D Colombo

S Cantournet

F Lequeux

H Montes

K Le Gorju

Towards constitutive equation for non-linear mechanics of PDMS-nanosilica

Nanosilica reinforced PDMS is a performing material, but with a complex mechanical response. In our project we aim at modelling its behavior through a multi-scale approach. This paper presents the main advancements in the development of a mechanical constitutive equation from a microscopic physical law. Experimental results prove the modification of the polymer chain dynamics. Mobility reduction is reproduced by gradients in the viscoelastic properties of confined matrix. A wide spectrum of relaxation times allows the representation of a large span of different behaviors by means of one single function.

under multi-axial loadings (Cantournet & Besson 2009[START_REF] Amin | Nonlinear dependence of viscosity in modelling the rate-dependent response of natural and high damping rubbers in compression and shear: Experimental identification and numerical verification[END_REF]. Macroscopic phenomenological approaches are limited by the amount of parameters which increase with the number of non-linearities. On the other hand, physical interpretations lead to a qualitative description of the overall phenomena, but not a quantitative one [START_REF] Papon | Unique nonlinear behaviour of nano-filled elastomers: from the onset of strain softening to large amplitude shear deformations[END_REF]). Thus, they cannot be used for structural prediction.

In our work, we aim at overcoming the limits of the mechanical and of the physical approach by combining them in a multiscale model, linking the microscopic behaviour to the macroscopic one. Firstly, we focus on the experimental characterisation of pure and silica filled PDMS in the linear and non-linear domain. Secondly, a local behaviour based on the modification of the matrix dynamic is set at a microscopic scale. In further developments, the assembly of confined domains to a mesoscopic network is investigated for the homogenisation of the material response.

EXPERIMENTAL SECTION

Materials

Pure PDMS and 40phr Silica filled PDMS from Hutchinson are tested on cyclic solicitation. An Anton Paar Physica MCR 501 rheometer with plate-plate configuration is used on cylindrical 1 INTRODUCTION Silica reinforced silicone rubbers are performing materials for many engineering applications. Because of their high chain flexibility and their chemical stability and tailoring, Polydimethilsiloxans (PDMS) are preferred for wide temperature and frequency range specifications. Their mechanical properties are improved by adding nano-silica aggregates, with strong compatibility with the silicone matrix.

At the same time, silica fillers introduce nonlinearities and long time evolutions in the PDMS response. Under cyclic solicitation, dependences on the strain amplitude-Payne effect [START_REF] Clement | Investigation of the Payne effect and its temperature dependence on silica-filled Polydimethylsiloxane networks. Part I: Experimental Results[END_REF])-and on the maximum stretch-Mullins effect [START_REF] Clement | Study of the reinforcement mechanisms in silica-filled polydimethylsiloxane networks[END_REF]-are observed on filled PDMS, as on other reinforced rubbers [REF]. Stress-strain Lissajous curves show non-linearities resulting from static-strain/precycling-strain couplings [START_REF] Vincent | response for a 2 nm glassy bridge under constant rate stretch (10 -4 s -1 ) and T = T g + 7K: no mobility reduction (Δ); mobility reduction and dissipation ( ); stress induced loss in the mobility reduction[END_REF]. Moreover, reinforcement is proven to be temperature dependent and, when mechanically modified, long time recovery is experienced [START_REF] Vincent | response for a 2 nm glassy bridge under constant rate stretch (10 -4 s -1 ) and T = T g + 7K: no mobility reduction (Δ); mobility reduction and dissipation ( ); stress induced loss in the mobility reduction[END_REF]. Those strong nonlinearities are depending not only on the external solicitation and environmental conditions but also on the morphological structure and filler-matrix interaction [START_REF] Montes | Particles in model filled rubber: Dispersion and mechanical properties[END_REF]).

To our knowledge, existing mechanical models fail in predicting all these multifaceted behaviours specimens (diameter 8mm-thickness 2mm). DSC measurements with a Q800 TA Instrument at 20°C/ min increase rate shows a glass transition temperature T g at -123°C for both materials, while the fusion temperature T f is -39°C for the pure PDMS and -41°C for the filled PDMS. Stabilised cycles are studied and moduli are computed with first order linear approximation. Temperature sweeps at 1 Hz are performed between -30°C and 130°C, within the linear domain-small strain. Increasing strain amplitude sweeps are performed at temperature 30°C at the same frequency and compared for both materials.

Results

In the linear domain, filled PDMS exhibit a remarkable enhancement in the storage modulusmore than one order of magnitude-typical of reinforced elastomers.

Figure 1 shows the evolution of the storage modulus with respect to the temperature for filled and unfilled PDMS. A clear difference in the temperature dependence is observed: filled elastomer follows a decreasing trend, while pure silicone rubber shows an increasing storage modulus as predicted by classical entropic elasticity [START_REF] Treolar | The physics of rubber elasticity[END_REF].

Fillers modify the viscous response as well. The dissipative factor, in Figure 2, is 3 times larger for filled material. The latter is also more sensible to temperature variation.

In the non-linear domain PDMS-nanosilica composite sample are affected by Payne effect (Fig. 3).

At increasing strain amplitude, filled material shows a drop in storage modulus with respect to the linear behavior observed on unfilled PDMS. This behavior is proven to be temperature dependent. In particular, filled silicone rubber has a large strain trend which is the opposite of small strain one-linear domain. It is rather similar to the one observed on the unfilled matrix.

Discussion of the experimental results

The empirical analysis put into evidence the temperature dependence of the reinforcement phenomenon. Filler addition has an influence other than the simple hydrodynamic effect [START_REF] Guth | Theory of filler reinforcement[END_REF], since it affects the dynamical response of the matrix. At a microscopic level, physico-chemical interactions induce a reduction of polymer chain mobility, thus, leading to an increase of the elastic and viscous responses. Confined zones have reasons to be regions where the presence of silica has a stronger effect.

The mobility reduction is proven to depend on many factors. Figure 4 sums up the outlines of the experienced dependences. In the following section, we aim at building a microscopic scale model that takes the observed behaviors as key element of the mechanical description. At the current state, a quantitative comparison is not developed yet, though the main principles may be applied to other filled systems, other than silicone rubber based ones.

MICROSCOPIC MODEL

Local viscoelastic behavior

In the previous section, silica filler is proven to modify the dynamical response of the matrix. Filler-matrix interactions cause a reduction in chain mobility of the matrix surrounding the aggregate [START_REF] Berriot | Evidence for the shift of the glass transition near the particles in silica-filled elastomers[END_REF][START_REF] Papon | Low-field NMR investigations of nanocomposites: polymer dynamics and network effects[END_REF]). This modification is modeled by means of a glass transition temperature gradient of few nanometers wide, represented by the formula:

T T z g g T T T T ( ) z z = + ⎛ ⎝ ⎜ ⎛ ⎛ ⎝ ⎝ ⎞ ⎠ ⎟ ⎞ ⎞ ⎠ ⎠ ⎡ ⎣ ⎢ ⎡ ⎡ ⎢ ⎣ ⎣ ⎢ ⎢ ⎤ ⎦ ⎥ ⎤ ⎤ ⎥ ⎦ ⎦ ⎥ ⎥ ∞ 1 1 δ ν (1)
where T g ∞ = glass transition temperature at the bulk (without solicitation); δ = filler-matrix interaction coefficient [in nm]; ν = gradient exponent; and z i = distance from the particle surface within the glassy bridge.

Wherever aggregates are sufficiently close, a confinement condition is attained, with thermodynamic properties different from the bulk polymer, typically an enhanced glass transition temperature. These regions are modeled as overlaps of multiple glass transition gradients: a rigid connection, called glassy bridge, establishes a preferential path for force transmission between aggregates. The set of glassy bridges forms a temperature depending percolation network, responsible of the reinforcement effect at the macroscopic scale [START_REF] Montes | Influence of the glass transition temperature gradient on the nonlinear viscoelastic behaviour in reinforced elastomers[END_REF].

