Diversity of dinoflagellate assemblages in coastal temperate and offshore tropical waters of Australia.

BACKGROUND: Dinoflagellates are a ubiquitous and ecologically important component of marine phytoplankton communities, with particularly notable species including those associated with harmful algal blooms (HABs) and those that bioluminesce. High-throughput sequencing offers a novel approach compared to traditional microscopy for determining species assemblages and distributions of dinoflagellates, which are poorly known especially in Australian waters. RESULTS: We assessed the composition of dinoflagellate assemblages in two Australian locations: coastal temperate Port Phillip Bay and offshore tropical waters of Davies Reef (Great Barrier Reef). These locations differ in certain environmental parameters reflecting latitude as well as possible anthropogenic influences. Molecular taxonomic assessment revealed more species than traditional microscopy, and it showed statistically significant differences in dinoflagellate assemblages between locations. Bioluminescent species and known associates of HABs were present at both sites. Dinoflagellates in both areas were mainly represented by the order Gymnodiniales (66%-82% of total sequence reads). In the warm waters of Davies Reef, Gymnodiniales were equally represented by the two superclades, Gymnodiniales sensu stricto (33%) and Gyrodinium (34%). In contrast, in cooler waters of Port Phillip Bay, Gymnodiniales was mainly represented by Gyrodinium (82%). In both locations, bioluminescent dinoflagellates represented up to 0.24% of the total sequence reads, with Protoperidinium the most abundant genus. HAB-related species, mainly represented by Gyrodinium, were more abundant in Port Phillip Bay (up to 47%) than at Davies Reef (28%), potentially reflecting anthropogenic influence from highly populated and industrial areas surrounding the bay. The entire assemblage of dinoflagellates, as well as the subsets of HAB and bioluminescent species, were strongly correlated with water quality parameters (R2 = 0.56-0.92). Significant predictors differed between the subsets: HAB assemblages were explained by salinity, temperature, dissolved oxygen, and total dissolved solids; whereas, bioluminescent assemblages were explained only by salinity and dissolved oxygen, and had greater variability. CONCLUSION: High-throughput sequencing and genotyping revealed greater diversity of dinoflagellate assemblages than previously known in both subtropical and temperate Australian waters. Significant correlations of assemblage structure with environmental variables suggest the potential for explaining the distribution and composition of both HAB species and bioluminescent species.

A comparison of the antifouling performance of air plasma spray (APS) ceramic and high velocity oxygen fuel (HVOF) coatings for use in marine hydraulic applications.

Maritime hydraulic components are often exposed to harsh environmental conditions which can lead to accelerated deterioration, reduced function, equipment failure and costly repair. Two leading causes of maritime hydraulic failure are biofouling accumulation and corrosion. This study examined the antifouling performance of three candidate replacement high velocity oxygen fuel (HVOF) coatings relative to the performance of the current baseline air plasma spray (APS) ceramic coating for protection of hydraulic actuators. Following 20 weeks immersion at tropical and temperate field exposure sites, the control APS ceramic accumulated significantly greater levels of biofouling compared to the HVOF coatings. More specifically, the magnitude of growth of real-world nuisance hard fouling observed on in-service hydraulic components (eg calcareous tubeworms and encrusting bryozoans) was significantly greater on the APS ceramic relative to HVOF coatings. Possible explanations for the observed patterns include differences in surface topography and roughness, the electrochemical potential of the surfaces and the colour/brightness of the coatings.

Assessments of quaternary ammonium compounds (QAC) for in-water treatment of mussel fouling in vessel internals and sea chests using an experimental seawater pipework system.

