Effects of simulated ship transport on the susceptibility of the Green Mussel Perna viridis to heat stress
EFFECTS OF SIMULATED SHIP TRANSPORT ON THE
SUSCEPTIBILITY OF THE GREEN MUSSEL Perna viridis
TO HEAT STRESS
YASSER AHMED
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
i
ii
ISSUES RELATED TO THIS THESIS AND THE SOURCE
OF INFORMATION
With this I declare that this thesis with title Effects of Simulated Ship
Transport On The Susceptibility Of The Green Mussel Perna viridis To Heat
Stress is my own work under the direction of an advisory committee. It has not yet
been presented in any form to any Education institution. The sources of
information which is published or not yet published by other researchers have
been mentioned and listed in the references of this thesis.
Bogor, July 2013
Yasser Ahmed
C551100021
Copyright delegation on the thesis of research collaboration with outside parties
other from IPB must based on related cooperation agreement
iii
RINGKASAN
YASSER AHMED. Pengaruh simulasi transportasi kapal terhadap kerentanan
Kerang Hijau Perna viridis pada stress suhu yang meningkat. Dibawah bimbingan
NEVIATY PUTRI ZAMANI and KAREN VON JUTERZENKA.
Pergerakan organisme dipengaruhi oleh penyebaran alami dan aktivitas
manusia. Spesies yang terbawa memiliki sebaran yang melebihi distribusi batas
normal oleh pengaruh tertentu (vector). Pergerakan organisme secara alami
memiliki pergerakan yang terbatas. Spesies yang terbawa dapat mempengaruhi
kerentanan pada stress yang baik disengaja maupun tidak disengaja. Spesies yang
terbawa karena pengaruh alam seperti organisme yang bergerak akibat pergerakan
arus dan beberapa organisme ada yang dapat bermigrasi ke area yang lain yang
dapat melalui pergerakan arus; spesies yang terbawa akibat pengaruh aktivitas
manusia baik disengaja seperti perdagangan ikan hias, aktivitas budidaya atau
kegiatan manusia yang tidak disengaja seperti masukan air ballast dalam kapal
(Baker et al. 2007). Air ballast digunakan untuk menjaga keseimbangan kapal dan
beberapa organisme memiliki kemampuan untuk menempel pada permukaan atau
dinding dari tanki air ballast kapal tersebut (Mead et al., 2011). Pada umumnya
spesies yang terbawa oleh kapal sering terjadi dan ini terkait dengan padatnya
aktivitas perjalanan kapal kargo yang ukurannya lebih dari 10.000 GT selama
tahun 2007 (Kaluza et al., 2010). Menurut data dari Global cargo ship network
(GCSN), pergerakan kapal mencapai lebih dari 80% yang membawa barangbarang ke seluruh dunia. Masalahnya bahwa pergerakan kapal ini tidak hanya
membawa barang-barang, akan tetapi juga membawa organisme yang ada dalam
badan atau tanki air ballast kapal (Kölzsch and Blasius, 2011). Pertanyaannya
adalah apa yang terjadi pada spesies yang terbawa selama transportasi kapal?
Spesies yang terbawa terdapat berbagai mekanisme (vector) yang tersedia untuk
membawa organisme yang bersifat invasi akibat pengaruh perjalanan kapal dan
menyebar dalam jumlah yang besar; akan tetapi hanya dalam jumlah kecil yang
dapat bertahan dan menjadi pesaing (competitor) dengan spesies asli dalam
mendapatkan makanan di lingkungan yang baru, sehingga ini disebut sebagai
spesies yang invasif (invasive species). Spesies yang berinvasif dianggap sebagai
spesies pendatang atau tidak asli yang dapat menyebabkan kerusakan ekosistem,
sehingga berdampak pada perekonomian setempat, dan juga kesehatan masyarakat
(Bax et al., 2003; ISAC, 2006).
Menurut Lenz et al (2011), yang membandingkan spesies asli dan tidak
asli pada invertebrata yang dianggap memiliki toleransi terhadap stress. Enam
Bivalvia (diawali dengan spesies asli): Brachidontes exustus dan Perna viridis
(Trinidad), P. perna dan Isognomon bicolor (Brazil), Saccostrea glomerata dan
Crassostrea gigas (New Zealand); dua Ascidians: Diplosoma listerianum dan
Didemnum vixellum (Wales); dan dua Crustaceans: Gammarus zaddachi dan G.
tigrinus (Finland) yang diinvestigasi. Mereka menemukan bahwa spesies tidak
asli lebih resistan terhadap stress seperti stress salinitas (desalination), stress suhu
(warming) dan stress oksigen (hypoxia) yang sesuai dengan taksonomi dan
ekologi yang tersedia. Menurut Kleunen et al. (2010), bahwa toleransi terhadap
abiotik stress adalah kunci yang penting dalam menentukan spesies yang invasif.
iv
Tentunya hasil yang didapat oleh Lenz et al (2011) menarik dan oleh karena itu
perbedaan yang menonjol pada masing-masing spesies memiliki toleransi
terhadap stress yang berbeda pula. Satu peluang dalam menjelaskan observasi ini
adalah apa yang terjadi pada kondisi yang kritis bagi spesies selama transportasi
yang terdapat pada badan dan tanki air ballast kapal.
Spesies yang terbawa oleh kapal semakin meningkat sejak abad yang lalu
(Mead et al., 2011), dan tentunya perlu meniru atau simulasi transportasi kapal
untuk mengetahui apakah spesies yang terbawa dapat meningkat ketahanannya
selama transportasi. Tujuan dari penelitian ini untuk menginvestigasi apakah P.
viridis dapat meningkatkan toleransi terhadap suhu selama transportasi.
Pendekatan eksperimen ini sebaiknya memiliki keterkaitan dengan transportasi
kapal dan dibuat secara sederhana. Desain penelitian ini disesuaiakan dengan
kondisi di laboratorium. Hipothesis dari penelitian ini adalah grup pre stress dapat
meningkatkan toleransinya selama transportasi.
Berdasarkan hasil yang didapat dari penelitian ini menunjukkan bahwa P.
viridis tidak dapat meningkatkan toleransinya terhadap suhu selama simulasi
transportasi kapal, dan juga terdapat penurunan jumlah benang byssus, serta
penurunan berat badan selama transportasi.
Penelitian ini dilakukan sebagai bagian dari proyek GAME-X (Global
Approach by Modular Experiment) yang bekerja sama dengan Geomar Jerman
dan beberapa universitas dunia (Jerman, Brasil, Chili, Portugal), penelitian
dilakukan untuk membandingkan kerentanan spesies terhadap stress.
v
SUMMARY
YASSER AHMED. Effects Of Simulated Ship Transport On The Susceptibility
Of The Green Mussel Perna viridis To Heat Stress. Under supervised by
NEVIATY PUTRI ZAMANI and KAREN VON JUTERZENKA.
The introduced species are species which exceed their natural distribution
limits (native range) by a certain mode of introduction (vector). Natural
movements are responsible to limited extent. These introduction of a species
could influence the susceptibility of a species to environmental stress no matter if
introduced purposely or unpurposely. Introduction of species can happen caused
by natural movement i.e. organism can movement influenced by current and some
organism can migrate to one area to other area; and human activity either
purposely i.e. aquarium trade, aquaculture or unpurposely such as accidentally
through fouling on ship hulls (Baker et al., 2007). But dominant vector which
have large contribution on species introduction is transport by ship hulls and
ballast water tanks (Mead et al., 2011). Ballast water tank used to keep the
balancing of the ship and some organism has ability to attach on the wall of ship
hulls. Introduction species by ships mostly happened where a lot of traffic journey
of cargo ship bigger than 10.000 GT during 2007 (Kaluza et al., 2010). Based on
data from Global cargo ship network (GCSN), shipping moves over 80 % of the
world the goods. The problem is that ships do not only bring the goods but also
bring species in the ship hulls and ballast water tank (Kölzsch and Blasius, 2011).
The question is what happened to transport condition? For introduction of species,
there are a wide variety of transfer mechanisms (vectors) available for hitchhiking
of marine invaders to travel and spread in large amount of individuals; but only
small amount of individuals can be robust and act as competitors to get food
supply in the new environment, it called invasive species. Marine invasive species
are defined as non-native species that cause or are likely to cause harm to
ecosystems, economies, and or public health (Bax et al., 2003; ISAC, 2006).
Based on Lenz et al (2011), they compare native and non-native marine
invertebrates regarding their tolerance to stress. Six Bivalve (native named first):
Brachidontes exustus and Perna viridis (Trinidad), P. perna and Isognomon
bicolor (Brazil), Saccostrea glomerata and Crassostrea gigas (New Zealand); two
Ascidians: Diplosoma listerianum and Didemnum vixellum (Wales); and two
Crustaceans: Gammarus zaddachi and G. tigrinus (Finland) has been investigated.
They found that non native are more resistant towards environmental stress, such
as desalination, warming and hypoxia, than taxonomically related and
ecologically similar native species. Kleunen et al. (2010) said that tolerance
towards abiotic stress is a key trait that determines the invasiveness of species.
Furthermore, this was surprise and therefore pronounced differences in tolerant
towards stress exist between different species. One possible explanation for this
observation are adverse condition during transport, e.g. in the ship hulls and
ballast water tanks of cargo vessels.
The introduction of species more increased since past century (Mead et al.,
2011), and needed mimicking ship transport to know whether that species can
vi
increase their tolerance during transport. The objective for this experiment is to
investigate whether P. viridis can increase their tolerance during transport. The
experimental approach have connection with simulated ship transport and be
rather simple. This experimental design is design to mimicking transport
condition in short term lab experiments. Hypothesis for this experiment is pre
stress group can increase their tolerance during transport.
Based on the result shows that P. viridis could not increase their tolerance
during simulated ship transport, and they decrease amount of byssus threads, also
they decrease their weight during transport.
This research was conducted as part of the Global Approach by Modular
Experiment (GAME-X) project, to do a latitudinal comparison of susceptibility to
stress.
vii
Copyright © 2013 Bogor Agricultural University
Copyright are Protected by Law
1.
2.
It is a prohibited to cite all or part of this thesis without referring to and
mentioning the source.
a. Citation only permitted for the sake of education, research, scientific
writing, report writing, critical writing, or reviewing scientific problem.
b. Citation doesn’t inflict the name and honor of Bogor Agricultural
University.
It is prohibited to republish and reproduce all or part of this thesis without
the writer permission from Bogor Agricultural University.
viii
EFFECTS OF SIMULATED SHIP TRANSPORT ON THE
SUSCEPTIBILITY OF THE GREEN MUSSEL Perna viridis
TO HEAT STRESS
YASSER AHMED
Thesis
as one of the requirements for achieving
Master of Science degree in
Marine Science Program
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
ix
Outside Committee Examiner on Thesis Examination:
Prof. Dr. Ir. Dedi Soedharma, DEA
x
Title
Name
NIM
: Effects of simulated ship transport on the susceptibility of the
Green Mussel Perna viridis to heat stress
: Vasser Ahmed
: C5511 00021
Approved by
Advisory Commiteee
tL/)!(
Dr. Ir. Neviatv P. Zamani. M.Sc.
(Head)
Dr. Karen von Juterzenka
(Member)
Known by
Head of Marine Science Program
Dr. Ir. Neviatv P. Zamani. M.Sc.
Date of Examination:
July
Date of Graduation:
2013
2 6 JUL 2613
Title
Name
NIM
: Effect of simulated ship transport on the susceptibility of the
Green Mussel Perna viridis to heat stress.
: Yasser Ahmed
: C551100021
Approved by
Advisory Commiteee
Dr. Ir. Neviaty P. Zamani, M.Sc.
(Head)
Dr. Karen von Juterzenka
(Member)
Known by
Head of Marine Science Program
Dean of Graduate School
Dr. Ir. Neviaty P. Zamani, M.Sc.
Dr. Ir. Dahrul Syah, M.Sc.Agr.
