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The Veterinary Medicine International Conference (VMIC)

KnE Life Sciences / The Veterinary Medicine International Conference (VMIC) VMIC—The Veterinary Medicine International Conference—is one of the world's leading conference focusing on a wide array of topics including Veterinary Medicine and Biomedical Science. It offers a stimulating venue for scientists, researchers, lecturers, general practitioners, and others to broaden their social scientific network. This conference aimed at improving human and public health by improving agricultural and food systems, advancing biomedical and comparative medical research, preventing and addressing zoonotic diseases, kit diagnostic, enhancing environmental and ecosystem health, and helping manage the 21st-century public health challenges.

PUBLISHED

2017-12-03

Conference date: 12–14 July 2017 Location: Surabaya, East Java, Indonesia Editors: Sri Agus Sudjarwo, Fedik A. Rantam, rer. nat. Gunawan Indrayanto, Muchammad Yunus, Rimayant, Wiwik Misaco, Tita Damayanti Lestari, Mustofa Helmi Effendi, Dikky Eka Mandala Putranto, and Shafia Khairani Organizer: Faculty of Veterinary Medicine, Universitas Airlangga, Indonesia Sponsors: Ministry of Research, Technology and Higher Education of the Republic of Indonesia, Universitas Airlangga, ROMINDO PRIMAVETCOM, Gadjah Mada University, Bogor Agriculture Institute, ILRI (International Livestock Research Institute), Kagoshima University, Miyazaki University, Faculty of Veterinary Science, Chulalongkorn University, College of Veterinary Medicine, Tarlac Agricultural University, Yamaguchi University, Erasmus MC, Kasetsart University, Wyndham Veterinary Clinic, GERBU – Germany, USAID Published: 29 November 2017 ISSN: 2413-0877

Toll-Like Receptors (TLRs) Play Role in Adaptive Immunity in Rabbits Immunized by Sarcoptes scabiei Proteins

THE VETERINARY MEDICINE INTERNATIONAL CONFERENCE (VMIC)

Pages 1-9 Nunuk Dyah Retno Lastuti, Fedik Abdul Rantam, Poedji Hastutiek, Dony Chrismanto

151 VIEWS | 53 DOWNLOADS

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Preservation Effect of Grouper (Epinephelus sp) Fillet Against Survival of Anisakidae


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Pages 10-20 Hartanto M. Raharjo, Setiawan Koesdarto, A.T Soelih Estoepangestie, Kusuma

Wardhani L.D


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Acanthocephalan in Xenochrophis piscator Snake in Sidoarjo Indonesia


Pages 21-27 Inggarsetya Syah Audini, Lucia Tri Suwanti, Setiawan Koesdarto, Emmanuel Djoko

Poetranto


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The Identification Blood Parasites On Pig (Susdomesticus) In Polewali Mandar District


Pages 28-33 Silvana Arfin, Lucia Muslimin, Adryani ris


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Spirometra in Ptyas mucosus Snake in Sidoarjo, Indonesia


Pages 34-40 Garindra Tiara Pranashinta, Lucia Tri Suwanti, Setiawan Koesdarto, Emmanuel

Djoko Poetranto


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Toxicity of Citrus mitis, Citrus aurantifolia, and Citrus maxima leaf extract toward mortality of Aedes aegypti larvae (Diptera: Culicidae)


Pages 41-47 Hamidah Hamidah, Hebert Adrianto


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An In Vitro Antibacterial Activity Test of Meniran Herbs’(Phyllanthus Niruri L.) Ethanol ExtractAgainst Mycoplasma gallisepticum causes Chronic Respiratory Disease (CRD) in Broiler Chickens


Pages 48-61 Emy Koestanti Sabdoningrum, Sri Hidanah, Retno Sri Wahjuni, Sri Chusniati,

Arimbi Arimbi


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Bioremediation of Mercury (II) Contaminated Seawater Using the Diatom Skeletonema costatum


Pages 62-68 Thin Soedarti, Tini S., Sucipto H., Eko P. Kuncoro


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BMP-2 Expression of Post Tooth Extraction that Catfish Oil Application


Pages 69-76 Theresia Indah B., Bambang Sumaryono, Ketut Suardita, Amelia Putri R.


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Inhibition of Apoptosis in Retinal of Newborn Mice Due to Congenital Toxoplasmosis


Pages 77-83 Lucia Tri Suwanti, Mufasirin Mufasirin, Hani Plumeriastuti


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Effect of Spirulina Platensis on The Number of Spermatogenic Cells in The Seminiferous Tubules of Rat (Rattus Norvegicus) with Excessive Physical Exercise


Pages 84-92 Rahmah Wahyu Rosidawati, Rimayanti Rimayanti, Koesnoto Supranianondo

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Phytochemicals, Antioxidant and Antifungal Properties of Acorus calamus, Curcuma mangga, and Allium sativum


Pages 93-104 Bayyinatul Muchtaromah, Mujahidin Ahmad, Emy Koestanti S, Yuni Ma’rifatul A,

