GENETIKA FORENSIK DAN DILENIASI POPULASI HIU DAN PARI DI INDONESIA: IMPLIKASI PADA PENGELOLAAN DAN KONSERVASI

Hawis Madduppa, Sutanto Hadi, Nurlita Putri Anggraini, Djumadi Parluhutan, Budi Raharjo, Nurmila Anwar, Fitrian Dwi Cahyo, Dani Dasa Permana, Yunita, Effin Muttaqin, Benaya M. Simeon, Muhammad Ichsan, Abraham Sinaipar, Toufan Phardana, Beginer Subhan

Pendahuluan

Permen KP No. 5/PERMEN-KP/2018tentang Larangan Pengeluaran Hiu Koboidan Hiu Martil dari Indonesia.

Sphyrna lewiniCarcharhinus falciformis

Alopias pelagicus

© The IUCN Red List of Threatened Species: Manta alfredi – published in 2011http://dx.doi.org/10.2305/IUCN.UK.2011-2.RLTS.T195459A8969079.en

KEPUTUSAN MENTERI KELAUTAN DAN PERIKANAN REPUBLIK INDONESIA NOMOR 4/KEPMEN-KP/2014 TENTANG PENETAPAN STATUS PERLINDUNGAN PENUH IKAN PARI MANTA

Manta alfredi (Reef Manta Ray)Status: Vulnerable/RawanPop. trend: decreasing CITES : Appendik II

Produksi perikanan hiu di dunia tahun 2002-2011 (Mundy et al. 2013)

Indonesia memproduksi 10.762 ton atau sekitar 13 % dari penangkapan hiu di dunia.

• all parts of the rays have high economic value

• The stingrays are exploited intensively but unregulated in Southeast Asia

(Blaber et al., 2009; White and Kyne, 2010)

Hiu berbeda dengan komoditas perikanan laut lainnya

Sumber : www.PewEnvironment.org

DNA Barcoding

Mitocondrial DNA (COI):

Conserve region

Maternal

Dapat identifikasi sampai species level

Dapat membedakan pada species level

Dapat mengkode Protein

Membedakan spesies yang punya vasiasi tinggi (Hanson et al. 1992)

Mitochondrial DNA

DNA Sampling

Muara Baru

Tanjung Luar

Aceh

Makassar

Cilacap

Raja Ampat

KomodoNusa Penida

DNA Sampling

Prosedur penanganan sampel genetik hiu

DNA Sampling

Prosedur penanganan sampel genetik pari

Foto ID

Pengambilan sampel jaringan (biopsi)

Underwater tissue collection(Robbins 2006)

Tentukan Kelamin

DNA Analisis

7654321

DNA Extraction

PCR gen COI

Electrophoresis

CycleSequencing

Sequence edit

Blast

DataAnalysis

DNA Forensik

0 1 2 3 4 5 6 7

Alopias pelagicus

Carcharhinus dussumieri

Alopias superciliosus

Carcharhinus falciformis

Carcharhinus amblyrhynchos

Carcharhinus amblythynchoides

Carcharhinus coatesi

Carcharhinus melanopterus

Carcharhinus obscurus

Carcharhinus sorrah

Glyphis fowlerae

Hemigaleus australiensis

Prionace glauca

Sphyrna lewini

Lamiopsis tephrodes

Triaaenodon obesus

Muara Baru

Carcharhinus brevipinnaCarcharhinus macloti

Carcharhinus sorrah Galeocerdo cuvier

Hemipristis elongata

Rhynchobatus australiae

Carcharhinus dussumieri

Carcharhinus amblyrhynchoides

Hemigaleus australiensis

Carcharhinus melanopterus

Prionace glauca

Glyphis fowlerae Carcharhinus amblyrhynchos

Carcharhinus falciformis

Alopias pelagicus

Alopias superciliosus

DNA Forensik

2%

4%2%

6%

24%

17%3%

12%

11%

9%

1% 9%

Daging Glondongan tanpasirip dan kepala

Kulit frozeen

Kulit kering

Sirip Beku

Sirip Kering

Tulang

Tulang Punggung Sayap

Tulang Sirip

Tulang Sirip Sedang

Muara Baru

Aceh

45 ekor (98-100% of BLAST)