Date of Examination:
July 8th 2013
Date of Graduation:
xi
PREFACE
Praise and thanksgiving the author to Allah SWT for the bounty so that
this thesis succesfull to done. Choosen of theme on this experiment was done on
April to September 2012 and related with the survival of Green Mussel Perna
viridis, the title are Effect of Simulated Ship Transport On The Susceptibility Of
Green Mussel Perna Viridis To Heat Stress.
The author want to say thanks to Dr. Neviaty P. Zamani, Dr. Karen von
Juterzenka, and Dr. Mark Lenz as supervisor. On other hand, the author give
thanks to my partner for the experiment in Indonesia are Armin Fabritzek and
other participant of GAME-X Project.
Hopefully this thesis could be useful.
Bogor, July 2013
Yasser Ahmed
xii
ACKNOWLEDGEMENT
The author want to say thank you to all the parties who supports and help,
so that this research and thesis can be done.
1.
Parents (Nasrun Tasila and Juliani also Djarot Soedarsono and Sri Ulfah
Hartati), Lettu (Inf) Nasser Khaled and Leila Nabila (Brother and Sister)
and all the family who patiently supports, inspires, motivate, help, and pray
for keep reminding to do the best, on time, always learn, humble, and work
hard and smart for the better and the best to reach the goals.
2.
Wife (Dian Respati Widianari) who support, inspires, motivate, help, as
best friend to discuss and share, and pray to achieve our dreams also for keep
reminding to do the best, on time, always learn, humble, and work hard and
smart for the better and the best to reach the goals.
3.
Dr. Ir. Neviaty Putri Zamani, M.Sc. as the head of advisory committee and
head of Marine Science Program who patiently give support, advice, help,
and time through finishing of this thesis.
4.
Dr. Karen Von Juterzenka as member of advisory committee who gave a
lot of support, help, time, advice, new experience (for me), and share
knowledge and fun through finishing of this thesis.
5.
Dr. Mark Lenz and Prof. Martin Wahl as project coordinator Global
Approach by Modular Experiment (GAME X) from Geomar – Germany who
gave the opportunity to join their project and broaden my knowledge,
experience, and network.
6.
GAME-X Team: Armin Fabritzek (my partner), Felipe Ribeiro and Lisa
Oberschelp (Team Brazil), Sandra Eichorn and Miguel Angel Penna (Team
Chile), Marie Garcia and Filipa Antunes (Team Portugal)
7.
Prof. Dr. Ir. Dedi Soedharma, DEA as outside examiner committee who
help a lot giving advise thoroughly through this thesis so that this thesis can
be better.
8.
Prof. Dr. Ir. Utomo Kartosuwondo, MS. as seminar moderator and
examiner who give time in supporting and positive comment during the
seminar.
xiii
9.
Friends in Marine Science Program class 2010 Bogor Agricultural
University (Abd Muthalib Angkotasan, Adi Purwandana, (Alm) Yuida
Labetubun, Evangelin Kadmaer, Rezi Apri, Princy (for all the listening, help,
and many moments together)
10. Friends in Marine Habitat Lab (Dr. Michael K. Schmid, Mareike Huhn,
Nurina Ayu, Rebecca Mueller, Giannina Hattich, Titan, Shelly Tutupoho,
Andre Wizemann, Veronica, David, Wira, Nora Hase (for helping and
accompanying study) thank you for all the good times, laugh, advise, critics,
info, support, and help during we study and working together.
11. Dondy Arafat, thank you for helping in providing seawater to the lab.
12. Aris and Pak Baso as Fisherman in Muara Kamal who helped us to take
sample in Jakarta Bay.
13. All parties and everybody who not yet mentioned, thank you for all supports,
help, and time through finishing this thesis.
xiv
CONTENTS
LIST OF TABLES
xvii
LIST OF FIGURES
xvii
1. INTRODUCTION
Background
Objectives
Hypothesis
1
1
2
2
2. LITERATURE REVIEW
Transport of organism
Natural dispersal
Human mediated dispersal
Vector of marine introduction
Stressors
Bivalves
Perna viridis
The spread of Green Mussel (P. viridis)
Introduction of species
3
3
3
3
6
8
9
10
11
12
3. RESEARCH METHODOLOGY
Time and Location
Materials
Methods
Organism choice and sample sites
Sampling and transport the organism
Acclimation and feeding
Experimental design and data analysis
Laboratorium experiment
Water quality control
Temperature
Problematics
Experiment
Pilot study
Set up and main experiment
Response variables
Survivorship, Byssus, Body condition index (BCI)
Position of mussels
Response behaviour
Daily routine
14
14
14
17
17
19
20
21
23
23
24
24
24
24
25
26
26
27
27
28
4. RESULTS AND DISCUSSION
Results
Species identification
Acclimation
Experiment
29
29
29
29
29
xv
CONTENTS (continued)
Survivorship of the mussels during the pilot study
Performance of the mussels during the main experiment
Survival
Byssus
Body condition index (BCI)
Position of mussels
Response behaviour
Discussion
Adverse condition during transport
Number of byssus during transport
Growth performance during transport
Position of mussels during transport
Response behaviour during transport
29
30
30
32
32
33
36
40
40
41
41
41
42
5. CONCLUSION AND SUGGESTION
Summary
Suggestion
43
43
43
REFERENCES
44
xvi
LIST OF TABLES
1.
2.
Anthropogenic vectors for marine introduction (Bax et al, 2003)
Materials that was used to perform this experiment
7
14
LIST OF FIGURES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
(a) Cumulative number of marine introduced species recorded
since 1840. (b) Liner regression indicating rate of discovery of
marine introduced species since 1840 (Mead et al., 2011)
The trajectories of all cargo bigger than 10.000 GT during 2007.
The colour scale indicates the number of journeys along each routes.
Ships are assumed to travel along the shortest (geodesic)
paths on water (Kaluza et al., 2010)
Habitat distribution of marine introduction (Mead et al., 2011)
Main vectors of marine introduction (Mead et al., 2011)
Marine introduced species across taxonomic groups (Mead et al., 2011)
Laterally compressed body is the most distinctive feature of
bivalves (Castro and Huber, 2007).
(a). Perna viridis were measured in a ruler; (b) Perna viridis
on the glass tank; (c) Fresh harvest of Perna viridis
Approximate native and introduced range of the Asian Green Mussel
P. viridis, base on the literature review (Baker et al. 2007)
Mytillus galloprovincialis dominating fouling assemblages in the
sea chest of the SA “Agulhas” (Lee and Chown, 2007)
Asian Green Mussel, P. viridis (dark object) and Eastern Oyster,
Crassostrea virginica (white object) on a bridge pier in Tampa Bay,
Florida, USA, at low tide in 2001. The majority of mussels in this
photo are 2-3 cm in shell length. (Baker et al. 2007)
(a). P. viridis; (b). Fresh harvest of mussels from Jakarta Bay
(a). Muara Kamal, Jakarta Bay; (b). Sub-district Muara Kamal;
(c). Muara Kamal in Java Island (red colour)
(Created by: Annisa Indah Sari, 2012)
Sampling organisms and measured the size;
(b) Mussels in the cool box; (c) Cleaned the mussels
Acclimation tanks in the lab.
Simulated ship transport from Europe to Australia.
Mimicking transport condition for this experiment
Main groups for this experiment
Scheme of pilot study
(a). Single containers on the fiber tanks for pilot study;
(b) Each containers fixed in the frame for pilot study
(a). Wooden frame for thermal stress phase; (b). Rubber
frame for control group; (c). Tank to pre heat water
xvii
4
5
5
8
8
10
11
12
13
13
18
18
19
20
22
22
23
24
25
26
LIST OF FIGURES (continued)
21. Measuring BCI of the mussels.
22. (a) Water exchanged; (b) Feeding the mussels
23. Survivorship of the mussels connected with
temperature during pilot study.
24. Survivorship of the mussels for pre stress group without
recovery
25. Survivorship of the mussels for pre stress group with
recovery
26. Byssus of the mussels for each groups
27. Body condition index (BCI) of the mussels each period
28. Position of mussels for group B (stress group without
recovery) (S: surface; M: middle; B: bottom; Ai: air stone;
NA: not attach)
29. Position of mussels for group BC (non stress group
without recovery) (S: surface; M: middle; B: bottom; Ai: air
stone; NA: not attach)
30. Position for control group (S: surface; M: middle; B: bottom;
Ai: air stone; NA: not attach)
31. Position of mussels for group A (stress group with recovery),
(S: surface; M: middle; B: bottom; Ai: air stone; NA: not attach)
32. Position of mussels for group C (non stress group with recovery)
(S: surface; M: middle; B: bottom; Ai: air stone; NA: not attach)
33. Position for control group (S: surface; M: middle;
B: bottom; Ai: air stone; NA: not attach)
34. Response behaviour group B (stress group without
recovery) (F: Fast; C: Closed; S: Slow)
35. Response behaviour group BC (non stress group
without recovery) (F: Fast; C: Closed; S: Slow)
36. Response behaviour for control (F: Fast; C: Closed; S: Slow)
37. Response behaviour group A (stress group with recovery)
(F: Fast; C: Closed; S: Slow)
38. Response behaviour group C (non stress group with recovery)
(F: Fast; C: Closed; S: Slow)
39. Response behaviour for control (F: Fast; C: Closed; S: Slow)
xviii
27
28
30
31
31
32
33
33
34
34
35
35
36
37
37
38
38
39
39
1
1 INTRODUCTION
1.1 Background
The introduced species are species which exceed their natural distribution limits
(native range) by a certain mode of introduction (vector). Natural movements are
responsible to limited extent. These introduction of a species could influence the
susceptibility of a species to environmental stress no matter if introduced
purposely or unpurposely. Introduction of species can happen caused by natural
movement i.e. organism can movement influenced by current and some organism
can migrate to one area to other area; and human activity either purposely i.e.
aquarium trade, aquaculture or unpurposely such as accidentally through fouling
on ship hulls (Baker et al. 2007). But dominant vector which have large
contribution on species introduction is transport by ship hulls and ballast water
tanks (Mead et al. 2011). Ballast water tank used to keep the balancing of the ship
and some organism has ability to attach on the wall of ship hulls. Introduction
species by ships mostly happened related with a lot of traffic journey of cargo ship
bigger than 10.000 GT during 2007 (Kaluza et al. 2010). Based on data from
Global Cargo Ship Network (GCSN), shipping moves over 80 % of the world the
goods. The problem is that ships do not only bring the goods but also bring
species in the ship hulls and ballast water tank (Kölzsch and Blasius, 2011). The
question is what happened to transport condition? For introduction of species,
there are a wide variety of transfer mechanisms (vectors) available for hitchhiking
of marine invaders to travel and spread in large amount of individuals; but only
small amount of individuals can be robust and act as competitors to get food
supply in the new environment, it called invasive species. Marine invasive species
are defined as non-native species that cause or are likely to cause harm to
ecosystems, economies, and or public health (Bax et al. 2003; ISAC, 2006).
Based on Lenz et al (2011), they compare native and non-native marine
invertebrates regarding their tolerance to stress. Six Bivalve (native named first):
Brachidontes exustus and Perna viridis (Trinidad), P. perna and Isognomon
bicolor (Brazil), Saccostrea glomerata and Crassostrea gigas (New Zealand); two
Ascidians: Diplosoma listerianum and Didemnum vixellum (Wales); and two
Crustaceans: Gammarus zaddachi and G. tigrinus (Finland) has been investigated.
They found that non native are more resistant towards environmental stress, such
as desalination, warming and hypoxia, than taxonomically related and
ecologically similar native species. Kleunen et al. (2010) said that tolerance
towards abiotic stress is a key trait that determines the invasiveness of species.
Furthermore, this was surprise and therefore pronounced differences in tolerant
towards stress exist between different species. One possible explanation for this
observation are adverse condition during transport, e.g. in the ship hulls and
ballast water tanks of cargo vessels.
The introduction of species more increased since past century (Mead et al.