Velayati Labone A


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Shark Species on Export Products from East Java and Bali by Dna Barcoding Based on Internal Transcribed Spacer-2 (Its-2) Locus in Mitochondrial


Pages 105-110 Eduardus Bimo Aksono


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Bioactivity of human Menopausal Gonadotrophin (hMG) and Deglycosylated hMG (hMGdG) from Urine of Post-Menopausal Women On invitro Bovine Embryonic cleavage


Pages 111-124 Herry Agoes Hermadiv


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The Effect of Frequency Acoustic Stimulation Sound on Intrauterine Weakening of Pregnant Sheep


Pages 125-138 Djamil Suherman, Hermanto Tri Joewono, I Komang Wiarsa Sardjana


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The Potency of ∆F 508-T Gen Mutant the Coding of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) As Prototype at the

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Congenital Bilateral Absence of Vas Deferens (CBAVD) Disease in Indonesia


Pages 139-152 Maslichah Mafruchati, Mas’ud Harijadi, Widjiati Widjiati, Boerhan Hidayat


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A Retrospective Study of Canine Pyometra in Segar Veterinary Hospital, Kuala Lumpur, Malaysia Year 2012-2016


Pages 153-165 Ng Xin Hui, Mas’ud Hariadi, Hardany Primarizky


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Oocyte Quality and Subsequent In Vitro Maturation of Sheep Oocyte-Cumulus Complex from Ovary with Presence and Absence of Corpus Luteum


Pages 166-174 Rini Widyastuti, Mas Rizky A.A. Syamsunarno, Takdir Saili, Arief Boediono


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The Effect of Insulin-Like Growth Factor-I of Pregnant Crossbred Mare Serum and Insulin-Like Growth Factor-I Recombinant Mouse on Estrous Cycles and Litter Sizes of Mice (Mus musculus)


Pages 175-188 Tjuk Imam Restiadi, Imam Mustofa, Suzanita Utama, Sri Mulyati


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Adding of L-Arginin Amino Acidin Skim Milk Diluent to Maintain Quality of Buck Sperm in Cold Temperature


Pages 189-196 Tri Wahyu Suprayogy, Suherni Susilowati, Tatik Hernawati


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Profile of Crude Protein Tyrosine Kinase on Plasma Membrane of Merino Sheep Spermatozoa Using the Method of SDS-Page (Sodium Dodecyl Sulphate-Polyacrilamide Gel Electrophoresis)


Pages 197-204 Vilda Carlenia Wardani, Sri Pantja Madyawati, Poedji Hastutiek


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Comparison of Morula and Blastula Embryo Vitrification by Using Cryoprotectant Ethylene Glycol, Propanediol, DMSO and Insulin Transferrin Selenium


Pages 205-211 Widjiati Widjiati, Epy Muhammad Luqman, Portia Sumarsono


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Effect of L-Arginine on the Thickness Iliac Arteries Wall Post Fogarty Balloon Embolectomy Catheter in Rabbit (Oryctolagus cuniculus)


Pages 212-223 Gavrila Amadea Puspitarani, Ngakan Made Rai Widjaja, Hardany Primarizky


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Ellagic Acid Activity in Healing Process of Incision Wound on Male Albino Rats (Rattus norvegicus)


Pages 224-233 Hardany Primarizky, Wiwik Misaco Yuniarti, Bambang Sektiari Lukiswanto


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Control and Preventive Study of Brucellosis by Using


Pages 234-240 Rahmahani J, Handijatno D, Tyaningsih W, Suwarno Suwarno

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Hispathology of Coronary artery of male rat (Ratus Norvegicus) with high fat diet after being given ethanol extract of Indian acalypha (Acalipha indica. L)


Pages 241-251 Kurnijasanti R, Winarti D, Wahyuni R.S., Puguh K, Setyabudi Setyabudi,

Sukmanadi Sukmanadi, Sugihartuti R, Damayanti R, Hidayati N, Rahmawati K, Sudjarwo A.S.


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Immunohistochemical Detection of Porcine Reproductive and Respiratory Syndrome Virus Antigen in Formalin-Fixed, Paraffin-Embedded Tissues with Correlation to Clinicopathologic Data


Pages 252-265 Lavina Gracia G. Manzano


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Anthelmintic Activity of Ocimum sanctum Linn. Leaves Ethanol Extract Against Fasciola gigantica in vitro


Pages 266-277 Mesia Margi Mahardika, Sri Agus Sudjarwo, Setiawan Koesdarto


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Scabiosis in Rabbit


Pages 278-285 Miyayu Soneta Sofyan, Doni Chrismanto


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Detection of Newcastle Disease Virus by Immunohistochemistry on the Brains of Laying Birds with Clinical Signs Torticollis and Curled Toe Paralysis

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Pages 286-295 Ocie Harum Wulan, Niken Yunita, Hastari Wuryastuty, Raden Wasito


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Implementation of Meniran Extract (Phyllanthus Niruri Linn) on the Performance of Broiler Chickens Infected by Mycoplasma gallisepticum Caused Chronic Respiratory Disease


