Neohexangitrema spp. (Trematoda: Microscaphidiidae) in Indo-West Pacific Acanthuridae: Richness, distribution, diet and contemporary naming issues.

As part of a broad survey of the trematodes of damselfishes (Pomacentridae) in the tropical Indo-West Pacific, zoogonids were collected from multiple localities in Australia, New Caledonia, and French Polynesia. All zoogonid specimens collected were consistent with the subfamily Lecithostaphylinae, and morphological and molecular data (ITS2 and 28S rDNA, and cox1 mtDNA) were generated for most host-locality combinations to enable an integrative species delimitation. The collection comprised three species: Deretrema stratiotes n. sp. from four species of Abudefduf Forsskål from Ningaloo Reef in Western Australia, and two species consistent with the genus Lecithostaphylus Odhner, 1911 for which Innuptacola n. gen. is proposed based on phylogenetic and morphological distinction, the type-species I. gibsoni (Cribb, Bray & Barker, 1992) n. comb. (= L. gibsoni) from six species of Abudefduf in Ningaloo Reef, Queensland and New Caledonia, and I. torquata n. sp. from 12 pomacentrid species in Ningaloo Reef, the Great Barrier Reef in Queensland, and the Gambier Islands in French Polynesia. The new collection demonstrates that some zoogonid species are geographically widespread (from the Pacific Ocean to the Indian Ocean) and can infect a broad range of hosts (multiple genera within a family), whereas others are apparently geographically restricted and exhibit higher host-specificity (fishes within a single genus).

We report eight species of Lintonium from tetraodontiform fishes from Australian waters and describe six of them as new. Two species are described from tetraodontids from the Great Barrier Reef (GBR): Lintonium kostadinovae n. sp. from Arothron nigropunctatus (Bloch & Schneider) and Arothron hispidus (Linnaeus); and Lintonium droneni n. sp. from A. nigropunctatus. Two species are described from temperate monacanthids: Lintonium crowcrofti n. sp. from Meuschenia hippocrepis (Quoy & Gaimard) and Meuschenia freycineti (Quoy & Gaimard) off Tasmania and from M. hippocrepis off Glenelg, South Australia and off Fremantle, Western Australia; and Lintonium blendi n. sp. from M. hippocrepis off Stanley, Tasmania. The final two new species are described from tropical monacanthids: Lintonium currani n. sp. from Cantherhines pardalis (Rüppell) from Ningaloo Reef, Western Australia; and Lintonium madhaviae n. sp. from Amanses scopas (Cuvier) from the southern GBR. Two previously described species are reported from tetraodontids: Lintonium pulchrum (Johnston, 1913) Yamaguti, 1954 from Arothron stellatus (Anonymous), A. hispidus, A. manilensis (Marion de Procé) and Lagocephalus lunaris (Bloch & Schneider) from the GBR and southern Queensland; and Lintonium consors (Lühe, 1906) Crowcroft, 1950 from A. nigropunctatus from the southern GBR. Sequence data for three markers (ITS2 and 28S rDNA and cox1 mtDNA) for six of the eight species (L. crowcrofti n. sp., L. currani n. sp., L. droneni n. sp., L. kostadinovae n. sp., L. madhaviae n. sp. and L. pulchrum) are the first for the genus and distinguish each species unambiguously. Many records of species of Lintonium, especially widespread records of the type species, L. vibex (Linton, 1900) Stunkard & Nigrelli, 1930, remain to be clarified. A key finding of the present study is that three fish species (A. hispidus, A. nigropunctatus and M. hippocrepis) are identified as harbouring either two or three species of Lintonium at individual localities.

The taxonomy of species of Bivesicula Yamaguti, 1934 is analysed for samples from holocentrid, muraenid and serranid fishes from Japan, Ningaloo Reef (Western Australia), the Great Barrier Reef (Queensland), New Caledonia and French Polynesia. Analysis of three genetic markers (cox1 mtDNA, ITS2 and 28S rDNA) identifies three strongly supported clades of species and suggests that Bivesicula as presently recognized is not monophyletic. On the basis of combined morphological, molecular and biological data, 10 species are distinguished of which five are proposed as new. Bivesicula Clade 1 comprises seven species of which three are effectively morphologically cryptic relative to each other; all seven infect serranids and four also infect holocentrids. Bivesicula Clade 2 comprises three species of which two are effectively morphologically cryptic relative to each other; all three infect serranids and one also infects a muraenid. Bivesicula Clade 3 comprises two known species from apogonids and a pomacentrid, and forms a clade with species of Paucivitellosus Coil, Reid & Kuntz, 1965 to the exclusion of other Bivesicula species. Taxonomy in this genus is made challenging by the combination of low resolving power of ribosomal markers, the existence of regional cox1 mtDNA populations, exceptional and unpredictable host-specificity and geographical distribution, and significant host-induced morphological variation.

