Phylogeography and Species Limits in the Papuan White Snake (Micropechis ikaheka) (Reptilia Serpentes Elapidae)

PHYLOGEOGRAPHY AND SPECIES LIMITS IN THE
PAPUAN WHITE SNAKE (Micropechis ikaheka)
(REPTILIA: SERPENTES: ELAPIDAE)

KELIOPAS KREY

GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2015

STATEMENT LETTER

I hereby declare that dissertation with title: Phylogeography and species
limits in the Papuan White Snake (Micropechis ikaheka) (Reptilia: Serpentes:
Elapidae) is the original result of my own research supervised by supervisory
committee and has never been submitted in any form at any institution before. All
information from other authors cited here are mentioned in the text and listed in
the reference at the end part of the dissertation.

Bogor, January 2015

Keliopas Krey
Student ID G362110051

SUMMARY
KELIOPAS KREY. Phylogeography and Species Limits in the Papuan White
Snake (Micropechis ikaheka) (Reptilia: Serpentes: Elapidae). Supervised by
BAMBANG SURYOBROTO, ACHMAD FARAJALLAH, and DEDE SETIADI
Papua white snake, Micropechis ikaheka, has been distingushed into two
subspecies M.i. ikaheka and M.i. fasciatus. However, due to lack of data on their
morphology, distribution and ecology, both subspecies often ignored in scintific
discussions and refers only byas , Micropechis ikaheka. This study was designed
to document the differences between M.i. ikaheka and M.i fasciatus by using
scales characteristics and reproductive organ or hemipenes. The study also
examine, zoogeography, habitat, diet and cannibalism of both subspecies to better
understanding the the ecology of one of the most venomous snakes in the world.
This study consists of three topics with method and also different in the size
samples, respectively. The first study is the histology and microtopography of the
skin were studied using paraffin method and Scanning Electron Microscope
(SEM). The second is the hemipenis were examined by following Myers and

Cadle (2003), and Zaher and Prudente (2003). The third is the abdominal surgery
was carried out to document diet type and the investigate phenomenon of
cannibalism in the snake. Specimens for this study came from the collection from
Museum Zoologicum Bogoriense (MZB) and Universitas Negeri Papua’s
Laboratory (LZU).
The research result also indicated that the markers of skin and hemipenis
described the same structure and microornamentation in all M. ikaheka groups
being compared. Based specimens examined morphological descriptions do not s
support the further differentiation in Micropechis. In short, two markers that used
in this study could not sufficiently indicate the differences in the level of species
and subspecies of M. ikaheka. The study, however, were able to delineate
zoogeography and skin color patterns, which fall into four groups: M. i. ikaheka;
M. i. fasciatus. M. i. ssp. “intermediate”; and M. i. ssp “black”. This study also
revealed that color variations did not overlap in their geographic distribution. It is
therefore, it suggested that environmental factors may contribute to the skin color
variations. It also assumed that habitat discontinuous and biogeography factors
may also cause the loss of the connection between the original populations
(vicarians) or follow colonization events (dispersal).
The examined of 22 stomach contents of M. ikaheka shows this species is
carnivorous, and prey on vertebrates such as amphibians, fish, reptiles, and

Mammals. These snakes are opportunistic predators since they prey on all that can
be captured, but mainly reptiles (61.5%), followed by mammals and fish (both
15.4%), and amphibians (7.7%). Although categorized as terrestrial animal, the
diet study showed that M. ikaheka snakes often go into the water to hunt for fish
(the sample from Yapen island), including eel (the sample from Aru island).
Based on prey items indentified, there is evidence that M. ikaheka is cannibal. At
this time, there still no record on reason for cannibalism, but it is assumed that

cannibalism was due to competition between populations and means to control
population.
Key words: geographic variation, white snake, papua

RINGKASAN
KELIOPAS KREY. Filogeografi dan Batasan Spesies dalam Ular Putih Papua
(Micropechis ikaheka) (Reptil: Serpentes: Elapidae). Dibimbing oleh BAMBANG
SURYOBROTO, ACHMAD FARAJALLAH, dan DEDE SETIADI
Ular putih Papua, Micropechis ikaheka, telah diperkenalkan menjadi dua
subspecies M.i. ikaheka dan M.i. fasciatus. Namun demikian, kedua subspesies
tersebut diabaikan dalam pembahasan-pembahasan ilmiah karena keterbatasan
data morfologi, distribusi dan ekologi mereka sehingga hanya merujuk pada satu

nama saja, Micropechis ikaheka. Penelitian ini mencoba menguraikan dan
membuktikan perbedaan intraspesies M. ikaheka lebih jauh dengan pendekatan
karakteristik kulit sisik dan alat reproduksi hemipenis. Selain itu, zoogeografi,
habitat, diet dan kanibalisme juga dipelajari guna menambah pengetahuan ekologi
tentang salah satu ular berbisa tinggi di dunia ini.
Penelitian ini terdiri dari tiga topik dengan metode masing-masing dan juga
ukuran sampel yang berbeda. Penelitian pertama terkait Histologi dan
microtopografi kulit, dipelajari menggunakan metode parafin dan Scaning
Electron Microskop (SEM). Yang kedua adalah kajian hemipenis menggunakan
metode preparasi mengikuti Myers dan Cadle (2003), dan Zaher dan Prudente
(2003). Yang terakhir adalah penelitian tentang diet dan fenomena kanibalisme
berdasarkan isi perut. Seluruh sample M. ikaheka yang diamati dan diukur dalam
penelitian ini bersumber dari koleksi spesimen yang telah diformalin dan
disimpan di MZB dan LZU.
Hasil penelitian menunjukkan bahwa keduanya, penanda kulit maupun
hemipenis, menggambarkan struktur dan mikroornamentasi yang sama pada
semua kelompok M. ikaheka yang diperbandingkan. Seluruh data dan deskripsi
morfologi yang diperoleh tidak menunjang diferensiasi takson Micropechis yang
telah diperkenalkan menjadi dua subspesies tersebut. Prinsipnya bahwa kedua
penanda yang telah digunakan dalam penelitian ini tidak dapat membuktikan