Moreover, external solicitation modifies matrix properties within the confined region. Glass transition temperature is proven to depend on the stress field [START_REF] O'connel | Large deformation response of Polycarbonate: time-temperature, time-aging time and time-strain superposition[END_REF]. For a glassy bridge scale under applied strain, viscous flow phenomena take place, leading to energy dissipation and Payne effect at macroscopic scale [START_REF] Montes | Influence of the glass transition temperature gradient on the nonlinear viscoelastic behaviour in reinforced elastomers[END_REF]. Local glass transition temperature is, thus, expressed within a glassy bridge by:

T T z f g i T T g T T i δ ν ( ) z i z , σ = + ⎛ ⎝ ⎜ ⎛ ⎛ ⎝ ⎝ ⎞ ⎠ ⎟ ⎞ ⎞ ⎠ ⎠ ⎡ ⎣ ⎢ ⎡ ⎡ ⎢ ⎣ ⎣ ⎢ ⎢ ⎤ ⎦ ⎥ ⎤ ⎤ ⎥ ⎦ ⎦ ⎥ ⎥ -( ) eq σ ∞ 1 1 (2)
where z i = distance from the surface of the ith particle of the glassy bridge; and f(σ eq ) is a scalar function of an equivalent local stress.

In the glass transition gradient approximation, the mechanical behavior of the polymer surrounding the particle gives responses spanning from the pure rubber up to the glassy state. The local viscoelasticity is reproduced by means of a Zener model (Fig. 5): on one side a hyperelastic function describes the purely rubbery behaviorwith modulus of the order of few MPa-; on the other side a glassy elastic term-modulus of few GPa-is coupled with a locally variable relaxation time τ, whose expression is directly deduced from the William-Landell-Ferry law and the glass transition temperature gradient:

Log C T T C T T g g i T T g i T T τ τ g ( ) T i σ T z i , , i z i ⎡ ⎣ ⎢ ⎡ ⎡ ⎣ ⎣ ⎢ ⎢ ⎣ ⎣ ⎣ ⎣ ⎤ ⎦ ⎥ ⎤ ⎤ ⎦ ⎦ ⎥ ⎥ ⎦ ⎦ ⎦ ⎦ = - -( ) z i z σ , ⎡ ⎣ ⎤ ⎦ ⎤ ⎤ - T ( ) z i z σ , ⎡ ⎣ 1 C C 2 C C ⎤ ⎤ ⎦ ⎤ ⎤ ⎤ ⎤ (3)
where the local shift factor a T Tg is expressed as the ratio between the local relaxation time and the relaxation time at the glass transition temperature; C 1 and C 2 = WLF material parameters; and T = temperature.

The present assumption allows the description with a unique and simple model of all the behaviors between two antithetical trends. Moreover, it includes a wide relaxation spectrum of relaxation time which may be able to reproduce long time recovery phenomena.

Previous equations have been implemented in the Finite Element code Z-set (http://www.zset-software. com) in large strain formulation. The axis of the glassy bridge between two silica particles has been reproduced by a stick of concatenated cubic c3d20 elements, where the glassy bridge height-minimum particle to particle distance-is denoted h 0 (Fig. 6).

Preliminary results

In the first instance, cyclic stretching simulations are performed from an unsolicited configuration with a glass transition temperature gradient law, with no stress dependence (Equation 1), at temperature T g + 20 K. Figure 7 shows the profile of the relaxation times along the axis, for a 2 nm glassy bridge. Filler-matrix interaction parameters are chosen δ = 0.1 nm and ν = 0.88. Similar behaviors are observed at different glassy bridge height h 0 , for different temperature.

At the unsolicited state, relaxation time profile is much higher than the relaxation time of the bulk matrix. As a consequence, the glassy bridge behaves as a rigid element.

Under stretching, relaxation time undergoes a smooth reduction till the point of viscoelastic transition between the glassy and rubbery behavior. The bridge behavior is soft at maximum stretch.

Finally, if no stress dependence in present the initial relaxation time profile and stiffness is established.

In the case of a stress dependent glass transition temperature (Equation 2), the stiffness reduction is anticipated by a viscous flow phenomenon. Figure 8 shows the traction force under uniaxial stretch at a constant strain rate of 10 -4 s -1 and at temperature T g + 7K.

The behavior in absence of mobility reduction corresponds to the one at high temperature: the mechanical response is the one of a pure rubber matrix. The addition of a mobility reduction by mean of a glass transition temperature gradient (Equation 1) increase of 3 orders of magnitude the mechanical response and introduce a dissipative term within the system. The stress dependence glass transition temperature (Equation 2) leads to a yielding-like phenomenon, with a viscous flow behavior induced by the implicit interdependence of stress and relaxation time.

Discussion of the numerical results

The strong non-linearities are induced by both confinement and stress dependence f(σ eq ). The reduction of chain dynamics observed by the experiments is reproduced by taking into consideration the filler-matrix interaction. Rigid-like behavior is simulated at temperature higher than the glass transition temperature. Hence, the local stiffness is temperature dependent.

The application of an external solicitation modifies the relaxation times profiles on two aspects. On one side it changes the glassy layer overlap, affecting the scope of the filler-matrix interaction. On the other side it forces a viscous behavior, leading to a softening of the glassy bridge and strong dissipation within the material.

In conclusion, the dynamical dependencies experimentally observed are here reproduced with one single physical law at the microscopic level.

CONCLUSION

In the present paper we discuss the actual advancement in the definition of a mixed physical-mechanics approach, which aims at describing the complex macroscopic behaviour of reinforced elastomers. The experimental characterisation brings us to the conclusion that the reinforcement observed for silica filled PDMS is driven by the modification of the dynamical response of the matrix. Environmental parameters, external solicitation and morphological organisation are main factors on the composite response.

Chain mobility reduction is reproduced by means of a function of glass transition temperature within confined regions, called glassy bridges. From a glass transition temperature gradient approximation, we develop a local mechanical model. One single law is used to represent the set of dependencies observed at the empirical level.

From this microscopic physical law we aim at building a multi-scale homogenisation approach. In future perspective the computed law will be applied on a Representative Elementary Volume of the filler-elastomer morphology [START_REF] Jean | A multiscale microstructure model of carbon black distribution in rubber[END_REF][START_REF] Jean | Large-scale computation of effective elastic properties of rubber with carbon black fillers[END_REF]. The network of glassy bridges is used to obtain a macroscopic response.

Figure 1 .

 1 Figure 1. Storage modulus vs. temperature-linear domain: filled PDMS (•); pure PDMS ( ); entropic elasticity fit (dashed line).

Figure 2 .

 2 Figure 2. Dissipation factor vs. temperature-linear domain: filled PDMS (•) and pure PDMS ( ).

Figure 3 .

 3 Figure 3. Storage modulus vs. strain amplitude: pure PDMS ( ); filled PDMS (•) at different temperatures.

Figure 4 .

 4 Figure 4. Fillers modify matrix dynamical response: chain mobility reduction: sum up scheme of the chain mobility reduction observations.

Figure 5 .

 5 Figure 5. Chain mobility reduction is modeled by a local Zener large strain viscoelasticity: a relaxation time function depending on filler-matrix interaction, stress tensor and temperature is applied.

Figure 6 .

 6 Figure 6. The axis of a glassy bridge confined between two particles is represented by a stretched stick of cubic elements and tested by Finite Element Method.

Figure 7 .

