Tetraodon+nigroviridis+Marion+de+Procé,+1822

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 * Spotted Green Pufferfish** 🐡 ** //Tetraodon nigroviridis// Marion de Procé, 1822 **

=Introduction = The pufferfish, also known as the blowfish, bubblefish, swellfish, toadfish, globefish, toadies, honey toads, sugar toads and sea squab, belongs to the Tetraodontidae family which consists of 190 pufferfish species in 26 genera. 1] Despite its adorable look when it puffs up into a ball-like shape, pufferfishes are believed to be the second most poisonous vertebrate in the world. 2] In Singapore, a total of ten different species of pufferfish have been recorded. 3] 

=**Name** = Scientific name: //Tetraodon nigroviridis// Marion de Procé, 1822 Common name: Spotted green pufferfish (墨绿凹鼻豚)

**Etymology**
//**Tetraodon**//: Greek, __tetra__ = **four** + Greek, __odous__ = **teeth** 5] //**nigroviridis**//: Latin, __niger__ = **black** (black spots) + Latin, __viridis__ = **<span style="color: #00821a; font-family: Georgia,serif; font-size: 15px;">green **<span style="color: #535353; font-family: Georgia,serif; font-size: 15px;"> (colour of upper body) <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">21]

=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Biology =

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Life Cycle**
<span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Mature females lay around 200 eggs, which are attached to a hard, flat submerged surface in shallow water. A parent (usually the male) guards the eggs until they hatch. Adults have been observed tending schools of juveniles. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">6] <span style="color: #935177; display: block; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; line-height: 1.3; vertical-align: super;"> <span style="color: #935177; display: block; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; line-height: 1.3; vertical-align: super;">

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Diet**
<span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">The pufferfish is a predatory fish. It uses its small mouth, which resembles a parrot’s beak to suck its prey into it before crushing them to bits with its large strong teeth. It usually feeds on shrimps, worms, other finfish, benthic algae, crustaceans and mollusks. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">7]

media type="youtube" key="BCGmrCD2wGA" width="420" height="315" align="center" Video 1: A spotted green pufferfish is able to attack snails with its strong teeth. "Green spotted puffer attacking ramshorn snail" by smallyellowdevil by Youtube Channel 20 July 2008. URL: [] (accessed on 21 Nov 2014)

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Self Defense Mechanism**
<span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">Using a combination of fins, the pufferfish is able to attain hi<span style="font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">gh mane <span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">uverability. As such, it is unable to swim fast – which makes it an easy target for predators. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">8] <span style="font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Fortunately, the pufferfish possesses both mechanical and chemical defenses to protect it from predators. When the pufferfish is threatened by a predator, it quickly puffs its body up by two to three times of its normal size, forming a ball-like shape which makes it difficult to handle. media type="youtube" key="xu47OTMsCg8" width="420" height="315" align="center" Video 2: Be amazed by how this pufferfish managed to avoid becoming a snack of the otter through its wonderful puffing ability. " Freaks of Nature: Self Inflating Fish " by National Geographic by Youtube Channel 5 February 2008. URL: [] (accessed on 21 Nov 2014)

<span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">**How is this possible?** <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">The secret to the pufferfish's amazing ability is attributed to a unique oral valve which is attached to the floor of its mouth. After filling its mouth with water, the pufferfish flexes a large muscle at the base of the oral valve which then catapults forward against the entire front of the mouth, forming a tight seal against the back of the front teeth. This prevents the water from escaping! <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">At the same time, the flexing action of the large muscle propels water into the fish's esophagus and then its stomach. With every gulp, the stomach of the pufferfish becomes bigger and so does its body! This amazing ability is made possible by the high elasticity of its stomach and skin. Can you imagine how much the stomach has to expand? <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">During this process, the small body spines hidden under the skin at the back, sides and belly of the pufferfish protrude outwards. This gives it a spine-chilling appearance which makes it impossible to be swallowed by its predators. Also notice how the spine of the pufferfish bends when its stomach expands!

<span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">However, this tactic is only used as the last line of defense, such as when it is actually grabbed by the predator. This is because it is unable to swim well when it is inflated and it becomes helpless.