The primary in-water emergency treatment method for mussel fouling of internal seawater systems of Royal Australian Navy vessels is to flush with a 1% detergent solution containing quaternary ammonium compounds (QAC). Parameters for application of this treatment are based on previous research; however, much of the research has been conducted at small-scales under controlled laboratory conditions. This study examined the efficacy of QAC solutions for treating mussel biofouling under realistic field conditions using experimental seawater piping systems. The efficacy of QAC solutions was highly dependent on the size of mussels present. Chemical treatments comprising 1, 2 and 5% v v-1 QAC solution were effective at killing large (50-92 mm) mussels in the pipework and sea chest of the system following 24 h exposure. In contrast, small mussels (10-30 mm) appeared resilient to the majority of treatment regimes. Differences in water temperature, DO and pH during dosing had no discernible impact on treatment efficacy.

Factors influencing the en route survivorship and post-voyage growth of a common ship biofouling organism, Bugula neritina.

The likelihood that viable non-indigenous biofouling species will survive a voyage on a vessel is influenced by a range of factors, including the speed, duration, and route of the voyage and the amount of time the vessel spends in port. In this study, a land-based dynamic flow device was used to test the effect of recruit age, vessel speed and voyage duration on the survivorship and growth of the bryozoan Bugula neritina. In the experiment, one-week-old recruits had a higher likelihood (100%) of surviving voyages than older (one-month-old, 90%) or younger (one-day-old, 79%) recruits, but survival was not influenced by vessel speed (6 and 18 knots) or voyage duration (two and eight days). The results suggest that the non-indigenous species B. neritina can be effectively transferred at a range of ages but one-week-old recruits are more likely to survive the translocation process and survive in the recipient environment.

Predation limits spread of Didemnum vexillum into natural habitats from refuges on anthropogenic structures.

Non-indigenous species can dominate fouling assemblages on artificial structures in marine environments; however, the extent to which infected structures act as reservoirs for subsequent spread to natural habitats is poorly understood. Didemnum vexillum is one of few colonial ascidian species that is widely reported to be highly invasive in natural ecosystems, but which in New Zealand proliferates only on suspended structures. Experimental work revealed that D. vexillum established equally well on suspended artificial and natural substrata, and was able to overgrow suspended settlement plates that were completely covered in other cosmopolitan fouling species. Fragmentation led to a level of D. vexillum cover that was significantly greater than was achieved as a result of ambient larval recruitment. The species failed to establish following fragment transplants onto seabed cobbles and into beds of macroalgae. The establishment success of D. vexillum was greatest in summer compared with autumn, and on the underside of experimental settlement plates that were suspended off the seabed to avoid benthic predators. Where benthic predation pressure was reduced by caging, D. vexillum establishment success was broadly comparable to suspended treatments; by contrast, the species did not establish on the face-up aspect of uncaged plates. This study provides compelling evidence that benthic predation was a key mechanism that prevented D. vexillum's establishment in the cobble habitats of the study region. The widespread occurrence of D. vexillum on suspended anthropogenic structures is consistent with evidence for other sessile invertebrates that such habitats provide a refuge from benthic predation. For invasive species generally, anthropogenic structures are likely to be most important as propagule reservoirs for spread to natural habitats in situations where predation and other mechanisms do not limit their subsequent proliferation.

Marine biosecurity: the importance of awareness, support and cooperation in managing a successful incursion response.

Shipping is almost certainly the most prevalent human-mediated transport vector for non-indigenous species (NIS) within the marine environment. The Royal Australian Navy (RAN) has long acknowledged the importance of sound environmental management and in recent years has taken a proactive approach to addressing risks associated with marine biosecurity. primarily as a result of biofouling on Navy vessel returning from overseas operations. This paper describes two case studies that highlight the effectiveness of the RAN marine biosecurity management framework in identifying an unwanted marine species on Navy vessels, and the successful biosecurity management program that ensued. In particular, the early detection and identification of a suspect NIS, the quick response to the discovery and the collaborative approach adopted between the RAN and the Government regulatory agency (Western Australian Department of Fisheries) charged with coordinating the incursion response serves as a model for how future incursion responses should be reported and managed.

Thermal treatment as a method to control transfers of invasive biofouling species via vessel sea chests.