perbedaan level spesies maupun subspesies dalam M. ikaheka di Papua. Penelitian
ini, bagaimanapun, mampu menggambarkan zoogeografi dan pola warna kulit
yang terbagi mejadi empat kelompok: M.i. ikaheka; M.i. fasciatus. M.i. ssp.
"Intermediate" ; dan M.i. ssp " Hitam". Penelitian ini juga mengungkapkan bahwa
variasi warna tidak tumpang tindih dalam distribusi geografis mereka. Oleh
karena itu, meyakinkan bahwa faktor lingkungan dapat berkontribusi pada variasi
warna kulit. Hal ini juga diasumsikan bahwa faktor diskontinyu habitat dan
biogeografi juga dapat menyebabkan hilangnya hubungan antara populasi asli
(vicarians) atau mengikuti kejadian kolonisasi (dispersal).
Hasil bedah 22 perut spesimen M. ikaheka menunjukkan bahwa ular
beracun ini merupakan karnifor sejati terhadap vertebrata amphibi, ikan, reptil,
dan mamalia. Ular ini adalah pemangsa yang oportunistik, mereka memangsa
semua yang dapat ditangkap, namun utamanya reptil (61.5%), diikuti oleh
mamalia dan ikan (masing-masing 15.4%), dan amphibi (7.7%). Walaupun
teresterial, petunjuk terbaru diperoleh dalam penelitian ini bahwa M. ikaheka
sering masuk ke dalam sungai untuk berburu ikan (sampel dari Pulau Yapen)

termasuk sidat (sampel dari pulau Aru). Berdasarkan identifikasi item mangsa,
terdapat petunjuk bahwa M. ikaheka adalah kanibal. Pada saat ini, masih ada
catatan tentang alasan kanibalisme, tetapi diasumsikan bahwa kanibalisme ini

karena persaingan antara populasi dan berarti untuk mengendalikan populasi.
Kata kunci: variasi geografis, ular putih, papua

Copyright©2015, Bogor Agricultural University
Copyright are protected by law
Prohibited to cite all or a part of this dissertation without referring to and mentioning the
source. Citation permits to the purposes of education, research, scientific paper, report,
or critism writing only; and it does not defame the name and honor of Bogor
Agricultural University.
Prohibited to republish and reproduce all or a part of dissertation without the written
permission from Bogor Agricultural University.

PHYLOGEOGRAPHY AND SPECIES LIMITS
IN THE PAPUAN WHITE SNAKE (Micropechis ikaheka)
(REPTILIA: SERPENTES: ELAPIDAE)

KELIOPAS KREY

Dissertation
submitted in partial fulfillment of the requirements for a Doctoral Degree in

Animal Biosience Major in Graduate School of Bogor Agricultural University

GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2015

Examiners in the close examination: Dr. Ir. Mirza D. Kusrini
Yance de Fretes, MES, PhD

Examiners in the open examination: Yance de Fretes, MES, PhD
Dr. Evy Ayu Arida

Title

: Phylogeography and Species Limits in the Papuan White
Snake (Micropechis ikaheka) (Reptilia: Serpentes:
Elapidae)

Name

: Keliopas Krey

Register Number

: G362110051

Endorsed by,
Supervisory committee:

Dr. Bambang Suryobroto
Chairman

Dr. Ir. Achmad Farajallah, M.Si
Member

Prof. Dr. Ir. Dede Setiadi, MS
Member

Chair of Major in Animal Biosciences

Dean of Graduate School

Dr. Ir. RR Dyah Perwitasari

Examination Date : 24 December 2014

Dr. Ir. Dahrul Syah, M.Sc.Agr.

Date of Graduation:

FOREWORD
This dissertation was developed from three topics articles that have
submitted to scientific journals. The first topic entitled: Skin histology and
microtopography of Papuan White Snake (Micropechis ikaheka) in relation to
their zoogeographical distribution, has been published on March 2013 in Hayati J
Biosci 20(1): 7-14). The second topic entitled: The hemipenis of Micropechis
ikaheka (Reptilia: Squamata: Elapidae) from Papua, Indonesia, with comments on
the phylogeography of the species, has been received in journal of herpetology.
The third topic entitled: Micropechis ikaheka (Elapidae) in Papua, Indonesia: a

study of diet and cannibalism has been accepted in Herpetology Notes.
On July 15, 2011 the author enrolled into doctoral program student at
Institut Pertanian Bogor (IPB) Graduate School, Majoring in Animal Bioscience,
with financial support from BPPS (Beasiswa Pendidikan Pascasarjana).
I would express my sincere thank to Rector of IPB and the Dean of
Graduate School; in particular to Dr. Bambang Suryobroto, Dr. Ir. Achmad
Farajallah, M.Si, and Prof. Dr. Ir. Dede Setiadi, MS, who was served as
Supervisory Committee.
This study was partially supported by the Ministry of National Education of
Indonesia, Government of Papua and West Papua Province, and University of
Papua. I thank all the supports which allow me to finish this study.
During my study, I have the most wonderful colleagues, associates, friends,
and family, which. Without their supports and more contributions I may never
finish this dissertation. I would take this opportunity sincerely thank:
1. Perhimpunan Herpetologi Indonesia (PHI), especially Dr. Mirza Kusrini, who
invited in series of workshop, seminar and of course writing and the
publishing of short news of Papuan snake in Warta Herpetofauna.
2. Colleague at the Conservation International Indonesia (CI), BP-Tangguh, PT.
Ekologika Consultant, and PT. Freeport Indonesia for some accommodation
and traveling during the field work in Papua.

3. Burhan Tjaturadi, and David Price for the willingness to provide some
specimens of M. ikaheka.
4. Irvan Sidik who allowed me to access to the MZB collections and Dr. Sri
Hartini for the assistance of the hemipenial line drawing on camera lucida at
MZB Cibinong.
5. Tini Wahyuni who helps me to prepare of all of the equipment of the skin
histology and hemipenial works in Laboratory of Micro Technique IPB.
6. Dr. Daisy Wowor and Ir. Ristiyanti Marwoto, M.Si, the researchers from
MZB who observed, photographed and documented the MicropechisAnguilla interaction on Aru Island.
7. Prof. Eric Smith and Dr. Michael Harvey, who help in discussions and
assistances for preparation methods on the snake hemipenial.
8. Mark O’Shea, who help in discussion and also send some of snake article
9. Devi Manuhua and Tri Setiadi who make the distribution map of M. ikaheka.
10. I am very grateful to local guides who contributed and facilitated for the all of
stages in of my filed works in Papua and Aru.