2011), and needed mimicking ship transport to know whether that species can
increase their tolerance during transport. The objective for this experiment is to
investigate whether P. viridis can increase their tolerance during simulated ship
2
transport. The experimental approach have connection with simulated ship
transport and be rather simple. This experimental design is design to mimicking
transport condition in short term lab experiments. Hypothesis for this experiment
is pre stress group can increase their tolerance during transport.
This research was conducted as part of the Global Approach by Modular
Experiment (GAME-X) project, to do a latitudinal comparison of susceptibility to
stress with theme “Do adverse condition during transport select for stress tolerant
genotypes in founder population of native and non-native marine invertebrates”.
1.2 Objectives
The objective for this experiment is to investigate whether P. viridis can
increase their tolerance during transport.
1.3 Hypothesis
Pre-stress can increase their tolerance during transport.
3
2 LITERATURE REVIEW
2.1 Transport of organisms
Transportation network play crucial role in human mobility, distributed the
goods and spread species. Transport of organisms has two factors was natural
movements and human activity. Large contribution to transport of organism was
caused transportation of ships where most of world trade is hauled by ships and
7,4 billion tons of goods were spread to the worldwide (International Maritime
Organization (IMO), 2006) and also spread of species.
2.1.1 Natural dispersal
At first movement of organisms influenced by natural dispersal. Some
organism has limited movements so that they follow natural influence by current
(plankton) and other organism can migrates through the ocean current such as sea
turtle, whales etc. Perna viridis are similar to those of Mytilus species but no one
has determined whether P. viridis has a planktonic postlarval phase like some
other members of the family (Baker and Mann, 1997). P. viridis has byssus to
attach on the surface of substrata so that they was easily can invasive to other
area. One of possibility is influenced by natural dispersal where driftwood
movement, plastic and other human debris was increased important factors for
introduction of organisms (Winston 1982, Aliani and Molcard 2003, Barnes and
Fraser 2003 in Baker et al. 2007). Other possibility natural objects which can
found anywhere was occurred in globalization times because of anthropogenic
factors (Baker et al. 1999).
Marine invertebrates specially for P. viridis has large distribution. P.
viridis was native species and distributed in Indo-Pacific such as Philippines,
Sumatra, Borneo, Bali, Java, Vietnam, China (Siddal, 1980) and South Asia
(Baker et al. 2007). Secondary sources P. viridis was native range in island of
New Guinea (FAO fisheries, 2006).
2.1.2 Human mediated dispersal
One of factor organism transport and spread is human mediated dispersal.
Marine invertebrates can cross to different latitudes or other places by ship
transport. Some marine invertebrates have the ability to attach on the wall of ship
hulls and some larvae inside in ballast water tank. Most of the commodities or the
goods in the world are transported by cargo ships and transferred species to other
places (Kölzsch and Blasius 2011). Since industrial development of transported
was established, humans increased the size of ship because trade commodity. The
ballast water tank used to get balanced of ships so that secure to bring the goods.
The impact of species introduction with deballasting can be pass through ballast
water intakes are known to survive transportation over long distances (Flagella et
al. 2007). One of case the number of species introduction increased in South
4
African coast since 1840 because industrial development of ship (Mead et al.
2011) (Figure 1).
Figure 1 (a) Cumulative number of marine introduced species recorded since
1840. (b) Liner regression indicating rate of discovery of marine
introduced species since 1840 (Mead et al. 2011)
Transportation network play a crucial role in human mobility to transfer
and movement goods and the impact would spread the species. Around 90% of
world trade is hauled by ships (International Maritime Organisation, 2006). Two
major pathways for marine bioinvasion are discharged water from ships ballast
water tank and hull fouling. Ballast water in cargo ship contribute large transport
of organisms around 3 to 4 billion tons per year in the ports of the world (Kölzsch
and Blasius, 2011). European ports, American, Asian, African and Australian also
contain among of the high number invasive species. The financial was loss in
estimated to be $120 billion in US alone (Kaluza et al. 2010). The transportation
network can be seen in Figure 2. Based on the Figure 2, large size of ship more
traffic with the transportation network and would be give impact to marine
introduction. Dominant the distribution for marine introduction was in the harbour
where the ship docked (Mead et al. 2011) (Figure 3).
5
Figure 2 The trajectories of all cargo bigger than 10.000 GT during 2007. The
colour scale indicates the number of journeys along each routes. Ships
are assumed to travel along the shortest (geodesic) paths on water
(Kaluza et al. 2010)
Figure 3 Habitat distribution of marine introduction (Mead et al. 2011)
6
2.1.3 Vector of marine introduction
Increasingly the introduction of alien species is a major threat to marine
biodiversity and a contributor to environmental change. As these marine
introduction, intentional and accidental can result from numerous human mediated
activities, management responses need to cover a diverse range of human activity.
Based on historical data, a new marine or estuarine species establishes itself every
32-85 weeks in each of six ports studied in the US, New Zealand and Australia, a
rate that appears to be increasing. While many of the alien species become part of
the background flora and fauna, others become invasive and come to dominate the
native flora and fauna. There are 15 broad categories of vectors that transport
marine organisms from shallow coastal waters to similar habitats outside the
species home range which can be seen in Table 1 (Bax et al. 2003).
The main economic and social impacts of invasive alien marine species are
negative impacts on human health and decrease in economic production of
activities based on marine environments and resources such as fisheries,
aquaculture, tourism and marine infrastructure. These affect have related social
impacts through decreases in employment in economic activities directly affected
by invasive alien species but also through decreases in people’s welfare from the
reduced quality of their environments and natural surroundings. Some vectors for
alien marine species was happened caused human activity by vessel for trading
commodities and vessel for recreation. In the modern era, increase size of vessel
will give effect to introduction of organisms. Some case in Australia ports, one of
kind mussels P. viridis was introduce to Darwin harbour (Bax et al. 2003).
Characteristics such as dispersal, life history strategies, growth rates,
distributional ranges, association with humans, and tolerance towards
environmental stress are determinants of invasiveness for animal and plant species
(Braby and Somero, 2006; Lenz et al. 2011). One of possibility is that marine
organisms have the ability to move by ship hulls and ballast water. Mead et al
(2011), observed marine bioinvasion in South African and show that some vectors
marine organism introduce to the environment and most of the vectors dominant
to marine introduction was ship hulls and ballast water (Figure 4). Ballast water
released in the environment has lead to damage of establishment hundreds of
introduction organism to Europe and North America (Mills et al. 1996; Bij de
Vaate et al. 2002). One of factor was the ships of ballast water have potential to
transfer coastal marine species to the other area and this marked a shift from the
centuries-old practice of using rocks and sand from the shore to provide balance
for ships (Davidson and Simkanin, 2012). P. viridis is mussels where including
molluscs and marine organism could movement or introduction to other place
(Mead et al. 2011) (Figure 5).
7
Table 1 Anthropogenic vectors for marine introductions (Bax et al. 2003)
Source
Commercial shipping
Vector
Ballast water
Target taxa
Plankton,
nekton,
benthos in sediment
Hull fouling
Encrusting, nestling, and
some mobile species
Solid ballast (rocks, sand, Encrusting,
benthos,
etc)
meiofauna and flora
Aquaculture
and Intentional release for
Single species
fisheries
stock enhancement
Gear, stock or food
Various
movement
Discarded nets, floats,
Various
traps, trawls, etc
Discarded live packing
Various
materials
Release of transgenic
Single species
species
Drilling platforms
Ballast water
Plankton, nekton,
benthos in sediment
Hull fouling
Encrusting, nestling, and
some mobile species
Canals
Movements of species
Various
through locks due to
water motion or active
swimming
Aquarium industry
Accidental or intentional Aquarium fauna and
release
flora
Recreational boating
Hull fouling
Encrusting, nestling, and
some mobile species
Dive practices
Snorkeling and scuba Algal spores, bacteria,
gear
some mobile species
Floating debris
Discarded plastic debris
Encrusting, and some
mobile species
8
Figure 4 Main vectors of marine introduction (Mead et al, 2011)
Figure 5 Marine introduced species across taxonomic groups (Mead et al, 2011).
2.2 Stressors
Temperature as one of suitable stressor with condition in ballast water tank
and give effect to introduction species. Most of the ship material was metal and it
act as conductor which easily influenced by temperature. On other hand,
temperature (heat) is a relevant stressors for benthic, shallow water organism and
they also act on organisms shuttled in ballast water tanks or on ship hulls. This
9
stressor (temperature) also related with condition in ship hulls and ballast water
tank because when some species come to inside ballast water tanks and they
influenced by temperature; then, when some species attach in the ship hulls and
the journey has different lattitude and of course influenced different temperature
as well (Schneider, 2008; Seiden et al. 2011).
Temperature is one of important factor to control geographical range and
seasonal cycles of marine organism. As general, organism that maintain their body
temperature by absorbing heat from the environment (ectotherm). Temperature
too high can act as stressor in two ways: First, it may cause the denaturing of
sensitive proteins; This damage can be minimized by the actions of heat shock
proteins; which increase the thermostability of proteins and chaperone cellular
processes. Second, high temperatures may cause oxygen limitation (Jansen et al.
2007). Temperature also has a linier correlation with heart rate; high temperature
lead to fall in heart rate (Akberali and Trueman 1985).
Species “biogeographical” has tolerance to environmental extremes.
Intertidal species is one of organism has tolerance to environmental stress such as
temperature and salinity (Castro and Huber 2007).
2.3 Bivalves
Class Bivalvia consist of clams, mussels, and oysters. The body of bivalvia
is laterally compressed and enclosed in a shell with two parts or valves. The umbo,
the upper hump near the hinge of each shell is the oldest part of the shell. The
form of concentric growth lines is growth from the umbo. The inner surface of the
shell is lined by the mantle, so that the whole body lies in the mantle cavity. The
mantle cavity is a large space between the two halves of the mantle. Special for
the Perna viridis has ability attach to the rocks and other surface by byssal threads
or byssus (Castro and Huber, 2007). Structure the body of bivalve can be seen in
Figure 6.
10
Figure 6 Laterally compressed body is the most distinctive feature of bivalves
(Castro and Huber, 2007).
2.3.1 Perna viridis
Perna viridis has the common name Asian Green Mussel or Green Mussel
and they live in intertidal zone. They has ability to attach in the wall, so they can
live on a variety of structures such as vessels, wharves, mariculture equipment,
buoys and other hard substrate. This species is filter feeder, feeding on small
zooplankton, phytoplankton and other suspended organic material (Castro and
Huber 2007; National Introduced Marine Pest Information System (NIMPIS). P.
viridis can be seen in Figure 7A, 7B and 7C.
11
a.
b.
c.
Figure 7 (a). Perna viridis were measured in a ruler; (b) Perna viridis on the
glass tank; (c) Fresh harvest of Perna viridis.
The taxonomical name of Perna viridis (Linnaeus, 1758) is:
Phylum: Mollusca
Class: Bivalvia
Subclass: Pteriomorphia
Order: Mytiloida
Superfamily: Mytiloidea
Family: Mytilidae
Genus: Perna
Species: Perna viridis
(Poutiers 1987; Carpenter and Niem 1998; Leal 2002)
2.3.2 The Spread of Green Mussel (P. viridis)
Native distribution of P. viridis extends from the Persian Gulf to all of
Philippines, Sumatra, Borneo, Bali, Java, Sulawesi, Northeast Vietnam (Siddal,
1980). Siddal (1980) reported that presence of P. viridis in Hongkong as
introduced species. The island in New Guinea also include native range (FAO,
2006). Gindy et al (2001) reported, distribution of P. viridis also spread in
Musandam Peninsula of Oman. Most of P. viridis distributed in Indo-Pacific.
12
Green Mussel also one of fouling organism and ships have been sailing to many
area (Kaluza et al. 2010), it is possible P. viridis have been introduced to Florida,
Jamaica, Trinidad, China, Japan (Figure 8). Some evidence that P. viridis
dominating in fouling and introduced to the coast of Florida (Figure 9 and Figure
10).