DNA Barcoding

Tanjung Luar

32 ekor (99-100% of BLAST)

Muara Baru

3 ekor (99% of BLAST)

Muara Baru2 ekor (100% of BLAST)Tanjung Luar

41 ekor (99-100% of BLAST)

Aceh

4 ekor (99-100% of BLAST)

Carcharinus falciformis

Aceh

4 ekor (99-100% of BLAST)

Tanjung Luar

3 ekor (99-100% of BLAST)

Alopias pelagicus

Sphyrna lewini

Sebaran Populasi

2973877_T

L_251017_1_14_S

L_B

CL

29

79

71

6_

ITK

_M

B_

23

11

17

_A

L_

48

_B

CL

2973876_T

L_261017_1_21_S

L_B

CL

2973875_T

L_061117_1_1_A

P_B

CL

2973874_T

L_261017_1_46_S

L_B

CL

2973873_T

L_091117_1_1_A

P_B

CL

2973869_TL_061117_1_2_A

P_B

CL

2973

868_

TL_

2610

17_1

_26_

SL_

BC

L

2973

867_

TL_

2610

17_1

_31_

SL_

BC

L

2973

866_

TL_0

3111

7_1_

1_AP_B

CL

2973

865_

TL_26

1017

_1_4

2_SL_

BCL

2973864_TL_261117_1_29_SL_BCL

2973862_TL_231017_2_23_SL_BCL

2973860_TL_261017_1_18_SL_BCL

2973857_TL_261017_1_20_SL_BCL

2973856_TL_261017_1_37_SL_BCL

2973854_TL_221017_5_1_SL_BCL

2973853_TL_261017_1_23_SL_BCL

2973852_TL_261017_1_19_SL_BCL

2973851_TL_261017_1_34_SL_BCL

2973850_TL_261017_1_15_SL_BCL2973848_TL_261017_2_32_SL_BCL2973847_TL_261017_1_35_SL_BCL

2973846_TL_261017_1_13_SL_BCL

2973845_TL_261017_1_39_BCL

2973844_TL_231017_2_27_BCL

2973768_TL_171017_1_47_BCL

2973835_MBO_50_BCL

2973836_MBO_54_BCL

2973832_MBO

_48_BCL

2973834_MBO

_45_BCL

2973799_261017_02_09_AJ_B

CL

2973804_261017_02_07_AJ_B

CL

2973790_261017_02_02_AJ_B

CL

2973750_M

BO

_191017_2_B

CL

2973749_M

BO

_171022_19_B

CL

2973748_M

BO

_57_B

CL

2973742_M

BO

_171022_18_B

CL

2973741_M

BO

_14_B

CL

29

73

73

2_

MB

O_

47

_B

CL

29

73

72

8_

MB

O_

11

10

21

_0

3_

BC

L29

73

72

7_

MB

O_

17

10

24

_2

2_

BC

L

2973726_M

BO

_171024_23_B

CL

2973715_M

BO

_44_B

CL

Sphyr

na le

win

i KU

366619.1

57

2973716_M

BO

_33_B

CL

2973798_261017_02_03_B

CL

2973

830_

MB

O_5

5_BC

L

2973

840_

MBO

_51_

BC

L

2973

855_

TL_2

6101

7_1_

16_S

L_BCL

2979

712_

TL_22

1017

_2_6

_SL_

BCL

59 86

2973717_MBO

_9_BCL

2973718_MBO_171025_28_BCL

2973729_MBO_41_BCL

2973730_MBO_171021_07_BCL

2973731_MBO_171022_16_BCL

2973735_MBO_40_BCL

2973737_MBO_171021_04_BCL

2973738_MBO_29_BCL2973739_MBO_171021_06_BCL2973744_MBO_42_BCL

2973746_MBO_171024_21_BCL

2973786_261017_02_01_AJ_BCL

2973792_261017_02_06_AJ_BCL

2973791_261017_01_01_AJ_BCL

2973801_261017_03_01_AJ_BCL

2973831_MBO_52_BCL

2973829_MBO_10_BCL

2981134_MBO_40_BCL

2980001_MBO

_53_BCL

2979709_MBO

_34_BCL

2973839_MBO

_32_BCL

2973838_MBO

_40_BC

L

2973797_RG

P91017_1_1_B

CL

2973871_TL_131117_1_25_A

P_B

CL

99

95

2973734_M

BO

_171023_20_B

CL

Rhin

codon typ

us F

J375725.1

FST = 0,356; P = 0

2979715_IT

K_M

B_231117_A

L_69_B

CL

Ca

rch

arh

inu

s f

alc

ifo

rmis

KX

78

95

08

.1

2973891_IT

K_M

B_291117_M

L_71_B

CL

2973759_061117_2_2_LP

_B

CL

2973743_M

BO

_171025_27_B

CL

2973725_M

BO

_35_B

CL

2973724_M

BO

_36_B

CL

2973

898_

TL_

2210