I am in debt for my family: Matias Marisan and her families, Martinus Krey,
Markus Krey, Arnol Krey, Frans Krey, Hendrikus Krey, Evert Burwos, Noak
Awi, Erwin Yenusi, and Ariel Taime. My dear wife, Valentina Mayabubun, and
my three boy Yores, Mambri and Fanar are always together in Bogor.

Bogor, January 2015

Keliopas Krey

TABLE OF CONTENTS
Page

I

II

III

IV

V

VI

LIST OF TABLES
LIST OF FIGURES
LIST OF APPENDIXES

x
xi
xi

GENERAL INTRODUCTION
Background
Research Urgensy
Aims of this Study

1
2
3

LITERATURE REVIEW
The Micropechis species
Colour and Structure of Snake Skin
Structure of Snake Hemipenial

5
5
7

MATERIALS AND METHODS
Field work
Skin Histology Preparation
Hemipenial Preparation
Diet Analysis

8
8
8
9

SKIN HISTOLOGY AND MICROTOPOGRAPHY OF PAPUAN
WHITE SNAKE (Micropechis ikaheka) IN RELATION TO THEIR
ZOOGEOGRAPHICAL DISTRIBUTION
Introduction
Materials and methods
Results
Discussion

10
11
13
17

THE HEMIPENIS OF Micropechis ikaheka (REPTILIA:
SQUAMATA: ELAPIDAE) FROM PAPUA, INDONESIA, WITH
COMMENTS ON THE PHYLOGEOGRAPHY OF THE SPECIES
Introduction
Materials and Methods
Results
Discussion

20
21
22
24

Micropechis ikaheka (ELAPIDAE) IN PAPUA, INDONESIA: A
STUDY OF DIET AND CANNIBALISM
Introduction
Materials and Methods
Result and Discussion

27
28
28

VII

GENERAL DISCUSSION
The Limitations of this Research and Future Study
Geographic Variation and Taxonomy Significant in M. ikaheka
Skin melanin and Sex Dimorphism of M. ikaheka
Diet and cannibalism phenomena with comment to conservation
The contributions of snakes for conservation, and the local wisdom

VIII GENERAL CONCLUSIONS AND SUGGESTION
Conclusion
Suggestion
REFERENCES
APPENDIX
CURRICULUM VITAE

35
35
37
38
39
40
40
41
46
51

LIST OF TABLES

1
2
3
4
5
6

Page
Study sites of M. ikaheka in Papua
11
Specimen materials of M. ikaheka used in this study
21
Selected character variation in M. ikaheka
25
Identifiable prey items from gastrointestinal tract (GI) of M. ikaheka
28
Identified prey items in male, female and juvinile of M. ikaheka
31
Comparison prey items identified from gastrointestinal tract of M. ikaheka
31

LIST OF FIGURES

1
2
3
4
5
6
7

Page
Generalized epidermis of a squamate reptile
6
Diagram of the cross section of the snake integument
6
Sulcate (left) and asulcate (right) morphology illustrate of snakes
hemipenis
7
Map of distribution of M. ikaheka in Papua, showing specific
zoogeographical
13
The arrangement and different color of M. ikaheka skin scales
14
The scales overlapping and pigmentation pattern of the dorsal caudal
region
15
Photomicrographs of the dorsal midbody skin
15

8
9
10
11
12
13
14
15
16
17
18
19

SEM images illustrating the microstructure of the oberhautchen on the
dorsal outer scale surface of the mid-body (A,B) and caudal (C,D) scales
SEM images illustrating the microtopography of the oberhautchen on the
ventral outer scale surface of the mid-body
Sulcate (A), asulcate (B) and lateral (C) view morphology of left
hemipenis of M.i. ikaheka (LZU 28)
Sulcate (A) and asulcate (B) morphology illustrate of left hemipenis of
M.i. ikaheka (LZU 28)
Distribution of the seven (1-7) specimen of M. ikaheka used in this study
with geographic variation in color morph of the species in Papua
Prey items identified as a freshwater fish, Cyprinidae, eat by a juvenile of
M. ikaheka from Yapen Island
A juvenile M. ikaheka (length from tail to upper body without head: 400
mm) from the gastrointestinal tract of an adult
The cannibalism of M. ikaheka at Manokwari, West Papua Province
The direction of ingestion of M. ikaheka is head first
M. ikaheka (fasciatus form) found ingesting an eel, Anguilla sp., at
Warialau Island, North Aru
Geographical variation of M. ikaheka in Papua
Map of the distribution of M. ikaheka based on specimens now examined
and from the literature

16
17
22
23
26
29
32
33
33
34
36
37

LIST OF APPENDIXEX

1
2
3
4

Page
Colour variation of the adult of M. ikaheka in Papua
46
The three distinct color patterns of the adult and juveniles of M. ikaheka
found in Papua
47
The guide to the snake hemipenis
48
Specimens and locality data of M. ikaheka used in this study
49