Figure 8 Approximate native and introduced range of the Asian Green mussel,
P. viridis, based on the literature review (Baker et al. 2007)
2.4 Introduction of species
Most of introduction of species happened by human activity such as
aquaculture, marine litter, oil rig, intentional, fisheries, solid ballast, ship boring,
ballast water, and ship fouling (Mead et al. 2011), but the dominant vector
organism introduced by ballast water and ship fouling. It is related with increase
number of ships which bring the good and distributed to worldwide. Ships carry
ballast water for vessel stability and trim, it is well established that ballast water
can contain some species such as mussels, metazoan, microorganism, some of
them are pathogenic and so on (Drake et al. 2002).
Species which introduced into new environment would survived because
ballast water tank support the survival of mussels and also they would attach on
the substrate and act as competitors to get food supply. Introduction of species
increased because no predatory in the new habitat and would alteration food chain
and impact to decreased native species (Baker et al. 2007).
13
Figure 9 Mytilus galloprovincialis dominating fouling assemblages in the sea
chests of the SA “Agulhas” (Lee and Chown, 2007).
Figure 10 Asian Green Mussel, P. viridis (dark object) and Eastern Oyster,
Crassostrea virginica (white object) on a bridge pier in Tampa Bay,
Florida, USA, at low tide in 2001. The majority of mussels in this
photo are 2-3 cm in shell length. (Baker et al. 2007)
14
3 RESEARCH METHODOLOGY
3.1 Time and Location
This research was conducted in the Marine Habitat Laboratory,
Department of Marine Science and Technology, Faculty of Fisheries and Marine
Sciences, Bogor Agricultural University (FPIK – IPB) from April to September
2012.
3.2 Materials
The experiment was used some materials and shown in Table 2. The
pictures of materials are shown in Appendix 1.
Table 2 Materials which used to perform this experiment
Material
s
Seawater
Air
pump
Brand
(Type)
Sonic
P-125
Small
hose and L,
I, T junction
Single
Jack
container
Family
Tank
Specification
Salinity: 33-35 ‰ from Teluk
Jakarta, filter processed sea water
in Gelanggang Samudera and
Seaworld, Ancol, Jakarta (Arafat,
pers.com., 2012)
AC 220-240 V
50 Hz
85 W
125 L/min
0.04 MPa
1.5 litre
Fiber tank
Functions
To supplyed
aeration
To
connected
aeration
Experiment
al container for
kept the
organisms
As water
bath for pilot
study and main
experiment
15
Table 2 Material which used to perform this experiment (continued)
Materials
Transparen
t pipe and
fishing net
Water
pump
Brand
(Type)
Specification
Amara AA
1800
Rubber
Heater
Thermo
logger
Aquatic
Nature and
Eheim Jager
Onset Hobo
Calliper
Scale
Thermome
ter Digital
“Lucky”
(pocket scale)
and Bosch
Krisbow
KW06-308
Temperature setting
Company: Aquatic
Nature and Eheim Jager
300 W
220-240 V
50-60 Hz
20-35 0C
Pendant temp
Part # UA-001-08
Patent 6.826.664
15 cm (max.)
0.05 cm (accuracy)
6 inches (max.)
1/128 inches (accuracy)
Digital scale
Capacity: 200 g x 0.01 g
400C – 2500C
400 – 4820 F
Resolution: 0.10
Functions
To build
frame (pilot
study)
Creating a
current in
waterbath so
that the heat
distributed
evenly
To build
frame
(experiment)
To
produced
heating for
increased and
decreased
temperature in
waterbath and
pre heat water
To recorded
the
temperature in
waterbath.
Measure
length and
width of
mussels
Measured
weight of
mussels
Measured
temperature in
single
container and
the tank
16
Table 2 Material which used to perform this experiment (continued)
Materials
Bamboo
stick
Brand (Type)
Specification
Bamboo
Functions
Tool to
checked
mussels
Camera
Canon and
Canon DSLR 600 and
Documentat
Casio
Casio QV-R100
ion
Food
Coral sands
Nanochloropsis
Food for the
(DT’s premium
aculata, Phaeodactylum
mussels
blend live marine tricornutum, Chlorella 2phytoplankton)
20 microns
Oven
Heraeus
To dry the
mussels
Data sheet
Paper
Notes
Refractomet
Atago
Salinity 0~100 ‰
Measure
er
(S/Mill-E)
salinity in the
(Hand-held
beginning and
refractometer,
in the end of
Made in Japan)
experiment
Nitrate and
API
Measure
For fresh and
Nitrite
(Aquarium
nitrite
and
saltwater aquarium
Aquarium test Pharmaceuticals)
Nitrite (NO2 ): 0–0.5– nitrate
strips
1–3–5–10
Nitrate (NO3-): 0–20–
40–80–160–200
Ammonia
API
Measure
For fresh and saltwater
Aquarium Test (Aquarium
ammonia
aquarium
Strips
Pharmaceuticals)
Ammonia (ppm
(mg/L)): 0 – 0.5 – 1.0
– 3.0 – 6.0
pH
API
Measure pH
For fresh and saltwater
Aquarium Test (Aquarium
aquarium
Strips
Pharmaceuticals)
PH freshwater: 6.0 –
6.5 – 7.0 – 7.5 – 8.0 –
8.5 – 9.0
PH saltwater: 6.0 – 6.5
– 7.0 – 7.5 – 8.0 – 8.5
– 9.0
17
Table 2 Material which used to perform this experiment (continued)
Materials
Brand (Type)
Specification
Tank filter and
pump system had
been provided by
Marine Habitat
Lab
Function
Filter and mix
seawater inside
the tank.
3.3 Methods
3.3.1 Organism choice and sample sites
The purpose to choose of mussels have some criteria for this experiment:
(1) the mussels native in Indo-pacific and was invasive to other area (Mead et al.
2011); (2) generally can found it in the ship hulls and ballast water tank (Lee and
Chown 2007). Thus, it is relevant for simulated ship transport; (3) it is easy to
collect P. viridis as it can be found in Jakarta Bay (Muara Kamal, Indonesia)
where it is cultured and harvested by local fishermen and has economic value
(Figure 11); (4) members of the same family, Mytilidae, can be found in other
countries participating in GAME X project, such as Brasil, Chile, Finland and
Portugal, so that we can compare our results to these countries.
Sample site was located in North Jakarta, Indonesia. P. viridis (Green
mussel) were taken from Jakarta Bay, Muara Kamal (Figure 12). The mussel was
taken on June 12th, 2012. The temperature on the sea water was 29 0C and salinity
was 32 ppt. Muara Kamal was the place to harvested the mussels which is
influenced by anthropogenic activity. The majority population of Indonesians live
in Java and therefore coastal resources are under considerable human pressure and
every year the Jakarta Bay show increase in eutrophication and sedimentation
levels, which had impact on marine communities (Damar, 2003; van der Meij et
al. 2009).
18
Figure 11 (a). P. viridis; (b). Fresh harvest of mussels from Jakarta Bay.
a
b
c
Figure 12 (a). Muara Kamal, Jakarta Bay; (b). Sub-district Muara Kamal; (c).
Muara Kamal in Java Island (red colour) (Created by: Annisa Indah
Sari, 2012).
19
3.3.2 Sampling and transport the organism
Samples for experiment and bring the samples from the site to the lab. The
mussels taken in Muara Kamal (Jakarta Bay) with some local fishermen and was
transported by boat to the field. The mussels taken in culture area and harvested
by local fishermen. Afterwards, the mussels were transported from Muara Kamal
to the lab by car. The organisms were put in plastic aquarium without seawater.
Ice was filled in the bottom of cool box and covered with sterofoam. Afterwards,
the plastic aquarium was put on the top of sterofoam so that the organisms stayed
cool and in low metabolism. The transport took two hours from Jakarta Bay to the
lab. After arrival in the lab, the organism were cleaned also from barnacles that
are attached to it (Figure 13). Afterwards, the organism were put into the glass
aquarium for acclimation.
a.
b.
c.
Figure 13 (a) Sampling organisms and measured the size; (b) Mussels in the cool
box; (c) Cleaned the mussels.
20
3.3.3 Acclimation and feeding
Acclimation is response organism to specific environmental change(s) e.g
temperature, salinity, oxygen and so on, that observed in the lab to distinguish
effect and response to these changes e.g. physiology, activity, growth, etc (Nielsen
1990; Braby and Somero 2006). So, the purpose of acclimation is that organism
get adapted to lab conditions (temperature, salinity, oxygen) until achieved 1%
mortality (GAME X). During acclimation, water exchange and feeding of the
mussels were done. Half of the sea water in glass aquarium (26 litre) was
exchanged every day. First to second day acclimation, water exchange was done
twice a day and next day water was changed once a day. Natural sea water was
provided by a 1000 liters tank system equipped with a mechanical and a biological
filter unit and is distributed to smaller glass aquaria. All organism must have been
acclimatized before use in the experiment. Acclimation tanks can be seen in
Figure 14.
The mussels were fed with coral sands DT’s premium blend live marine
phytoplankton (Nanochloropsis aculata, Phaeodactylum tricornutum, Chlorella 220 microns) which is a liquid phytoplankton food that had a concentration 1,58 x
107. The first day to second day of acclimation phase, the organism were not fed
due to stress from transportation. Afterwards, started to feed the mussels daily
with 0.0125 ml/ind with coral sands. They should adapt with the lab condition
first. The third day to next days, the organisms were fed.
Total number of organism for acclimation before pilot study was 168
mussels. Total number of organism for acclimation before main experiment was
480 mussels which is the mussels separated in 12 glass aquaria with aeration
inside. Each glass aquaria consist of 40 mussels.
Figure 14 Acclimation tanks in the lab.
21
3.3.4 Experimental design and data analysis.
Most of the ship material was metal and it act as conductor which easily
influenced by temperature. Therefore, heat stress was chosen since it is relevant
with condition inside ballast water tank or ship hulls. On other hand, heat stress is
a relevant stressors for benthic, shallow water organism where they also act on
organisms shuttled in ballast water tanks or on ship hulls. Species which attach in
the ship hulls and carried on the way which has different lattitude would
influenced by temperature as well (Schneider, 2008; Seiden et al. 2011).
This experiment was done to simulated ship transport from Europe to
Australia because they have a lot of traffic journey of cargo ship bigger than
10.000 GT in 2007 (Kaluza et al. 2010). It can be seen the traffic journey in
Europe to US; US to Asia Pacific; US to Australia; Europe to Austalia and so on.
Thus, the simulated took from Europe to Australia because the journey has
different temperature from temperate – tropical – temperate (Figure 15).
The experimental design is designed to mimicking transport conditions in the lab.
In laboratory experiments, organisms will be exposed to stress regimes likely to
occur during transport by ship and which cause moderate mortality until up to
80% around 3 weeks (pre stress). After pre-stress period, the survivors will be
given some time (~2 weeks) to recover under stress free conditions and the other
group exposed directly to the same stressor again (second stress). Survival rates
were measured as a response variable over the course around 3 weeks. Control
group, consisting of the same number of conspecifics which have not previously
been exposed to adverse condition but have been acclimatized to lab conditions,
will be treated the same way and serves as a reference or control. Finally, we
compared survival between pre-stressed and non stressed group to investigate
whether pre stress group can increase the robustness during transport (Figure 16).
This experiment have 4 groups which consist of pre-stress group with
recovery (group A); pre-stress group without recovery (group B); then, non stress
group (group D and group C). Thus, pre-stress group without recovery compared
with non stress group (group B and group D). Then, pre-stress group with
recovery compared with non stress group (group A and group C) (Figure 17).
22
Figure 15 Simulated ship transport from Europe to Australia.