17_1

_9_C

F_B

CL

2973

897_

TL_

2210

17_1

_13_

CF_B

CL

2973

825_

TL_2

2101

7_1_

4_CF_

BCL

2973

824_

TL_22

1017

_2_8

_CF_B

CL

2973821_TL_221017_3_16_CF_BC

L

2973820_TL_221017_1_5_CF_BCL

2973819_Tl_221017_1_23_CF_BCL

2973818_TL_221017_1_11_CF_BCL

2973817_TL_221017_3_11_CF_BCL

2973816_TL_221017_2_11_CF_BCL

2973815_TL_221017_3_8_CF_BCL

2973814_TL_221017_2_2_CF_BCL

2973813_TL_221017_1_3_CF_BCL

2973812_TL_221017_1_22_CF_BCL

2973811_TL_221017_3_10_CF_BCL2973810_TL_221017_1_2_CF_BCL2973809_TL_221017_1_6_CF_BCL

2973808_TL_221017_1_15_CF_BCL

2973807_TL_221017_2_4_CF_BCL

2973806_TL_2210171_2_9_CF_BCL

2973805_TL_221017_1_16_BCL

2973785_TL_181017_1_25_BCL

2973782_TL_181017_1_22_BCL

2973781_TL_181017_1_21_BCL

2973780_TL_181017_1_1_BCL

2973779_TL_181017_1_9_BCL

2973778_TL_181017_1_7_B

CL

2973777_TL_181017_1_2_B

CL

2973776_TL_181017_1_13_B

CL

2973774_TL_181017_1_24_B

CL

2973772_T

L_181017_1_4_B

CL

2973771_T

L_181017_1_19_B

CL

2973770_T

L_181017_1_18_B

CL

2973769_T

L_171017_2_5_B

CL

29

73

76

4_

TL

_1

81

01

7_

1_

3_

BC

L

29

73

76

5_

TL

_1

81

01

7_

1_

6_

BC

L

29

73

77

3_

TL

_1

81

01

7_

1_

8_

BC

L2973766_T

L_181017_1_23_B

CL

27

2973784_T

L_181017_1_14_B

CL

59

2973823_T

L_221017_1_10_C

F_B

CL

99

2973870_T

L_261017_1_30_S

L_B

CL

2973871_TL_131117_1_25_A

P_B

CL

2973

855_

TL_

2610

17_1

_16_

SL_

BC

L

2973

844_

TL_

2310

17_2

_27_

BC

L

2973

846_

TL_2

6101

7_1_

13_S

L_BCL

2973

847_

TL_26

1017

_1_3

5_SL_

BCL

2973848_TL_261017_2_32_SL_BCL

2973850_TL_261017_1_15_SL_BCL

2973851_TL_261017_1_34_SL_BCL

2973852_TL_261017_1_19_SL_BCL

2973853_TL_261017_1_23_SL_BCL

2973854_TL_221017_5_1_SL_BCL

2973856_TL_261017_1_37_SL_BCL

2973857_TL_261017_1_20_SL_BCL

2973860_TL_261017_1_18_SL_BCL2973861_TL_181117_1_23_BCL

2973862_TL_231017_2_23_SL_BCL

2973863_TL_061117_1_33_SL_BCL

2973864_TL_261117_1_29_SL_BCL

2973865_TL_261017_1_42_SL_BCL

2973866_TL_031117_1_1_AP_BCL

2973867_TL_261017_1_31_SL_BCL

2973868_TL_261017_1_26_SL_BCL

2973873_TL_091117_1_1_AP_BCL

2973874_TL_261017_1_46_SL_BCL

2973875_TL_061117_1_1_AP_BCL

2973876_TL_261017_1_21_SL_BCL

2973877_TL_251017_1_14_SL_B

CL

2973845_TL_261017_1_39_B

CL

2973869_TL_061117_1_2_A

P_B

CL

5720

00

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

02

87 96

93

67

80

2973872_TL_251017_1_29_A

P_B

CL

2973878_TL_231017_2_8__A

P_B

CL

99

Rhin

codon typ

us F

J375725

Carcharhinus falciformis

Sebaran Populasi

FST = 0,733; P = 0

Pelagic Thresher, Alopias pelagicus

Sebaran Populasi

2973711 M

BO

59 B

CL

Alopi

as p

elag

icus

KF02

0876

.1

17

2973761 181017 1 3 LP BCL

10

2973757 061117 2 3 LP B

CL

15

2973872 T

L 2

51017 1

29 A

P B

CL

67

2973

878

TL 23

1017

2 8

AP B

CL

2973896 TL 251017 1 27 AP BCL

66

Rhincodon typus FJ375725

Konektivitas Antar Populasi Manta alfredi Manta alfredi COI JQ765532.1(Ningaloo Reef)

Manta alfredi COI JQ765531.1(Ningaloo Reef)