1

I GENERAL INTRODUCTION

Background
Papua is Indonesia‟s second largest island, encompassing 416.129 km2.
Papua makes up the western half of the great sub continental island of New
Guinea (Beehler 2007). This region is known to support a diverse range of marine
and terrestrial ecosystems, as well as an array of endemic species. The habitat
mostly consists of tropical rainforest, providing ample opportunity for a diversity
of species to flourish.
Eighty-three snake species have been recorded on Papua (Allison 2007),
consisting of three percent of the worlds 2700 species (Taylor and O'Shea 2004).
These eighty-three species are classified into seven families, including
Acrochordidae (filesnakes), Boidae (boid), Pythonidae (pythons), Colubridae (tree
snakes, water snakes and ground snakes), Elapidae (rear-fanged venomous
snakes), and Typhlopidae (burrowing snakes). Within these families is the
Cylindrophiidae, represented by Cylindrophis aruensis, which is endemic to the
Aru Islands (McDowell 1975; O'Shea 1996; Allison 2007). Five of the seven
families (all except the Boidae and Cylindrophiidae) are represented in Australia
and six of the families (all except the Boidae) occur in Southeast Asia. Most
families appear to have had a long history in the Indo-Australian region (Allison
2007).
Only three hundred species of snakes in the world are dangerous to humans
(Taylor and O'Shea 2004), one of which is Micropechis ikaheka (local name
"white snake"; English name "New Guinea or the small-eyed snake ikaheka")
(O‟Shea 1996). This venomous snake is endemic to New Guinea and ranges from
the lowlands up to about 1.500 m in the mountains. This species is widespread
across the main island of New Guinea, as well as some of its smaller satellite
islands (O'Shea 1996). M. ikaheka is a widespread and medically important
venomous snake in New Guinea. A better understanding of its taxonomy will
underpin studies of venom variation, which may be of medical significance
(Wuster W 25 January 2012, personal communication).
Until now, the most widely accepted hypotheses of M. ikaheka relationships
are based on analyses of external scale morphology. The species was described by
Lesson in 1830, from a single specimen collected at Dorery (now known as
Manokwari, West Papua Province, Indonesia) (Lesson 1830). In 1884 J.G. Fischer
described M. i. fasciatus from the Aru Islands. The two subspecies, M. ikaheka
ikaheka and M. ikaheka fasciatus (Fischer 1884) have been recognised by some
authors, but their distributional limits are unclear. M. i. fasciatus has been
considered an Aru endemic by some (e.g., Klemmer 1963), but others listed it
from parts of Papua New Guinea (Loveridge 1948), restricting the M. i. ikaheka to
Papua. Due to lack of clear diagnostic characters, the subspecific designations
have often been largely ignored (e.g., O'Shea 1996).
This dangerous snake, M. ikaheka, displays extreme geographic variation in
colours. The most recent study conducted by Krey (2009), investigated colour
variation and zoogeography of the species in Papua. This study found three
distinct colour morph: Black, Brown and Yellow (Appendix 1 and 2). These are

2
the main characteristics distinguishing populations in different areas of the
distribution of M. ikaheka. This means that M. ikaheka exhibits prominent
geographical variation along the mainland of New Guinea as well as some of its
smaller satellite islands in scale color. The black colored group is the new group
found in northwestern Vogelkop Batanta and Waigeo islands. The yellow colored
population is distributed in Vogelkop region starting from Bintuni, Manokwari,
Sorong to the Salawati islands. Finally, the brown colored population is
distributed from the Yapen islands to the mainland region from Waropen,
Mamberamo, Jayawijaya, Mindiptana to Papua New Guinea. A hybrid color
morph appears between the brown and the yellow populations in southwest
Vogelkop, Bintuni. However, the taxonomic significance of this variation is not
clear because the existing data do not distinguish between local adaptation,
polymorphism or species-level differentiation.
Snakes serve an important role in nature, especially in a food chain. In Java,
for example, snakes serve as predators controlling pest rodents that often attack
rice plants (Whitten et al. 1999). However, human discrimination prevents snakes
from fully fulfilling this role in the ecosystem. On the other hand, carnivorous
birds may prey on snakes in nature. However, so far there has been no information
about predatory snakes in Papua. Some reports have indicated that a group of
elapid snakes may be the main predators of other snake species in New Guinea
(Shine and Keogh 1996; O'Shea 1994a), and, even more than that, they are
cannibals (O'Shea 1994b).
Very little is known about this venomous species, despite some available
data from Papua New Guinea (PNG). A number of papers and notes on the diet of
M. ikaheka have been published, but these were confined to the eastern half of
New Guinea, which comprises the independent country of PNG. In PNG the
snake had been reported to prey on lizards, snakes, frogs, and small mammals
(O'Shea 1994a, 1994b; O'Shea 1996; Shine and Keogh 1996).

Research Urgency
Although not well reported, there have been many cases of death of
mothers and children, students, and workers in the plantation due to bites and
exposure to venom spray by snakes in Papua, especially M. ikaheka. A 25 year
old man in Arso SP XI Jayapura was bitten by a male M. ikaheka (now deposited
at the British Museum of Natural History London, accession number BMNH
1994.525) although he recovered after spending two weeks in the hospital in
Jayapura (Warrell et al. 1996). In another case, an elementary school student and
a mother in Manokwari died after 12 and 18 hours due to the venom of M.
ikaheka (Krey observation). A young man on the island of Kar Kar only survived
for 6 hours after being bitten by M. ikaheka before he eventually died, and another
young man in Wewak also died after enduring 36 hours (Blasco and Hornabrook
1972). Finallly, a victim of M. ikaheka in Madang died 18 hours after the snake
attack (Hudson 1988).
The effect of M. ikaheka venom is very fatal and deadly because it
contains a neurotoxin to post-synaps, possibly pre-synaps (Geh et al. 1997), which

3
can also destroy blood cells (haemolytic) and poison muscle (myotoxic) (Hudson
1988; Warrell et al. 1996). Morphological features of this highly venomous
species are similar to the type of non-venomous Stegonotus spp. (O'Shea 1996),
which often confuses and endangers humans. In addition, the geographic variation
in the skin color makes it difficult to distinguish whether the species is venomous
or non-venomous, and the myth that says that the white snake venom (M. ikaheka)
is contained in its pointed tail, not in its bite, has also put humans in danger. Some
indigenous Papuans in Vogelkop believe that the white snake has a mystical
power that can cause rain if found or captured by humans.
Geographic variation in the skin color patterns of M. ikaheka in New
Guinea has never been fully understood. As a result, the taxonomic naming of the
snakes has been neglected, and zoogeographical information is lacking, including
information on certain endemic regions which are medically significant.
Additionally, the possibility of hybrid zones and distribution overlap between
populations of M. ikaheka in New Guinea is little understood. By comparing
morphological characters such as skin color and surface structure of oberthoucen
and the appearance of hemipenial morphology in conjunction with
phylogeography, one can investigate interspecific differences. Understanding the
biology of these snakes has implications for preventing bites to Papuans, and in
the identification of species before medical treatment.
Pigment cells (chromatophores) scattered in the dermis layer of the skin,
exclusively on ectotherms, play a role in maintaining color constancy (Hildebrand
and Goslow 2001). Adaptation to local conditions may be responsible for the
variation of M. ikaheka skin color between populations, which may be maintained
by chromatophores. The distribution of chromatophores on the skin dermis has
the implication on the distribution of melanin colors by melanin cells
(melanophores) that affect the variability of snake skin colors on the scales. Skin
histology, physiological phenomena and ecology have become important
knowledge that can be learned in this study.
Micropechis ikaheka have been recorded to live on land (O'Shea, 1996),
very close to other terrestrial and or aquatic vertebrates. A study of the diet of M.
ikaheka is important for our understanding of the inter and intraspecific
relationships of this species within the ecosystem. In addition, an increase in data
availability regarding snakes species within Papua-Indonesia supports and
strengthens the study of biology, ecology, evolution and natural history.