Figure 16 Mimicking transport condition for this experiment
23
Figure 17 Main groups for this experiment
The data for this experiment were analyzed by using the R program
(version 2.14.2). Kaplan Meier curve to analysed survivorship (log rank test), and
normality to analysed byssus and body condition index (Shapir
SUSCEPTIBILITY OF THE GREEN MUSSEL Perna viridis
TO HEAT STRESS
YASSER AHMED
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
i
ii
ISSUES RELATED TO THIS THESIS AND THE SOURCE
OF INFORMATION
With this I declare that this thesis with title Effects of Simulated Ship
Transport On The Susceptibility Of The Green Mussel Perna viridis To Heat
Stress is my own work under the direction of an advisory committee. It has not yet
been presented in any form to any Education institution. The sources of
information which is published or not yet published by other researchers have
been mentioned and listed in the references of this thesis.
Bogor, July 2013
Yasser Ahmed
C551100021
Copyright delegation on the thesis of research collaboration with outside parties
other from IPB must based on related cooperation agreement
iii
RINGKASAN
YASSER AHMED. Pengaruh simulasi transportasi kapal terhadap kerentanan
Kerang Hijau Perna viridis pada stress suhu yang meningkat. Dibawah bimbingan
NEVIATY PUTRI ZAMANI and KAREN VON JUTERZENKA.
Pergerakan organisme dipengaruhi oleh penyebaran alami dan aktivitas
manusia. Spesies yang terbawa memiliki sebaran yang melebihi distribusi batas
normal oleh pengaruh tertentu (vector). Pergerakan organisme secara alami
memiliki pergerakan yang terbatas. Spesies yang terbawa dapat mempengaruhi
kerentanan pada stress yang baik disengaja maupun tidak disengaja. Spesies yang
terbawa karena pengaruh alam seperti organisme yang bergerak akibat pergerakan
arus dan beberapa organisme ada yang dapat bermigrasi ke area yang lain yang
dapat melalui pergerakan arus; spesies yang terbawa akibat pengaruh aktivitas
manusia baik disengaja seperti perdagangan ikan hias, aktivitas budidaya atau
kegiatan manusia yang tidak disengaja seperti masukan air ballast dalam kapal
(Baker et al. 2007). Air ballast digunakan untuk menjaga keseimbangan kapal dan
beberapa organisme memiliki kemampuan untuk menempel pada permukaan atau
dinding dari tanki air ballast kapal tersebut (Mead et al., 2011). Pada umumnya
spesies yang terbawa oleh kapal sering terjadi dan ini terkait dengan padatnya
aktivitas perjalanan kapal kargo yang ukurannya lebih dari 10.000 GT selama
tahun 2007 (Kaluza et al., 2010). Menurut data dari Global cargo ship network
(GCSN), pergerakan kapal mencapai lebih dari 80% yang membawa barangbarang ke seluruh dunia. Masalahnya bahwa pergerakan kapal ini tidak hanya
membawa barang-barang, akan tetapi juga membawa organisme yang ada dalam
badan atau tanki air ballast kapal (Kölzsch and Blasius, 2011). Pertanyaannya
adalah apa yang terjadi pada spesies yang terbawa selama transportasi kapal?
Spesies yang terbawa terdapat berbagai mekanisme (vector) yang tersedia untuk
membawa organisme yang bersifat invasi akibat pengaruh perjalanan kapal dan
menyebar dalam jumlah yang besar; akan tetapi hanya dalam jumlah kecil yang
dapat bertahan dan menjadi pesaing (competitor) dengan spesies asli dalam
mendapatkan makanan di lingkungan yang baru, sehingga ini disebut sebagai
spesies yang invasif (invasive species). Spesies yang berinvasif dianggap sebagai
spesies pendatang atau tidak asli yang dapat menyebabkan kerusakan ekosistem,
sehingga berdampak pada perekonomian setempat, dan juga kesehatan masyarakat
(Bax et al., 2003; ISAC, 2006).
Menurut Lenz et al (2011), yang membandingkan spesies asli dan tidak
asli pada invertebrata yang dianggap memiliki toleransi terhadap stress. Enam
Bivalvia (diawali dengan spesies asli): Brachidontes exustus dan Perna viridis
(Trinidad), P. perna dan Isognomon bicolor (Brazil), Saccostrea glomerata dan
Crassostrea gigas (New Zealand); dua Ascidians: Diplosoma listerianum dan
Didemnum vixellum (Wales); dan dua Crustaceans: Gammarus zaddachi dan G.
tigrinus (Finland) yang diinvestigasi. Mereka menemukan bahwa spesies tidak
asli lebih resistan terhadap stress seperti stress salinitas (desalination), stress suhu
(warming) dan stress oksigen (hypoxia) yang sesuai dengan taksonomi dan
ekologi yang tersedia. Menurut Kleunen et al. (2010), bahwa toleransi terhadap
abiotik stress adalah kunci yang penting dalam menentukan spesies yang invasif.
iv
Tentunya hasil yang didapat oleh Lenz et al (2011) menarik dan oleh karena itu
perbedaan yang menonjol pada masing-masing spesies memiliki toleransi
terhadap stress yang berbeda pula. Satu peluang dalam menjelaskan observasi ini
adalah apa yang terjadi pada kondisi yang kritis bagi spesies selama transportasi
yang terdapat pada badan dan tanki air ballast kapal.
Spesies yang terbawa oleh kapal semakin meningkat sejak abad yang lalu
(Mead et al., 2011), dan tentunya perlu meniru atau simulasi transportasi kapal
untuk mengetahui apakah spesies yang terbawa dapat meningkat ketahanannya
selama transportasi. Tujuan dari penelitian ini untuk menginvestigasi apakah P.
viridis dapat meningkatkan toleransi terhadap suhu selama transportasi.
Pendekatan eksperimen ini sebaiknya memiliki keterkaitan dengan transportasi
kapal dan dibuat secara sederhana. Desain penelitian ini disesuaiakan dengan
kondisi di laboratorium. Hipothesis dari penelitian ini adalah grup pre stress dapat
meningkatkan toleransinya selama transportasi.
Berdasarkan hasil yang didapat dari penelitian ini menunjukkan bahwa P.
viridis tidak dapat meningkatkan toleransinya terhadap suhu selama simulasi
transportasi kapal, dan juga terdapat penurunan jumlah benang byssus, serta
penurunan berat badan selama transportasi.
Penelitian ini dilakukan sebagai bagian dari proyek GAME-X (Global
Approach by Modular Experiment) yang bekerja sama dengan Geomar Jerman
dan beberapa universitas dunia (Jerman, Brasil, Chili, Portugal), penelitian
dilakukan untuk membandingkan kerentanan spesies terhadap stress.
v
SUMMARY
YASSER AHMED. Effects Of Simulated Ship Transport On The Susceptibility
Of The Green Mussel Perna viridis To Heat Stress. Under supervised by
NEVIATY PUTRI ZAMANI and KAREN VON JUTERZENKA.
The introduced species are species which exceed their natural distribution
limits (native range) by a certain mode of introduction (vector). Natural
movements are responsible to limited extent. These introduction of a species
could influence the susceptibility of a species to environmental stress no matter if
introduced purposely or unpurposely. Introduction of species can happen caused
by natural movement i.e. organism can movement influenced by current and some
organism can migrate to one area to other area; and human activity either
purposely i.e. aquarium trade, aquaculture or unpurposely such as accidentally
through fouling on ship hulls (Baker et al., 2007). But dominant vector which
have large contribution on species introduction is transport by ship hulls and
ballast water tanks (Mead et al., 2011). Ballast water tank used to keep the
balancing of the ship and some organism has ability to attach on the wall of ship
hulls. Introduction species by ships mostly happened where a lot of traffic journey
of cargo ship bigger than 10.000 GT during 2007 (Kaluza et al., 2010). Based on
data from Global cargo ship network (GCSN), shipping moves over 80 % of the
world the goods. The problem is that ships do not only bring the goods but also
bring species in the ship hulls and ballast water tank (Kölzsch and Blasius, 2011).
The question is what happened to transport condition? For introduction of species,
there are a wide variety of transfer mechanisms (vectors) available for hitchhiking
of marine invaders to travel and spread in large amount of individuals; but only
small amount of individuals can be robust and act as competitors to get food
supply in the new environment, it called invasive species. Marine invasive species
are defined as non-native species that cause or are likely to cause harm to
ecosystems, economies, and or public health (Bax et al., 2003; ISAC, 2006).
Based on Lenz et al (2011), they compare native and non-native marine
invertebrates regarding their tolerance to stress. Six Bivalve (native named first):
Brachidontes exustus and Perna viridis (Trinidad), P. perna and Isognomon
bicolor (Brazil), Saccostrea glomerata and Crassostrea gigas (New Zealand); two
Ascidians: Diplosoma listerianum and Didemnum vixellum (Wales); and two
Crustaceans: Gammarus zaddachi and G. tigrinus (Finland) has been investigated.
They found that non native are more resistant towards environmental stress, such
as desalination, warming and hypoxia, than taxonomically related and
ecologically similar native species. Kleunen et al. (2010) said that tolerance
towards abiotic stress is a key trait that determines the invasiveness of species.
Furthermore, this was surprise and therefore pronounced differences in tolerant
towards stress exist between different species. One possible explanation for this
observation are adverse condition during transport, e.g. in the ship hulls and
ballast water tanks of cargo vessels.
The introduction of species more increased since past century (Mead et al.,
2011), and needed mimicking ship transport to know whether that species can
vi
increase their tolerance during transport. The objective for this experiment is to
investigate whether P. viridis can increase their tolerance during transport. The
experimental approach have connection with simulated ship transport and be
rather simple. This experimental design is design to mimicking transport
condition in short term lab experiments. Hypothesis for this experiment is pre
stress group can increase their tolerance during transport.
Based on the result shows that P. viridis could not increase their tolerance
during simulated ship transport, and they decrease amount of byssus threads, also
they decrease their weight during transport.
This research was conducted as part of the Global Approach by Modular
Experiment (GAME-X) project, to do a latitudinal comparison of susceptibility to
stress.
vii
Copyright © 2013 Bogor Agricultural University
Copyright are Protected by Law
1.
2.
It is a prohibited to cite all or part of this thesis without referring to and
mentioning the source.
a. Citation only permitted for the sake of education, research, scientific
writing, report writing, critical writing, or reviewing scientific problem.
b. Citation doesn’t inflict the name and honor of Bogor Agricultural
University.
It is prohibited to republish and reproduce all or part of this thesis without
the writer permission from Bogor Agricultural University.
viii
EFFECTS OF SIMULATED SHIP TRANSPORT ON THE
SUSCEPTIBILITY OF THE GREEN MUSSEL Perna viridis
TO HEAT STRESS
YASSER AHMED
Thesis
as one of the requirements for achieving
Master of Science degree in
Marine Science Program
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
2013
ix
Outside Committee Examiner on Thesis Examination:
Prof. Dr. Ir. Dedi Soedharma, DEA
x
Title
Name
NIM
: Effects of simulated ship transport on the susceptibility of the
Green Mussel Perna viridis to heat stress
: Vasser Ahmed
: C5511 00021
Approved by
Advisory Commiteee
tL/)!(
Dr. Ir. Neviatv P. Zamani. M.Sc.
(Head)
Dr. Karen von Juterzenka
(Member)
Known by
Head of Marine Science Program
Dr. Ir. Neviatv P. Zamani. M.Sc.
Date of Examination:
July
Date of Graduation:
2013
2 6 JUL 2613
Title
Name
NIM
: Effect of simulated ship transport on the susceptibility of the
Green Mussel Perna viridis to heat stress.
: Yasser Ahmed
: C551100021
Approved by
Advisory Commiteee
Dr. Ir. Neviaty P. Zamani, M.Sc.
(Head)
Dr. Karen von Juterzenka
(Member)
Known by
Head of Marine Science Program
Dean of Graduate School
Dr. Ir. Neviaty P. Zamani, M.Sc.
Dr. Ir. Dahrul Syah, M.Sc.Agr.