 7 Figure 7. Relaxation time vs. position on the axis for a 2 nm glassy bridge-mobility reduction induced from glass transition gradient overlap T g (z i ): unstretched configuration (dashed line); stretched configuration (full line); bulk behavior (point line).


Régis Hocdé

Indra Bayu Vimono

Amir M Suruwaki

Yosephine Tuti

Rizkie Satriya Utama

Abrar Mohammad

Paulus Boli

Hidayat Ashari

Hadi Wikanta

Jean-Baptiste Juhel

Christophe Cochet

Christophe Thébaud

Borja Milá

Gofir Abdul

Julien Leblond

Eric Bahuet

Gilles Diraimondo

Bruno Fromento

Jean Chevallard

Bernard Sumanta

Pouyaud

Laurent Kadarusman

Kaimana Regency

West Papua

Indonesia Régis

Indra Bayu

Sumanta

Bernard Pouyaud

Laurent Pouyaud

Mission report: LENGGURU 2017 Expedition 'Biodiversity assessment in reef twilight zone and cloud forests'

Keywords: birds, BRUV, capacity building, DNA barcoding, echinoderms, environmental DNA, gorgonians, hard corals, marine biodiversity, marine invertebrates, oceanographic cruise, reef fishes, scientific expedition

établissements d'enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Table of contents

Summary : Keywords: Table of contents

I BACKGROUND AND AIMS

I-1 Background

Indonesia is the world's largest archipelago, lying between the Indian and Pacific Oceans and encompassing the heart of the Coral Triangle (CT) [START_REF] Veron | Delineating the Coral Triangle[END_REF]. Indonesia is home to the richest marine biodiversity on Earth, with nearly 20% of the world's coral reefs, 10% for seagrass and 15% for mangroves ecosystems. The Bird's Head Isthmus, also called the Bird's Neck, connecting the Bird's Head Peninsula (BHP) with the rest of New Guinea, is one of the last pristine areas remaining in Southeast Asia. The Lengguru limestone karsts have evolved through complex tectonic uplift movements. For the last 10 million years, they have formed "islands within the island" which is today a major biodiversity reservoir with high levels of endemism.

In many areas of Southeast Asia, karst systems have formed "islands within islands" [START_REF] Clements | Limestone Karsts of Southeast Asia: Imperiled Arks of Biodiversity[END_REF].

The high species diversity and endemism generally observed in karst landscapes is explained by a large array of ecosystems, complex and fragmented terrains, variable local climatic conditions, tectonic and eustatic histories, variable degrees of isolation, and incidence of random events. Karsts cover large sections of New Guinea but their exploration is the least advanced of any ecosystems in the Australasian region. In a review, [START_REF] Clements | Limestone Karsts of Southeast Asia: Imperiled Arks of Biodiversity[END_REF] stated that karsts contribute just 1% of the global and regional biodiversity research output from terrestrial and freshwater ecosystems conducted over a 20-year period (1985)(1986)(1987)(1988)(1989)(1990)(1991)(1992)(1993)(1994)(1995)(1996)(1997)(1998)(1999)(2000)(2001)(2002)(2003)(2004). Given that karsts cover around 10% of the land area in Southeast Asia [START_REF] Day | An assessment of protected karst landscapes in Southeast Asia, Cave and Karst[END_REF] and given that they encompass significant proportions of the surrounding region's biodiversity and endemism [START_REF] Yong | Sarawak Bau Limestone Biodiversity[END_REF], more studies need to be devoted to these ecosystems.

The Lengguru karsts are biogeographically located at the heart of the Coral Triangle (CT) (Veron J. (Charlie) E. N. et al., 2011). Distributed on six Southeast Asian countries (Indonesia, Malaysia, Philippines, Papua-New-Guinea, Solomon, Timor-Leste), the CT is the marine mega diversity centre of the planet with more than 76% of world's coral reef species and more than 52% of Indo-pacific reef fish diversity. The richest diversities for soft corals, molluscs, crustaceans and many other biotas are also observed in the CT. The CT biodiversity epicentre is located in West Papua, an area still unexplored.

The Coral Triangle significantly sustains the world's fisheries, tuna industry and tourism sectors and direct activities for more than 200 million peoples. Nevertheless, intensification of environmental pressures coupled with impacts of global warming have already degraded around 40% of global reefs and mangroves during the last 40 years.

The largest archipelago in the world, Indonesia extends across more than 5000 km from west to east and encompasses more than half of the CT. Aware of the environmental and societal challenges faced by people and economic development, the Indonesian President Bapak Joko Widodo gave the sustainable management of marine resources one its major priority for the country.

Increasing the characterization of marine genetic resources by training and providing schorlarships to young researchers is a prerequisite for meeting this crucial and interesting challenge. The general methodology is based on field surveys conducted in a large array of marine and mountain ecosystems. Specific ecosystems were selected according to the preliminary data obtained during the Franco-Indonesian expedition Lengguru 2014. Besides conducting an inventory of animal communities based on DNA barcoding and traditional taxonomy, biologists will also infer the phylogenetic relationships of the taxa collected in Lengguru with those originating from the peripheral regions.

The confrontation between the chronologies of geomorphologic processes with molecular clocks will enable researchers to understand their diversification processes and will test the karst region's capacity as a reservoir of old lineages, speciation hotspots or arks of biodiversity.

This project also will provide an opportunity to increase significantly the barcoding inventory of marine resources in the Coral Triangle.

1-3 Objectives

The "Lengguru 2017" scientific expedition aims to assess the functional, genetic and morphological diversity of several marine biotas (echinoderms, hard corals, gorgonians and reef fishes). Exploration and sampling effort were concentrated for the marine part on several reef slopes located in front of the Kumawa and Lengguru ranges, in and off shore of Triton Bay (from -100 m to the surface). Species collection and sampling were performed by teams of divers in open circuit and closed circuit rebreather (CCR). Nets were also used. Barcoding of sampled species was systematically performed. Environmental DNA and baited cameras were used to investigate the marine biodiversity and physico-chemical variables (CTD, other variables) were measured to characterize the abiotic environment. The project Lengguru 2017 Expedition also aims to inventory the morphological and genetic diversity of mountain avifauna.

The study area mainly focused on the '540 -SW coast of Papua' ecoregion of the Coral Triangle defined by [START_REF] Veron | The Coral Triangle[END_REF].

II-7

List of participants: scientific staff, crew, logistics

The project comprises 19 Indonesians and 13 foreigners distributed in 2 research teams (ornithology and marine biology), 1 security and health team and 1 crew team (Airaha2 vessel). 120m). Submerged anticlines form complex islands networks, rias and shallow rocky banks. For example, in Triton Bay, faults and rock collapses are numerous and represent original submarine landscape similar to natural labyrinths with high potentialities for exploring various assemblages of organisms. On the contrary, the southwest extremity of Kumawa is exposed to the strong Torres' east-west surface current, and consists of a sharp slope plunging to the abyssal pit of the Seram Sea and facing a permanent upwelling.

Our preliminary explorations on the Lengguru coastline show the following:

-Presence of a large sedimentary cone (terrigenous inputs from continental erosion) in the Arguni Bay and extending until the south cape of Pulau Adi; accumulation of deep and unstable submarine dunes alternating with numerous scattered coral reefs, -A mesotidal regime characterized by asymmetric tides (2,6 m maximal range), tidal propagation extending far upstream (up to 100 km) in Arguni Bay (Sewiki Lake) and sometime forming tidal bore (observed on Lengguru River in October 2014),

-An important hydrodynamic regime with a westward-flowing current particularly strong on exposed capes (south extremity of Nusa Ulan Cape) with the presence of submarine hydraulic sand dunes with depths ranging between -20 and -50 m, -A rugged and tormented geomorphology at south extremity of Nusa Ulan Cape with absence of fouling and presence of scarce benthic fauna on substrate exposed to current until 90-100 m depths, -A disjoint abundance for many organisms compared to other areas in Indonesia with generally abundant species becoming rare in Lengguru and conversely rare species elsewhere becoming more abundant in the area, -An important species diversity within each studied group but characterized by low intraspecific abundance at a given locality, -A spectacular species diversity of soft corals and associated organisms in Triton Bay, -A stratification of species assemblages according to depth with some species characterized by an unusually shallow occurrence (20-40 m) that are generally confined to depths below about 50-100m at most locations in the Indo-Pacific; an ascent of deep species probably explained by the permanent turbidity of the surface section of the water-column.