Aims of this Study
This study will address the following aims:
1) Geographic variation on skin colour patterns of M. ikaheka in New Guinea is
not clearly known until now. This study aims to identify the distributional
patterns, including endemic regions, hybrid zones or overlapping distributions
among populations within M. ikaheka in Papua.
2) The dermis of M. ikaheka contains pigments implicated in the morphological
variability of skin coloration. This study will attempt to better understand the

4
section of M. ikaheka skin, and its influence on maintaining melanin
distributions.
3) All of the meristic and color pattern data available is not enough to define the
differences between populations. This study attempts to re-diagnose this
species based on hemipenal morphology, a study which has not been done
since Lesson 184 years ago. Snake hemipenes show great variation in
structural ornamentation among different species, and can be of great
taxonomic importance.
4) Ecological knowledge of this highly venomous snake is very limited, despite
available data from PNG. The collection at Laboratorium Zoology Unipa
(LZU) and Museum Zoologicum Bogoriense (MZB) provide a unique
opportunity to study the diet and cannibalism in M. ikaheka from the western
half of the great sub-continental island of New Guinea.

5

II LITERATURE REVIEW

The Micropechis species
Micropechis ikaheka, a monotypic species, is a highly venomous elapid
species native to New Guinea (Papua Indonesia and Papua New Guinea). Lesson
first described it in 1830, from a single specimen collected at Dorery (Manokwari,
West Papua Province, Indonesia) (Lesson 1830).
Micropechis ikaheka has a fairly stocky body with a relatively short tail.
The head is narrow and only slightly boarder and distinct from the neck, with very
small eyes and a round pupil (O‟Shea 1996). Coloration of M. ikaheka indicates
variability by locally (O‟Shea 1996; Krey and Farajallah 2013). In PNG, the
northern population exhibits different coloration from the southern. The northern
population has a light to dark grey head and distinct from the yellow or cream
neck, while the southern population has a dark pigment scales, with the dark
crossbands which increase in width and frequency posteriorly until the tail
(O‟Shea 1996).
Micropechis ikaheka can grow up to 2 m in length (O‟Shea 1996). The
number of dorsal at midbody (DMB) scales 15, all smooth; 174-223 of ventral
scales; subcaudals 37-55, diveded; anal plate divided; 6 supralabials with 3th and
4 th contacting eye and temporolabial between 5 th and 6 th; loreal and subocular
scale absent (Lesson 1830; Fischer 1884; Rooij 1917; O‟Shea 1996; Krey 2009).

Colour and Structure of Snake Skin
Reptilian scales are generally very colorful and organized in an attractive
pattern. The colour of terrestrial reptiles can be diverse and their pigmentation
complex, but less so in fossorial groups (Withers and O‟Shea 1993). The colours
of snakes come from the pigments in the scales and from the way light reflects of
the scales (Tylor and O'Shea 2004). Many elapids show great variation in the
ground color (Shea et al. 1993), including the New Guinea elapid Micropechis
ikaheka.
Every part of a snake‟s body is covered by a layer of skin scales. The scales,
like reptiles in general, grow from the top layer or the epidermis (Abdel-Aal et al.
2011), and usually made of a horny substance called keratin (Hildebrand and
Goslow 2001). The skin plays a particularly important role in protecting them
from desiccation, abrasion dehydration, ultraviolet radiation, and in providing an
impermeable barrier to exogenous organism, including potential pathogens
(Cooper 2006).
Like many vertebrates, reptilian skin has two principal layers i.e. the dermis
and the epidermis. The deeper layer, the dermis, provides connective tissue with a
rich supply of blood and lymphatic vessels, nerves and chromatophores (Cooper
2006), while the epidermis has no blood supply and forms the outer protective
coating of the body (Rossi 1996). There are seven epidermal layers (Figure 1), the

6
“stratum germinativum”, the deepest layer lining cells which have the capacity
for rapid cell division, and the six layers which form each “epidermal
generation”, the old and the new skin layers (Abdel and Mansori 2009; Abdel-Aal
et al. 2011). The epidermis of snakes (Figure 2) now consists of six main layers
(Klein and Gorb 2012). From the outer scale surface towards the dermis are
known the “oberhautchen” layer, (β)-layer, mesos-layer, (α)−layer, lacunar layer
and the clear layer (Hildebrand and Goslow 2001; Abdel-Aal et al. 2011; Klein
and Gorb 2012).

Figure 1 Generalized epidermis of a squamate reptile.
Source: Abdel-Aal and Mansori (2009).

Figure 2 Diagram of the cross section of snake integument.
Source: Klein and Gorb (2012).

7
Structure of Snake Hemipenial
The structure of the male genitalia of snakes was first used for systematics
by Edward Drinker Cope (Cope 1894) 120 years ago. Hemipenes are used
regularly to study taxonomy, as new techniques of preparation are regularly
proposed (Myers and Cadle 2003; Zaher and Prudente 2003). Recent papers have
described hemipenial morphology from different snake species from many areas
of the world (e.g. Keogh 1999; King et al. 2009; Jadin et al. 2010; Jadin and
Parkhill 2011; Smith et al. 2012).
The complete description of a hemipenis (Figure 3, Appendix 3) for
systematic purposes includes recording information on its length, shape, condition
of sulcus spermaticus, hooks, nature and pattern of ornamentation (spin, calyces,
papillae) and type of apex. This technique follow Cope (1894), Dowling and
Savage (1960) and the most useful and comprehensive for the Australia-New
Guinean region snakes, Keogh (1999).