Pages 296-307 Sri Hidanah, Emy Koestanti Sabdoningrum, Retno Sri Wahjuni, Arimbi Arimbi


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Blood Parasite Infection Prevalence in Kampong Chicken Breeder’s Group in Garut


Pages 308-315 Djoko Legowo, Syifa Husnul Khotimah, Lucia Tri Suwanti


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Amino Acid Analysis of Fusion (F) Gene and Prediction of Epitope B-Cell Newcastle Disease Surabaya Isolate As Vaccine Candidate


Pages 316-326 Indah Laili Rahmawati, Fedik Abdul Rantam, Wiwik Tyasningsih


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Mini Review : Liver Fibrosis Mechanism


Pages 327-337 Wiwik Misaco Yuniarti, Hardany Primarizky


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The Nutrients Contents, Dry Matter Digestibility, Organic Matter Digestibility, Total Digestible Nutrient, and NH3 Rumen Production of Three Kinds of Cattle Feeding Models


Pages 338-343 M. Anam Al-Arif, Lucia Tri Suwanti, AT Soelih Estoepangestie, Mirni Lamid


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Immunogenicity of Bone Graft Using Xenograft Freeze-Dried Cortical Bovine, Allograft Freeze-Dried Cortical New Zealand White Rabbit, Xenograft Hydroxyapatite Bovine, And Xenograft Demineralized Bone Matrix Bovine In Bone Defect Of Femoral Diaphysis White Rabbit Experimental Study In Vivo


Pages 344-355 Ferdiansyah Ferdiansyah, Dwikora Novembri Utomo, Heri Suroto


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Genotyping Analysis of Mycobacterium leprae isolated in Water Environment of Leprosy Endemic Places in Lamongan, East Java


Pages 356-368 Cita Rosita Sigit Prakoeswa, Nanny Herwanto, Ratna Wahyuni, Iswahyudi

Iswahyudi, Dinar Adriaty, Indropo Agusni, Shinzo Izumi


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Iron Overload Reduces Cholestrol and Triglyceride Serum of Mice


Pages 369-377 Devi Agustin Setiawati, Mas Rizky A.A. Syamsunarno, Pandji Irani Fianza, Nur

Atik, Neni Anggraeini, Mohammad Ghozali, Ratu Safitri, Ramdan Panigoro


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Measurement of Alkaloids Achyranthes Aspera Linn Level Using Thin Layer Chromatography Method and High-Performance Liquid Chromatography

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Pages 378-385 Dewa Ketut Meles, Wurlina Wurlina, Dewa Putu Anom Adnyana


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Propolis Potential Toward the Amount of Limphoblast and Spleen Diameter of Male Mice (Mus musculus)


Pages 386-394 Werstant Adhityananda Rinaldhi, Eka Pramyrtha Hestianah, Sri Mumpuni

Sosiawati, Lita Rakhma Yustinasari


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Identification the Gene Nucleotide Sequence of Outer Membrane Protein Aeromonas Hydrophilla Bacteria from East Java Local Isolates Using Polimerase Chain Reaction


Pages 395-403 M. Gandul Atik Yuliani, Didik Handijatno, Sri Pantja Madyawati


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Postmortem Interval Estimation Time from Algormortis Temperature of Rats Expressed by MARS Model Approach


Pages 404-412 Dwi M. Syabani, Hana Eliyani, Suharsono Suharsono, Fedik A. Rantam, Anwar

Ma’ruf


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Aquoeous Extract of Neem Leaves (Azadirachta Indica) Decrease Expression of Immunoglbulin E (IgE) and Interleukin 4 (IL-4) in Gingiva Tissue of BALB/c Mice Injected by Ovalbumine


Pages 413-421 I Dewa Ayu Ratna Dewanti, I Dewa Ayu Susilawati, Pujiana Endah Lestari,

Roedy Budirahardjo, Erawati Wulandari, Ristya Widi, Sunlip Wibisono

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Experimental Models Point Mutations In Plasmodium falciparum pfatpase6 Gene Exposed to Recuring Artemisinin In Vitro


Pages 422-435 Lilik Maslachah, Yoes Prijatna Dachlan, Chairul A. Nidom, Loeki Enggar Fitr


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Sinensetin-Rich Fraction Solid Dispersion Inhibits Cancer Cell Cycle


Pages 436-442 Lusiana Arifianti, Sukardiman Sukardiman, Mulja Hadi Santosa


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Blunted Expression of PPARa in Mice with FABP-4 and -5 Deficiency under Acute Cold Exposure


Pages 443-449 Mas Rizky A.A Syamsunarno, Mirasari Putri, Tatsuya Iso, Rini Widyastuti,

Ramdan Panigoro, Masahiko Kurabayashi


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Increased Iron in Pediatric β-Thalassaemia Major Associates with CD3+, Not γδ Lymphocytes


Pages 450-459 Mohammad Ghozali, Ulrike Panjaitan, Adi Imam Cahyadi, Reni Ghrahani, Lelani