Date of Examination:
July 8th 2013
Date of Graduation:
xi
PREFACE
Praise and thanksgiving the author to Allah SWT for the bounty so that
this thesis succesfull to done. Choosen of theme on this experiment was done on
April to September 2012 and related with the survival of Green Mussel Perna
viridis, the title are Effect of Simulated Ship Transport On The Susceptibility Of
Green Mussel Perna Viridis To Heat Stress.
The author want to say thanks to Dr. Neviaty P. Zamani, Dr. Karen von
Juterzenka, and Dr. Mark Lenz as supervisor. On other hand, the author give
thanks to my partner for the experiment in Indonesia are Armin Fabritzek and
other participant of GAME-X Project.
Hopefully this thesis could be useful.
Bogor, July 2013
Yasser Ahmed
xii
ACKNOWLEDGEMENT
The author want to say thank you to all the parties who supports and help,
so that this research and thesis can be done.
1.
Parents (Nasrun Tasila and Juliani also Djarot Soedarsono and Sri Ulfah
Hartati), Lettu (Inf) Nasser Khaled and Leila Nabila (Brother and Sister)
and all the family who patiently supports, inspires, motivate, help, and pray
for keep reminding to do the best, on time, always learn, humble, and work
hard and smart for the better and the best to reach the goals.
2.
Wife (Dian Respati Widianari) who support, inspires, motivate, help, as
best friend to discuss and share, and pray to achieve our dreams also for keep
reminding to do the best, on time, always learn, humble, and work hard and
smart for the better and the best to reach the goals.
3.
Dr. Ir. Neviaty Putri Zamani, M.Sc. as the head of advisory committee and
head of Marine Science Program who patiently give support, advice, help,
and time through finishing of this thesis.
4.
Dr. Karen Von Juterzenka as member of advisory committee who gave a
lot of support, help, time, advice, new experience (for me), and share
knowledge and fun through finishing of this thesis.
5.
Dr. Mark Lenz and Prof. Martin Wahl as project coordinator Global
Approach by Modular Experiment (GAME X) from Geomar – Germany who
gave the opportunity to join their project and broaden my knowledge,
experience, and network.
6.
GAME-X Team: Armin Fabritzek (my partner), Felipe Ribeiro and Lisa
Oberschelp (Team Brazil), Sandra Eichorn and Miguel Angel Penna (Team
Chile), Marie Garcia and Filipa Antunes (Team Portugal)
7.
Prof. Dr. Ir. Dedi Soedharma, DEA as outside examiner committee who
help a lot giving advise thoroughly through this thesis so that this thesis can
be better.
8.
Prof. Dr. Ir. Utomo Kartosuwondo, MS. as seminar moderator and
examiner who give time in supporting and positive comment during the
seminar.
xiii
9.
Friends in Marine Science Program class 2010 Bogor Agricultural
University (Abd Muthalib Angkotasan, Adi Purwandana, (Alm) Yuida
Labetubun, Evangelin Kadmaer, Rezi Apri, Princy (for all the listening, help,
and many moments together)
10. Friends in Marine Habitat Lab (Dr. Michael K. Schmid, Mareike Huhn,
Nurina Ayu, Rebecca Mueller, Giannina Hattich, Titan, Shelly Tutupoho,
Andre Wizemann, Veronica, David, Wira, Nora Hase (for helping and
accompanying study) thank you for all the good times, laugh, advise, critics,
info, support, and help during we study and working together.
11. Dondy Arafat, thank you for helping in providing seawater to the lab.
12. Aris and Pak Baso as Fisherman in Muara Kamal who helped us to take
sample in Jakarta Bay.
13. All parties and everybody who not yet mentioned, thank you for all supports,
help, and time through finishing this thesis.
xiv
CONTENTS
LIST OF TABLES
xvii
LIST OF FIGURES
xvii
1. INTRODUCTION
Background
Objectives
Hypothesis
1
1
2
2
2. LITERATURE REVIEW
Transport of organism
Natural dispersal
Human mediated dispersal
Vector of marine introduction
Stressors
Bivalves
Perna viridis
The spread of Green Mussel (P. viridis)
Introduction of species
3
3
3
3
6
8
9
10
11
12
3. RESEARCH METHODOLOGY
Time and Location
Materials
Methods
Organism choice and sample sites
Sampling and transport the organism
Acclimation and feeding
Experimental design and data analysis
Laboratorium experiment
Water quality control
Temperature
Problematics
Experiment
Pilot study
Set up and main experiment
Response variables
Survivorship, Byssus, Body condition index (BCI)
Position of mussels
Response behaviour
Daily routine
14
14
14
17
17
19
20
21
23
23
24
24
24
24
25
26
26
27
27
28
4. RESULTS AND DISCUSSION
Results
Species identification
Acclimation
Experiment
29
29
29
29
29
xv
CONTENTS (continued)
Survivorship of the mussels during the pilot study
Performance of the mussels during the main experiment
Survival
Byssus
Body condition index (BCI)
Position of mussels
Response behaviour
Discussion
Adverse condition during transport
Number of byssus during transport
Growth performance during transport
Position of mussels during transport
Response behaviour during transport
29
30
30
32
32
33
36
40
40
41
41
41
42
5. CONCLUSION AND SUGGESTION
Summary
Suggestion
43
43
43
REFERENCES
44
xvi
LIST OF TABLES
1.
2.
Anthropogenic vectors for marine introduction (Bax et al, 2003)
Materials that was used to perform this experiment
7
14
LIST OF FIGURES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
(a) Cumulative number of marine introduced species recorded
since 1840. (b) Liner regression indicating rate of discovery of
marine introduced species since 1840 (Mead et al., 2011)
The trajectories of all cargo bigger than 10.000 GT during 2007.
The colour scale indicates the number of journeys along each routes.
Ships are assumed to travel along the shortest (geodesic)
paths on water (Kaluza et al., 2010)
Habitat distribution of marine introduction (Mead et al., 2011)
Main vectors of marine introduction (Mead et al., 2011)
Marine introduced species across taxonomic groups (Mead et al., 2011)
Laterally compressed body is the most distinctive feature of
bivalves (Castro and Huber, 2007).
(a). Perna viridis were measured in a ruler; (b) Perna viridis
on the glass tank; (c) Fresh harvest of Perna viridis
Approximate native and introduced range of the Asian Green Mussel
P. viridis, base on the literature review (Baker et al. 2007)
Mytillus galloprovincialis dominating fouling assemblages in the
sea chest of the SA “Agulhas” (Lee and Chown, 2007)
Asian Green Mussel, P. viridis (dark object) and Eastern Oyster,
Crassostrea virginica (white object) on a bridge pier in Tampa Bay,
Florida, USA, at low tide in 2001. The majority of mussels in this
photo are 2-3 cm in shell length. (Baker et al. 2007)
(a). P. viridis; (b). Fresh harvest of mussels from Jakarta Bay
(a). Muara Kamal, Jakarta Bay; (b). Sub-district Muara Kamal;
(c). Muara Kamal in Java Island (red colour)
(Created by: Annisa Indah Sari, 2012)
Sampling organisms and measured the size;
(b) Mussels in the cool box; (c) Cleaned the mussels
Acclimation tanks in the lab.
Simulated ship transport from Europe to Australia.
Mimicking transport condition for this experiment
Main groups for this experiment
Scheme of pilot study
(a). Single containers on the fiber tanks for pilot study;
(b) Each containers fixed in the frame for pilot study
(a). Wooden frame for thermal stress phase; (b). Rubber
frame for control group; (c). Tank to pre heat water
xvii
4
5
5
8
8
10
11
12
13
13
18
18
19
20
22
22
23
24
25
26
LIST OF FIGURES (continued)
21. Measuring BCI of the mussels.
22. (a) Water exchanged; (b) Feeding the mussels
23. Survivorship of the mussels connected with
temperature during pilot study.
24. Survivorship of the mussels for pre stress group without
recovery
25. Survivorship of the mussels for pre stress group with
recovery
26. Byssus of the mussels for each groups
27. Body condition index (BCI) of the mussels each period
28. Position of mussels for group B (stress group without
recovery) (S: surface; M: middle; B: bottom; Ai: air stone;
NA: not attach)
29. Position of mussels for group BC (non stress group
without recovery) (S: surface; M: middle; B: bottom; Ai: air
stone; NA: not attach)
30. Position for control group (S: surface; M: middle; B: bottom;
Ai: air stone; NA: not attach)
31. Position of mussels for group A (stress group with recovery),
(S: surface; M: middle; B: bottom; Ai: air stone; NA: not attach)
32. Position of mussels for group C (non stress group with recovery)
(S: surface; M: middle; B: bottom; Ai: air stone; NA: not attach)
33. Position for control group (S: surface; M: middle;
B: bottom; Ai: air stone; NA: not attach)
34. Response behaviour group B (stress group without
recovery) (F: Fast; C: Closed; S: Slow)
35. Response behaviour group BC (non stress group
without recovery) (F: Fast; C: Closed; S: Slow)
36. Response behaviour for control (F: Fast; C: Closed; S: Slow)
37. Response behaviour group A (stress group with recovery)
(F: Fast; C: Closed; S: Slow)
38. Response behaviour group C (non stress group with recovery)
(F: Fast; C: Closed; S: Slow)
39. Response behaviour for control (F: Fast; C: Closed; S: Slow)
xviii
27
28
30
31
31
32
33
33
34
34
35
35
36
37
37
38
38
39
39
1
1 INTRODUCTION
1.1 Background
The introduced species are species which exceed their natural distribution limits
(native range) by a certain mode of introduction (vector). Natural movements are
responsible to limited extent. These introduction of a species could influence the
susceptibility of a species to environmental stress no matter if introduced
purposely or unpurposely. Introduction of species can happen caused by natural
movement i.e. organism can movement influenced by current and some organism
can migrate to one area to other area; and human activity either purposely i.e.
aquarium trade, aquaculture or unpurposely such as accidentally through fouling
on ship hulls (Baker et al. 2007). But dominant vector which have large
contribution on species introduction is transport by ship hulls and ballast water
tanks (Mead et al. 2011). Ballast water tank used to keep the balancing of the ship
and some organism has ability to attach on the wall of ship hulls. Introduction
species by ships mostly happened related with a lot of traffic journey of cargo ship
bigger than 10.000 GT during 2007 (Kaluza et al. 2010). Based on data from
Global Cargo Ship Network (GCSN), shipping moves over 80 % of the world the
goods. The problem is that ships do not only bring the goods but also bring
species in the ship hulls and ballast water tank (Kölzsch and Blasius, 2011). The
question is what happened to transport condition? For introduction of species,
there are a wide variety of transfer mechanisms (vectors) available for hitchhiking
of marine invaders to travel and spread in large amount of individuals; but only
small amount of individuals can be robust and act as competitors to get food
supply in the new environment, it called invasive species. Marine invasive species
are defined as non-native species that cause or are likely to cause harm to
ecosystems, economies, and or public health (Bax et al. 2003; ISAC, 2006).
Based on Lenz et al (2011), they compare native and non-native marine
invertebrates regarding their tolerance to stress. Six Bivalve (native named first):
Brachidontes exustus and Perna viridis (Trinidad), P. perna and Isognomon
bicolor (Brazil), Saccostrea glomerata and Crassostrea gigas (New Zealand); two
Ascidians: Diplosoma listerianum and Didemnum vixellum (Wales); and two
Crustaceans: Gammarus zaddachi and G. tigrinus (Finland) has been investigated.
They found that non native are more resistant towards environmental stress, such
as desalination, warming and hypoxia, than taxonomically related and
ecologically similar native species. Kleunen et al. (2010) said that tolerance
towards abiotic stress is a key trait that determines the invasiveness of species.
Furthermore, this was surprise and therefore pronounced differences in tolerant
towards stress exist between different species. One possible explanation for this
observation are adverse condition during transport, e.g. in the ship hulls and
ballast water tanks of cargo vessels.
The introduction of species more increased since past century (Mead et al.
