Sains Malaysiana 51(6)(2022): 1625-1634

http://doi.org/10.17576/jsm-2022-5106-02

Faecal Concentrations of Progesterone and 17β-Oestradiol of Female Malayan Tapir from Different Faecal Hormone Extraction Methods

(Kepekatan Najis yang Mengandungi Progesteron dan 17β-Oestradiol Tapir Betina Malaya daripada Kaedah Pengekstrakan Hormon Najis yang Berbeza)

MUHAMMAD NAJIB RUSLAN¹, MASHITAH SHIKHMAIDIN^1,2,3,*, SALFARINA RAMLI⁴, NALISHA ITHNIN⁵, INTAN NASUHA AZHAR¹, NUR HAFIZAH MOHAMMED¹ & SYAIZWAN ZAHMIR ZULKIFLI¹

¹Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang,

Selangor Darul Ehsan, Malaysia

²Institute of Tropical Agriculture and Food Security (ITAFos), Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

³Division of Life Sciences (Molecular Biology Major), Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Jeonbuk National University, 54896 Jeonju, Korea

⁴Department of Pharmaceutical Pharmacology and Chemistry, Faculty of Pharmacy, Integrative Pharmacogenomics Institute (iPROMISE), Universiti Teknologi MARA, Cawangan Selangor, 42300 Puncak Alam, Selangor Darul Ehsan, Malaysia

⁵Sime Darby Plantation Technology Centre Sdn Bhd, 1^st Floor Block B, UPM-MTDC TIC III, Jalan Lebuh Silikon, 43000 UPM Serdang, Selangor Darul Ehsan, Malaysia

Diserahkan: 10 Jun 2020/Diterima: 22 November 2021

ABSTRACT

Non-invasive hormone monitoring of endangered wildlife provides an essential tool to optimize breeding strategies, mainly in captive management. However, the protocol of reproductive hormone monitoring on pregnancy status using non-invasive faecal samples in Malayan tapir is still inconclusive. Therefore, we compared the metabolites reproductive hormones; 17β-oestradiol and progesterone in the faecal samples extracted using methods of Schwarzenberger et al. (1996) (Methods A and B), Brown et al. (2001) (Method C) and Shutt et al. (2012) (Method D) from pregnant and non-pregnant captive Malayan Tapir. Faecal samples from four female of Malayan Tapir (Tapirus indicus) were collected for five months at Sungai Dusun Wildlife Reserve (n = 2), Zoo Taiping (n = 1) and Zoo Negara (n = 1), Malaysia. Analysis by liquid chromatography-mass spectrometry (LCMS) confirmed that progesterone was detected in all fecal sample extracted by extraction Method A, while 17β-oestradiol was undetectable in all methods. The measurement of reproductive hormones for pregnancy status via Enzyme-Linked Immunosorbent Assay (ELISA) analysis showed no significant difference (p > 0.05) for progesterone between the fecal extraction methods. Yet the data obtained were not able to validate the pregnancy status, due to similarity in concentration for both pregnant and non-pregnant tapirs. While for 17β-oestradiol, significant results were observed in all extraction methods and in pregnancy status (p < 0.05). Method C was found to be the most reliable extraction method (p < 0.05) to indicate pregnancy status. From the results, it showed that solvent used, boiling process and multiphase extraction plays important role in fecal extraction process in Malayan Tapir.

Keywords: Malayan tapir; non-invasive fecal; oestradiol; pregnancy; progesterone

ABSTRAK

Pemantauan hormon tak invasif terhadap hidupan liar yang terancam penting untuk mengoptimumkan strategi pembiakan terutamanya dalam pengurusan haiwan tawanan. Walau bagaimanapun, protokol pemantauan hormon pembiakan terhadap status pembiakan dengan menggunakan tak invasif daripada sampel najis tapir Malaya masih tidak muktamad. Oleh itu, dalam penyelidikan ini kami mengkaji perbezaan pengukuran hormon pembiakan metabolit; 17β-oestradiol dan progesteron dalam kaedah pengekstrakan najis Schwarzenberger et al. (1996) (Kaedah A dan Kaedah B), Brown et al. (2001) (Kaedah C) dan Shutt et al. (2012) (Kaedah D). Sampel najis daripada empat betina tapir Malaya (Tapirus indicus) dikutip selama lima bulan di Hutan Simpan Sungai Dusun (n = 2), Zoo Taiping (n = 1) dan Zoo Negara (n = 1), Malaysia. Analisis dengan spektrometri kromatografi gas cecair (LCMS) mengesahkan bahawa semua sampel najis menunjukkan progesteron hanya dari Kaedah Pengekstrakan A, sementara 17β-oestradiol tidak dapat dikesan daripada mana-mana Kaedah Pengekstrakan najis. Pengukuran hormon pembiakan untuk status kehamilan melalui analisis ujian imunosorben berkaitan enzim (ELISA) tidak menunjukkan sebarang perbezaan yang signifikan bagi progesteron antara Kaedah Pengekstrakan tinja. Namun data yang ditunjukkan dalam kajian ini tidak dapat mengesahkan status kehamilan kerana nilai progesteron untuk hamil dan tidak hamil adalah hampir sama. Sementara untuk 17β-oestradiol, ia menunjukkan hasil yang signifikan dalam semua kaedah pengekstrakan dan status kebuntingan (p < 0.05) dengan kepekatan tertinggi 17β-oestradiol dengan menggunakan Kaedah C, ini menunjukkan bahawa pelarut memainkan peranan penting dalam proses pengekstrakan najis dan nampaknya kaedah ini boleh digunakan untuk mendiagnos kehamilan tapir Malaya (p < 0.05). Kami mencadangkan untuk progesteron, pengesahan immunoasei diperlukan mungkin disebabkan oleh merendahkan kepekatan progesteron. Keputusan kajian menunjukkan jenis pelarut yang digunakan, proses pendidihan dan pengekstrakan multifasa memainkan peranan penting dalam proses pengekstrakan najis tapir Malaya.

Kata kunci: Kehamilan; najis tak invasif; oestradiol; progesteron; tapir Malaya

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^*Pengarang untuk surat-menyurat; email: mashitah@upm.edu.my