Reniarti, MRAA. Syamsunarno, Ramdan Panigoro


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Low Serum Cholesterol in Mice Pre-treated with Imperata cylindrica L. after Acute Olive Oil Gavage


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Pages 460-467 Neni Anggraeni, Mas Rizky A.A Syamsunarno, Ghina Rahmadianti

Mukarromah, Almira Zada, Rima Destya Triatin, Yunisa Pamela, Diah Dhianawaty


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The Role Of IL-6 In TMPD-Treated Lupus Arthritis Mice


Pages 468-475 Niken Indriyanti


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Immunopathological Approach for Avian Influenza Virus Detection in Brain of Laying Bird with Clinical Signs of Torticollis and Curled Toe Paralysis


Pages 476-485 Niken Yunita, Ocie Harum Wulan, Hastari Wuryastuty, Raden Wasito


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The Efficacy of Permot (Passiflora Foetida Linn.) Leaves Crude Extract Ointment on the Healing of Skin of Rabbit with Scabies


Pages 486-496 Poedji Hastutiek, Hana Eliyani


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The Effect of Sappan Wood Extract (Caesalpinia sappan), Wheat grass and Vitamin E Treatment on the Liver Structure of Iron overload of Rat (Rattus norvegicus)


Pages 497-512 Ratu Safitri, Lelani Reniarti, Madihah Madihah, Lila Delia, Mas Rizky A.A

Syamsunarno, Ramdan Panigoro


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The Effectiveness of Antibiotics and Hematopoietic Stem Cell Treatment in Periodontitis Rat Model Toward TNF α Expression


Pages 513-519 Retno Indrawati R, Indeswati Diyatri, Dwi Rahmawati


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Construction Hybrid immunoglobulin All Four Dengue serotype Using Mesenchymal Stem


Pages 520-527 Rofiqul A’la, Rahaju Ernawati, Nunuk Dyah Retno L, Mufasirin Mufasirin, Anwar

Ma’ruf, Fedik A. Rantam


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The Potential of Black Gluten and Red Rice in Rations on the Biological Values and Ideally Body Score of Healthy “Mini Rex Rabbit”


Pages 528-535 Romziah Sidik


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Antiviral Activity Effect of Silver Nanoparticles (Agnps) Solution Against the Growth of Infectious Bursal Disease Virus on Embryonated Chicken Eggs with Elisa Test


Pages 536-548 Rosa Pangestika, Rahaju Ernawati


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Identification and Characterization Indigenous of Lactobacillus spfrom Bovine Rumen Fluidof Slaughterhouse


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Pages 549-556 Tri Nurhajati, Koesnoto Soepranianondo, Widya Paramita Lokapirnasari,

Adriana Monica Sahidu


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Protective Effect of Propolis Extract Against Lead Acetate Toxicity in Mice (Mus Musculus) Testes


Pages 557-565 Tuti Widawati, Sri Agus Sudjarwo, Herry Agoes Hermadi


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Detection of Antibiotic Residues and Concentration in Raw Milk from Lembang Small Holder Dairy Farm


Pages 566-578 Virgianty Vivi


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Potency of Bacillus cereus WPL 415 to Increase Crude Protein and Decrease Crude Fiber of Animal Feed Stuff


Pages 579-587 Widya Paramita Lokapirnasari, Adriana Monica Sahidu, Tri Nurhajati, Koesnoto

Soepranianondo, Andreas Berny Yulianto


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Acute Toxicity Tests of Alkaloid Pare (Momordica Charanthia) Fruit on The Histopathology of Liver


Pages 588-695 Wurlina Wurlina, Dewa Ketut Meles, Sunarni Zakaria, Imam Mustofa, Suherni

Susilowati, I Dewa Putu Anom Adnyana


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Teratogenic Effect of Congenital Toxoplasmosis in Chicken Embryo


Pages 696-602 Lucia Tri Suwanti, Mufasirin Mufasirin, Hani Plumeriastuti, Erma Safitri


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Case Study : Dystocia on Beef Cattle in Kunir Regency of Lumajang District, East Java, Indonesia in 2015 and 2016


Pages 603-608 Rosiana Febrianila, Widya P Lokapirnasari, Tjuk I Restiadi, Imam Mustofa,

Herry A Hermadi, Erma Safitri


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The Effectiveness of Honey in Physiological Nacl to Maintain of Viability and Motility of Spermatozoa


Pages 609-618 Elsa Agustina, Herry Agoes Hermadi, Hario Puntodewo S, Tatik Hernawati,

Indah Norma Triana, Erma Safitri


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Utilization of Sumbawa Tropical Forest Honey Apis Dorsata to Improve Fertility of Indonesia Oriental Magpie Robin (Copsychus saularis) as Effort Animal Population Increasement


Pages 619-626 Abdullah Hasib, Risaldi Muhamad, Talita Yuanda Reksa, Alvina Ulimaz Artha,

Erma Safitri


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Utilization of Honey Apis dorsata as Antiosteoporosis on Requirements of Bone Calcium Ash Density on Ovariohysterectomized White Rat (Ratus norvegicus)