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Tetrodotoxin**
<span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">More often than not, bright and beautiful colors of animals indicate danger. It is a way of saying ‘Stay away from from me!’ Similarly, the brightly coloured pufferfish contains tetrodotoxin, a type of neurotoxin, which plays a crucial role in protecting it against predators. This toxin makes its muscular tissue and viscera extremely toxic. This toxin is probably the most lethal form in nature, where it is more than 1000 times more potent than cyanide. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">9]

<span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Tetrodotoxin in the pufferfish is produced by certain symbiotic bacteria found in their natural environment, which is accumulated from its diet. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">10] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> To humans, the tetrodotoxin is so deadly that just one pufferfish alone can kill 30 adult humans and there is no known antidote. This acts as a defense mechanism for the pufferfish as the toxin makes it unpalatable. A small bite by a predator can mean death!

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=<span style="color: #00821c; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Habitat = <span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">The spotted green pufferfish can be found in a wide range of salinity. The adults can be found in freshwater streams, rivers and flood plains while the juveniles can be found in brackish water in coastal areas such as mangrove forests. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">7] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">In Singapore, the <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">spotted green pufferfish is seen <span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> hovering in the brackish water in Sungei Buloh. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">11]



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=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Distribution = <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">The <span style="font-family: Georgia,serif; font-size: 15px;">spotted green pufferfish <span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> can be found in coastal freshwater and brackish water habitats in tropical regions (24°C - 28°C) across South and Southeast Asia. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">[9] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> It is native to Thailand, Philippines, Malaysia, Vietnam, Cambodia, Myanmar, India, Sri Lanka, Indonesia and Singapore. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">12]



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=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">General Description =

<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Adult spotted green pufferfish usually span between 3 to 6 cm but it can reach a maximum of 14 cm. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">13] <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">The length of the fish is measured from the tip of its mouth to the posterior end of the last vertebra.
 * <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Size **

<span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">Like its name suggests, the spotted green pufferfish has a bright green body with black spots <span style="font-family: Georgia,serif; font-size: 15px;">of variable sizes and forms which may coalesce or be encircled by lighter rings. The belly is white and the lateral line across the body is mostly indistinct. Its caudal fin has between 1 to 8 dark transverse bands and sometimes its dorsal fin has small dark spots near the base. The other fins are plain. <span style="font-family: Georgia,serif; font-size: 8pt; vertical-align: super;">21] <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">The body of the spotted green pufferfish is oblong and the posterior part is laterally compressed. Its dorsal profile is arched and the highest point is at the middle of the back. Its mouth is terminal, which allows it to feed on other fishes. The nasal organ is a tentacle and at least half of it is divided into two flattened and broadened lobes. The sides of the lobes have a spongy tissue.<span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">21] <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Back to Top
 * <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Colour **
 * <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Morphology **

=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Diagnosis =

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Pufferfish vs Porcupinefish
<span style="color: #535353; font-family: Georgia,serif;"> Pufferfish from the family Tetraodontidae is often confused with the morphologically similar and closely related porcupinefish of family Diodontidae. However, they can be easily distinguished by the spines on their body. The porcupine fish has large external spines unlike the smaller, hidden spines of the pufferfish from family Tetraodontidae. Also, the porcupinefish can be found in both temperate and tropical waters but the pufferfish is mostly found in the tropics.



<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Spotted Green Pufferfish vs Green Pufferfish
<span style="color: #535353; font-family: Georgia,serif;"><span style="color: #935177; font-family: Trebuchet MS',Helvetica,sans-serif; font-size: 11pt; line-height: 1.3; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">The spotted green pufferfish (//Tetraodon nigroviridis// Marion de Procé, 1822) is easily mistaken for its close relative, the green pufferfish (//Tetraodon fluviatilis// (Hamilton, 1822)) due to the spots on their green bodies. However, they can be distinguished by the differences in their spots. Both have spots and blotches of variable sizes on their upper bodies, but on closer look, green pufferfish (//T. fluviatilis//) can be distinguished by the three large dark patches on its back! 7] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">The spotted green pufferfish can also be better distinguished by its more rounded shape when compared to the //T. fluviatilis//. However, due to difficulties in defining the two closed related species through morphology alone, DNA analysis is usually used to distinguish the //Tetraodon// species. <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; vertical-align: super;">14]

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Spotted Green Pufferfish vs Spotted Scat
<span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;">At first glance, the spotted green pufferfish may also be confused with another similar looking fish - the spotted scat (//Scatophagus argus//) from the the family Scatophagidae. These two species co-exist in brackish mangrove habitats but they play very different ecological roles. Find out why here.

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=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Commercial Importance =

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Genetic Research**


<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">The spotted green pufferfish is important in comparative genomic research, which involves the studying of our evolutionary history. This is due to its extremely compact genome – it contains the same number of genes as humans but its genome length is only about ten percent of other vertebrates, which is about 340 million base pairs. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">17] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">This makes it a model organism for comparative genomics where human genes can be identified quickly and reliably.