2011), and needed mimicking ship transport to know whether that species can
increase their tolerance during transport. The objective for this experiment is to
investigate whether P. viridis can increase their tolerance during simulated ship
2
transport. The experimental approach have connection with simulated ship
transport and be rather simple. This experimental design is design to mimicking
transport condition in short term lab experiments. Hypothesis for this experiment
is pre stress group can increase their tolerance during transport.
This research was conducted as part of the Global Approach by Modular
Experiment (GAME-X) project, to do a latitudinal comparison of susceptibility to
stress with theme “Do adverse condition during transport select for stress tolerant
genotypes in founder population of native and non-native marine invertebrates”.
1.2 Objectives
The objective for this experiment is to investigate whether P. viridis can
increase their tolerance during transport.
1.3 Hypothesis
Pre-stress can increase their tolerance during transport.
3
2 LITERATURE REVIEW
2.1 Transport of organisms
Transportation network play crucial role in human mobility, distributed the
goods and spread species. Transport of organisms has two factors was natural
movements and human activity. Large contribution to transport of organism was
caused transportation of ships where most of world trade is hauled by ships and
7,4 billion tons of goods were spread to the worldwide (International Maritime
Organization (IMO), 2006) and also spread of species.
2.1.1 Natural dispersal
At first movement of organisms influenced by natural dispersal. Some
organism has limited movements so that they follow natural influence by current
(plankton) and other organism can migrates through the ocean current such as sea
turtle, whales etc. Perna viridis are similar to those of Mytilus species but no one
has determined whether P. viridis has a planktonic postlarval phase like some
other members of the family (Baker and Mann, 1997). P. viridis has byssus to
attach on the surface of substrata so that they was easily can invasive to other
area. One of possibility is influenced by natural dispersal where driftwood
movement, plastic and other human debris was increased important factors for
introduction of organisms (Winston 1982, Aliani and Molcard 2003, Barnes and
Fraser 2003 in Baker et al. 2007). Other possibility natural objects which can
found anywhere was occurred in globalization times because of anthropogenic
factors (Baker et al. 1999).
Marine invertebrates specially for P. viridis has large distribution. P.
viridis was native species and distributed in Indo-Pacific such as Philippines,
Sumatra, Borneo, Bali, Java, Vietnam, China (Siddal, 1980) and South Asia
(Baker et al. 2007). Secondary sources P. viridis was native range in island of
New Guinea (FAO fisheries, 2006).
2.1.2 Human mediated dispersal
One of factor organism transport and spread is human mediated dispersal.
Marine invertebrates can cross to different latitudes or other places by ship
transport. Some marine invertebrates have the ability to attach on the wall of ship
hulls and some larvae inside in ballast water tank. Most of the commodities or the
goods in the world are transported by cargo ships and transferred species to other
places (Kölzsch and Blasius 2011). Since industrial development of transported
was established, humans increased the size of ship because trade commodity. The
ballast water tank used to get balanced of ships so that secure to bring the goods.
The impact of species introduction with deballasting can be pass through ballast
water intakes are known to survive transportation over long distances (Flagella et
al. 2007). One of case the number of species introduction increased in South
4
African coast since 1840 because industrial development of ship (Mead et al.
2011) (Figure 1).
Figure 1 (a) Cumulative number of marine introduced species recorded since
1840. (b) Liner regression indicating rate of discovery of marine
introduced species since 1840 (Mead et al. 2011)
Transportation network play a crucial role in human mobility to transfer
and movement goods and the impact would spread the species. Around 90% of
world trade is hauled by ships (International Maritime Organisation, 2006). Two
major pathways for marine bioinvasion are discharged water from ships ballast
water tank and hull fouling. Ballast water in cargo ship contribute large transport
of organisms around 3 to 4 billion tons per year in the ports of the world (Kölzsch
and Blasius, 2011). European ports, American, Asian, African and Australian also
contain among of the high number invasive species. The financial was loss in
estimated to be $120 billion in US alone (Kaluza et al. 2010). The transportation
network can be seen in Figure 2. Based on the Figure 2, large size of ship more
traffic with the transportation network and would be give impact to marine
introduction. Dominant the distribution for marine introduction was in the harbour
where the ship docked (Mead et al. 2011) (Figure 3).
5
Figure 2 The trajectories of all cargo bigger than 10.000 GT during 2007. The
colour scale indicates the number of journeys along each routes. Ships
are assumed to travel along the shortest (geodesic) paths on water
(Kaluza et al. 2010)
Figure 3 Habitat distribution of marine introduction (Mead et al. 2011)
6
2.1.3 Vector of marine introduction
Increasingly the introduction of alien species is a major threat to marine
biodiversity and a contributor to environmental change. As these marine
introduction, intentional and accidental can result from numerous human mediated
activities, management responses need to cover a diverse range of human activity.
Based on historical data, a new marine or estuarine species establishes itself every
32-85 weeks in each of six ports studied in the US, New Zealand and Australia, a
rate that appears to be increasing. While many of the alien species become part of
the background flora and fauna, others become invasive and come to dominate the
native flora and fauna. There are 15 broad categories of vectors that transport
marine organisms from shallow coastal waters to similar habitats outside the
species home range which can be seen in Table 1 (Bax et al. 2003).
The main economic and social impacts of invasive alien marine species are
negative impacts on human health and decrease in economic production of
activities based on marine environments and resources such as fisheries,
aquaculture, tourism and marine infrastructure. These affect have related social
impacts through decreases in employment in economic activities directly affected
by invasive alien species but also through decreases in people’s welfare from the
reduced quality of their environments and natural surroundings. Some vectors for
alien marine species was happened caused human activity by vessel for trading
commodities and vessel for recreation. In the modern era, increase size of vessel
will give effect to introduction of organisms. Some case in Australia ports, one of
kind mussels P. viridis was introduce to Darwin harbour (Bax et al. 2003).
Characteristics such as dispersal, life history strategies, growth rates,
distributional ranges, association with humans, and tolerance towards
environmental stress are determinants of invasiveness for animal and plant species
(Braby and Somero, 2006; Lenz et al. 2011). One of possibility is that marine
organisms have the ability to move by ship hulls and ballast water. Mead et al
(2011), observed marine bioinvasion in South African and show that some vectors
marine organism introduce to the environment and most of the vectors dominant
to marine introduction was ship hulls and ballast water (Figure 4). Ballast water
released in the environment has lead to damage of establishment hundreds of
introduction organism to Europe and North America (Mills et al. 1996; Bij de
Vaate et al. 2002). One of factor was the ships of ballast water have potential to
transfer coastal marine species to the other area and this marked a shift from the
centuries-old practice of using rocks and sand from the shore to provide balance
for ships (Davidson and Simkanin, 2012). P. viridis is mussels where including
molluscs and marine organism could movement or introduction to other place
(Mead et al. 2011) (Figure 5).
7
Table 1 Anthropogenic vectors for marine introductions (Bax et al. 2003)
Source
Commercial shipping
Vector
Ballast water
Target taxa
Plankton,
nekton,
benthos in sediment
Hull fouling
Encrusting, nestling, and
some mobile species
Solid ballast (rocks, sand, Encrusting,
benthos,
etc)
meiofauna and flora
Aquaculture
and Intentional release for
Single species
fisheries
stock enhancement
Gear, stock or food
Various
movement
Discarded nets, floats,
Various
traps, trawls, etc
Discarded live packing
Various
materials
Release of transgenic
Single species
species
Drilling platforms
Ballast water
Plankton, nekton,
benthos in sediment
Hull fouling
Encrusting, nestling, and
some mobile species
Canals
Movements of species
Various
through locks due to
water motion or active
swimming
Aquarium industry
Accidental or intentional Aquarium fauna and
release
flora
Recreational boating
Hull fouling
Encrusting, nestling, and
some mobile species
Dive practices
Snorkeling and scuba Algal spores, bacteria,
gear
some mobile species
Floating debris
Discarded plastic debris
Encrusting, and some
mobile species
8
Figure 4 Main vectors of marine introduction (Mead et al, 2011)
Figure 5 Marine introduced species across taxonomic groups (Mead et al, 2011).
2.2 Stressors
Temperature as one of suitable stressor with condition in ballast water tank
and give effect to introduction species. Most of the ship material was metal and it
act as conductor which easily influenced by temperature. On other hand,
temperature (heat) is a relevant stressors for benthic, shallow water organism and
they also act on organisms shuttled in ballast water tanks or on ship hulls. This
9
stressor (temperature) also related with condition in ship hulls and ballast water
tank because when some species come to inside ballast water tanks and they
influenced by temperature; then, when some species attach in the ship hulls and
the journey has different lattitude and of course influenced different temperature
as well (Schneider, 2008; Seiden et al. 2011).
Temperature is one of important factor to control geographical range and
seasonal cycles of marine organism. As general, organism that maintain their body
temperature by absorbing heat from the environment (ectotherm). Temperature
too high can act as stressor in two ways: First, it may cause the denaturing of
sensitive proteins; This damage can be minimized by the actions of heat shock
proteins; which increase the thermostability of proteins and chaperone cellular
processes. Second, high temperatures may cause oxygen limitation (Jansen et al.
2007). Temperature also has a linier correlation with heart rate; high temperature
lead to fall in heart rate (Akberali and Trueman 1985).
Species “biogeographical” has tolerance to environmental extremes.
Intertidal species is one of organism has tolerance to environmental stress such as
temperature and salinity (Castro and Huber 2007).
2.3 Bivalves
Class Bivalvia consist of clams, mussels, and oysters. The body of bivalvia
is laterally compressed and enclosed in a shell with two parts or valves. The umbo,
the upper hump near the hinge of each shell is the oldest part of the shell. The
form of concentric growth lines is growth from the umbo. The inner surface of the
shell is lined by the mantle, so that the whole body lies in the mantle cavity. The
mantle cavity is a large space between the two halves of the mantle. Special for
the Perna viridis has ability attach to the rocks and other surface by byssal threads
or byssus (Castro and Huber, 2007). Structure the body of bivalve can be seen in
Figure 6.
10
Figure 6 Laterally compressed body is the most distinctive feature of bivalves
(Castro and Huber, 2007).
2.3.1 Perna viridis
Perna viridis has the common name Asian Green Mussel or Green Mussel
and they live in intertidal zone. They has ability to attach in the wall, so they can
live on a variety of structures such as vessels, wharves, mariculture equipment,
buoys and other hard substrate. This species is filter feeder, feeding on small
zooplankton, phytoplankton and other suspended organic material (Castro and
Huber 2007; National Introduced Marine Pest Information System (NIMPIS). P.
viridis can be seen in Figure 7A, 7B and 7C.
11
a.
b.
c.
Figure 7 (a). Perna viridis were measured in a ruler; (b) Perna viridis on the
glass tank; (c) Fresh harvest of Perna viridis.
The taxonomical name of Perna viridis (Linnaeus, 1758) is:
Phylum: Mollusca
Class: Bivalvia
Subclass: Pteriomorphia
Order: Mytiloida
Superfamily: Mytiloidea
Family: Mytilidae
Genus: Perna
Species: Perna viridis
(Poutiers 1987; Carpenter and Niem 1998; Leal 2002)
2.3.2 The Spread of Green Mussel (P. viridis)
Native distribution of P. viridis extends from the Persian Gulf to all of
Philippines, Sumatra, Borneo, Bali, Java, Sulawesi, Northeast Vietnam (Siddal,
1980). Siddal (1980) reported that presence of P. viridis in Hongkong as
introduced species. The island in New Guinea also include native range (FAO,
2006). Gindy et al (2001) reported, distribution of P. viridis also spread in
Musandam Peninsula of Oman. Most of P. viridis distributed in Indo-Pacific.
12
Green Mussel also one of fouling organism and ships have been sailing to many
area (Kaluza et al. 2010), it is possible P. viridis have been introduced to Florida,
Jamaica, Trinidad, China, Japan (Figure 8). Some evidence that P. viridis
dominating in fouling and introduced to the coast of Florida (Figure 9 and Figure
10).