This study examined the efficacy of heated seawater for the treatment and remediation of fouled vessel sea chest habitats. In laboratory trials, three temperature regimes (37.5°C for 60 min, 40°C for 30min and 42.5°C for 20 min) were tested on a range of temperate taxa commonly found in sea chests. Field validation trials further assessed the efficacy of heat treatment within a replica sea chest environment. During laboratory trials, 100% mortality was achieved across all three treatments for the majority of taxa; the exceptions being the barnacle Elminius modestus and the oyster Crassostrea gigas. Temperature tolerance limits observed in the laboratory were successfully replicated under simulated sea chest conditions; however, a failure to achieve even heat distribution was an obstacle to achieving uniform mortality. This study provides guidance on the temperature/exposure parameters required for vessels plying temperate latitudes, and demonstrates that heated seawater has potential for controlling biofouling in vessel sea chests.

The effect of vessel speed on the survivorship of biofouling organisms at different hull locations.

This study used a specially designed MAGPLATE system to quantify the en route survivorship and post-voyage recovery of biofouling assemblages subjected to short voyages (< 12 h) across a range of vessel speeds (slow, medium, fast; in the range 4.0-21.5 knots). The effect of hull location (bow, amidships and stern) was also examined. While no significant differences were evident in en route survivorship of biofouling organisms amongst hull locations, biofouling cover and richness were markedly reduced on faster vessels relative to slower craft. Therefore, the potential inoculum size of non-indigenous marine species and richness is likely to be reduced for vessels that travel at faster speeds (> 14 knots), which is likely to also reduce the chances of successful introductions. Despite this, the magnitude of introductions from biofouling on fast vessels can be considered minor, especially for species richness where 90% of source-port species were recorded at destinations.

Effect of vessel voyage speed on survival of biofouling organisms: implications for translocation of non-indigenous marine species.

This study experimentally determined the effect of different vessel voyage speeds (5, 10 and 18 knots = 2.6, 5.1 and 9.3 ms(-1), respectively) and morphological characteristics including growth form (solitary or colonial), profile (erect or encrusting) and structure (soft, hard or flexible) on the survival of a range of common biofouling organisms. A custom built hydrodynamic keel attached to the bottom of a 6 m aluminium powerboat was used to subject pre-fouled settlement plates for this purpose. Vessel speeds of 5 and 10 knots had little effect on the species richness of biofouling assemblages tested, however richness decreased by 50% following 18 knots treatments. Species percentage cover decreased with increasing speed across all speed treatments and this decrease was most pronounced at 10 and 18 knots, with cover reduced by 24 and 85% respectively. Survival was greatest for organisms with colonial, encrusting, hard and/or flexible morphological characteristics, and this effect increased with increasing speed. This study suggests that there is predictive power in forecasting future introductions if we can understand the extent to which such traits explain the world-wide distributions of non-indigenous species. Future introductions are a certainty and can only provide an increasing source of new information on which to test the validity of these predications.

Assessing the efficacy of spray-delivered 'eco-friendly' chemicals for the control and eradication of marine fouling pests.

Despite its frequent use in terrestrial and freshwater systems, there is a lack of published experimental research examining the effectiveness of spray-delivered chemicals for the management of non-indigenous and/or unwanted pests in marine habitats. This study tested the efficacy of spraying acetic acid, hydrated lime and sodium hypochlorite for the control of marine fouling assemblages. The chemicals are considered relatively 'eco-friendly' due to their low toxicity and reduced environmental persistence compared to synthetic biocides, and they were effective in controlling a wide range of organisms. Pilot trials highlighted acetic acid as the most effective chemical at removing fouling cover, therefore it was selected for more comprehensive full-scale trials. A single spray of 5% acetic acid with an exposure time of 1 min effectively removed up to 55% of the invertebrate species present and 65% of the cover on fouled experimental plates, while one application of 10% acetic acid over 30 min removed up to 78% of species present and 95% of cover. Single-spray treatments of 5% acetic acid reduced cover of the tunicate pest species Didemnum vexillum by up to 100% depending on the exposure duration, while repeat-spraying ensured that even short exposure times (1 min) achieved approximately 99% mortality. Both 5 and 10% acetic acid solutions appeared equally effective at removing the majority of algal species. This technique could be used for controlling the introduction of unwanted species or preventing the spread of pests, and is applicable to use on a variety of natural and artificial substrata, or for the treatment of structures that can be removed from the water.