Emprostiotrema contains just 3 species: E. fusum, E. kuntzi and E. sigani. As adults, all 3 species infect rabbitfishes (Siganidae: Siganus). New collections from 11 species of Siganus from northern Australia, Indonesia, New Caledonia, French Polynesia, Palau and Japan enabled an exploration of species composition within this genus. Phylogenetic analyses demonstrate a deep distinction between 2 major clades; clade 1 comprises most of the sequences of specimens from Australia as well as all of those from Japan, Palau and New Caledonia and clade 2 comprises all sequences of specimens from French Polynesia, 2 sequences from Australia and the single sequence from Bali. In all analyses, both major clades have genetic structuring leading to distinct geographic lineages. Morphologically, specimens relating to clades 1 and 2 differ but overlap in body shape, oral sucker and egg size. Principle component analysis shows a general (but not complete) separation between specimens relating to the 2 clades. We interpret the 2 clades as representing 2 species: clade 1 is identified as E. fusum and is reported in this study from 10 species of siganids from Australia, Japan, Palau and New Caledonia; clade 2 is described as E. gotozakiorum n. sp., for all specimens from French Polynesia and rare specimens from Australia and Indonesia. We recognize E. sigani as a junior synonym of E. fusum. Although species of Emprostiotrema occur widely in the tropical Indo-Pacific, they have not been detected from Ningaloo Reef (Western Australia), the southern Great Barrier Reef or Moreton Bay (southern Queensland).

Five species of the genus Multitestis are described, figured or discussed: Multitestis pyriformis from Platax orbicularis off Lizard Island, northern Great Barrier Reef, Australia and Platax teira off New Caledonia; Multitestis coradioni n. sp. (syn. Multitestis pyriformis Machida, 1963 of Bray et al. (1994)) from Coradion chrysozonus off Heron Island, which differs from M. pyriformis in its oval body-shape, the more posteriorly situated testicular fields and larger eggs, Multitestis elongatus from Platax pinnatus off Lizard Island, Multitestis magnacetabulum from P. teira off Heron Island, southern Great Barrier Reef, Australia, and New Caledonia, Multitestis paramagnacetabulum n. sp. from P. orbicularis off Ningaloo Reef, Western Australia, which differs from M. magnacetabulum in the more posteriorly situated testicular fields.

