Towards better management of Australia’s shark fishery: genetic analyses reveal unexpected ratios of cryptic blacktip species Carcharhinus tilstoni and C. limbatus

The common blacktip shark (Carcharhinus limbatus) and the Australian blacktip shark (C. tilstoni) are morphologically similar species that co-occur in subtropical and tropical Australia. In striking contrast to what has been previously reported, we demonstrate that the common blacktip shark is not rare in northern Australia but occurs in approximately equal frequencies with the Australian blacktip shark. Management of shark resources in northern Australia needs to take account of this new information. Species identification was performed using nucleotide sequences of the control, NADH dehydrogenase subunit 4 (ND4) and cytochrome oxidase I (COI) regions in the mitochondrial genome. The proportion of overall genetic variation (FST) between the two species was small (0.042, P < 0.01) based on allele frequencies at five microsatellite loci. We confirm that a third blacktip species (C. amblyrhynchoides, graceful shark) is closely related to C. tilstoni and C. limbatus and can be distinguished from them on the basis of mtDNA sequences from two gene regions. The Australian blacktip shark (C. tilstoni) was not encountered among 20 samples from central Indonesia that were later confirmed to be common blacktip and graceful sharks. Fisheries regulators urgently need new information on life history, population structure and morphological characters for species identification of blacktip shark species in Australia.

Characterization of highly informative cross-species microsatellite panels for the Australian dugong (Dugong dugon) and Florida manatee (Trichechus manatus latirostris) including five novel primers

The Australian dugong (Dugong dugon) and Florida manatee (Trichechus manatus latirostris) are threatened species of aquatic mammals in the order Sirenia. Sirenian conservation and management actions would benefit from a more complete understanding of genetic diversity and population structure. Generally, species-specific microsatellite markers are employed in conservation genetic studies; however, robust markers can be difficult and costly to isolate. To increase the number of available markers, dugong and manatee microsatellite primers were evaluated for cross-species amplification. Furthermore, one manatee and four dugong novel primers are reported. After polymerase chain reaction optimization, 23 (92%) manatee primers successfully amplified dugong DNA, of which 11 (48%) were polymorphic. Of the 32 dugong primers tested, 27 (84%) yielded product in the manatee, of which 17 (63%) were polymorphic. Dugong and manatee primers were compared and the most informative markers were selected to create robust and informative marker-panels for each species. These cross-species microsatellite marker-panels can be employed to assess other sirenian populations and can provide beneficial information for the protection and management of these unique mammals.

The extent of population genetic subdivision differs among four co-distributed shark species in the Indo-Australian archipelago

Background: The territorial fishing zones of Australia and Indonesia are contiguous to the north of Australia in the Timor and Arafura Seas and in the Indian Ocean to the north of Christmas Island. The area surrounding the shared boundary consists of a variety of bio-diverse marine habitats including shallow continental shelf waters, oceanic trenches and numerous offshore islands. Both countries exploit a variety of fisheries species, including whaler (Carcharhinus spp.) and hammerhead sharks (Sphyrna spp.). Despite their differences in social and financial arrangements, the two countries are motivated to develop complementary co-management practices to achieve resource sustainability. An essential starting point is knowledge of the degree of population subdivision, and hence fisheries stock status, in exploited species. Results: Populations of four commercially harvested shark species (Carcharhinus obscurus, Carcharhinus sorrah, Prionace glauca, Sphyrna lewini) were sampled from northern Australia and central Indonesia. Neutral genetic markers (mitochondrial DNA control region sequence and allelic variation at co-dominant microsatellite loci) revealed genetic subdivision between Australian and Indonesian populations of C. sorrah. Further research is needed to address the possibility of genetic subdivision among C. obscurus populations. There was no evidence of genetic subdivision for P. glauca and S. lewini populations, but the sampling represented a relatively small part of their distributional range. For these species, more detailed analyses of population genetic structure is recommended in the future. Conclusion: Cooperative management between Australia and Indonesia is the best option at present for P. glauca and S. lewini, while C. sorrah and C. obscurus should be managed independently. On-going research on these and other exploited shark and ray species is strongly recommended. Biological and ecological similarity between species may not be a predictor of population genetic structure, so species-specific studies are recommended to provide new data to assist with sustainable fisheries management.