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Pages 627-632 Muhammad Huda Ramadhan Ibrahim, Abdullah Hasib Hasib, Siti Nur Rohmah,

Salsabilla Abani, Samsi Yordan, Ira Sari Yudaniayanti


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Increased Integrity of Plasma Membrane and Acrosome Cap Spermatozoa Limousin Cattle at Post Thawing in Frozen Media by adding Seawater Extract


Pages 633-641 Nur Faidah, Tatik Hernawati, Mirni Lamid, Ismudiono Ismudiono, Tri Wahyu

Suprayogi, Sri Mulyati


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The Relation of Body Temperature and Vaginal Cytology Examination in Time Artificial Insemination Rate Fat-tailed Sheep (Ovis Aries) in The District Sidoarjo East Java


Pages 642-649 Rhendyka Prasetya Anggriawan, Suzanita Utama, Hana Eliyani


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Effect of Laser Acupuncture Shoot on Ova Point of Male Mojosari Duck (Anas plathyrhynchos) on The Number of Sertoli and Leydig Cells


Pages 650-657 Yuanara AR Adikara, Abdul Samik, Ira S Yudaniayanti, Tatang S Adikara, Eka P

Hestianah, Suzanita Utama


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Insulin-Like Growth Factor-I (IGF-I) from Crossbred Pregnant MareSerum to Increase Follicle Number of Mice (Mus musculus)


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Pages 658-667 Abdullah Abdullah, Tjuk IRestiadi, Nunuk DR Lastuti, Tita Damayanti, Wurlina

Wurlina, Erma Safitri


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Morphological Identification Nematodes Tanqua tiara Found on Gastric Varanus salvator at East Java


Pages 668-676 Alfiana Laili Dwi Agustin, Setiawan Koesdarto, Bambang Sektiari Lukiswanto,

Lucia Tri Suwanti, Zainal Arifin, Emmanuel Djoko Putranto


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Effect of Propolis on Spermatogenic Cells Number and Diameter of Seminiferous Tubules in Male Mice (Mus musculus)


Pages 677-683 Dona Astari Nurkarimah, Eka Pramyrtha Hestianah, Retno Sri Wahjuni, Mas’ud

Hariadi, Suryo Kuncorojakti, Herry Agoes Hermadi


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The Effect of Blue Green Algae (Spirulina platensis) Extract in White Rat (RattusNorvegicus) Treated with Excessive Physical Exercise on Leydig Cell Number and Seminiferous Tubules Diameter


Pages 684-693 Dimas Yuzrifar Rhavindra Lazuardi, Rimayanti Rimayanti, Hardany Primarizky,

Sri Agus Sudjarwo, Suzanita Utama, Kadek Rachmawati


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The Effect of Mahkota Dewa (Phaleria macrocarpa) Pulp Extract by Peroral Administration Toward The Percentage of Capacitation and Acrosome Reaction in Rat (Rattus norvegicus)


Pages 694-701 Dhanang Estu Bagyo, Budi Utomo, Rudy Sukamto Setiabudi

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Protection of Dayak Onion Tuber Extract (Eleutherine Palmifolia) Against Kidney Histopathological Appearence of Albino Male Rat Strain Wistar which was Induced by Alloxan


Pages 702-711 Dwi Gayatri Nurcahyawati, Hani Plumeriastuti, Lilik Maslachah


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Test Various Estrus Detection Device Against Pregnancy Rates on Dairy Cows in Cooperative Tunas Setia Baru Kabupaten Pasuruan


Pages 712-717 Silvia Rani Andriyanti, Mas’ud Hariadi, Roesno Darsono, Pudji Srianto


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Effect of Laserpuncture Shoot on Reproduction Point of Male Mojosari Duck (Anas Platyrhynchos) on The Numbers of Spermatogonium Cells and Seminiferous Tubules Diameter


Pages 718-726 Melyandari Ayu Qomar, Rimayanti Rimayanti, Tri Nurhajati


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Cases of Reproduction Disorder in Beef Cattle of Modo District, Lamongan in 2015


Pages 727-733 Azharuddin Anshoria, Tri Nurhajati, Budi Utomo


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The Application of Equine Chorionic Gonadotropin (Ecg) and Prostaglandin F2α to Increase the Rate of Pregnancy in Bali Cattle at Buleleng, Bali

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Pages 734-741 Yugenthri A/P Chandran, Herry Agoes Hermadi, Eka Pramyrtha Hestianah


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Morfometry Study of Hemipenis Biawak Air Varanus Salvator on Length Measurement of Snouth Vent Length (Svl) Andbody Weight


Pages 742-752 Ilham Adi Kusuma, Dicky Beo Alfiyanto, Pudji Srianto, Nusdianto Triakoso,

Djoko Legowo


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Effect of Polygonum Minus (Knotweed) Leaves Extract on the Histopathological Changes of Kidney in Mice (Mus Musculus) Induced by Mercuric Chloride


