Kesimpulan
Dari hasil penelitian yang telah dilakukan diperoleh beberapa kesimpulan sebagai berikut:
1. Organ reproduksi Echymipera kalubu memiliki ciri khas pada glans penis yang berbentukbhipid(bercabang) dan tidak memiliki kelenjar vesikularis. 2. Spermatozoa yang berasal dari kauda epididimis memiliki nilai motilitas dan
konsentrasi yang meningkat sejalan dengan peningkatan kedewasaan. Tingkat abnormalitas kepala terbanyak pada kelompok prapubertas dan ekor pada kelompok dewasa muda. Ukuran panjang keseluruhan spermatozoa 162.51±5.12 μm dengan panjang kepala 2.91±0.40 μm, panjang midpiece 13.99±0.87μm danprincipal pieceadalah 145.59±5.38μm.
3. Tahapan spermatogenesis pada E. kalubu adalah sembilan tahap dengan sepuluh langkah transformasi (perkembangan) spermatid.
Saran
Perlu adanya kajian lebih lanjut mengenai pola reproduksi dari E. kalubu jantan yang hidup bersama dengan betina.
Spermatogenesis and Spermatogenic Efficiency in the Wild Boar (Sus scrofa scrofa). J Biol Reprod.75:792–799.
Allen J., Cantab MB. 1893. The Function of The Urethral Bulb. J Anat Physiol 27: 235–236.
Anonim. 2010. Human Embryology: Gametogenesis. http://www.embryology. ch/anglais/cgametogen/spermato03.html [10 Desember 2010]
Arifiantini RI, Wresdiyati T, Retnani EF. 2006. Kaji Banding Morfometri Spermatozoa Sapi Bali (Bos sondaicus) Menggunakan Pewarnaan Williams, Eosin, Eosin Nigrosin dan Formol Saline. J Sains FKH UGM 24: 65-70.
Ax RL, Didion BA, Lenz RW, Love CC, Varner DD, Hafez B, Bellin ME. 2000. Semen Evaluation. In: Hafez B, Hafez ESE, editor. Reproduction in Farm Animals. Seventh Ed. South Carolina: Baltimore, Lippincott Williams & Wilkins. Pp 365-375.
Ballantyne K, Matson P, Noakes N, Nicolson V, Johnston SD. 2009. Reproductive Physiology of the Female Greater Bilby (Macrotis lagotis Thylacomidae: Evidence for a Male-Induced Luteal Phase). J Repro, Fertil and Develop.21:274-282.
Barham B, Pennington JA. 2009. Breeding Soundness Evaluation for Beef and Dairy Bulls. http://www.uaex.edu/other_areas/publications/pdf/fsa-3046.pdf [14 Oktober 2010].
Biscoe RT, Renfree M. 1987. Reproductive Physiology of Marsupials. Cambridge University Press. New York. USA. Pp 124-171.
Bissett C, Bernard RTF. 2010. The Effect of Prolonged Cold Storage of Eland (Taurotragus oryx) Cauda Epididymides on the Spermatozoa: Possible Implications for the Conservation of Biodiversity. http://eprints.ru.ac.az/760/1/The-effect-of prolonged. pdf. [27 Juni 2010].
Bonet S, Briz M, Fradera A. 1993. Ultrastructural Abnormalities of Boar Spermatozoa.J Theriogenelogy63:383-396.
Cocchia Net al.2009. Cryopreservation of Feline Epididymal Spermatozoa from Dead and Alive Animals and Its Use in Assisted Reproduction. Cambridge. J Online. 18:1-8.
Comizzoli P, Mermillod P, Mauget R. 2000. Reproductive Biotechnologies for Endangered Mammalian Species.J Repr Nutr Dev40: 493-504.