LC/MS/MS analysis of the daphnane orthoester simplexin in poisonous Pimelea species of Australian rangelands.

Liquid chromatography/mass spectrometry (MS)/MS was used to analyse toxins in P. trichostachia, P. simplex subsp. continua, P. simplex subsp. continua and P. elongata samples (flowers, seeds, branches, main stem, leaves and roots) collected from various locations in Queensland, Saskatchewan and New South Wales, Australia. Simplexin was the major analyte in all taxa, with varying minor levels of huratoxin. Simplexin levels in P. trichostachia and P. elongata were higher (580 and 540 mg/kg in flowering foliage, respectively) than in P. simplex (255 mg/kg). Levels of huratoxin were higher in P. simplex (relative to simplexin) than in P. trichostachia or P. elongata. P. simplex flower heads and roots contained similar simplexin levels, with very small amounts of toxins detected in branches, stems and leaves. In P. trichostachia, simplexin levels were high in flower heads but low in the the other plant parts. The simplexin levels in aerial parts were generally higher from the pre-flowering to the flowering stage, decreasing towards the post-flowering stage; similar trends were recorded for P.elongata samples collected from a site near Bollon and P. trichostachia samples collected from a site near Jericho (both sites in Queensland). The simplexin concentration in roots was much less variable. Flowers and seeds had much higher simplexin levels than the foliage. The breakdown of the toxin in litter was more rapid compared to seeds under the same weathering conditions. Unlike the results from the litter samples, no significant decrease occurred in seed samples after 18 months of exposure.

Productivity of plantation hardwood tree species in north-eastern Australia: A report from the Forest Adaptation and Sequestration Alliance

The report summarises data from a large number of trials of species with potential for use by the plantation forest industry in north-eastern Australia and provides information aimed at improving the understanding of growth rates, pest and disease risks and carbon sequestration. Data is summarised and presented at a regional level as opposed to individual trial or plot level. As well, nutritional impediments to tree growth and impacts on forest health are also reported. This report is intended to contribute to policy deliberations about developing forestry opportunities that can that can be integrated into the landscape, with particular consideration given to lower rainfall regions. There are several examples in north-eastern Australia where production forests have developed sub-optimally; this has often been due to poor selection of tree species as little information has been available. This report helps address this deficiency.

Bivalves for the remediation of prawn farm effluent: identification of some potentially useful species in Southern Queensland

Several species of oysters, clams and mussels are currently being used around the world to create extra profits and help remediate waste-waters from mariculture operations. To identify opportunities and potentially suitable species of bivalves for remediation of prawn farm effluent in Australia, recent literature dealing with bivalve filtration is reviewed, and species occurring naturally in a banana prawn, Penaeus (Fenneropenaeus) merguiensis, grow-out pond and effluent streams at the Bribie Island Aquaculture Research Centre (BIARC) were collected, identified and assessed in terms of their tolerance of high silt loadings over 3 months. Three bivalve species predominated in the BIARC case study. These were the mud ark, Anadara trapezia, the rock oyster, Dendostrea folium, and the pearl shell, Pinctada maculata. The mud ark demonstrated the highest tolerance of silt loading (99% survival), followed by pearl shells and rock oysters (88 and 63% survival respectively).

Contagious dispersal of seeds of synchronously fruiting species beneath invasive and native fleshy-fruited trees

In subtropical Australia, many native and invasive plant species rely on a shared suite of frugivores, largely birds, for seed dispersal. Many native plants fruit during summer in this region, whereas most invasive plants fruit during winter, thus providing the opportunity for contagious dispersal of seeds beneath synchronously fruiting species. We sampled invasive and native seed rain beneath the canopy of a native summer-fruiting tree Guioa semiglauca and an invasive winter-fruiting tree Cinnamomum camphora, in three study sites over the course of a year. In July, during peak fruiting season for C. camphora and other invasive species, seed rain of invasive species was higher beneath C. camphora than G. semiglauca. This was partly due to the invasive tree Ligustrum lucidum, whose seed rain was three times higher beneath C. camphora than beneath the native tree. In February, seed rain of native species was more abundant beneath the canopy of G. semiglauca than beneath C. camphora, despite the fact that C. camphora was also fruiting at this time. This was probably due to the larger fruit crop produced by G. semiglauca at this time of year. Our study provides evidence that the presence of invasive bird-dispersed plants may facilitate contagious seed dispersal of other invaders, and likewise native species may facilitate seed spread of other native plants.