Figure 8 Approximate native and introduced range of the Asian Green mussel,
P. viridis, based on the literature review (Baker et al. 2007)
2.4 Introduction of species
Most of introduction of species happened by human activity such as
aquaculture, marine litter, oil rig, intentional, fisheries, solid ballast, ship boring,
ballast water, and ship fouling (Mead et al. 2011), but the dominant vector
organism introduced by ballast water and ship fouling. It is related with increase
number of ships which bring the good and distributed to worldwide. Ships carry
ballast water for vessel stability and trim, it is well established that ballast water
can contain some species such as mussels, metazoan, microorganism, some of
them are pathogenic and so on (Drake et al. 2002).
Species which introduced into new environment would survived because
ballast water tank support the survival of mussels and also they would attach on
the substrate and act as competitors to get food supply. Introduction of species
increased because no predatory in the new habitat and would alteration food chain
and impact to decreased native species (Baker et al. 2007).
13
Figure 9 Mytilus galloprovincialis dominating fouling assemblages in the sea
chests of the SA “Agulhas” (Lee and Chown, 2007).
Figure 10 Asian Green Mussel, P. viridis (dark object) and Eastern Oyster,
Crassostrea virginica (white object) on a bridge pier in Tampa Bay,
Florida, USA, at low tide in 2001. The majority of mussels in this
photo are 2-3 cm in shell length. (Baker et al. 2007)
14
3 RESEARCH METHODOLOGY
3.1 Time and Location
This research was conducted in the Marine Habitat Laboratory,
Department of Marine Science and Technology, Faculty of Fisheries and Marine
Sciences, Bogor Agricultural University (FPIK – IPB) from April to September
2012.
3.2 Materials
The experiment was used some materials and shown in Table 2. The
pictures of materials are shown in Appendix 1.
Table 2 Materials which used to perform this experiment
Material
s
Seawater
Air
pump
Brand
(Type)
Sonic
P-125
Small
hose and L,
I, T junction
Single
Jack
container
Family
Tank
Specification
Salinity: 33-35 ‰ from Teluk
Jakarta, filter processed sea water
in Gelanggang Samudera and
Seaworld, Ancol, Jakarta (Arafat,
pers.com., 2012)
AC 220-240 V
50 Hz
85 W
125 L/min
0.04 MPa
1.5 litre
Fiber tank
Functions
To supplyed
aeration
To
connected
aeration
Experiment
al container for
kept the
organisms
As water
bath for pilot
study and main
experiment
15
Table 2 Material which used to perform this experiment (continued)
Materials
Transparen
t pipe and
fishing net
Water
pump
Brand
(Type)
Specification
Amara AA
1800
Rubber
Heater
Thermo
logger
Aquatic
Nature and
Eheim Jager
Onset Hobo
Calliper
Scale
Thermome
ter Digital
“Lucky”
(pocket scale)
and Bosch
Krisbow
KW06-308
Temperature setting
Company: Aquatic
Nature and Eheim Jager
300 W
220-240 V
50-60 Hz
20-35 0C
Pendant temp
Part # UA-001-08
Patent 6.826.664
15 cm (max.)
0.05 cm (accuracy)
6 inches (max.)
1/128 inches (accuracy)
Digital scale
Capacity: 200 g x 0.01 g
400C – 2500C
400 – 4820 F
Resolution: 0.10
Functions
To build
frame (pilot
study)
Creating a
current in
waterbath so
that the heat
distributed
evenly
To build
frame
(experiment)
To
produced
heating for
increased and
decreased
temperature in
waterbath and
pre heat water
To recorded
the
temperature in
waterbath.
Measure
length and
width of
mussels
Measured
weight of
mussels
Measured
temperature in
single
container and
the tank
16
Table 2 Material which used to perform this experiment (continued)
Materials
Bamboo
stick
Brand (Type)
Specification
Bamboo
Functions
Tool to
checked
mussels
Camera
Canon and
Canon DSLR 600 and
Documentat
Casio
Casio QV-R100
ion
Food
Coral sands
Nanochloropsis
Food for the
(DT’s premium
aculata, Phaeodactylum
mussels
blend live marine tricornutum, Chlorella 2phytoplankton)
20 microns
Oven
Heraeus
To dry the
mussels
Data sheet
Paper
Notes
Refractomet
Atago
Salinity 0~100 ‰
Measure
er
(S/Mill-E)
salinity in the
(Hand-held
beginning and
refractometer,
in the end of
Made in Japan)
experiment
Nitrate and
API
Measure
For fresh and
Nitrite
(Aquarium
nitrite
and
saltwater aquarium
Aquarium test Pharmaceuticals)
Nitrite (NO2 ): 0–0.5– nitrate
strips
1–3–5–10
Nitrate (NO3-): 0–20–
40–80–160–200
Ammonia
API
Measure
For fresh and saltwater
Aquarium Test (Aquarium
ammonia
aquarium
Strips
Pharmaceuticals)
Ammonia (ppm
(mg/L)): 0 – 0.5 – 1.0
– 3.0 – 6.0
pH
API
Measure pH
For fresh and saltwater
Aquarium Test (Aquarium
aquarium
Strips
Pharmaceuticals)
PH freshwater: 6.0 –
6.5 – 7.0 – 7.5 – 8.0 –
8.5 – 9.0
PH saltwater: 6.0 – 6.5
– 7.0 – 7.5 – 8.0 – 8.5
– 9.0
17
Table 2 Material which used to perform this experiment (continued)
Materials
Brand (Type)
Specification
Tank filter and
pump system had
been provided by
Marine Habitat
Lab
Function
Filter and mix
seawater inside
the tank.
3.3 Methods
3.3.1 Organism choice and sample sites
The purpose to choose of mussels have some criteria for this experiment:
(1) the mussels native in Indo-pacific and was invasive to other area (Mead et al.
2011); (2) generally can found it in the ship hulls and ballast water tank (Lee and
Chown 2007). Thus, it is relevant for simulated ship transport; (3) it is easy to
collect P. viridis as it can be found in Jakarta Bay (Muara Kamal, Indonesia)
where it is cultured and harvested by local fishermen and has economic value
(Figure 11); (4) members of the same family, Mytilidae, can be found in other
countries participating in GAME X project, such as Brasil, Chile, Finland and
Portugal, so that we can compare our results to these countries.
Sample site was located in North Jakarta, Indonesia. P. viridis (Green
mussel) were taken from Jakarta Bay, Muara Kamal (Figure 12). The mussel was
taken on June 12th, 2012. The temperature on the sea water was 29 0C and salinity
was 32 ppt. Muara Kamal was the place to harvested the mussels which is
influenced by anthropogenic activity. The majority population of Indonesians live
in Java and therefore coastal resources are under considerable human pressure and
every year the Jakarta Bay show increase in eutrophication and sedimentation
levels, which had impact on marine communities (Damar, 2003; van der Meij et
al. 2009).
18
Figure 11 (a). P. viridis; (b). Fresh harvest of mussels from Jakarta Bay.
a
b
c
Figure 12 (a). Muara Kamal, Jakarta Bay; (b). Sub-district Muara Kamal; (c).
Muara Kamal in Java Island (red colour) (Created by: Annisa Indah
Sari, 2012).
19
3.3.2 Sampling and transport the organism
Samples for experiment and bring the samples from the site to the lab. The
mussels taken in Muara Kamal (Jakarta Bay) with some local fishermen and was
transported by boat to the field. The mussels taken in culture area and harvested
by local fishermen. Afterwards, the mussels were transported from Muara Kamal
to the lab by car. The organisms were put in plastic aquarium without seawater.
Ice was filled in the bottom of cool box and covered with sterofoam. Afterwards,
the plastic aquarium was put on the top of sterofoam so that the organisms stayed
cool and in low metabolism. The transport took two hours from Jakarta Bay to the
lab. After arrival in the lab, the organism were cleaned also from barnacles that
are attached to it (Figure 13). Afterwards, the organism were put into the glass
aquarium for acclimation.
a.
b.
c.
Figure 13 (a) Sampling organisms and measured the size; (b) Mussels in the cool
box; (c) Cleaned the mussels.
20
3.3.3 Acclimation and feeding
Acclimation is response organism to specific environmental change(s) e.g
temperature, salinity, oxygen and so on, that observed in the lab to distinguish
effect and response to these changes e.g. physiology, activity, growth, etc (Nielsen
1990; Braby and Somero 2006). So, the purpose of acclimation is that organism
get adapted to lab conditions (temperature, salinity, oxygen) until achieved 1%
mortality (GAME X). During acclimation, water exchange and feeding of the
mussels were done. Half of the sea water in glass aquarium (26 litre) was
exchanged every day. First to second day acclimation, water exchange was done
twice a day and next day water was changed once a day. Natural sea water was
provided by a 1000 liters tank system equipped with a mechanical and a biological
filter unit and is distributed to smaller glass aquaria. All organism must have been
acclimatized before use in the experiment. Acclimation tanks can be seen in
Figure 14.
The mussels were fed with coral sands DT’s premium blend live marine
phytoplankton (Nanochloropsis aculata, Phaeodactylum tricornutum, Chlorella 220 microns) which is a liquid phytoplankton food that had a concentration 1,58 x
107. The first day to second day of acclimation phase, the organism were not fed
due to stress from transportation. Afterwards, started to feed the mussels daily
with 0.0125 ml/ind with coral sands. They should adapt with the lab condition
first. The third day to next days, the organisms were fed.
Total number of organism for acclimation before pilot study was 168
mussels. Total number of organism for acclimation before main experiment was
480 mussels which is the mussels separated in 12 glass aquaria with aeration
inside. Each glass aquaria consist of 40 mussels.
Figure 14 Acclimation tanks in the lab.
21
3.3.4 Experimental design and data analysis.
Most of the ship material was metal and it act as conductor which easily
influenced by temperature. Therefore, heat stress was chosen since it is relevant
with condition inside ballast water tank or ship hulls. On other hand, heat stress is
a relevant stressors for benthic, shallow water organism where they also act on
organisms shuttled in ballast water tanks or on ship hulls. Species which attach in
the ship hulls and carried on the way which has different lattitude would
influenced by temperature as well (Schneider, 2008; Seiden et al. 2011).
This experiment was done to simulated ship transport from Europe to
Australia because they have a lot of traffic journey of cargo ship bigger than
10.000 GT in 2007 (Kaluza et al. 2010). It can be seen the traffic journey in
Europe to US; US to Asia Pacific; US to Australia; Europe to Austalia and so on.
Thus, the simulated took from Europe to Australia because the journey has
different temperature from temperate – tropical – temperate (Figure 15).
The experimental design is designed to mimicking transport conditions in the lab.
In laboratory experiments, organisms will be exposed to stress regimes likely to
occur during transport by ship and which cause moderate mortality until up to
80% around 3 weeks (pre stress). After pre-stress period, the survivors will be
given some time (~2 weeks) to recover under stress free conditions and the other
group exposed directly to the same stressor again (second stress). Survival rates
were measured as a response variable over the course around 3 weeks. Control
group, consisting of the same number of conspecifics which have not previously
been exposed to adverse condition but have been acclimatized to lab conditions,
will be treated the same way and serves as a reference or control. Finally, we
compared survival between pre-stressed and non stressed group to investigate
whether pre stress group can increase the robustness during transport (Figure 16).
This experiment have 4 groups which consist of pre-stress group with
recovery (group A); pre-stress group without recovery (group B); then, non stress
group (group D and group C). Thus, pre-stress group without recovery compared
with non stress group (group B and group D). Then, pre-stress group with
recovery compared with non stress group (group A and group C) (Figure 17).
22
Figure 15 Simulated ship transport from Europe to Australia.
Figure 16 Mimicking transport condition for this experiment
23
Figure 17 Main groups for this experiment
The data for this experiment were analyzed by using the R program
(version 2.14.2). Kaplan Meier curve to analysed survivorship (log rank test), and
normality to analysed byssus and body condition index (Shapir