Cummins JM, Temple-Smith PD, Renfree MB. 1986. Reproduction in the Male Honey Possum (Tarsipes rostratus: Marsupialia): The Epididymis. Am J Anat177:385–401
46
Dawson TJet al. 1999. Morphology and Physiology of Metatheria. In: Fauna of Australia. http://www.environment.gov.au/biodiversity/abrs/publication /fauna-of-australia/pubs/volume1b/17-ind.pdf. [14 Oktober 2010] Durrant BS. 1990. Semen Collection, Evaluation and Cryopreservation on in Exotic
Animals Species: Maximing Reproductive Potential. J. ILAR 32(1). http://dels-old.nas.edu/ilar_n/ilarjournal/32_1/V32_1Semen.shtml [10 Juni 2011].
Dreef HC, Van Esch E, De Rijk EPCT. 2007. Spermatogenesis in the Cynomolgus Monkey (Macaca fascicularis): A Practical Guide for Routine Morphological Staging. J Tox Pathol.35:395-404.
Flannery T. 1995. Mammals of New Guinea. Robert Brown and Associates. The Australian Museum. Sidney. Pp 104-121.
Fowler ME. 2008. Restraint and Handling of Wild and Domestic Animals. First Ed. Blackwell Publishing. State avenue IOWA. USA. Pp 248-261. Gallardo MH, Mondaca FC, Ojeda RA, Kohler N, Garrido N. 2004.
Morphological Diversity In The Sperms Of Caviomorph Rodents. J Neotrop. Mammal.9:159-170.
Ganan N, Gomendio M, Roldan ERS. 2009. Effect of Storage of Domestic Cat (Felis catus) Epididymides at 50C on Sperm Quality and Cryopreservation. J Theriogenology. 72:1268–1277.
Garner DL, Hafez ESE. 2000. Spermatozoa and Seminal Plasma. In: Hafez B, Hafez ESE, editor. Reproduction in Farm Animals. Seventh Ed. South Carolina: Baltimore, Lippincott Williams & Wilkins. Pp 96-109. Gordon G, Hulbert AJ. 1989. Peramelidae. In: Fauna of Australia.
http://www.environment.gov.au/biodiversity/abrs/publications/fauna-of-australia/pubs/volume1b/24-ind.pdf [5 Mei 2010].
Gordon G, Hall LS, Antherton RG. 1990. Status Bandicoots in Queensland. In: Seebeck JH, Brown PR, Wallis RL, Kemper CM, editor. Bandicoots and Bilbies. Surrey Beatty and Sons. Australia. Pp 37-42.
Hafez ESE. 2000. Anatomy of Male Reproduction. In: Hafez B, Hafez ESE, editor. Reproduction in Farm Animals. Seventh Ed. South Carolina: Baltimore, Lippincott Williams & Wilkins. Pp 3-11.
Hafez B, Hafez ESE. 2000. Micromanipulation of Gametes and Embryos: In Vitro Fertilization and Embryo Transfer (IVF/ET). In: Hafez B, Hafez ESE, editor. Reproduction in Farm Animals. Seventh Ed. South Carolina: Baltimore, Lippincott Williams & Wilkins. Pp 443-463. Harding HR, Shorey CD, Cleland KW. 1990. Ultrastructure of Spermatozoa and
Epididymal Sperm Maturation in Some Perameloids. In: Seebeck JH, Brown PR, Wallis RL, Kemper CM, editor. Bandicoots and Bilbies. Surrey Beatty and Sons. Australia. Pp 107-116.
Haron AW, Yong M, Zainudin ZZ. 1999. Evaluation of Semen Collected by Electro Ejaculation From Captive Lesser Mouse Deer Malay Chevrotain (Tragulus javanicus). J Zoo Wild Med.31:164-167.
Hess RA. 1999. Spermmatogenesis Overview. J Ency of Reprod.4:539-545. Holt WV dan Pickard AR. 1999. Role of Reproductive Technologies and
Genetic Resource Banks in Animal Concervation.J Rev Reprod. 4: 143-150.