Relative host plant species use by the lantana biological control agent Aconophora compressa (Membracidae) across its native and introduced ranges

Aconophora compressa Walker (Hemiptera: Membracidae) was released in 1995 against the weed lantana in Australia, and is now found on multiple host plant species. The intensity and regularity at which A. compressa uses different host species was quantified in its introduced Australian range and also its native Mexican range. In Australia, host plants fell into three statistically defined categories, as indicated by the relative rates and intensities at which they were used in the field. Fiddlewood (Citharexylum spinosum L.: Verbenaceae) was used much more regularly and at higher densities than any other host sampled, and alone made up the first group. The second group, lantana (Lantana camara L.: Verbenaceae; pink variety) and geisha girl (Duranta erecta L.: Verbenaceae), were used less regularly and at much lower densities than fiddlewood. The third group, Sheena’s gold (another variety of D. erecta), jacaranda (Jacaranda mimosifolia D. Don: Bignoniaceae) and myoporum (Myoporum acuminatum R. Br.: Myoporaceae), were used infrequently and at even lower densities. In Mexico, the insect was found at relatively low densities on all hosts relative to those in Australia. Densities were highest on L. urticifolia, D. erecta and Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae), which were used at similar rates to one another. It was found also on a few other verbenaceous and non-verbenaceous host species but at even lower densities. The relative rate at which Citharexylum spp. and L. urticifolia were used could not be assessed in Mexico because A. compressa was found on only one plant of each species in areas where these host species co-occurred. The low rate at which A. compressa occurred on fiddlewood in Mexico is likely to be an artefact of the short-term nature of the surveys or differences in the suites of Citharexylum and Lantana species available there. These results provide further incentive to insist on structured and quantified surveys of non-target host use in the native range of potential biological control agents prior to host testing studies in quarantine.

Teratosphaeria pseudoeucalypti, new cryptic species responsible for leaf blight of Eucalyptus in subtropical and tropical Australia

Sub-tropical and tropical plantations of Eucalyptus grandis hybrids in eastern Australia have been severely affected by anamorphs of Teratosphaeria (formerly Kirramyces) causing a serious leaf blight disease. Initially the causal organism in Queensland, Australia, was identified as Teratosphaeria eucalypti, a known leaf parasite of endemic Eucalyptus spp. However, some inconsistencies in symptoms, damage and host range suggested that the pathogen in Queensland may be a new species. Isolates of T. eucalypti from throughout its known endemic range, including Queensland and New Zealand, where it is an exotic pathogen, were compared using multiple gene phylogenies. Phylogenetic studies revealed that the species responsible for leaf blight in Queensland represents a new taxon, described here as Teratosphaeria pseudoeucalypti. While the DNA sequence of T. pseudoeucalypti was more similar to T. eucalypti, the symptoms and cultural characteristics resembled that of T. destructans. The impact of this disease in central Queensland has increased annually and is the major threat to the eucalypt plantation industry in the region.

Species within Mycosphaerellaceae and Teratosphaeriaceae from eucalypts in eastern Australia

Twenty eight species within Mycosphaerellaceae and Teratosphaeriaceae (includes Mycosphaerella, Teratosphaeria, Pseudocercospora and Sonderhenia) are reported from Eucalyptus and Corymbia in New South Wales and Queensland, Australia, based on field surveys and examination of herbarium specimens and published reports. Teratosphaeria cryptica was the most commonly recorded species, with the widest host range and distribution, followed by Mycosphaerella marksii. Six new species are described: T. keanei, T. coolabuniensis, T. crispata, M. medusae, M. nootherensis and T. praelongispora. New or interesting records for known species are reported, including new records for Australia (T. pluritubularis and T. verrucosiafricana) and new records for Queensland (T. excentrica, T. multiseptata, T. nubilosa, T. suberosa and Ps. pseudoeucalyptorum).