Pages 753-762 Winni Aprianti, Thomas Valentinus Widiyatno, Sri Agus Sudjarwo


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Skin Stem Cell Resource Potential for Peripheral Nerve Repair Due to trauma of post regional anasthesia


Pages 763-775 Sumartono Christrijogo, Fedik A Rantam, Eddy Rahardjo, Martia R Tacharina


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Public Awareness in ensuring Animal Originated Food Safety : A Review on “One Health” Approach in Veterinary Medicine


Pages 776-781 A. T. Soelih Estoepangestie


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VMIC 2017The Veterinary Medicine International Conference 2017Volume 2017

Conference Paper

The Effect of Mahkota Dewa (Phaleriamacrocarpa) Pulp Extract by PeroralAdministration Toward The Percentage ofCapacitation and Acrosome Reaction in Rat(Rattus norvegicus)Dhanang Estu Bagyo1, Budi Utomo2, and Rudy Sukamto Setiabudi3

1Student of Veterinary Medicine Faculty Surabaya, 60115, Indonesia2Veterinary Reproduction Department of Veterinary Medicine Faculty, Surabaya, 60115,Indonesia3Veterinary Anatomy Department of Veterinary Medicine Faculty, Surabaya, 60115, Indonesia

AbstractThis research was conducted to find outthe influence of mahkota dewa (Phaleriamacrocarpa) pulp extract against the percentage of capacitation and acrosomereaction of male rat (Rattus norvegicus). The experimental animals used are 20 malerats with the average body weight of 200g and were divided into four different groups.P0 group was the control group and given only CMCNa 0.5%. P1, P2,and P3 was givenmahkota dewa pulp extract with a dosage of 7.5 mg/200gr BW/ day, 15mg/200grBW/day,and 30 mg/200gr BW/day respectively through peroral administration for 14days. The research used Completely Randoized Design (CRD) and the data was thenanalyzed by one way ANOVA (Analysis of Variance) followed by Duncan test withthe significance of 0.5. The results show that the mahkota dewa pulp extract showssignificant impact on capacitation and acrosome reaction of rat. P2 group shows thehighest percentage capacitation significance the a mean of 30.00 ± 4.24 and 5.60 ±0.89 for acrosome reaction percentage.

Keywords: Phaleria macrocarpa pulp extrtact; capasitation; acrosome reaction;sperm potention.

1. Introduction

Indonesia’s meat consumption development per capita fluctuates and tends to risefrom 1993 to 2014 from 0.704kg/capita/year to 2.36kg/capita/year in 2014 [1]. Unfor-tunately, so far Indonesia has yet to supply all the needs of meat. To supply the risingmeat demands a large increase of cattle population is needed. Therefore technology

How to cite this article: Dhanang Estu Bagyo, Budi Utomo, and Rudy Sukamto Setiabudi, (2017), “The Effect of Mahkota Dewa (Phaleria macrocarpa)Pulp Extract by Peroral Administration Toward The Percentage of Capacitation and Acrosome Reaction in Rat (Rattus norvegicus)” in The VeterinaryMedicine International Conference 2017, KnE Life Sciences, pages 694–701. DOI 10.18502/kls.v3i6.1199

Page 694

Corresponding Author:

Dhanang Estu Bagyo

Received: 03 October 2017

Accepted: 10 October 2017

Published: 29 November 2017

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innovation development is direly in need to optimize and empower local resources. Toencourage beef cattle business the farm biotechnology development including IVF (InVitro Fertilization) needs to be acknowledged [2-3].

In Vitro Fertilization (IVF) is embryo reproduction technology in a media outside thebody [4]. In Vitro Fertilization can be defined as the process of fertilization by extractingeggs, retrieving a sperm sample, and the manually combining an egg and sperm in alaboratory dish with culturemedia [5-6]. Generally, fertilization involves two importantprocesses which are capacitation and acrosome reaction [7]. Those two processes aremutually sustainable. Without capacitation process, spermatozoa is unable to fertilized[8]. One of the main barrier in the application of in vitro fertilization technology is invitro spermatozoa capacitation [9-10]. Capacitation is meant to dismissed decapacita-tion factor (protects the stability of the spermatozoa plasma membrane) that is foundin semen plasma so that capacitation and acrosome reaction can occur [10].

Spermatozoa needs Reactive Species Oxygen (ROS) in low concentration to inducecapacitation and acrosome reaction [11]. If free radical production increases or antiox-idant production decreases, it will cause oxidative stress [12]. Oxidative stress is con-sidered one of the factors which causes infertility by giving negative effect to sperma-tozoa quality such as increasing motility loss, membrane damage, decreasing normalmorphology, viability and capacitation ability of spermatozoa.

To prevent those things above additional external antioxidant is needed. Herbal sub-stance which is potential as exogen antioxidant is mahkota dewa fruit which fluorishisin Indonesia. Mahkota dewa can be utilized as antioxidant to reduce the effect causedby free radicals [13].

Keeping those things in mind, this experiment is done to determine the effect ofmahkota dewa (Phaleria macrocarpa) pulp extract by peroral administration towardthe percentage of capacitation and acrosome reaction in rat (Rattus norvegicus).