Exploration of outer reef slopes were made at six locations (

IV METHODOLOGY, FIELD STUDIES AND DATA MANAGEMENT

IV-1 Sampling methodology and data acquisition

The sampling methodology for the marine biotas consisted of the manual collection of hard invertebrates (hard corals, gorgonians, echinoderms) and reef fishes along a vertical transect from -100m to the surface, and to determine their distribution according to depth. Each collected sample was photographed and labeled with a detailed description of its habitat (GPS coordinates, depth, salinity, etc.). The samples consisted of vouchers (Museum collections) and tissues for subsequent DNA barcoding analysis.

The specimens of reef fishes were mainly collected by hand or with 4 to 8 m long bottom gillnets deployed by open-circuit and closed-circuit divers in the 0-100 m depth range (Hocdé R. et al., 2017). Eugenol or spearguns have been used sometimes. Some brackish and estuarine fishes were also collected with 10 m beach purse seines, and pelagic fish with line fishing and spearfishing.

The inventory of reef fishes was also performed with environmental DNA (eDNA) in order to detect hidden diversity. This consisted of the filtration of 2L of sea water (with 1 replicate) at different depths for each sampling station.

Besides the implementation of DNA barcoding and eDNA for the evaluation of reef fish diversity, a complementary approach was performed with the use of Stereo Baited Remote Underwater Video Systems (stereo-BRUVS). This observation tool allows to monitor reef communities, detect elusive species and investigate their behavior.

IV-2 Collection of biological samples

The marine samples were preserved in 70% ethanol, with fragments in 95% absolute ethanol, and stored at -20°C for genetic analysis.

Hard corals skeletons were air dried and stored at ambient temperature.

Blood and tissue samples from bird species were stored in pure ethanol for genetic analysis. Some specimens were collected and prepared as study skins, and are deposited at the Museum Zoologicum Bogoriense (Cibinong, Indonesia), and others were released after measurements and analysis.

IV-3 Oceanographic observations

Physico-chemical variables (CTD, other variables) were measured with both surface and autonomous underwater sensors (vertical profiles, eDNA sampling, diving profiles).

Measurements were performed in different habitats: offshore, coastal zones, estuaries, lagoons, upper lakes.

IV-4 Sample management, data accessibility and data citation

All samples are managed by Indonesian institutions. All data will be shared through international databases (NCBI, BOLD, GBIF…).

IV-5 Research ethics and field studies

The Lengguru 2017 Expedition is based on a joint collaboration between Indonesian and European scientists and is covered by a MOU signed between LIPI and IRD on 5th April 2017. The project is co-organized by IRD (French Institution), LIPI-P2O (National counterpart) and Politeknik-KP-Sorong (Local counterpart). RISTEK Authorization: 3179/FRP/E5/Dit.KI/IX/2017.

V PRELIMINARY RESULTS AND FURTHER CONSIDERATION -MARINE TEAM

V-1 Brief description of the main habitats

Research sites

The Lengguru Range is located in the Kaimana Regency (West Papua, Indonesia). It is characterized by a series of parallel and oblique folds, with altitude ranging from 600 to 1500 m. These carbonate anticlines are separated by deep valleys sometimes closed and rifted with no outflow of collected water (i.e. endorheic).

Lengguru's topography consists of karst limestone shaped by uplift or subsidence geologic events and under the dissolving action of water on carbonate bedrocks since a dozen million years ago (cc. 10-12 Myr.). This rough and fragmented landscape displays unusual surface and marine subsurface features including sinkholes, vertical shafts, and multiple reef slopes.

These sedimentary outcrops are therefore characterized by a large array of ecological niches afforded by complex terrains, and promoting unexpected levels of biodiversity. Lengguru seafront consists of drowned karsts geologically built during successive marine regressions since the end of the Miocene. Preliminary investigations performed in 2014, revealed an important richness of ecosystems, such as submerged freshwater springs (Vruljas), flooded river canyons, stratified estuaries, coastal marine lakes, and reef slopes. We plan to explore these areas of interest with the help of a diving team equipped with conventional scuba systems (using Open Circuit with tanks and regulators) or with recycled-air scuba system (using some Closed Circuit Rebreathers -eCCR). Areas of investigation will be made at Nusa Ulan (Kumawa-Buruway), Pulau Venu (Arguni Bay), around Pulau Buasai, in & off shore of Triton Bay.

Montane cloud forests : Tops of Gunung Kumawa

Papuan biodiversity is known to increase with altitude. In Lengguru, the tops of anticlines are generally characterized by Lapies formations. Lapies are fissured limestone plateaus consisting of natural labyrinths and forming patchworks of habitats and forest types according to altitude, geological substrate, exposure to trade winds, rainfall and remoteness from seashore. These extreme ecosystems, never studied nor visited, were explored for the first time in 2014. We discovered that these original habitats host a number of differentiated avian taxa, with several potential new subspecies and species of birds.

V-2 Echinoderms

Echinoderms collected specimens:

-17 sea stars -7 sea cucumbers -7 sea urchins DNA samples :

(identical)

Results to date > The principal investigator for Echinoderms, I. B. Vimono (RCO-LIPI) has obtained a 3-year PhD scholarship (2019 -2022) from the French Foreign Office. The PhD will be registered at Montpellier University (France) under the supervision of L. Pouyaud (IRD) and P. Borsa (IRD). By the way, all material collected during 2014 and 2017 expeditions and additional material from other locations in Indonesia will be analyzed between the end of 2019 and the beginning of 2020 by I. B. Vimono.

V-3 Gorgonians

Gorgonians All material was sequenced in France between the end of 2018 and the beginning of 2019.

At the present time, the molecular database of 1500 reef fishes has been achieved. Morphological and sequences data reveal the existence of around 450 species. This represents near half of expected species in the area following Allen G. R. & Erdmann M. V. (2012). To our knowledge, this is the largest reef fish molecular barcoding database available in the Coral Triangle. This database will be used as reference database for assignment of sequence data obtained from the eDNA operation.

COI DNA barcode revealed the presence of several new species which will be described soon from voucher specimens stored at RCO-LIPI.

Several collaborations were initiated to valorize these data and currently several papers are being prepared.

V-6

Stereo Baited Remote Underwater Video Systems (stereo-BRUVS)

Underwater video devices consisted of two Gopro© Hero 5 camcorders oriented horizontally toward a bait with an inward convergent angle of 8°. The bait was a mix of dried sardine powder, flour and sardine oil placed in a PVC canister suspended approximatively 1.4m in front of the cameras. The stereo-BRUVS were deployed from the boat at 6-20m deep (mean ± SD: 12.8 ± 3.8m) for one-hour recording. For each site, the stereo-BRUVS were set at least 400m apart. A total of 53 stereo-BRUVS were deployed.

Stereo-BRUVs 53 records image analysis in progress

Video analyses > The footage will be hosted on an interactive platform named Webfish currently under development at the university of Montpellier. This open access platform will be connected to internet and will allow all Lengguru collaborators to process the footage. This platform will allow to measure species richness and abundance of marine organisms. It will also store thumbnails of each species to provide an image database for the development of automated species identification.