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">**Aquarium Trade**


<span style="color: #535353; font-family: Georgia,serif;"> <span style="color: #935177; font-family: 'Trebuchet MS',',Helvetica,sans-serif; font-size: 11pt; font-size: 11pt; line-height: 1.3; line-height: 1.3; text-align: left; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Unlike some pufferfish species, the spotted green pufferfish is not sold commercially as a delicacy. Instead, it is a popular fish in the live aquarium trade due to its vivid colours. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">18] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Although it is not bred in captivity for commercial uses due to its high mortality rate, scientific breakthrough has allowed the spotted green pufferfish to be successfully bred in captivity at University of Florida using a new variation of the ovarian lavage technique in February 2009. This will definitely benefit the tropical fish industry and genetic researchers! <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">17]

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=<span style="color: #00821c; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Con <span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">servation Status = <span style="color: #535353; font-family: Georgia,serif;"><span style="color: #935177; font-family: 'Trebuchet MS',',Helvetica,sans-serif; font-size: 11pt; font-size: 11pt; line-height: 1.3; line-height: 1.3; text-align: left; text-align: left;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">The pufferfishes in Singapore are not listed among the threatened animals. However, like other creatures in their intertidal habitat, they are affected by human activities such as land reclamation projects and water pollution. <span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 8px; vertical-align: super;">11] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">Currently, there are about 6.59km <span style="color: #535353; font-family: Georgia,serif; font-size: 8pt; line-height: 1.3; vertical-align: super;">2 <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;">of mangrove forest left in Singapore, which makes up 0.95% of the total land area. Due to increasing demands for land, the spotted green pufferfish may face further habitat loss as mangrove forests continue to be lost to urban development. <span style="color: #535353; font-family: Georgia,serif; font-size: 8px; vertical-align: super;">22]

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=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Taxonomy and Phylogeny =

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Synonym
<span style="color: #535353; font-family: Georgia,serif;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">//Arothrom// //simula// //<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">ns //<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;"> (Cantor, 1849) <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">//Che lonodo n nigroviridis// (Marion de Procé, 1822) <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">//Tetraodon potamophilus// Bleeker, 1849 <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">//Tetraodon simulans// Cantor, 1849

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Type Information
<span style="color: #535353; font-family: Georgia,serif;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">The holotype of the //Tetraodon nigroviridis// <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">Marion de Procé, 1822 <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">, which was collected and described by Marion de Procé <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">during <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;"> a trip from France to Manila, Philippines is <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">believed to have been lost in a revolt. <span style="font-family: Georgia,serif; font-size: 8pt; vertical-align: super;">21]

<span style="color: #535353; font-family: Georgia,serif;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">A neotype which is designated by Dekkers in 1975 is now housed in the Institute of Taxonomic Zoology (Zoölogisch Museum), University of Amsterdam, The Netherlands. 21]

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Taxonomic Classification
<span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Kingdom: Animalia <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Phylum: Chordata <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Class: Actinopterygii <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Order: Tetraodontiformes <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Family: Tetraodontidae <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 11pt;">Genus: //Tetraodon// <span style="color: #535353; font-family: Georgia,serif;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">Species: //Tetraodon nigroviridis// <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">Marion de Procé, 1822 <span style="color: #535353; font-family: Georgia,serif; font-size: 11px; vertical-align: super;">15]

<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 16px;">Phylogenetic Relationship of //Tetraodon// pufferfish
<span style="color: #535353; font-family: Georgia,serif;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">In the study by Igarashi //et al//., 2013, the phylogenetic relationship of 17 //Tetraodon// species were analyzed by sequencing their mitochondrial genes encoding 16S rRNA and cytochrome b. 16] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;"> The sequencing results were then used to construct the following molecular phylogenetic tree using the maximum likelihood (ML) method and supermatrix analysis.



<span style="color: #535353; font-family: Georgia,serif;"> <span style="color: #535353; font-family: Georgia,serif;"> Through the phylogenetic tree, we can see that the genus //Tetraodon// contains pufferfish which do not form a monophyletic group. Instead, they are nested in four different clades in the family in Clade D (shown in yellow). Within the four clades, t <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">hree distinct groups (Asian freshwater, Asian brackish water and African freshwater groups), which correlated to their habitats, were identified. These three //Tetraodon// groups form a well-supported cluster (bootstrap probability 100%) with each sister group.