Oral Actinomyces Species in Health and Disease: Identification, Occurence and Importance of Early Colonization

The genus Actinomyces consists of a heterogeneous group of gram-positive, mainly facultatively anaerobic or microaerobic rods showing various degrees of branching. In the oral cavity, streptococci and Actinomyces form a fundamental component of the indigenous microbiota, being among initial colonizers in polymicrobial biofilms. The significance of the genus Actinomyces is based on the capability of species to adhere to surfaces such as on teeth and to co-aggregate with other bacteria. Identification of Actinomyces species has mainly been based on only a few biochemical characteristics, such as pigmentation and catalase production, or on the use of a single commercial kit. The limited identification of oral Actinomyces isolates to species level has hampered knowledge of their role both in health and disease. In recent years, Actinomyces and related organisms have attracted the attention of clinical microbiologists because of a growing awareness of their presence in clinical specimens and their association with disease. This series of studies aimed to amplify the identification methods for Actinomyces species. With the newly developed identification scheme, the age-related occurrence of Actinomyces in healthy mouths of infants and their distribution in failed dental implants was investigated. Adhesion of Actinomyces species to titanium surfaces processed in various ways was studied in vitro. The results of phenotypic identification methods indicated a relatively low applicability of commercially available test kits for reliable identification within the genus Actinomyces. However, in the study of conventional phenotypic methods, it was possible to develop an identification scheme that resulted in accurate differentiation of Actinomyces and closely related species, using various different test methods. Genotypic methods based on 16S rRNA sequence analysis of Actinomyces proved to be a useful method for genus level identification and further clarified the species level identification with phenotypic methods. The results of the study of infants showed that the isolation frequency of salivary Actinomyces species increased according to age: thirty-one percent of the infants at 2 months but 97% at 2 years of age were positive for Actinomyces. A. odontolyticus was the most prominent Actinomyces colonizer during the study period followed in frequency by A. naeslundii and A. viscosus. In the study of explanted dental implants, Actinomyces was the most prevalent bacterial genus, colonizing 94% of the fixtures. Also in the implants A. odontolyticus was revealed as the most common Actinomyces species. It was present in 84% of Actinomyces -positive fixtures followed in frequency by A. naeslundii, A. viscosus and A. israelii. In an in vitro study of titanium surfaces, different Actinomyces species showed variation regarding their adhesion to titanium. Surface roughness as well as albumin coating of titanium had significant effects on adhesion. The use of improved phenotypic and molecular diagnostic methods increased the accuracy of the identification of the Actinomyces to species level. This facilitated an investigation of their occurrence and distribution in oral specimens in both health and disease.

