the vulva, and swelling of the external reproductive tract. Furthermore, the estrus intensity examination was conducted at the appearance time of the estrus. The table indicates that the estrus intensity is on a 2 (++) scale. Table 1 shows the average estrus intensity. Statistical testing results with Mann-Whitney showed no difference (P>0.05) between the groups with and without leptin.

(Furqon, 2015) stated that BCS influences estrus intensity, the better the BCS the estrus intensity will increase. A good BCS will increase the intensity of estrus and artificial insemination (AI) so that pregnancy can occur. This is because livestock that have high BCS experience complete uterine involution more quickly (return to normal estrus) so that their fertility reaches optimal. The parent’s Body Condition Score (BCS) is closely related to the status of the animal’s body energy reserves, while the energy reserves are closely related to the nutrition consumed. The nutrition and nutritional status of livestock can influence the intensity of lust because it is related to reproductive hormones (Winugroho, 2002).

According to (Handayani, 2014), the first postpartum estrus was 3.8 ± 1.55 months, which was classified as slow with postpartum anestrus. This is because the interval between parturition and the first estrus in cattle varies from 50 to 60 days or 1.5–2 months. Estrogen increases and gives rise to both postpartum estrus and luteinizing hormones for ovulation due to the dominant follicular growth and maturation of the dominant follicle. (Dewi et al., 2011) increasing estradiol levels cause an increase in the amount of blood supply to the genital tract, resulting in increased activity of cells in the vaginal area, which increases vaginal temperature. (Purwasih et al., 2014) added that increasing estrogen levels in the blood will increase the hormone adrenaline and cause it to trigger heartbeats and contractions, which can increase blood circulation. Estrogen also influences the amount of mucus that occurs during estrus, which is produced by the cervical glands. Primiparas provide a longer post-partum second estrus performance than pluriparas (Hadisutanto, 2012). Furthermore, there is a delay in the appearance time of estrus in the sixth and eighth births. This is because an increase in birth rates leads to a decline in reproductive function and a delay in the onset of postpartum estrus.

CONCLUSIONS AND RECOMMENDATIONS

Leptin administration can accelerate postpartum estrus, namely 7.56 ± 0.81 days compared to without giving it, namely 31.81 days in primiparous Bali cattle in several SIPADUs, Sobangan village, Mengwi sub-district, Badung regency. Leptin can be used as a hormone preparation to induce estrus to treat cases of postpartum anestrus in Bali cattle. Further research needs to be done to see the fertility level of cows by looking at the success of pregnancy after this hormone administration.

ACKNOWLEDGMENTS

The authors thank the Institute of Research and Community Service at Udayana University for the grant.

NOVELTY STATEMENT

Leptin, as a metabolic signal, not only maintains energy balance but also regulates the reproductive process via the hypothalamic-pituitary-ovary axis.

AUTHOR’S CONTRIBUTION

The authors contributed according to their respective duties. DNDIL and IGNBT created the research concept and design. IWS, IWNF, and INOW worked on the research and data analysis. DNDIL and IMM carried out literacy and wrote papers.

Conflict of Interest

The authors declare there is no conflict of interest.

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