<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;"> This relationship may be associated with the salinity tolerance of the Tetraodon species which was caused by the the molecular evolution associated with osmotic regulation. Therefore, in the Asian brackish water group*, //T. erythrotaenia// diverged from the common ancestor first followed by //T. biocellatus//. Finally, two remaining and very closely-related species formed, namely //T. fluviatilis// and //T. nigroviridis// (spotted green pufferfish).

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=<span style="color: #00821d; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Did You Know? =

<span style="color: #535353; font-family: Georgia,serif;"><span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">Despite the long history of toxic effects, the flesh of some pufferfish species such as the //<span style="color: #535353; font-family: Georgia,serif;"> Takifugu radiatus //<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">( **<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">excluding **<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;"> the spotted green pufferfish) is considered as a delicacy in Japan. <span style="color: #535353; font-family: Georgia,serif; font-size: 8px; vertical-align: super;">18] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">Although the toxins are accumulated in its skin, ovaries, intestines and liver, consuming its flesh still comes with some risk. <span style="color: #535353; font-family: Georgia,serif; font-size: 8px; vertical-align: super;">19] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">Known as //fugu//, the delicacy is only prepared by licensed chefs who have undergone years of training. However, one wrong cut may mean the death the consumer. In fact, up to 50 deaths from pufferfish poisoning are reported in Japan annually. This is due to the potent tetrodoxin, which is 1000 times more lethal than cyanide. Due to this reason, the //fugu// is the only food which the Emperor of Japan is forbidden to eat.

<span style="color: #535353; font-family: Georgia,serif;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">In Singapore, //fugu// is partially banned and special permits are required to import the flesh while other parts of the fish is forbidden. Unfortun <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">ately, accidents are still reported. <span style="color: #535353; font-family: Georgia,serif; font-size: 8px; vertical-align: super;">20] <span style="color: #535353; font-family: Georgia,serif; font-size: 11pt;">Symptoms of poisoning include vomiting, diarrhea and paralysis. In severe cases, death can result from respiratory paralysis. Indeed, it is a dish to die for.

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=<span style="color: #00821c; font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">Glossary of Terms = **Bootstrap probability**: A computer based technique used to assess the accuracy of the statistical estimate.

<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> **Holotype**: A single physical example (or illustration) of an organism, known to have been used when the species was formally described.

<span style="color: #535353; font-family: Georgia,serif;"> ** Monophyletic group / clade ** : A group consisting of an ancestor and all its descendants, a single branch on the tree of life.

<span style="color: #535353; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3;"> **Neotype**: A specimen later selected to serve as the single type specimen when an original holotype has been lost or destroyed or where the original author never cited a specimen.

<span style="color: #535353; font-family: Georgia,serif;"> ** Phylogenetic relationship: ** The relative times in the past that species shared common ancestors. Two species are more closely related to one another than either one is to a third species if, and only if, they share a more recent common ancestor with one another than they do with the third species.

**Phylogenetic tree:** A branching diagram showing the inferred evolutionary relationships of various biological species or their phylogeny. Each branching shows successive events of speciation, in which one species gives rise to two.

<span style="color: #535353; font-family: Georgia,serif;"> **<span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">Terminal **<span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">: A protrusible mouth design which helps the fish to feed on other fish efficiently.

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=<span style="font-family: 'Trebuchet MS',Helvetica,sans-serif; font-size: 20px;">References = <span style="color: #535353; display: block; font-family: Georgia,serif; font-size: 15px; line-height: 1.3; text-align: left;">[1] Nelson, J. S., 1994. Family Tetraodontidae - Puffers. FishBase. URL: http://www.fishbase.org/Summary/FamilySummary.php?ID=448 (Accessed on 1 Nov 2014) <span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[2] Puffer Fish (Tetraodontidae). A-Z Animals. URL: http://a-z-animals.com/animals/puffer-fish (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[3] Ria, T. 2008. Pufferfishes (Tetraodontidaes) on the Shores of Singapore. WildFactsheets, 13 Dec 2008. URL: http://www.wildsingapore.com/wildfacts/vertebrates/fish/tetraodontidae/tetraodontidae.htm (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[4] Tetraodon nigroviridis Marion de Procé, 1822 – Checklist View. GBIF. URL: http://www.gbif.org/species/5213566 (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[5] Romero, P., 2002. //An etymological dictionary of taxonomy//. Madrid. unpublished.

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[6] Pethiyagoda, R., 1991. //Freshwater fishes of Sri Lanka.// The Wildlife Heritage Trust of Sri Lanka, Colombo. 362 p.