2. Materials and methods

2.1. Treatment of Experimental Animals

Rat aged 2-3 months with an approximate weight of 200 grams is adapted for 7 daysand given treatment for 14 days. Experimental animals are grouped consists one groupof negative control and 3 treatment groups. Each group consists of five rats chosenrandomly. Control group (P0) consists of five rats given 1 ml of CMCNa 0.5%. P1 groupwas given 1 ml of mahkota dewa pulp extract 7.5 mg/200 gr BW. P2 group was given

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T 1: Experimental result of capacitation and acrosome reaction of rat (Rattus norvegicus) spermatozoa.

Treatment Capacitation Percentage (𝑋± SD, %) Acrosome Reaction Percentage (𝑋± SD, %)

P0 19.20𝑎± 4.39 2.00𝑎± 2.00

P1 24.80𝑏± 3.35 3.60𝑎𝑏± 1.67

P2 30.00𝑏± 4.24 5.60𝑏± 0.89

P3 28.00𝑏± 4.00 4.00𝑎𝑏± 1.41

1 ml of mahkota dewa pulp extract 15 mg/200 gr BW. P3 group was given 1 ml ofmahkota dewa pulp extract 30 mg/200 gr BW.

2.2. Capacitation and Acrosome Reaction Examination

Semen was fixated with formaldehyde 4% then washed by adding PBS 3ml and cen-trifuged 1500 rpm for 10 minutes. Supernatant was then thrown away and added in0.3ml FITC con A (Sigma) with concentration of 10µg/ml in PBS dulbecos. Staining isdone 25 minute at room temperature then washed twice by centrifuged 1500 rpm for10 minutes. Supernatant was thrown away and the sediment was stroked to flow labsslide (specimen), andwas added griserol 90% drops. Then the specimenwas observedwith epifluorescent microscope (Nikon Japan) with B excitation (490 rpm excitationwith 525 nm emission) to determine fluorescence of FITC spermatozoa results [14].

3. Results

Experimental result of capacitation and acrosome reaction of rat (Rattus norvegicus)which was given with mahkota dewa pulp (Phaleria macrocarpa) after stained withFITC (Fluorescent Isotiocianat) and was observed with epifluorescent microscope 400xmagnification as shown in Tabel 1 and Figure 1-2.

Different superscripts in one column shows significant difference (p

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Figure 1: Experimental result of capacitation and acrosome reaction of rat Rattus norvegicus spermatozoain bar diagram.

Figure 2: Rat (Rattus norvegicus) spermatozoa colored with FITC (Fluorescent Isotiocianat). A. Capacitation(there are acrosome intact and the top of the head of spermatozoa is fluorescent). B. Acrosome reaction(there are no acrosome intact and the aquatorial part of the spermatozoa head is fluorescent).

function [15] and ultimately reducing fertility. In normal conditions, spermatozoa pro-duces ROS in small quantities for sperm regulations this is because spermatozoa needsReactive Oxygen Species (ROS) in a small concentration to induce capacitation processand acrosome reaction [11].

Spermatozoa is easily damaged by induction of oxidative stress because its sellmembrane contains a lot of polyunsaturated fatty acids [16]. This statement isstrengthened by [17] dalam [18] Spermatozoa is really sensitive to oxidation because itconsists of polyunsaturated fatty acids (PUFA) in high concentration so ROS can cause

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the decrease of spermatozoa quality. Sperm parameter changes like motility, viability,capacitation and acrosome reaction can lower the fusion ability of spermatozoa andovum which causes decrease in fertility.

P0 group shows lowest average result of acrosome reaction, it was assumed thatthe low average result of acrosome reaction was caused by spermatozoa membranewhich mostly consists of unsaturated fatty acid that has been damaged by excessivefree radicals, considering the free radicals characteristic which easily oxidize unsat-urated fatty acid while there is no antioxidant protecting it. The plasma membraneof spermatozoa consist a massive amount of phospholipid and unsaturated fatty acidwhere unsaturated fatty acid is very vulnerable to free radicals strike mainly hydroxylradical, so that ROS easily penetrated into plasma membrane. Hydroxyl radical wouldinitiate chained reaction called lipid peroxidation [19]. The result of this chain was thebroken chain of fatty acid became toxic substance to the spermatozoa cell [19-20]. Fur-thermore, there will be a damage of cell membrane which caused membrane fluidityandmembrane integrity increased so it will interrupted the process of capacitation andacrosome reaction.