Johnson LA, Weitze KF, Fiser P, Maxwell WMC. 2000. Storage of Boar Semen. J Anim Sci62:143-172.
Johnston SD, McGowan MR, Philips NJ, O’Callaghan P. 2000. Optimal Physicochemical Conditions for the Manipulation and Short-Term Preservation of Koala (Phascolarctos cinereus). J Repro and Fert. 118:273-281.
Kiernan JA. 1990. Histological & Histochemical Methods. Theory & Practice. Second Ed. England: Pergamon Pr. Canada. Pp 96-186.
Larcombe AN. 2003. Activity Rithms of Sourthen Brown Bandicoots Isoodon obesulus (Marsupialia: Peramelidae) in Captivity. J Aust Mammal. 25:81-86.
Leary et al. 2008. Echymipera kalubu. http://www.iucnredlist.org/apps/redlist/ details/7018/0 [14 Oktober 2010]
Lindenmayer. 1997. Australian Metatherien: Dasyuromorphis, Notoryctemorphia, Peramelemorpha, Diprotodontia. http://voyager.uvm.edu [12 Maret 2010].
Long K. 2009. Burrowing Bandicoots an Adaptation to Life in Fire Prone Environment?. J Aust. Mammal.31:57-59.
Manufandu JS. 2000. Pola Tingkah Laku Harian Bandikut Hidung Panjang (Echymipera rufescens) di Penangkaran. Faperta Uncen. Manokwari. Menzies J, 1991. A Handbook of New Guinea Marsupials and Monotremes.
Kristen Inc. Papua New Guinea. Pp 51-60.
Neves ES, Chiarini-Garcia H, Franca LR. 2002. Comparative Testis Morphometry and Seminiferous Ephitelium Cycle Length in Donkeys and Mules. J Biol Reprod.64:247-255.
Onyango DW, Wango EO, Otiang’a-Owiti GE, Oduor-Okelo D, Werner G. 2000. Morphological Characterization of The Seminiferous Cycle in The Goat (Capra Hircus): A Histological and Ultrastructural Study. J. Ann Anat. 182:235-241
Parapanov RN et al. 2009. Testis Size, Sperm Characteristics and Testosterone Concentrations in Four Species of Shrews (Mammalia, Soricidae). J Ani Repro Sci 114:269–278.
Paris DBBP, Taggart DA, Shaw G, Temple-Smith PD, Renfree MB. 2005. Changes in Semen Quality and Morphology of the Reproductive Tract of the Male Tammar Wallaby Paralalel Seasonal Breeding Activity in the Female.J Soc Reprod and Fert. 130:367-378.
Petocz RG. 1994. Mamalia Darat Irian Jaya. Penerbit Gramedia Pustaka Utama. Jakarta. Hlm 69-77.
48
Renfree MB. 1993. Ontogeny, Genetic Control, and Phylogeny of Female Reproduction in Monotreme and Therian Mammals. http://www.eebweb.arizona.edu/Courses/Ecol485_585/Readings/Renfre e_1993.pdf [20 Mei 2010].
Russel EM dan Renfree MB. 1989. Tarsipedidae. In: Fauna of Australia.Vol.Ib http://www.environment.gov.au/biodiversity/abrs/publications/fauna-of-australia/pubs/volume1b/33-ind.pdf. [7 Juli 2010].
Russell LD, Ettlin RA, Hikim SAP, Clegg ED. 1990. Histological and Histopathological Evaluation of The Testis. Cache River Press, Clearwater Florida.
Salisbury GW, VanDemark NL. 1985. Fisiologi Reproduksi dan Inseminasi Buatan pada Sapi. R Djanuar, Penerjemah. Yogyakarta: Gajah Mada University Pr. Terjemahan dari: Physiologi of Reproduction and Artificial Insemination of Cattle.