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[7] Rainboth, W.J,. 1996. //Fishes of the Cambodian Mekong.// FAO Species Identification Field Guide for Fishery Purposes. FAO, Rome, 265 p.

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[8] Frederick, L. C, 2001. Puffernet: Anatomy and Physiology. URL: http://puffernet.tripod.com/ap.html (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[9] Tetraodon nigroviridis (Green Spotted Puffer) – Seriously Fish. URL: http://www.seriouslyfish.com/species/tetraodon-nigroviridis (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[10] Tetrodoxin: The Neurotoxin. University of Maryland. URL: http://www.life.umd.edu/grad/mlfsc/zctsim/Tetrodo.html (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px; line-height: 1.5;">[11] Ria, T. 2008. Spotted Green Pufferfishes (Tetraodon nigroviridis) on the Shores of Singapore. WildFactsheets, 13 Dec 2008. URL: http://www.wildsingapore.com/wildfacts/vertebrates/fish/tetraodontidae/nigroviridis.htm (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[12] Froese, R. and D. Pauly (eds). 2012. FishBase. World Wide Web electronic publication. URL: http://www.fishbase.org (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[13] Talwar, P. K. and A. G. Jhingran. 1992. Inland fishes of India and adjacent countries. Volume 2. A. A. Balkema, Rotterdam.

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #935177; font-family: 'Trebuchet MS',',Helvetica,sans-serif; font-size: 11pt; font-size: 11pt; line-height: 1.3; line-height: 1.3; text-align: left; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[14] <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">Tetraodon nigroviridis. Genoscope - Centre National de Séquençage. URL: http://www.genoscope.cns.fr/spip/spip.php?lang=en (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #935177; font-family: 'Trebuchet MS',',Helvetica,sans-serif; font-size: 11pt; font-size: 11pt; line-height: 1.3; line-height: 1.3; text-align: left; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[15] <span style="color: #535353; font-family: Georgia,serif; font-size: 15.600000381469727px;">Tetraodon nigroviridis. ITIS Standard Report Page. URL: http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=646446. (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[16] Igarashi, Y., H. Doi, Y. Yamanoue, S. Kinoshita, T. Ishibashi, H. Ushio, S. Asakawa, M. Nishida, S. Watabe. 2013. Molecular phylogenetic relationship of Tetraodon pufferfish based on mitochondrial DNA analysis. //The Japanese Society of Fisheries Science//. 79:243–250

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[17] Nordlie, T., 2009. UF experts breed puffer fish in captivity; pet trade and genetics research could benefit. University of Florida News. URL: http://news.ufl.edu/archive/2009/02/uf-experts-breed-puffer-fish-in-captivity-pet-trade-and-genetics-research-could-benefit.html (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[18] Foreign Fisheries Developments. "Japan's "Fugu" or Puffer Fish Market" Marine Fisheries Review. URL: http://spo.nmfs.noaa.gov/mfr511/mfr51112.pdf (Accessed on 1 Nov 2014)

<span style="color: #935177; font-family: Georgia,serif; font-size: 11pt; line-height: 1.3; text-align: left;"> <span style="color: #535353; font-family: Georgia,serif; font-size: 15px;">[19] Yong, Y. S., L. S. Quek, E. K. Lim, and A. Ngo. 2013. A Case Report of Puffer Fish Poisoning in Singapore. //Case Reports in Medicine//. vol. 2013, Article ID 206971, 4 pages, 2013.

<span style="color: #535353; font-family: Georgia,serif; font-size: 15px; line-height: 1.3; text-align: left;"> [20] Dekkers, W. J., 1975. Review of the Asiatic freshwater puffers of the genus Tetraodon Linnaeus, 1758 (Pisces, Tetraodontiformes, Tetraodontidae). Bijdragen tot de Dierkunde. 45(1), 123

<span style="color: #535353; font-family: Georgia,serif; font-size: 15px; line-height: 1.3; text-align: left;"> [21] Nijssen, H., L. van Tuijl, J. H. Isbrücker, 1993. Revised catalogue of the type specimens of recent fishes in the Institute of Taxonomic Zoology (Zoölogisch Museum), University of Amsterdam, The Netherlands. Bulletin Zoologisch Museum. 13(18), 235

<span style="color: #535353; font-family: Georgia,serif; font-size: 15px; line-height: 1.3; text-align: left;"> [22] Yee, A. T. K., W. F. Ang, S. Teo, S. C. Liew and H. T. W. Tan, 2010. The present extent of mangrove forests in Singapore. //Nature in Singapore 2010//. 3: 139–145