Ultimately, the ichthyological assemblages obtained using Stereo-BRUVS will be compared to the environmental DNA sequences to confirm identifications and evaluate the performance of both methods. The marine component of the Lengguru expedition aimed to investigate marine biodiversity in the Bird's Head region of West Papua. This objective was built around three axes: 1) Create a genetic reference database of sampled marine organisms, 2) Retrieve environmental DNA from water to implement a metabarcoding biodiversity assessment, and 3) Deploy baited underwater camera to record marine biodiversity on video.

Figure 13. CCR diver collecting water for eDNA.

Environmental DNA > The environmental DNA (eDNA) is an innovative tool allowing to detect the presence of organisms without directly seeing them. This method is based on the principle that organisms leave genetic traces in their surroundings through organic material including feces, mucus, gametes, shed skin or carcasses. The retrieved eDNA is then amplified, sequenced and compared (sequence assignment) to the genetic sequence databases (Lengguru reference database, GenBank).

Sampling > In the Lengguru expedition, eDNA was collected from water using different methods according to the sampled depth. For surface samples, the water was collected in DNA-free plastic bags from a dinghy or during snorkeling facing the current. For mid-water samples (10 -100 m deep), the water was sampled in DNA-free plastic bags during close circuit rebreather (CCR) diving. For deepwater samples (100 -300m deep), the water was sampled using Niskin bottles. eDNA 106 0.22µm filters 10 0.45µm filters Filtration > Two types of filter were used according to the water turbidity to avoid clogging. Marine and clear river/lakes samples were filtered using 0.22µm Sterivex filter units. A volume of 2L was filtered for each point with a duplicate (2L x 2 filters) at different depth (from surface to 270m). A total of 106 0.22µm filters were used.

Estuaries, bays and rivers samples were filtered using 0.45µm Sterivex filter units. A volume of 5L was filtered for each point with a duplicate (5L x 2 filters). A total of 10 0.45µm filters were used for surface sampling only. The DNA sequences obtained with the 0.45µm filters and 10 of the 0.22µm filters were amplified with the vertebrate 12S primer V05 to evaluate the diversity of vertebrate and optimize the laboratory analyses protocol.

The DNA sequences from all the filters were amplified with a teleost specific primer Teleo2 to evaluate the diversity of fishes. This approach is more precise and allows to get a clearer picture of the ichthyological assemblages.

Results to date > A big corpus of DNA sequences was obtained. Species assignment to online reference databases and custom database from species barcoding indicates the occurrence of around 393 marine fish species. Both 12S online reference databases and custom database cover 40% of the checklist of coastal fish (1,610 species). Thus, a large fraction of fish species is not sequenced underestimating the diversity found in our samples.

Two scientific articles have now been submitted. The first article reveals the incompleteness of online reference databases and put forwards new approached to overcome it. The second paper uses eDNA to estimate the hidden diversity left undetected by both traditional census methods and checklists.

VI PRELIMINARY RESULTS AND FURTHER CONSIDERATION -TERRESTRIAL TEAM

VI-1 Ornithology

Scientific priorities > During the Lengguru 2014 expedition, the ornithological team captured at 1200m elevation in the Kumawa Mountains a male bird that belongs to the berrypecker genus Melanocharis but appears to be an undescribed taxon.

The examination of the bird in the field quickly revealed that it shared similarities with M. longicauda. However, it became also quite clear that this bird belongs to a population that may represent a distinct taxon, be it a different subspecies of longicauda or even a different species of Melanocharis : while tail pattern and upperparts were like nominate longicauda, its underparts were shiny white with a lemon wash on throat, breast, and flanks. A specimen was collected by LIPI personnel and deposited at the MZB. Comparisons between this specimen and other Melanocharis specimens revealed further differences, the most obvious being a difference in body size, the Kumawa individual being larger than any other Melanocharis in the collection.

Altogether, the substantial differences between the Kumawa berrypecker and other berrypeckers were pointing towards a putatively undescribed species.

We proposed, as a first priority for the Lengguru Expedition 2017, to increase sampling effort in the Kumawa mountains to provide a robust scheme into which to determine whether the undescribed Kumawa Melanocharis population represents a new species or a subspecies of Melanocharis longicauda.

The mountains of New Guinea consist of the Central Range plus a number of smaller outlying ranges. Seven of these outlying ranges have been recognized in western Papua (Papua Barat), including the Kumawa Mts. A number of montane bird species or superspecies are distributed across these mountain ranges and available data suggest that population differentiation has occurred between populations inhabiting the different ranges [START_REF] Diamond | New Distributional Records and Taxa from the Outlying Mountain Ranges of New Guinea[END_REF][START_REF] Beehler | [END_REF]. Therefore, we proposed as a second priority to identify the relationship between the Kumawa Mts populations and populations from other mountain ranges, since there is some indication in the taxonomic literature that in some species these populations may be more closely related to populations from the Wandammen, van Rees, and Foja than to populations from the Arfak/Tamrau system. During the Lengguru Expedition 2017 we then sought to obtain samples, amenable to population genetic analyses, from a number of species with challenging taxonomy and distribution: Heteromyias albispecularis, Tregellasia leucops, Phylloscopus poliocephalus, Peneothello bimaculata, Crateroscelis murina, Sericornis beccari/nouhusy?, Sericornis spilodera, and Meliphaga orientalis.

Sampling > Our sampling strategy consisted in capturing birds in several localities between 1100 and 1200m elevation on the southern slopes of Kumawa Mts., in a habitat that can be described as cloud forest. Birds were captured using standard mist nets, targeting primarily understory birds. Individuals were aged using plumage characteristics, and the degree of skull ossification. They were weighed and marked permanently with numbered metal rings to avoid resampling the same individuals. Wing rulers and dial calipers were used to acquire on each individual a set of standard morphometric measurements. Approximately 10 microliters of blood were collected by venipuncture from each bird and immediately conserved in 100% ethanol for further genetic analyses. Every captured bird has been photographed from various angles according to a standard procedure. Species were identified by eye in the field based on the team's expertise, the new field guide "Birds of New Guinea" (Pratt T. K. & Beehler B. M., 2015), and unpublished documents kindly provided by Dr. Beehler.

Results > We did 15 days of netting at 1200 m and 4 at 1100 m, with an average of 14 nets (12 meters each) and 6 hours of operation per day for a total of about 1400 net hours, between October 22 and November 9, 2017.

A total of 215 birds were captured, belonging to 26 species (Table 9).

We were successful at capturing three additional individuals of the undescribed Melanocharis as well as individuals of all other target species with the exception of Peneothello bimaculata.

VII POST-EXPEDITION VALORIZATION

VII-1 Capacity building, research fellowships

More than ten different events were organized in Indonesia and in France during the post-Lengguru Expedition for promoting research acquisition and valorization and for strengthening research capacities.

VII-2 Ongoing publications

The work in progress concerns all groups. Several publications are expected from this year 2020: on eDNA, on the genetic sequence database of reef fish, on gorgonian and hard coral taxonomy... These Lengguru 2017 results will also allow the production of other publications in ecology or in the context of larger-scale inter-site comparisons in the future.

VII-3 Medias, communications, scientific dissemination

Since the end of the expedition, several actions were made including :

-TV documentaries broadcasted on ARTE, Ushuaia TV, TV5 Monde channels -Press, medias -Productions dedicated to young people (seminars in school, articles to young public) -Scientific dissemination (seminars in several festivals)

  investigators of the research project and the scientific expedition & contacts II-4 Marine operations management II-5 Collaborative Institutions / Research Units at IRD II-6 Institutional support and corporate sponsorship II-7 List of participants: scientific staff, crew, logistics II-8 Network of experts III Operations timetable & list of stations....................................................................................... III-1 Metadata of the oceanographic cruise III-2 Planning of Marine Biology and Ornithology teams III-3 General description of research areas and visited locations, Cruise tracks III-4 Statistics of scientific dives III-5 List of stations IV Methodology, field studies and data management .................................................................... IV-1 Sampling methodology and data acquisition IV-2 Collection of biological samples IV-3 Oceanographic observations IV-4 Sample management, data accessibility and data citation

Figure 1 .