2516654 ITK RAM MA 30 R12516653 ITK RAM MA 29 R12516648 ITK RAM MA 23 R12516642 ITK RAM MA 15 R1

2516638 ITK RAM MA 11 R1

2516633 ITK RAM MA 04 R1

2516632 ITK RAM M

A 03 R1

2516629 ITK NPN

MA 30 R

1

2516628 ITK N

PN M

A 29 R

1

2516626 ITK N

PN

MA 27 R

1

2516619 ITK N

PN

MA 20 R

1

2516617 IT

K N

PN

MA

18 R

1

2516616 IT

K N

PN

MA

17 R

1

2516615 IT

K N

PN

MA

16 R

1

2516612 IT

K N

PN

MA

13 R

1

2516611 IT

K N

PN

MA

12 R

1

2516610 IT

K N

PN

MA

11 R

1

25

16

60

9 IT

K N

PN

MA

10

R1

25

16

60

7 IT

K N

PN

MA

08

R1

2516604 IT

K N

PN

MA

05 R

12516600 IT

K N

PN

MA

01 R

1

2516599 IT

K K

OM

MA

24 R

1

2516598 IT

K K

OM

MA

23 R

1

2516597 IT

K K

OM

MA

22 R

1

2516592 IT

K K

OM

MA

17 R

1

2516590 ITK

KO

M M

A 15 R

1

2516585 ITK K

OM

MA 10 R

1

2516583 ITK K

OM

MA 08 R

1

2516579 ITK KOM

MA 04 R

1

2516578 ITK KOM

MA 03 R1

2516577 ITK KOM MA 01 R1

65

2516652 ITK RAM MA 28 R1

1

2516651 ITK RAM MA 26 R1

2516580 ITK KOM MA 05 R1

2516581 ITK KOM MA 06 R1

2516584 ITK KOM MA 09 R1

2516586 ITK KOM MA 11 R1

2516587 ITK KOM MA 12 R12516588 ITK KOM MA 13 R1

2516591 ITK KOM MA 16 R1

2516593 ITK KOM MA 18 R1

2516594 ITK KOM MA 19 R1

2516595 ITK KOM MA 20 R1

2516596 ITK KOM MA 21 R1

2516601 ITK NPN MA 02 R1

2516602 ITK NPN MA 03 R1

2516603 ITK NPN

MA 04 R

1

2516608 ITK N

PN M

A 09 R1

2516613 ITK N

PN

MA 14 R

1

2516614 ITK N

PN

MA 15 R

1

2516618 IT

K N

PN

MA

19 R

1

2516620 IT

K N

PN

MA

21 R

1

2516621 IT

K N

PN

MA

22 R

1

0

2516622 IT

K N

PN

MA

23 R

1

0

2516623 IT

K N

PN

MA

24 R

1

0

2516625 IT

K N

PN

MA

26 R

1

0

25

16

62

7 IT

K N

PN

MA

28

R1

0

25

16

63

0 IT

K R

AM

MA

05

R1

0

25

16

63

1 IT

K R

AM

MA

06

R1

0

2516634 IT

K R

AM

MA

07 R

1

0

2516635 IT

K R

AM

MA

08 R

1

0

2516636 IT

K R

AM

MA

09 R

10

2516637 IT

K R

AM

MA

10 R

1

0

2516639 IT

K R

AM

MA

12 R

1

0

2516640 ITK

RAM

MA

13 R1

0

2516641 ITK R

AM

MA 14 R

1

0

2516643 ITK R

AM

MA 16 R

1

0