Stock structure of exploited shark species in north-eastern Australia

The project has provided management and other stakeholders with information necessary to make informed decisions about the management of four of the key exploited shark species caught in the Queensland inshore net fishery and northern New South Wales line fishery. The project has determined that spatial management of milk sharks within Queensland, and scalloped hammerhead, common black tip and Australian black tip sharks within Queensland and New South Wales is appropriate. The project has determined that both black tip shark species are likely to require co-operative management arrangements between Queensland and New South Wales. For scalloped hammerheads separate stocks between the two jurisdictions were identified from the fisheriesdependent samples, however genetic exchange across borders is likely to be facilitated by movement of adult females and perhaps larger males to a lesser extent. This information will greatly assist compliance with the Commonwealth Environment Protection and Biodiversity Conservation Act (1999) for shark fisheries in north-eastern Australia by providing the necessary basis for robust assessment of the status of stocks of the study species, thereby helping to deliver their sustainable harvest. It also helps to achieve objectives of the Australian National Shark Plan. The project provides the appropriate spatial framework for future monitoring and assessment of the study species. This is at a time when shark fisheries are receiving close attention from all sectors and when monitoring programs are being implemented, aimed at better assessment of stock status. This project has provided the crucial information for developing an appropriate monitoring design as well as the necessary basis for making statements about stock status. The project has addressed research priorities identified by the Queensland Fisheries Research Advisory Board, Great Barrier Reef Marine Park Authority and Queensland Fisheries. Previously management has assumed a single stock for each species on the east coast of Queensland, and management of shark fisheries in New South Wales (NSW) and Queensland has been independent of one another. The project has been able to enhance and develop links between research, management and industry. Strong positive relationships with commercial fishers were crucial in the collection of samples throughout the study area and fisheries managers were part of the project team throughout the study period. During the project the study area was extended to include both Queensland and NSW waters, creating mutualistic and positive links between the States’ research and management agencies. Extension of project results included management representatives from NSW and Queensland, as well as the Northern Territory where similar shark fisheries operate and similar species are targeted. The project was able to provide significant human capital development opportunities providing considerable value to the project outcomes. Use of vertebral microchemistry and life history characteristics as stock determination methods provided material for two PhD students based at James Cook University: Ron Schroeder, vertebral chemistry; and Alastair Harry, life history characteristic. The project has developed novel research methods that have great capacity for future application, including: • Development of a simple and rapid genetic diagnostic tool (RT-HRM-PCR assay) for differentiating among the black tip shark species, for which no simple morphological identifier exists; and • Development of laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) methods for analysing and interpreting microchemical composition of shark vertebrae. The study has provided further confirmation of the effectiveness of using a holistic approach in stock structure studies and justifies investment into such studies.

Eliciting and encoding expert knowledge on variable selection into classical or Bayesian species distribution models

The quality of species distribution models (SDMs) relies to a large degree on the quality of the input data, from bioclimatic indices to environmental and habitat descriptors (Austin, 2002). Recent reviews of SDM techniques, have sought to optimize predictive performance e.g. Elith et al., 2006. In general SDMs employ one of three approaches to variable selection. The simplest approach relies on the expert to select the variables, as in environmental niche models Nix, 1986 or a generalized linear model without variable selection (Miller and Franklin, 2002). A second approach explicitly incorporates variable selection into model fitting, which allows examination of particular combinations of variables. Examples include generalized linear or additive models with variable selection (Hastie et al. 2002); or classification trees with complexity or model based pruning (Breiman et al., 1984, Zeileis, 2008). A third approach uses model averaging, to summarize the overall contribution of a variable, without considering particular combinations. Examples include neural networks, boosted or bagged regression trees and Maximum Entropy as compared in Elith et al. 2006. Typically, users of SDMs will either consider a small number of variable sets, via the first approach, or else supply all of the candidate variables (often numbering more than a hundred) to the second or third approaches. Bayesian SDMs exist, with several methods for eliciting and encoding priors on model parameters (see review in Low Choy et al. 2010). However few methods have been published for informative variable selection; one example is Bayesian trees (O’Leary 2008). Here we report an elicitation protocol that helps makes explicit a priori expert judgements on the quality of candidate variables. This protocol can be flexibly applied to any of the three approaches to variable selection, described above, Bayesian or otherwise. We demonstrate how this information can be obtained then used to guide variable selection in classical or machine learning SDMs, or to define priors within Bayesian SDMs.

Assessment of novel gear designs to reduce interactions between species

This project tested modified gillnets designed by commercial net fishers in the Queensland East Coast Inshore Finfish Fishery (ECIFF) to try and identify gears that would mitigate and/or improve interactions between fishing nets and Species of Conservation Interest (SOCI). The study also documents previously unrecognised initiatives by pro-active commercial net fishers that reflect a conservation-minded approach to their fishing practices, which is the opposite of what is perceived publicly. Between 2011 and 2014, scientists from James Cook University and the Queensland Department of Agriculture and Fisheries teamed with commercial fishers representing the Queensland Seafood Industry Association and the Moreton Bay Seafood Industry Association to conduct field trials of various modified net designs under normal fishery conditions. Trials were conducted in Moreton Bay (southern part of the fishery) and Bowling Green Bay (northern) and tested different net designs developed by fishers to improve the nature of interactions between net fishing gear and SOCI.