 1 Figure1. Global map with the reef fish species distribution. Location of the coral triangle(Veron et al. 2007)

Figure 2 .

 2 Figure 2. The 'Lengguru 2017' study area (yellow) among the 16 ecoregions of the Coral Triangle (Veron et al. 2011)

  Figure 4. Airaha2 R/V (© G. Diraimondo -Lengguru 2017)

Figure 5 .

 5 Figure 5. Lengguru 2017 team front Airaha2 vessel (© G. Diraimondo -Lengguru 2017)

  Figure 20): (1) inner part of Triton Bay; (2) eastern part of Pulau Buasai; (3) outer part of Triton Bay; (4) Bitsyari Bay; (5) Pulau Venue; (6) southern part of Nusa Ulan.

Figure 6 :

 6 Figure 6: Research area (yellow lines) with localities explored by diving teams (red triangles), eDNA locations (yellow lozenges) and BRUVs locations (orange circles)

Figure 8 :

 8 Figure 8: Geographic location of study sites

Figure 11 .

 11 Figure 11. Calibration of Stereo BRUVS for 3D measurements

Figure 14 .

 14 Figure 14. Map of the Bird's Head of Papua showing the location of eDNA samples using 0.22µm (white) and 0.45µm filters (red).

Table 1

 1

		-List of foreign participants -Lengguru 2017
	Nb: IRD** non official staff invited and under the responsibility of IRD
		Foreigner	Nationality Institution	Activities	Period in
		name/surnam				Indonesi
	1	POUYAUD Laurent	French	IRD (ISEM)	Reef Fish Genetics Taxonomy	01/10 to 29/11
	2	HOCDE Régis	French	IRD (MARBEC)	CCR Diver Logistics Marine Biodiversity 01/10 to 29/11
	3	LEBLOND Julien	French	IRD**	CCR Diver Sampling	01/10 to 27/11
	4	BAHUET Eric	French	IRD**	CCR Diver Head security, photograph 08/10 to 04/11
	5	DIRAIMONDO Gilles	French	IRD**	CCR Diver, video & photograph	01/10 to 27/11
	6	JUHEL Jean-Baptiste	French	IRD (MARBEC)	Diver, Reef Fish sampling	01/10 to 27/11
	7	FROMENTO Bruno	French	IRD**	Basecamp logistic ornithology	01/10 to 27/11
	8	AURELLE Didier	French	AMU (IMBE)	Gorgonian & Coral Genetics	08/10 to 21/11
	9	MILA Borja	Spain	MNCN CSIC	Avian Genetics & Taxonomy	01/10 to 16/11
	10 THEBAUD Christophe	French	UPS Toulouse	Avian Genetics & Taxonomy	01/10 to 27/11
	11 COCHET Christophe	French	IRD (ISEM)	Fish sampling & Taxonomy	01/10 to 27/11
	12 POUYAUD Bernard	French	IRD**	Logistics	01/10 to 27/11
	13 CHEVALLARD Jean	French	IRD**	Doctor	01/10 to 27/11
	Table 2 -List of indonesian participants -Lengguru 2017

Indonesian counterpart name/surname Institution Activities Period on Field 1

	11 Kadarusman	Politeknik-KP-Sorong Reef Fish Genetics Taxonomy	15/10 to 26/11
	12 Amir M. Suruwaky	Politeknik-KP-Sorong Diver Marine biology Sampling	12/10 to 23/11
	13 Abdul Gofir	Politeknik-KP-Sorong Diver Marine biology Sampling	12/10 to 23/11
	14 Anjas A.S. Komboe	Politeknik-KP-Sorong Airaha2 Captain	12/10 to 23/11
	15 Dewa Muditha	Politeknik-KP-Sorong Airaha2 Crew	12/10 to 23/11
	16 Nataniel Kalagison	Politeknik-KP-Sorong Airaha2 Crew	12/10 to 23/11
	17 Khairil Umasugi	Politeknik-KP-Sorong Airaha2 Crew	12/10 to 23/11
	18 Anton A. Orisu	Politeknik-KP-Sorong Airaha2 Crew	12/10 to 23/11
	19 Amos Ajambua	DISKAN Manokwari	Marine sampling	07/10 to 26/11
		Hidayat Ashari	RCB-LIPI	Ornithology Genetics Taxonomy	07/10 to 26/11
	2	Hadi Wikanta	RCB-LIPI	Ornithology Sampling	07/10 to 26/11
	3	Indra Bayu Vimono	RCO-LIPI	Diver, Echinoderm Genetics Taxonomy 28/10 to 20/11
	4	Yosephine Tuti	RCO-LIPI	Diver, Gorgonians Taxonomy	15/10 to 20/11
	5	Mohammad Abrar	RCO-LIPI	Diver, Hard corals Genetics Taxonomy	07/10 to 20/11
	6	Rizkie Satriya Utama	RCO-LIPI	Gorgonians Genetics Taxonomy	07/10 to 20/11
	7	Sumanta	IRD (local staff)	Cooking and logistic	07/10 to 26/11
	8	Paulus Boli	UNIPA Manokwari	Marine biology	15/10 to 03/11
	9	Napoleon Lemauk	DISKAN Kaimana	Guide and logistic	17/10 to 19/11
	10 Petrus Ruwe	DISKAN Kaimana	Guide and logistic	17/10 to 19/11

Table 3 -Lengguru 2017 provisional timetable Date Location -ornithology team Location -marine biology team

 3

	Sa	30/09/2017	Trip Europe -Indonesia
	Su	01/10/2017	Arrival of European team in Jakarta
		02-06/10/2017 Administrative formalities (immigration and RISTEK research visas)
	Sa-Su	07-08/10/2017 Trip Jakarta -Sorong
		09-11/10/2017 Administrative formalities, logistics
		12-14/10/2017 Trip Sorong -Kaimana
	Su	15/10/2017	Logistics Kaimana
	Mo	16/10/2017	Logistics Kaimana and meeting with Kaimana's Bupati
	Tu	17/10/2017	Transit Kaimana -Nusa Ulan (basecamp installation)
		18/10/2017	Dinas base camp east of Nusa Ulan /	Diving work in Pulau Nusa Ulan / Pulau Venu /
		To	Pegunungan Kumawa mountains	Pulau Buasai (North and West) / Pulau Lauzara
		18/11/2017
	Su	19/11/2017	Trip Nusa Ulan -Kaimana
	Mo	20/11/2017	BUPATI restitution meeting, conditioning of the samples
		21-23/11/2017 Trip Kaimana -Sorong
	Fr	24/11/2017	Packing of equipment in Sorong
	Sa	25/11/2017	Restitution meeting Politeknik-KP-Sorong -Loading container
	Su	26/11/2017	Trip Sorong -Jakarta
	Mo	27/11/2017	Trip Jakarta -Europe (European participants)

Table 4 :

 4 Statistics of scientific dives, by range of depths and by method

	Total	CCR	OC

Areas and depths of intervention were organized according to capacity and level of experience of each diver. Deep dives (up to -100m) were investigated by 5 divers (R. Hocdé, J. Leblond, E. Bahuet, G. Di Raimondo) with electronical-closed circuit rebreathers (e-CCR), while conventional dives (up to 40m) were organized by other members (Y. Tuti, M. Abrar, I.B. Vimono, A. Suruwaki, Gofir, P. Boli, R. Satriva Utama, C. Cochet).