2516644 ITK RAM

MA 17 R

1

0

2516645 ITK RAM M

A 19 R1

0

2516647 ITK RAM MA 22 R1

0

2516649 ITK RAM MA 24 R1

1

2516650 ITK RAM MA 25 R1

Manta birostris COI JF493866.1

Fst 0.035 (p=0.17009)

Kesimpulan• Deteksi spesies menggunakan pendekatan

molekular berdasarkan marka mtDNA lokus COI.

• Hasil genetika forensik secara keseluruhan mencapai tingkat deteksi 99-100%.

• Spesies yang terdeteksi, seperti C. falciformis, S. Lewini, A. pelagicus, Carcharhinus sorrah, dan C. amblyrhynchoides, serta # spesius lainnya.

• Pada spesies penting dideteksi terdapat struktur populasi yang tinggi dan signifikan (C. falciformis: FST = 0,733, S. Lewini: FST = 0,356; P = 0).

Rekomendasi

• Kegiatan pengelolaan dapat dilakukan secara terpadu dan berdasarkan karakteristik dari lokasi tersebut.

• Metode ini dapat digunakan untuk deteksi cepat ikan hiu dan pari yang dijual dipasaran, serta

• Karakteristik genetik dapat dipakai untuk mengetahui daerah “hotspot” untuk setiap jenis hiu dan pari

Publikasi hiu dan pariMadduppa H, Ayuningtyas RU, Subhan B, Arafat D. 2016. Exploited

but unevaluated: DNA barcoding reveals skates and stingrays(Chordata, Chondrichthyes) species landed in the Indonesian fishmarket. ILMU KELAUTAN: Indonesian Journal of MarineSciences. 21(2): 77-84.

Prehadi P, Sembiring A, Kurniasih EM, Rahmad R, Arafat D, Subhan B,Madduppa H. 2015. DNA barcoding and phylogeneticreconstruction of shark species landed in Muncar fisherieslanding site in comparison with Southern Java fishingport. Biodiversitas Journal of Biological Diversity. 16(1).

Sembiring A. Pertiwi NPD, Mahardini A, Wulandari R, Kurniasih EM,Kuncoro AW, Cahyani ND, Anggoro AW, Ulfa M, Madduppa H,Carpenter KE. 2015. DNA barcoding reveals targeted fisheries forendangered sharks in Indonesia. Fisheries Research. 164: 130-134.

Toha AH, Widodo N, Subhan B, Himawan MR, Tania C, Noor BA,Stewart BS, Madduppa H. 2016. Close genetic relatedness ofwhale sharks, Rhincodon typus in the Indo-Pacificregion. Aquaculture, Aquarium, Conservation & Legislation-International Journal of the Bioflux Society (AACL Bioflux). 9(3).