Santos MRF, Esteso MC, Montoro V, Soler AJ, Garde JJ. 2006. Cryopreservation of Iberian Red Deer (Cervus elaphus hispanicus) Epididymal Spermatozoa: Effects of Egg Yolk, Glycerol and Cooling Rate. J Theriogenelogy 66:1931–1942.
Schaeffer DO. 1997. Anesthesia and Analgesia in Nontraditional Laboratory Animal Species. In: Kohn DF, Wixson SK, White WJ, Benson GJ, editor. Anesthesia and Analgesia in Laboratory Animals. First Ed. Academic Press. New York USA. Pp 339-340.
Seebeck JH. 2001. Mammalian Species: Perameles gunnii. J Am Soc Mammal 654: 1-8.
Segatelli TM, Almeida CCD, Pinheiro PFF, Martinez M, Padovani CR, Martinez FE. 2002. Kinetic of Spermatogenesis in the Mongolian Gerbil (Meriones unguiculatus).J Tiss Cell34:7-13.
Senger PL. 2005. Pathways to Pregnancy and Parturition. Second Rev. Ed. Washington: Current Conception.
Setiadi MA, Suprayogi A, Yulnawati. 2006. Viabilitas dan Integritas Membran Plasma Spermatozoa Epididymis Anjing Selama Penyimpanan Pada Pengencer yang Berbeda. J Med Kedok Hewan. 22:118-123.
Suarni NMRS, Ermayanti IGAM. 2009. Viabilitas Spermatozoa Petaurus breviceps papuanusT. J Biologi. 8: 57 – 59.
Sumeidiana I, Wuwuh S, Mawarti E. 2007. Volume Semen dan Kosentrasi Sperma Sapi Simmental, Limousin dan Brahman di Balai Inseminasi Buatan Ungaran. J Indon. Trop. Anim. Agric. 32:131-137.
Taggart DA, Temple-Smith PD. 1990. Effects of Breeding Season and Mating on Total Number and Distribution of Spermatozoa in the Epididymis of the Brown Marsupial Mouse,Antechinus stuartii. J Reprod Fert. 88:81-91.
Tedman RA. 1990. Some Observations on the Visceral Anatomy of the BandicootIsoodon macrourus (Marsupialia: Peramelidae). In: Seebeck JH, Brown PR, Wallis RL, Kemper CM, editor. Bandicoots and Bilbies. Surrey Beatty and Sons. Australia. Pp 107-116.
Thodunter R, Gemmel RT. 1987. Seasonal Changes in the Reproductive Tract of the Male Marsupials Bandicoot, Issodon macrourus. Great Britain. J Anat.154:173-186.
Toelihere MR. 1993.Inseminasi Buatan pada Ternak. Bandung : Angkasa. Tourmente M, Gomendio M, Roldan ERS. 2011. Mass-Specific Metabolic Rate
and Sperm Competition Determine Sperm Size in Marsupial Mammals. J Plos One.6:1-7.
Tohda A et al. 2001. Testosterone Suppresses Spermatogenesis in Juvenile Spermatogonial Depletion (jsd ) Mice. J Biol of Reprod. 65:532-537. Toshimori K. 2009. Dinamycs of the Mammalian Sperm Head. Springer-Verlag
Berlin Heidelberg. Pp 6.
Vernes K, Pope LC. 2009. Reproduction in the Northern Brown Bandicoot (Isoodon macrourus) in the Australian Wet Tropics. J Zool. 57:105-109.
Warsono IU. 2009. Sifat Biologis dan Karakteristik Karkas dan Daging Bandikut (Echymipera kalubu) [disertasi]. Bogor : Sekolah Pascasarjana, Institut Pertanian Bogor.