This thesis explores the taxonomy, systematics and phylogenetic status of the family Faustulidae from corallivorous fishes in the Tropical Indo-west Pacific (TIWP). The main study is based on the parasitological examination of 2,901 individual coral reef fishes from six major TIWP localities: Ningaloo Reef, Western Australia; Palau; Lizard Island, Swain Reefs complex and Heron Island (all Queensland); and French Polynesia.Faustulidae are mainly parasites of the intestine of marine teleosts and are characterised by a spiny tegument, a canalicular seminal receptacle and a posterior opening Laurer’s canal. Of the 12 genera that belong to this family, the largest, and the main focus of this study, is Paradiscogaster Yamaguti, 1934. Before this study, Paradiscogaster comprised 24 species distributed in the Indo-west Pacific and infecting 13 families of fish. Of these, corallivorous fishes from the family Chaetodontidae had the largest number of Paradiscogaster species, followed by fishes, with a diverse diet, from Monacanthidae and Ostraciidae.The results from this study are structured first as three taxonomic chapters describing new species from this genus; I described and published six new Paradiscogaster species from Chaetodontidae and Ephippidae, and a further five new species not yet published: two new species corresponding to the Paradiscogaster eniwetokensis complex on the GBR, two from Palau, two from Ningaloo Reef and one from French Polynesia. This is followed by an overall phylogenetic study of the family, a study of trematodes from non-chaetodontid corallivorous fishes from the Great Barrier Reef (including the description of a new species from Fellodistomidae) and finally an account of a search for larval trematodes in coral.For the analysis of the phylogeny of Faustulidae, 28S rDNA sequences were obtained from ten Faustulidae species. These were aligned and analysed relative to other faustulids and species of key families of interest. The complete analysis included sequences for 35 species of Faustulidae, Fellodistomidae, Gymnophallidae, Tandanicolidae, and Zoogonidae (with Hemiuridae as an outgroup) and showed that some Faustulidae and Zoogonidae form a clade, but that relationships within the clade are not well-resolved. More sequences will be required to determine whether Faustulidae should be restructured relative to the Zoogonidae as previously suggested, and also to determine the taxonomic position of three species of Faustulidae, Pseudobacciger cheneyae and two species of Bacciger, which formed a clade quite separate from all other Faustulidae. This chapter also analyses the ITS2 rDNA sequences of the family, incorporating new sequences and published sequences available from GenBank. The genus Paradiscogaster incorporates a clade formed by species that infect corallivorous chaetodontids and another clade from non-chaetodontids.To determine the implications of corallivory on the trematode fauna of non-chaetodontid corallivores, I studied several obligate corallivores that have not been previously sampled, including species of Blenniidae (Exallias brevis), Gobiidae (Gobiodon citrinus), Labridae (Labropsis australis and Labrichthys unilineatus), Monacanthidae (Amanses scopas and Oxymonacanthus longirostris) and Tetraodontidae (Arothron nigropunctatus). No infections of Paradiscogaster species were found but a new species (Fellodistomidae: Prudhoeus) is described from Amanses scopas and Exallias brevis from Heron Island, Queensland. Clearly, exploitation of these corallivores by trematodes has been much less extensive than in the chaetodontids.Many species of chaetodontid fishes reported as obligate corallivores are heavily infected by faustulids, suggesting that the ingestion of the parasites in intermediate stages is through the diet, coral polyps or a symbiont associated with coral polyps. I thus looked for metacercariae in acroporid corals (the most abundant dietary item in obligate corallivores, representing 97–98% of the diet), examining between 4500-9000 coral polyps from 90 fragments, utilizing varied techniques to extract the polyps, but without a positive finding. This outcome means that the transmission mechanisms of these parasites remain unexplained. In terms of biogeography and host-specificity, the results show that the highest known concentration of Faustulidae species is found on the GBR, where the host-specificity is higher than in other localities. The richness of Paradiscogaster species varied among TIWP sites, having the GBR as the richest known area and lower in Palau, New Caledonia, Ningaloo Reef and French Polynesia. However, the host-specificity (specifically of P. sasali) reduced in the extremities partially isolated and remote areas of the TIWP and Paradiscogaster species were found infecting a wide variety of hosts, perhaps increasing the chance of reproduction and dispersion.A better understanding of the life cycles of these parasites, particularly the intermediate host specificity, will contribute to an understanding of the biogeography of these parasites and will contribute to an explanation of why these parasites are not present in all localities in the TIWP, even when appropriate definitive hosts are present.

Biogeography of tropical Indo-West Pacific parasites: A cryptic species of Transversotrema and evidence for rarity of Transversotrematidae (Trematoda) in French Polynesia

Diploproctodaeum monstrosum sp. nov. is described from Arothron stellatus and A. mappa from off Lizard Island, northern Great Barrier Reef. It differs from its congeners in having a body-length ventral scoop. Diploproctodaeum triodoni sp. nov. is described from Triodon macropterus off New Caledonia. It is distinguished by the extensive vitelline fields usually reaching to the ventral sucker and the folded scoop margins. Other related species are reported from new hosts or localities and dimensions are supplied for: Diploproctodaeum haustrum from Aluterus monoceros off New Caledonia; Diploproctodaeum arothroni from Arothron hispidus off Lizard Island and Ningaloo Reef, northern Western Australia, A. nigropunctatus off Lizard Island and Arothron manilensis off New Caledonia; Diploproctodaeum macracetabulum from Abalistes stellatus on the Swain Reefs, southern Great Barrier Reef and off New Caledonia; Diploproctodaeum momoaafata from Ostracion cubicus off Lizard Island; Diploproctodaeum rutellum from Platax teira off Heron Island, southern Great Barrier Reef; Diploproctodaeoides longipygum from Abalistes stellatus on the Swain Reefs and off New Caledonia; Diplocreadium tsontso from Balistoides conspicillum off Heron Island; Bianium arabicum from Lagocephalus sceleratus off New Caledonia. Attention is drawn to apparent convergent evolution in the body form of several families of trematodes infecting tetraodontids and especially species of Arothron.