Figure 3 Sulcate (left) and asulcate (right) morphology illustrate of snakes
hemipenis. This hemipenis structure to illustrate the relevant
morphological features found among Australian terrestarial elapid
snakes. AL=apical lobes, P=papillae, C=calyses, S=spines,
FP=fleshy protuberances, R=ridges, SS=sulcus spermaticus,
SL=spine line, H=basal hooks, BA=base. Source: Keogh (1999).

8

II MATERIALS AND METHODS
Field Work
Field work focused on the mainland of Papua and its satellite islands. All
samples and information of M. ikaheka were obtained from throughout its
geographical distribution within Papua as well as Aru Island. A total of 47
specimens of M. ikaheka were partially collected in this research between 1967 2012 (Appendix 4). The snake specimens were deposited in the Museum
Zoologicum Bogoriense (MZB) LIPI in Cibinong, West Java and Zoology
Laboratory University of Papua (LZU) in Manokwari, West Papua.
Observations of snakes were made day and night in a variety of habitats
such as riverine, wetland, forest, and agricultural land. All individuals of M.
ikaheka found in the field were captured by hand and preserved with 10%
formalin, and subsequently stored in 70% ethanol. Data was recorded, including
locality, habitat, altitude and GPS coordinate.

Skin Histology Preparation
All materials used in this study are based on available formalin-preserved
adult specimens in the collections of LZU. The paraffin method and Scan Electron
Microscopy (SEM) were used for snake skin preparation. The paraffin method
was used to study and analyze the internal structure of the snake skin layers. Only
one specimen (assession number: SJR 07803 from Salawati Island) was used in
this analyze as a model and the basic assumptions for the other color morph of M.
ikaheka. The patterns of distribution of the pigments that affect skin color on M.
ikaheka were also analyzed with a visual approach, although the paraffin method
was found to be ineffective. While SEM is useful in studying the outer layer
surface “oberthoutchen” texture of scales, microornamentation and also the
microtopography variation between the dorsal and ventral sides. Three specimens
with assession number SJR 07721 from Batanta Island, SJR 08092 from Sentani,
and LZU 30 from Bintuni was used in SEM analyze. All data obtained with this
method were used to analyze differentiation between scales surface of M. ikaheka
in relation to their zoogeography in Papua.

Hemipenial Preparation
The hemipenis of seven males from three different groups representing
population color morph (yellow, brown, black and one intermediate color morph)
were investigated. All materials used in this study are based on available
formalin-preserved adult specimens in the collections of LZU and MZB.

9
Hemipenial organs were prepared with the methods described in detail by Myers
and Cadle (2003), and Zaher and Prudente (2003). We also made the
morphological illustration of M. ikaheka hemipenis by using a camera lucida
(Nikon SMZ 800) stereo microscope. Hemipenial descriptions and terminology
follows Cope (1894), Dowling and Savage (1960), and Keogh (1999).

Diet Analysis
Diets of Micropechis ikaheka are based on stomach contents of the
museum specimen collection. A total of 22 specimens were examined in this
study. Stomach contents were removed and preserved in 70% ethanol. We opened
the whole abdomen to ensure the retrieval of all prey items. We record the
location of the prey item (stomach, intestine), and the number and condition of
prey items. Because of break down caused by partial digestion, we were only able
to identify some of prey items to Class or Family. Classifications and
identification of prey taxa was based on Menzies (1975) for frogs, Brown (1991)
for lizards, Rooij (1917) and O‟Shea (1996) for snakes, Allen (2000) for
Osteichtyes, Menzies and Dennis (1979) for mammals. The data of snakes diet
used for some analyses i.e. prey composition, comparison prey items, direction of
ingestion, and cannibalism.

10

IV SKIN HISTOLOGY AND MICROTOPOGRAPHY OF
PAPUAN WHITE SNAKE (Micropechis ikaheka) IN
RELATION TO THEIR ZOOGEOGRAPHICAL
DISTRIBUTION

Introduction
A scaly, keratinized integument is one of the distinctive features of
Reptilian, to make their skin airproof (Hildebrand and Goslow 2001). Snake
scales, like other reptiles, grow from top layer or the epidermis (Abdel-Aal et al.
2011). In lepidosaurians (lizards, snakes, and sphenodontids) germinal layer of the
epidermis spinosus-like keratinocytes alternate to hard (beta) and soft (alpha)
layers (Toni et al. 2007; Chang et al. 2009). The whole skin of lepidosaurians is
covered by overlapping epidermal scales that protect them from abrasion and
dehydration. The epidermis of lepidosaurs is of particular complexity and interest
(Hildebrand and Goslow 2001), moreover with very colorful scales and organized
in an attractive pattern. The color of terrestrial reptiles can be diverse and their
pigmentation complex, marked with colorful patches.
Reptilian skin has two principal layers i.e. the dermis and the epidermis.
The epidermis of snakes now consists of six main layers (Klein and Gorb 2012).
From the outer scale surface towards the dermis are known the “oberhautchen”
layer, (β)-layer, mesos-layer, (α)−layer, lacunar layer and the clear layer
(Hildebrand and Goslow 2001; Abdel-Aal et al. 2011; Klein and Gorb 2012).
A number of the larger elapids (venomous snake) show great variation in
the ground color as shown as a New Guinea elapid Micropechis ikaheka. These
colour patterns are the main characteristics distinguishing populations of the
distribution of M. ikaheka.
Micropechis ikaheka (local name "white snake"; English name "New
Guinea or the small-eyed snake ikaheka") is endemic to New Guinea ranges from
lowlands to about 1.500 m in the mountains. The species is a venomous snake
widespread across the mainland island of New Guinea as well as some of its
smaller satellite islands (O'Shea 1996).
Two subspecies, M.i. ikaheka and M.i. fasciatus, have been recognised by
some authors, but their distributional limits are unclear. Due to lack of diagnostic
characters, the subspecific designations have often been ignored (O'Shea 1996).
For the further analysis on this research used species level. The most recent study
by Krey (2009) on colour variation and zoogeography of the species in Papua
found three distinct colour patterns i.e. black, brown and yellow. A unique
opportunity to study the histology and microtophography skin scales of Papuan
white snake (M. ikaheka) because there has been no basic research previous is
crucial in exploring our knowledge. Therefore, this works were aimed to study the
internal histology of skin layer and the microtopography structure on the surface
of scales of M. ikaheka. It is also related to the existence of the snake on different
zoogeographical locations in Papua.