P1 and P2 group shows the increasing of capacitation and acrosome reaction per-centage. It is suitablewith the function of flavonoid substance as antioxidant. Flavonoidsubstance has proven to have many advantages which are as antioxidant, anticarcino-genic, antimicrobial, antimutagenic, anti inflammation and reduce the risk of cardio-vascular disease [21-22]. As one of antioxidant, flavonoid group acted through free rad-icals scavenger directly [23-26]. At first, flavonoid was oxidized by radical substance,then it changed to become stable because it reacted with other radical flavonoid sothat formed fenoxyl radical which less reactive then free radical reactivity could bemuted. Meanwhile, [27] said that acrosome reaction only occurred in spermatozoawhich has undamaged membrane. This was because antioxidant in the mahkota dewapulp extract (Flavonoid) could protect the plasma membrane of spermatozoa from thedamaged that caused by free radical oxidation (ROS) so that the membrane integritystay preserved acrosome reaction can occur. According to [28] that the existence ofantioxidant in flavonoid substance of mahkota dewa (Phaleria macrocarpa) pulp extractcould give protection to spermatozoa’s plasma membrane of the rat. The statementwas reinforced with the research by [29] who stated that flavonoid can protect plasmamembrane of spermatozoa from excessive ROS.

P3 group shows the decreasing of capacitation and acrosome reaction average per-centage. This was assumed that excessive administration of mahkota dewa (Phaleria

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macrocarpa) pulp extract caused toxicity to spermatozoa so that caused the morphol-ogy damage of the spermatozoa mainly the cell membrane. Cell membrane damagewould cause increasing of membrane fluidity, membrane integrity disturbance andinactivation of the membrane with enzyme and receptor. It would cause the rise ofcell damage including spermatozoa cell [14]. If the ATP production in mitochondriawas decreased and intracellular ATP was reduced rapidly, it would cause the damageof axonem, decreasing of spermatozoa viability, increasing of midpiece’s morphologydamage and the loss ability of capacitation and acrosome reaction [30].

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[2] Cunningham, E.P. 1999. Recent developments in biotechnology as they related toanimal genetic resources for food and aglriculture. Commision on Genetic Resourcesfor Food and Agriculture

[3] Diwyanto, Kusuma. 2008. Pemanfaatan Sumber Daya Lokal dan Inovasi Teknologidalam Mendukung Pengembangan Sapi Potong Di Indonesia. PengembanganInovasi Pertanian 1(3): 173-188

[4] Jaswandi, Boediono A., dan M.A. Setiadi. 2001. In Vitro maturation and fertilizationof ovine oocytes in system with absence of 5% CO2: 19-22.

[5] Sukra, Y., I. Djuwita, A. Boediono dan R. Golfiani. 1992. Studi tentang perkembanganteknik fertilisasi in vitro, kultur, pewarnaan kromosom dan penyayatan embriodalam pross perekayasaan embrio. Lapran Penelitian IPB-Bogor

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[7] Asmarinah. 2010. Peran Molekul Kanal Ion Pada Fungsi Spermatozoa. Maj KedoktIndon volum:60. Hal. 377.

[8] Bearden, H.J. and J.W. Fuquay, 2000. Applied Animal Reproduction. 5th Ed. PrenticeHall. New Jersey Bogor Agriculture University

[9] Kanagawa, H., O.A. Mazni and C.A. Valdez. 1989. Oocyte Maturation and in vitroFertilization in Far Animals. Biotechnology for Livestock Production. FAO. Romapp. 79-95.Sujoko, Heri., Setiadi, Muhammad Agus., Boediono, Arief. 2009. SeleksiSpermatozoa dengan Model Sentrifugasi Gradien Densitas Percoll. Jurnal VeterinerSeptember. Vol. 10 No. 3: 125-132.

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[10] Wattimena, J. 2006. Pengaruh Inkubasi Terhadap Pola Kapasitasi dan ReaksiAkrosom Spermatozoa Domba In Vitro. JITV 11(4); 295-301

[11] Sikka, S.C. 2004. Role of oxydative stress and antioxydants in andrology andassisted reproductive technology. J. Androl. 25:5-18.

[12] Widayati, Eni. 2011. Oxidasi Biologi, Radikal Bebas, dan Antioxidant. UniversitasIslam Sultan Agung. Semarang

[13] Guselli, A, Nardini, M., Baldi, A, and Scaccini, S, 1998. Antioxydant activity ofdifferent phenolic fraction separated from an Italian Red Wine. J. Aric Food Chem.48 (2)

[14] Utomo, Budi. 2011. Suplementasi Akrosin Pada Semen Kambing Peranakan EtawaPasca Thawing Terhadap Peningkatan Kualitas dan Potensi Spermatozoa

[15] Sikka, C.S. 1996. Oxydative stress and role of antioxydant in normal and abnormalsperm function. Department of urology, Tulane University School of Medicine, NewOrleans, Louisiana. USA

[16] Dahlan, M.S. dan Arjatmo Tjokronegoro (2002). Oxidative Stress andMale Infertility:Phatophisiology Clinical Implication. Jurnal Kedokteran yarsi, 10(1): 50-59.

[17] Soehadi, Koentjoro. 1996. Species Oxygen Reactive dan Kualitas Spermatozoa.Medika, 10:786-789

[18] Panghiyangani, Roselina dan Mashuri. 2009. Kualitas Spermatozoa dan AktivitasEnzim Katalase dala Darah Tikus Jantan Galuur Sprague Dawley (SD

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