Table 5 :

 5 List of scientific diving stations (red triangles)

	Diving	Longitude	Latitude
	stations	(East)	(North)	Localities
	S001	131,59549	-2,6779 Pulau Sabuda (Banana Island) -South West tanjung of the South island
	S002	132,97464	-4,11585 North (NE) wall of Pulau Nusaulang (big island front of Nusa Ulan)
	S003	133,02015	-4,11885 Papisol cap
	S004	133,05577	-4,08942 North face of the reef flat, before Pulau Derbi (Korowai ?)
	S005	133,05658	-4,09782 Pulau Derbi
	S006	133,02015	-4,11885 East face of Tanjung Papisol (Papisol Cap)
	S007	132,97754	-4,12528 Nusa Ulan Island -South East Tanjung (SE Cap)
	S008	132,97753	-4,12577 Nusa Ulan Island -South East Tanjung (SE Cap)
	S009	132,97753	-4,12577 Nusa Ulan Island -South East Tanjung (SE Cap)
	S010	132,97464	-4,11585 North wall of Pulau Nusaulang (big island front of Nusa Ula)
	S011	132,9751	-4,11723 Nusa Ulan Island -North East
	S012	132,97688	-4,12818 Nusa Ulan Island -South East
	S013	132,97925	-4,12829 Nusa Ulan Island -South East
	S014	132,93521	-4,09478 Nusa Ulan Island -West coast
	S015	132,97829	-4,12148 Nusa Ulan Island -East face
	S016	132,97829	-4,12148 Nusa Ulan Island -East face
	S017	132,9769	-4,12602 Nusa Ulan Island -East face
	S018	132,91273	-4,08471 Tanjung Suar (West cap of Nusa Ulan village)
	S019	132,91273	-4,08471 Tanjung Suar (West cap of Nusa Ulan village)

  Results to date > The principal investigator for Hard Corals, M. Abrar (RCO-LIPI) was invited by the Lengguru program for a 4-week visit at MIO Laboratory (Marseille, France) in May-June 2018. In collaboration with D. Aurelle, he performed DNA extraction, PCR and sequencing of all material collected during Lengguru 2017. All collected specimens were preliminary identified on a morphologically basis by C. Cochet and L. Pouyaud between January and February 2018. DNA was extracted from 1500 specimens in April 2018 in the RCO-LIPI genetic laboratory in Ancol. This operation was made on a large-scale extraction unit supported by the Lengguru program and involving specific material (INCU-Mixer MP4, Nucleovac 96 Vacuum Manifold, Vacuum Pump Chemker, microplate 96 centrifuge, and consumables). DNA extractions were performed in a collaborative work including H. Y. Sugeha, Kadarusman, and L. Pouyaud.

	V-4	Hard Corals
	Hard corals	> 500 collected specimens	603 DNA samples
	Data treatment and scientific valorization are presently under process.
	V-5	Reef fishes
	Reef fishes	2250 collected specimens	2250 DNA samples
	Results to date >
			800 collected specimens	800 DNA samples
	Results to date > The principal investigator for Gorgonians, R. Satriya Utama (RCO-LIPI) was invited by the
	Lengguru program for a 4-week visit at MIO Laboratory (Marseille, France) in May-June 2018. In collaboration
	with D. Aurelle, he performed DNA extraction, PCR and sequencing of all material collected during Lengguru
	2017.
	Data treatment and scientific valorization are presently under process.

DNA was amplified at two mitochondrial loci (12S and COI) for all 1500 extracted specimens between July and August 2018. The work was done by H. Y. Sugeha and Kadarusman during their visit in France in July 2018 and in collaboration with L. Pouyaud.

Table 6 -

 6 List of stereo BRUVs stations

	station	latitude	longitude	site	date	time	depth	rig	cam_left	cam_right	comments hard_drive
	nusa_ulan001	-04.10699	132.97247 nusa_ulan	22/10/2017	10:12	12	1	1		sandy substrat	01-févr
	nusa_ulan002	-04.10702	132.96887 nusa_ulan	22/10/2017	10:28	17	2	3		sandy substrat	01-févr
	nusa_ulan003	-04.10743	132.96507 nusa_ulan	22/10/2017	10:50	20	5	9		sandy substrat	01-févr
	nusa_ulan004	-04.10869	132.96053 nusa_ulan	22/10/2017	10:56	15	3	5		sandy substrat	01-févr
	nusa_ulan005	-04.10768	132.95721 nusa_ulan	22/10/2017	11:07	10	4	7		sandy substrat	01-févr
	nusa_ulan006	-04.12415	132.97322 nusa_ulan	23/10/2017	09:20	15	2	3			01-févr
	nusa_ulan007	-04.12249	132.97002 nusa_ulan	23/10/2017	09:35	20	1	1			01-févr
	nusa_ulan008	-04.11986	132.96834 nusa_ulan	23/10/2017	09:42	17	3	5			01-févr
	nusa_ulan009	-04.11554	132.96593 nusa_ulan	23/10/2017	09:52	8	5	9			01-févr
	nusa_ulan010	-04.11330	132.97028 nusa_ulan	23/10/2017	10:00	15	4	7			01-févr
	nusa_ulan011	-04.08527	132.91551 nusa_ulan	25/10/2017	09:10	10	4	7			01-févr
	nusa_ulan012	-04.08154	132.91228 nusa_ulan	25/10/2017	09:20	8	3	5			01-févr
	nusa_ulan013	-04.07690	132.91350 nusa_ulan	25/10/2017	09:30	12	5	9			01-févr
	nusa_ulan014	-04.07271	132.91530 nusa_ulan	25/10/2017	09:40	18	1	1			01-févr
	nusa_ulan015	-04.06686	132.91505 nusa_ulan	25/10/2017	09:55	12	2	3			01-févr
	pulau_venu01	-04.32162	133.49811 pulau_venu	28/10/2017	08:25	7	2	3			01-févr
	pulau_venu02	-04.32045	133.49670 pulau_venu	28/10/2017	08:35	10	5	9			01-févr
	pulau_venu03	-04.32272	133.49355 pulau_venu	28/10/2017	08:45	15	3	5			01-févr
	pulau_venu04	-04.32656	133.49469 pulau_venu	28/10/2017	08:55	15	1	1			01-févr
	pulau_venu05	-04.33015	133.49702 pulau_venu	28/10/2017	09:05	20	4	7			01-févr
	pulau_venu06	-04.33350	133.49947 pulau_venu	28/10/2017	15:25	12	4	7			01-févr
	pulau_venu07	-04.33677	133.50246 pulau_venu	28/10/2017	15:35	8	2	3			01-févr
	pulau_venu08	-04.33957	133.50630 pulau_venu	28/10/2017	15:45	15	3	5			01-févr
	pulau_venu09	-04.33644	133.50964 pulau_venu	28/10/2017	15:55	15	1	1			01-févr
	pulau_venu10	-04.33327	133.51300 pulau_venu	28/10/2017	16:00	15	5	9			01-févr
	pulau_venu11	-04.32546	133.49454 pulau_venu	29/10/2017	09:25	8	2	3			01-févr
	pulau_venu12	-04.32811	133.49591 pulau_venu	29/10/2017	09:35	15	3	5			01-févr
	pulau_venu13	-04.33194	133.49828 pulau_venu	29/10/2017	09:45	15	1	1			01-févr
	pulau_venu14	-04.33551	133.50125 pulau_venu	29/10/2017	09:55	12	4	7			01-févr
	pulau_venu15	-04.33846	133.50410 pulau_venu	29/10/2017	10:05	6	5	9			01-févr
	triton_bay01	-03.90978	134.07840 triton_bay	02/11/2017	08:00	8	1	1			01-févr
	triton_bay02	-03.90900	134.07951 triton_bay	02/11/2017	08:10	8	3	5			01-févr
	triton_bay03	-03.91250	134.04749 triton_bay	02/11/2017	08:30	10	2	3			01-févr
	triton_bay04	-03.91261	134.04140 triton_bay	02/11/2017	08:50	10	5	9			01-févr
	triton_bay05	-03.91194	134.03731 triton_bay	02/11/2017	09:00	15	4	7			01-févr
	triton_bay06	-03.90767	134.03619 triton_bay	02/11/2017	16:46	8	3	5			01-févr
	triton_bay07	-03.90939	134.03923 triton_bay	02/11/2017	17:00	8	5	9			01-févr
	triton_bay08	-03.90688	134.03381 triton_bay	02/11/2017	17:15	8	2	3			01-févr
	pulau_aiduma01 -03.91322	134.12190 pulau_aiduma 05/11/2017	09:35	8	4	7			01-févr
	pulau_aiduma02 -03.91017	134.11165 pulau_aiduma 05/11/2017	09:45	10	3	5			01-févr
	pulau_aiduma03 -03.90635	134.10788 pulau_aiduma 05/11/2017	10:05	15	1	1			01-févr
	pulau_aiduma04 -03.89861	134.10699 pulau_aiduma 05/11/2017	10:15	12	2	3			01-févr
	pulau_aiduma05 -03.92014	134.15126 pulau_aiduma 05/11/2017	11:00	12	5	9			01-févr
	kitikiti01	-03.93949	132.84041 kitikiti	16/11/2017	15:10	15	2	3			01-févr
	kitikiti02	-03.94732	132.83783 kitikiti	16/11/2017	15:20	15	4	7			01-févr
	kitikiti03	-03.94425	132.83363 kitikiti	16/11/2017	15:35	15	3	5			01-févr
	kitikiti04	-03.93735	132.82903 kitikiti	16/11/2017	15:45	15	1	1			01-févr
	kitikiti05	-03.93047	132.82614 kitikiti	16/11/2017	16:00	20	5	9			01-févr
	kitikiti06	-03.93632	132.80209 kitikiti	17/11/2017	07:50	10	2	3			03-avr
	kitikiti07	-03.94079	132.80408 kitikiti	17/11/2017	08:00	15	4	7			03-avr
	kitikiti08	-03.94334	132.80609 kitikiti	17/11/2017	08:10	15	1	1			03-avr
	kitikiti09	-03.94660	132.80783 kitikiti	17/11/2017	08:19	15	5	9			03-avr
	kitikiti10	-03.95314	132.81210 kitikiti	17/11/2017	08:30	15	3	5			03-avr