Wooley PA. 2001. Observations on the Reproductive Biology of Myoictis Wallacei, Neophasccogale Lorentzi, Dasyurus Albopunctatus and Dasyurus Spartacus, Dasyurid Marsupials Endemic to New Guinea. J Aust Mammal23:63-66
Yohanita AM. 2009. Keragaman Jenis Mamalia di Hutan Lahan Basah Dataran Rendah Kaliki Kabupaten Merauke. Dalam: Krey K, Kemp N editor. Survei Keanekaragaman Hayati Areal Konsesi HTI Medco Group di Kaliki-Merauke, Papua Indonesia [Laporan Akhir]. Manokwari: Universitas Negeri Papua. Hal. 80-85.
LAMPIRAN
Lampiran 1. Prosedur pembuatan preparat histologi
Fiksasi jaringan dengan paraformaldehid 4% selama 3x24 jam
Dehidrasi dengan alkohol bertingkat (70, 80, 90, 95 dan 100%)
Clearing dengan Xylol
Infiltrasi
Embbeding di paraffin
Sectioning dengan menggunakan mikrotom setebal 3 µm
Staining dengan pewarnaan PAS
Lampiran 2 Prosedur PewarnaanPeriodic Acid Schiff(PAS) PERIODIC ACID SCHIFF (PAS)
1. Deparafinisasi, air mengalir, DW sesuai dengan prosedur 2. Oksidasi dalam larutan 0.5-1% periodic acid RT, 5 menit
3. DW 1 kali RT, 5 menit
4. *DDW (akuabides) RT, 2 kali @ 5 menit
5. Schiff reagent RT, 15 menit
6. Air sulfit (selalu dibuat baru) RT, 3 kali @ 5 menit
7. DW RT, 3 kali @ 5 menit
8. Counterstain (missal: Mayer Hematoksilin) cek mikroskop
9. Air mengalir 10-60 menit
10. DW RT, 2 kali @ 5 menit
11. Dehidrasi, clearing, mounting
* atau 3 kali DW@ minimal 5 menit. Langkah ini penting untuk menjaga kebersihan dan keawetan Schiff Reagent.
0.5 Periodic acid: disimpan pada 40C, tahan beberapa bulan (buat catatan pemakaiannnya agar diketahui baru atau tidaknya).
Periodic acid atau ortho-periodic acid (HIO4) 0.5 gr
DW 100 ml
Schiff reagent
Rumus: 100 ml air (DW atau DDW), basic fuchsin 0.5 gr, HCl 1 N 11-20 ml, NaHSO30.4 gr dan karbon aktif 0.42 gr.
Cara membuat Schiff reagent:
1. Panaskan air 100 ml sampai air mendidih.
2. Masukkan basic fuchsin 0.5 gr sambil diaduk selama 1 menit (aduk rata, warna merah gelap).
3. Saring kemudian diamkan hingga suhu 500C.
4. Penting!!! Pada suhu 500C, tambahkan HCl 1N aduk, lalu NaHSO4 aduk hingga rata (stirrer ± 2 menit).
5. Biarkan dingin dengan merendam botol larutan dalam wadah berisi air dan hindarkan dari cahaya (tutup dengan aluminium foil).
6. Setelah dingin (larutan mulai nampak merah pink terang) masukkan ke botol gelap dan simpan dalam refrigerator 18-36 jam.
7. Setelah dari kulkas, hangatkan lalu tambahkan karbon aktif kemudian distrirer ± 1 menit.
8. Saring lagi sehingga reagent akan menjadi bersih dan tidak berwarna (colourless), jika belum colourless ulangi tahapan penambahan karbon. 9. Simpan Schiff reagent dalam kulkas dan bisa digunakan 48 jam kemudian. 10. Periksa larutan: 1. Jika larutan menjadi colourless, berarti siap pakai. 2. Uji
reagent, bila diteteskan dengan formaldehid berubah menjadi pink kemerahan dan ditetes dengan PBS warna tetap sama maka reagent bagus dan bisa dipakai.
Air sulfit: (selalu dibuat baru)
10% Larutan Na atau K Hidrogen Sulfit (HSO3) atau metabisulfit 6 ml
1N HCl (asam khlorida) 5 ml
53