11
Materials and Methods
Study Specimen
Field studies focus on the mainland of Papua and its satellite islands.
Micropechis ikaheka was captured by hand and preserved in the field with 10%
formalin, subsequently were stored in 70% ethanol. Field observation of the
snakes was carried out during day and night on some habitats such as along a
river, swamp, forest, and plantations as well. All field data such as locations of the
snake and their altitude were recorded.
Table 1 Study sites of M. ikaheka in Papua
Site number and
Locality
Black group

Latitude

Longitude

Biogeography
region

Altitude
(meter)

Collector

Waifoi
Lopintol
Urbinasopen
Waimnir
Wailebet

000 05.970‟
000 18.999‟
000 20.219‟
000 23‟50.6”
000 53.744‟

1300 45.642‟
1300 51.542‟
1310 15.544‟
1300 52‟36.5”
1300 38.498‟

Waigeo Is.
Waigeo Is.
Waigeo Is.
Waigeo Is.
Batanta Is.

50
5
15
24
13

Ps
Ps
Ps
Ps
Ps

Yellow group
6. Webya
7. Oransbari
8. Saukorem
9. Nuni
10. Gunung Meja
11. Andai
12. Prafi
13. Tanah Merah
14. Manokwari
15. Irian
16. Jamursbamedi
17. Terasai
18. Sorong

000 57,383‟
010 20‟55.3”
000 44‟45.3”
000 46‟09.2”
000 50‟51.5”
000 55‟ 902”
010 00‟ 03.0”
020 28‟ 31.4”
02°01‟54.0”
000 54‟ 40.3”

1300 47.060‟
1340 11‟19.8”
1330 23‟33.9”
1330 58‟57.0”
1340 4‟24.7”
1340 0‟ 557”
134o 00‟05.0”
1330 8‟23.2”

Salawati Is.
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop
Vogelkop

25
276
34
24
155
36
305
49
38
40

Hc
Ps
Ps
LZU
Ps
LZU
Ps
Ps
LZU
MZB
MZB
Ps
Ps

Brown group
19. Warironi
20. Noau
21. Kwerba
22. Cyclop
23. Kelila
24. Baliem Valley
25. Mindiptana
26. Kaimana
27. Kuala Kencana

010 50‟ 45.7”
020 04‟57.1”
02038‟28.46”
50 38‟ 33”
03036‟16.8”
04024‟11.3”

1360 32‟ 59.4”
1370 27‟32.1”
138024‟48.35”

Yapen Is.
Northern
Northern
Northeastern
Central
Central
Southeastern
Southwestern
South

32
22
64
98

Hc
Hc
Hc
Hc
MZB
MZB
MZB
Ps
Ps

1.
2.
3.
4.
5.

133°51‟01.8”
1310 18‟57.6”

1410
133043‟38.9”
136052‟12.4”

-: not available; Is.: Island; Ps; Present study; MZB: Muzeum Zoology Bogoriense; LZU: Zoology
Laboratory Unipa; Hc: Herpetologist collection

12
Specimens and locality data of M. ikaheka used in this study also consisted
of museum specimen collection from (i) Museum Zoology Bogoriense at
Cibinong, Bogor; (ii) Zoology Laboratory of the State University of Papua at
Manokwari; and (iii) the collections of several herpetologists. For the purpose of
analysis, specimens of M. ikaheka were chosen as representatives for variations of
the existing groups. There are at least three groups-yellow, brown and black used
in this study. Study sites have been surveyed as shown in Table 1.
Geographical location of M. ikaheka recorded with a Global Positioning System
(GPS). Most coordinates of latitude and longitude plus altitude were recorded
using GPS map 60csx (Garmin Co). The mapping of several locations without
recording the coordinates (site number 14, 15, 16, 22, 23, 24) was carried out
based on village names from the record of the specimens collected. The Map
Sources 6.9 and Arcview 3.3 software were used for mapping the snake existence
on different zoogeographical locations in Papua.

Internal Skin Histology Preparation
The skin histology of the M. ikaheka was analyzed for comparison with
external surfaces of scales using paraffin method. All M. ikaheka specimens were
preserved in 10% formalin for laboratory processing. Only one specimen from
yellow groups was used in this work as a model and the basic assumptions for the
other group of the M. ikaheka. There are two characteristics of the snake skin
were studied, i.e. internal structure of the skin layer and pigmentation pattern.
Two skin positions were choice for initial examination based on the color
pattern of the M. ikaheka (yellow group). Position I is a representative for the
dorsal caudal region, while position II represents of the dorsal mid-body region.
The skins were cut into + 0,5 cm pieces and fixed in FAAC (Formalin acetic acid
calcium chloride) solution for + 24 hours. Fixative solution was removed from the
tissues and washed several times with distilled water. The tissues were dehydrated
in a graded series of ethanol: 30, 50, 70, 80, 95, and 100% each 15-30 minutes.
After rarefaction in the xylene + 30 minutes, the tissues were infiltrated and
embedded in paraffin.
Skins were cut (1 + 2 μm) with a rotary microtome in longitudinal planes.
Several sections were deparaffined with xylene, dehydrate with ethanol,
subsequently were stained with double haematoxylin-eosin.