Two new species of the spongicolid shrimp genus Microprosthema Stimpson, 1860 are described and illustrated on the basis of material collected recently in Australia and Japan. Type specimens of Microprosthema ningaloo sp. nov. were collected on Ningaloo Reef, Western Australia, whereas type specimens of Microprosthema pallidum sp. nov. were collected on a coral reef off Ishigaki Island, Ryukyu Archipelago, southern Japan. Microprosthema ningaloo sp. nov. and M. pallidum sp. nov. differ from all other species of the genus by a combination of morphological characters, including the gill-exopod formulae, and by their diagnostic colour patterns. This study increases the total number of species described in the genus Microprosthema to 16, nine of them in the Indo-West Pacific. In addition, new records are provided for M. lubricum Saito & Okuno, 2011 (Guam), M. plumicorne (Richters, 1880) (Red Sea, Mariana and Marshall Islands, French Polynesia), M. scabricaudatum (Richters, 1880) (Red Sea, Glorieuses and Tuamotu Islands), and M. validum Stimpson, 1860 (Red Sea, Madagascar, Singapore, French Polynesia), significantly extending their previously known distribution ranges.

Background Coastal aggregations of whale sharks, typically dominated by juvenile males and occurring seasonally, are now documented in many places around the world, including Ningaloo Reef, Western Australia (WA). This aggregation occurs during the Austral autumn and winter and supports a lucrative tourism industry which operates during the whale shark “season” (generally April – July). Despite being one of the most highly studied aggregations of whale sharks in the world, their location and movements outside this period remain poorly understood, but are critical for understanding long term population dynamics and for effective management and conservation. An earlier study by ECOCEAN, combining satellite tracking and citizen science sightings of whale sharks outside the “season” at Ningaloo Reef and elsewhere along the WA coast, indicates that whale sharks may move north and south along the coast throughout the year, and/or make relatively short migrations, rather than undertaking long, trans-oceanic migrations. To test this further, a novel satellite tagging programme, supported by The Western Australian Department of Education (The Department), aims to discover where the whale sharks are when they are not at Ningaloo Reef and determine whether there are other unidentified “hotspots” for foraging and/or migration, i.e. areas of habitat critical for whale sharks. This programme also aims to provide innovative learning opportunities for school children, to make them more aware of the biology and ecology of WA's marine emblem and the marine environment, as well as to encourage engagement with STEM (Science, Technology, Engineering and Maths) learning. Approach Satellite tags (Wildlife Computers SPOT tags) were deployed on 12 whale sharks (identified using photo-identification through the Wildbook for Whale Sharks) at Ningaloo Reef in July 2015 using a custom-made clamp attachment on the first dorsal fin, designed to have minimal impact on the animals. Tags were funded by 16 schools from around WA participating in a joint ECOCEAN - Western Australian Department of Education science learning programme. Tracks of the sharks are displayed publically on the ZoaTrack website (www.zoatrack.org/projects/243/ analysis) and were used by participating schools in an eight week teaching programme. Results Data from satellite tags deployed in 2015 are still being collected and will be analysed as part of an Honours project in 2016, however, the teaching programme proved highly successful: The Department developed a comprehensive suite of resources addressing science, technology, engineering, mathematics and arts subjects to support students to engage in authentic learning experiences, and classes from pre-primary through to senior high school were involved. Preliminary results from the tags show that all have successfully reported positional data, for periods ranging from 15 to 156 days (mean 93.4 ± 13.1 SE) (as at 31/12/15). Homing migrations, to the southern part of Ningaloo Reef, have been recorded for 50% of all the sharks tagged, within periods ranging from 44 to 93 days (mean 65.7 ± 7.8 SE). These data show sharks making short forays away from Ningaloo Reef, and reveal areas important for commuting and foraging. Conclusions This innovative approach to funding allowed the satellite tracking programme to proceed and not only provided new information on the movements of whale sharks from Ningaloo Reef that can be used for conservation planning and management, but also provided novel learning experiences for school children and raised awareness for whale shark conservation and the marine environment. The data gathered support the hypothesis that whale sharks from the Ningaloo Reef aggregation do not typically undertake ocean basin scale migrations, rather they make shorter forays, perhaps to exploit food resources that vary spatially and temporally, and could be used to benchmark conservation outcomes for WA whale sharks and to provide pathways for more effective protection.

Species of the faustulid genus Antorchis Linton, 1911 of the tropical Indo-West Pacific are reviewed. We recognise five species in the region, including a novel form. Antorchis nasonis n. sp. is described from Naso annulatus (Quoy & Gaimard) and N. tonganus (Valenciennes) on the southern Great Barrier Reef (GBR). We interpret specimens reported from Naso hexacanthus (Bleeker) from Japan as the same species. This species appears to be the only faustulid known from acanthurid fishes and differs from all other species in the genus in having the prominent dorsal genital invagination close to the posterior end of the body. In addition, new host and locality records are reported for two described species of Antorchis, A. pomacanthi (Hafeezullah & Siddiqi, 1970) and A. tsushimaensis (Machida, 1971). The wide distribution of A. pomacanthi was further demonstrated by the generation of identical ITS2 rDNA sequences for specimens from Ningaloo Reef off Western Australia, off Lizard and Heron Islands (GBR) and off New Caledonia, localities separated by up to 5,300km. The host-specificity of the genus is considered.