Table 7 .

 7 Summary of environmental DNA sampling

	Type of filter Number of filters Depth
	0.45	10	Surface
	0.22	49	Surface
		28	3 -<40m
		19	40 -<100m
		10	>= 100m

Extraction, amplification & sequencing > The extraction, amplification and sequencing were performed in a collaborating laboratory, Spygen (http://www.spygen.com/fr/). This laboratory is specialized in environmental DNA treatment and equipped with cleanrooms to avoid contamination.

Table 8

 8 All images are of high quality. Tens of thousands images and hours of video are currently stored.

	1	-List of eDNA stations

Figure 15. Lengguru 2017 images database

Table 9 -

 9 Species and number of individuals capturedAll blood samples were taken by Borja Milá and have been deposited at the Museum Zoologicum Bogoriense (MZB)'s ornithological collection in Cibinong, West Java, Indonesia. Some specimens were collected and prepared as museum skins in the field by LIPI personnel and have been deposited at MZB.All fieldwork was conducted according to relevant guidelines by the government of the Republic of Indonesia and under research permits issued by RISTEK (Indonesia) (for 2014, 304/SIP/FRP/SM/X/2014; for 2017, 3179/FRP/E5/Dit.KI/IX/2017) and relevant Indonesian government collecting permits.

	Birds	215 captured specimens	215 DNA samples
	Species		English name	Captures
	Aerodramus nuditarsus (?)	Bare-legged swiftlet (?)
	Ailuroedus melanotis		Black-eared catbird
	Cacomantis castaneiventris	Chestnut-breasted cuckoo
	Chaetorhynchus papuensis	Drongo fantail
	Cicinnurus magnificus		Magnificent bird of paradise
	Colluricincla megarhynchus	Little shrike-thrush
	Craterocelis murina		Rusty mouse-warbler
	Drepanornis albertisi		Black-billed sicklebill
	Heteromyias albispecularis	Ashy robin
	Melanocharis sp. nov.		Undescribed berrypecker
	Melilestes megarhynchus		Long-billed honeyeater
	Meliphaga orientalis		Mountain meliphaga
	Myzomela cruentata		Red myzomela
	Myzomela rosenberghi		Red-collared myzomela
	Oedistoma iliolophus		Spectacled longbill
	Ornorectes cristatus		Piping bellbird
	Philemon buceroides		Helmeted friarbird
	Phylloscopus poliocephalus	Island leaf-warbler
	Pitohui dichrous		Hooded pithoui
	Sericornis nouhuysi (?)		Large scrubwren (?)
	Sericornis spilodera		Pale-billed scrubwren
	Symposiachrus axillaris		Fantailed monarch
	Toxorhamphus novaguinaea	Yellow-bellied longbill
	Tregellasia leucops		White-faced robin
	Zosterops novaeguineae		New Guinea white-eye
	Erythrura trichroa		Blue-faced parrotfinch

Acknowledgement

We thank the Indonesian Institute of Sciences (LIPI) for promoting our collaboration and the Sorong Polytechnic of Marine and Fisheries (Politeknik KP Sorong, West Papua) for providing the vessel Airaha 02 that we used in this campaign. We thank the crew of the Aihara 02 for assisting us during the operations and SPYGEN staff for the technical support in the eDNA laboratory. Fieldwork and laboratory activities were supported by the Lengguru 2017 Project (www.lengguru.org), conducted by the French National Research Institute for Sustainable Development (IRD), the Indonesian Institute of Sciences (LIPI) with the Research Center for Oceanography (RCO, the Politeknik KP Sorong), the University of Papua (UNIPA) with the help of the Institut Français in Indonesia (IFI) and with corporate sponsorship from the Total Foundation and TIPCO company. The sequencing was funded by Monaco Explorations.

VIII CONCLUSIONS AND PERSPECTIVES

The main objective of the Lengguru 2017 Expedition was to start and impulse an interdisciplinary international scientific cooperation on terrestrial and marine biodiversity domains at the heart of the Coral Triangle in West Papua (Indonesia). The Indonesian Institute of Sciences (LIPI) with the Research Centre for Oceanology (RCO) for marine biodiversity heads this program together with the Sorong Polytechnic School (Politeknik-KP-Sorong) and the French Research Institute for Development (IRD).

All the field objectives were fulfilled in terms of visited ecosystems panel, material collected, and international cooperation.

Preliminary results obtained from morphological observation and DNA barcoding are nearly achieved for all taxa enabling the valorization in international and Indonesian scientific journals.

The Lengguru team will continue in the coming months to acquire and to valorize jointly the results.

In that frame, it is expected to pursue the organization and support of research fellowships for Indonesian counterparts. This includes international midterm visits but also PhD degrees, which must be increased.

In term of capacity building we expect to continue to upgrade molecular capacities at RCO-LIPI especially for large-scale PCR amplification on 96 plates. We also plan to pursue training session in Indonesia and collaboration activities such as organization of new field trips and joint research network on marine biodiversity.

APPENDIX

A-1 Co-authored scientific publications and reports in the framework of the LENGGURU program since 2010