Preparation of Snake Skin for Scanning Electron Microscope (SEM)
Three individuals of M. ikaheka representing for the black, brown, and
yellow groups were chosen for SEM observations. The SEM method was used to
study the microtopography structure of the oberthoutchen layer surface between
the three groups of M. ikaheka. Skin characters between dorsal and ventral sides
of the snake body were observed were: surface texture of scales,
microornamentation and microtopography variation.
Three skin positions were identified for initial examination based on the
color pattern of the skin scales. Position I is a representative for the dorsal midbody region, position II represents of the dorsal caudal region, and position III
represents the ventral mid-body region. The skin region from each of the chosen

13
positions was examined at different magnifications (X35-X1000) in topographical
mode using scanning electron microscopy JEOL JSM 5310 LV. Several stages of
preparation were taken before snake skin SEM observation i.e. cleaning,
prefixation, fixation, dehydration, and drying. Dried samples were placed on the
stub and then coated by Au using 5-8 mA ion current for 5 minutes, with 400-500
Å thickness using ion coater IB-2.

Results
Distribution and Habitat of Micropechis ikaheka in Papua
Field observations in Papua and specimens study were carried out very
interesting specific pattern distribution of M. ikaheka (Figure 4). Three groups of
the snake i.e. brown, yellow, and black have specific zoogeography in mainland
of Papua and its satellite islands (such as Yapen Island, Batanta, and Waigeo
Island). All altitude observations showed the species spread from 5 to 305 m ASL.
However, specimen from Kelila and Baliem valley showed these snakes also live
in the sea level to approximately 1700 m ASL.

Figure 4 Map of distribution of M. ikaheka in Papua, showing specific
zoogeographical. The yellow group (yellow triangle), brown group
(reversed triangle), new group (black circle) and intermediate
individual (green pentagon).

14
Micropechis ikaheka that was found in the field observations confined to
terrestrial rainforest areas. The snakes also found in the monsoon areas and the
swamps. M. ikaheka was a nocturnal and secretive semi-fussorial snakes which
inhabits leaf litter, loose soil or piles of decaying vegetation, husks of cocoa,
coconut or palm oil. Several individuals were also observed in the hole of a fallen
palm tree, under the tree buttresses and also in the rock crevice.

Internal Skin Histology Analysis
Position I: dorsal caudal region. The specimen M. ikaheka from yellow
group showed a unique distinct color of skin scales in different parts of the snake
body (Figure 5). Paraffin method used in this work could not explain the
arrangement of pigment cells (chromatophore) in detail. However, some general
information about M. ikaheka skin coloration and pigmentation might be
described in these results. Dark colors on the entire surface of the dorsal caudal
skin were due to the pigment (Figure 5-A1 and A2). The dark skin might contain
melanophores, which only found within the dermis.
The other observation is the existence of overlapping scales and hinge
regions (h) (Figure 5-B1 and B2). The melanin pigment is not evenly spread on all
surfaces scales. There is a regular pattern of pigmentation in the M. ikaheka skin
scales. The scales overlapping caused the skin surface region covered by other
scales continued to the hinge region (h) to contain pigment (Figure 6).

Figure 5 The arrangement and different color of M. ikaheka skin scales.
A = skin samples from the dorsal caudal; B = skin samples from
the dorsal midbody; mp = melanin pigment; h = hinge. region.

15

Figure 6 The scales overlapping and pigmentation pattern of the dorsal caudal
region. mp = melanin pigment.

Figure 7 Photomicrographs of the dorsal midbody skin
Position II: dorsal midbody region. Approximately five layers were
identified in microscope observation (Figure 7). These were the oberhautchen, the
beta (β -layer, the mesos layer, the alpha (α)-layer, and the dermis. The β and α
layer consisted of cells which become keratinized with the production of two
types of keratin (β and α keratin). The oberhautchen did not show smooth
characteristics, followed the inner scale surface and hinge region (h) that
composed of thin beta-layer.

16
Microtopography Structure of M. ikaheka Skin Scales
Position I and II: dorsal midbody and caudal region. Based on SEM analysis
of the oberhautchen on the dorsal outer scale surface of the mid-body and caudal
of M. ikaheka were smooth and consisted none of microornamentation of pits/
pores and ridges (Figure 8-A and C). These structures were common to all of
specimen investigated in this work. However, dorsal scales with magnification of
x1000 and x500 showed many follicles on the entire surface of the boundary
scales (Figure 8-B and D).
The SEM observation of the dorsal scales surfaces also showed overlapping
scales, which is common in the reptile taxon. The dorsal midbody scales were
larger than those of dorsal caudal scales. Scales boundaries of the dorsal midbody
scales were covered by overlapping of large scales.
Position III: ventral midbody region. In general, the ventral mid-body scales
of M. ikaheka were larger than other body scales. Several ventral scales of the
snake have specialized ridge-like microornamentation (Figure 9-A, B, and C).
There were at least two ridges on each ventral scales with irregular arrangement.
As shown by the dorsal scales, the ventral boundary between scales showed many
follicles as well (see Figure 9-D). However, the follicles on the ventral scale lay
between the two scales, in contrast with the follicles on dorsal scales that were
located at the base of the scales.

Figure 8 SEM images illustrating the microstructure of the oberhautchen
on the dorsal outer scale surface of the mid-body (A,B) and
caudal (C,D) scales

17

Figure 9 SEM images illustrating the microtopography of the oberhautchen on
the ventral outer scale surface of the mid-body. Black group (A),
brown group (B), yellow group (C), and ventral scales boundary (D).

Discussion
Zoogeographical Distribution of M. ikaheka in Papua
According to the most generally accepted classification (O‟Shea 1996), the
yellow population is the subspecies of M.i. ikaheka with the distribution
throughout Vogelkop region, including Salawati Island in north-western Papua.
The brown group with dorsal banding pattern (M.i fasciatus) inhabits Yapen
Island in northern Papua, and most of the New Guinea mainland: northern,
southern and central region to Papua New Guinea. Surprisingly, an intermediate
color appears between both groups (sub-species) in southwest Vogelkop, Tanah
Merah, lowland forest of Bintuni, plus a new group (black group) from Batanta
and Waigeo Island (Krey 2009). The intermediate individual might indicated the
Bintuni as the hybrid zone of M.i. ikaheka from the Vogelkop region with M.i.
fasciatus from the other regions.
Coloration of M. ikaheka indicated a variable locally. This study has no
sufficient data to explain the variation in color pattern of the M. ikaheka in Papua.
However, it explained the possibility of the current population of M. ikaheka
founded following colonization events (dispersals) or the occurrence connections
between formerly populat