A precipitous increase in the abundance of the corallivorous snail Drupella cornus at Ningaloo Reef, Western Australia, raises fundamental questions about the population structure and genetics of this species. We examined genetic heterogeneity at ten polymorphic allozyme loci among samples of adult D. cornus from nine sites along 180 km of Ningaloo Reef, plus two sites from the Abrolhos Islands and one from the Dampier Archipelago, spanning a total distance of 1170 km. Variations in allelic frequencies were small (average FST=0.007), indicating that a high degree of planktonic dispersal is the norm. Nevertheless, some heterogeneity among samples was found at four of the loci. This heterogeneity occurred within Ningaloo Reef and did not increase with geographic distance. The local heterogeneity was not a function of habitat type but seemed to be associated with stage of outbreak. However, all outbreak populations came from within Ningaloo Reef and the non-outbreak populations were from outside Ningaloo Reef proper. Our results show peculiarities in the genetic structure of D. cornus on Ningaloo Reef, but the causes are not understood.

The first successful trials with stationary 'crest' nets to monitor the nocturnal fluxes of larval fish crossing reef margins in both eastern and western Australia are described. Lengthy deployments were possible on Ningaloo Reef, north-western Australia, because that system is topographically suitable: i.e. a fringing barrier reef where surf produces a constant flow into a coastal lagoon. Sampling on 85 nights between October 1994 and March 1995 revealed a rich larval fish fauna (56474 individuals) dominated by pelagic juveniles nearing settlement stage. Variations in the daily catches of replicate nets (200 m apart) were highly correlated, showing the suitability of this technique for monitoring larval supply. Another trial (five nights) was made at One Tree Reef, southern Great Barrier Reef. On nocturnal flood tides, when rising water first spilled into the lagoon, triplicate nets caught many presettlement fish (47797 individuals) in this flow. The behaviour of some taxa clearly assisted their transport through the surf. Despite the successful short-term deployments at One Tree, there may be limited potential to deploy this gear elsewhere on the Great Barrier Reef because of unsuitable flow regimes.

Environmental influences on the epidemiology of fibropapillomatosis in green turtles (Chelonia mydas) and consequences for management of inshore areas of the Great Barrier Reef

Strip-transect aerial surveys of Shark Bay, Ningaloo Reef and Exmouth Gulf were conducted during the winters of 1989 and 1994. These surveys were designed primarily to estimate the abundance and distribution of dugongs, although they also allowed sea turtles and dolphins, and, to a lesser extent, whales, manta rays and whale sharks to be surveyed. Shark Bay contains a large population of dugongs that is of international significance. Estimates of approximately 10000 dugongs resulted from both surveys. The density of dugongs is the highest recorded in Australia and the Middle East, where these surveys have been conducted. Exmouth Gulf and Ningaloo Reef are also important dugong habitats, each supporting in the order of 1000 dugongs. The estimated number of turtles in Shark Bay is comparable to the number in Exmouth Gulf plus Ningaloo Reef (7000–9000). The density of turtles in Ningaloo Reef and, to a lesser extent, Exmouth Gulf is exceptionally high compared with most other areas that have been surveyed by the same technique. Shark Bay supports a substantial population of bottlenose dolphins (2000–3000 minimum estimate). Exmouth Gulf and Ningaloo Reef were not significant habitats for dolphins during the winter surveys. Substantial numbers of whales (primarily humpbacks) and manta rays occur in northern and western Shark Bay in winter. Ningaloo Reef is an important area for whale sharks and manta rays in autumn and winter. The Shark Bay Marine Park excludes much of the winter habitats of the large vertebrate fauna of Shark Bay. In 1989 and 1994, more than half of all the dugongs were seen outside the Marine Park (57·4 and 50·7%, respectively). Approximately one-third to one-half of turtles and dolphins were seen outside the Marine Park (in 1989 and 1994 respectively: turtles, 43 and 27%; dolphins, 47 and 32%). Almost all the whales and most of the manta rays were seen outside the Marine Park. Expansion of the Shark Bay Marine Park, to bring it into alignment with the marine section of the Shark Bay World Heritage Area, would facilitate the appropriate management of these populations. This would also simplify the State– Commonwealth collaboration necessary to meet the obligations of World Heritage listing. The coastal waters of Western Australia north of the surveyed area (over 6000 km of coastline) are relatively poorly known and surveys of their marine megafauna are required for wise planning and management.