Comparing differential tolerance of native and non-indigenous marine species to metal pollution using novel assay techniques.

Recent research suggests anthropogenic disturbance may disproportionately advantage non-indigenous species (NIS), aiding their establishment within impacted environments. This study used novel laboratory- and field-based toxicity testing to determine whether non-indigenous and native bryozoans (common within marine epibenthic communities worldwide) displayed differential tolerance to the common marine pollutant copper (Cu). In laboratory assays on adult colonies, NIS showed remarkable tolerance to Cu, with strong post-exposure recovery and growth. In contrast, native species displayed negative growth and reduced feeding efficiency across most exposure levels. Field transplant experiments supported laboratory findings, with NIS growing faster under Cu conditions. In field-based larval assays, NIS showed strong recruitment and growth in the presence of Cu relative to the native species. We suggest that strong selective pressures exerted by the toxic antifouling paints used on transport vectors (vessels), combined with metal contamination in estuarine environments, may result in metal tolerant NIS advantaged by anthropogenically modified selection regimes.

The influence of antifouling practices on marine invasions.

Vessel hull-fouling is increasingly recognised as one of the major vectors for the transfer of marine non-indigenous species. For hundreds of years, copper (Cu) has been used as a primary biocide to prevent the establishment of fouling assemblages on ships' hulls. Some non-indigenous fouling taxa continue to be transferred via hull-fouling despite the presence of Cu antifouling biocides. In addition, several of these species appear to enjoy a competitive advantage over similar native taxa within metal-polluted environments. This metal tolerance may further assist their establishment and spread in new habitats. This review synthesises existing research on the links between Cu and the invasion of fouling species, and shows that, with respect to the vector of hull-fouling, tolerance to Cu has the potential to play a role in the transfer of non-indigenous fouling organisms. Also highlighted are the future directions for research into this important nexus between industry, ecology and environmental management.

The potential for translocation of marine species via small-scale disruptions to antifouling surfaces.

Vessel hull fouling is a major vector for the translocation of nonindigenous species (NIS). Antifouling (AF) paints are the primary method for preventing the establishment and translocation of fouling species. However, factors such as paint age, condition and method of application can all reduce the effectiveness of these coatings. Areas of hull that escape AF treatment (through limited application or damage) constitute key areas that may be expected to receive high levels of fouling. The investigation focused on whether small-scale (mm(2) to cm(2)) areas of unprotected surface or experimental 'scrapes' provided sufficient area for the formation of fouling assemblages within otherwise undamaged AF surfaces. Recruitment of fouling taxa such as algae, spirorbids and hydroids was recorded on scrapes as narrow as 0.5 cm wide. The abundance and species richness of fouling assemblages developing on scrapes > or =1 cm often equalled or surpassed levels observed in reference assemblages totally unprotected by AF coatings. Experiments were conducted at three sites within the highly protected and isolated marine park surrounding Lady Elliott Island at the southernmost tip of the Great Barrier Reef, Australia. Several NIS were recorded on scrapes of AF coated surfaces at this location, with 1-cm scrapes showing the greatest species richness and abundance of NIS relative to all other treatments (including controls) at two of the three sites investigated. Slight disruptions to newly antifouled surfaces may be all that is necessary for the establishment of fouling organisms and the translocation of a wide range of invasive taxa to otherwise highly protected marine areas.