Development of a DNA based aging technique for use in fisheries assessments. Final report, Australian Fisheries Research and Development Corporation Project 2007/033.

The productivity of a fisheries resource can be quantified from estimates of recruitment, individual growth and natural and fisheries-related mortality, assuming the spatial extent of the resource has been quantified and there is minimal immigration or emigration. The sustainability of a fisheries resource is facilitated by management controls such as minimum and maximum size limits and total allowable catch. Minimum size limits are often set to allow individuals the opportunity to reproduce at least once before the chance of capture. Total allowable catches are a proportion of the population biomass, which is estimated based on known reproduction, recruitment, mortality and growth rates. In some fisheries, however, management actions are put in place without quantification of the resource through the stock assessment process. This occurs because species-specific information, for example individual growth, may not be available. In these circumstances, management actions need to be precautionary to protect against future resource collapse, but this often means that the resource is lightly exploited. Consequently, the productivity of the resource is not fully realised. Australia’s most valuable fisheries are invertebrate fisheries (Australian Department of Agriculture Fisheries and Forestry, 2008). For example, Australian fisheries (i.e. excluding aquaculture) production of crustaceans (largely prawns, rock lobster and crab) was 41,000 tonnes in 2006/7, worth $778 million. Production from mollusc (largely abalone, scallops, oysters and squid) fisheries was 39,000 tonnes, worth $502 million. Together, in 2006/7 crustacean and mollusc fisheries represented 58% of the total value of Australian wild fisheries production. Sustainable management of Australia’s invertebrate fisheries is frustrated by the lack of data on species-specific growth rates. This project investigated a new method to estimate age, and hence individual growth rates, in invertebrate fisheries species. The principle behind the new aging method was that telomeres (i.e. DNA end-caps of chromosomes) get shorter as an individual gets older. We studied commercial crustacean and molluscan species. A vertebrate fish species (silver perch, Bidyanus bidyanus) was used as a control to standardise our work against the literature. We found a clear relationship between telomere length and shell size for temperate abalone (Haliotis rubra). Further research is recommended before the method can be implemented to assist management of wildharvested abalone populations. Age needs to be substituted for shell size in the relationship and it needs to be studied for abalone from several regions. This project showed that telomere length declined with increasing age in Sydney rock oysters (Saccostrea glomerata) and was affected by regional variation. A relationship was not apparent between telomere length and age (or size as a surrogate for age) for crustacean species (school prawns, Metapenaeus macleayi; eastern rock lobster, Sagmariasus verreauxi; southern rock lobster, Jasus edwardsii; and spanner crabs, Ranina ranina). For school prawns, there was no difference between telomere length in males and females. Further research is recommended, however, as telomeric DNA from crustaceans was difficult to analyse using the terminal restriction fragment (TRF) assay. Telomere lengths of spanner crabs and lobsters were at the upper limit of resolution of the assay used and results were affected by degradation and possible contamination of telomeric DNA. It is possible that telomere length is an indicator of remaining lifespan in molluscan and crustacean individuals, as suggested for some vertebrate species (e.g. Monaghan, 2010). Among abalone of similar shell size and among lobster pueruli, there was evidence of individuals having significantly longer or shorter telomeres than the group average. At a population level, this may be a surrogate for estimates of future natural mortality, which may have usefulness in the management of those populations. The method used to assay telomere length (terminal restriction fragment assay) performed adequately for most species, but it was too expensive and time-consuming to be considered a useful tool for gathering information for fisheries management. Research on alternative methods is strongly recommended.

Pleopod Cuticular Morphology as an Index of Moult Stage in the Ornate Rock Lobster, Panulirus ornatus (Fabricius 1789)

Seven discrete stages and substages of moulting in the ornate rock lobster, Panulirus ornatus, have been distinguished by microscopic examination of the cuticle and setae of the pleopods . The diagnostic features and the duration of each of the stages are described. Freezing did not visually alter the tissue features used to identify each moult stage. Pleopod morphology can reliably indicate whether a lobster has moulted within the previous 24 h or is within 72 h of the next ecdysis.

A spatially explicit multi-competitor coexistence model of penaeid (shrimp) distribution on the Australian Great Barrier Reef

A spatially explicit multi-competitor coexistence model was developed for meta-populations of prawns (shrimp) occupying habitat patches across the Great Barrier Reef, where dispersal was localised and dispersal rates varied between species. Prawns were modelled as individuals moving to and from patches or cells according to pre-set decision rules. The landscape was simulated as a matrix of cells with each cell having a spatially explicit survival index for each species. Mixed species prawn assemblages moved over this simplified spatially explicit landscape. A low level of chronic random environmental disturbance was assumed (cyclone and tropical storm damage) with additional acute spatially confined disturbance due to commercial trawling, modelled as an increase in mortality affecting inter-specific competition. The general form of the results was for increased disturbance to favour good-colonising "generalist" species at the expense of good-competitor "specialists". Increasing fishing mortality (local patch extinctions) combined with poor colonising ability resulted in low equilibrium abundance for even the best competitor, while in the same circumstances the poorest competitor but best coloniser could have the highest equilibrium abundance. This mimics the switch from high-value prawn species to lower-value prawn species as trawl effort increases, reflected in historic catch and effort logbook data and reported anecdotaly from the north Queensland trawl fleet. To match the observed distribution and behaviour of prawn assemblages, a combination inter-species competition, a spatially explicit landscape, and a defined pattern of disturbance (trawling) was required. Modelling this combination could simulate not only general trends in spatial distribution of each of prawn species but also localised concentrations observed in the survey data

Isolation and expression analysis of multiple isoforms of putative farnesoic acid O-methyltransferase in several crustacean species

Farnesoic acid O-methyltransferase (FaMeT) is the enzyme responsible for the conversion of farnesoic acid (FA) to methyl farnesoate (MF) in the final step of MF synthesis. Multiple isoforms of putative FaMeT were isolated from six crustacean species belonging to the families Portunidae, Penaeidae, Scyllaridae and Parastacidae. The portunid crabs Portunus pelagicus and Scylla serrata code for three forms: short, intermediate and long. Two isoforms (short and long) were isolated from the penaeid prawns Penaeus monodon and Fenneropenaeus merguiensis. Two isoforms were also identified in the scyllarid Thenus orientalis and parastacid Cherax quadricarinatus. Putative FaMeT sequences were also amplified from the genomic DNA of P. pelagicus and compared to the putative FaMeT transcripts expressed. Each putative FaMeT cDNA isoform was represented in the genomic DNA, indicative of a multi-gene family. Various tissues from P. pelagicus were individually screened for putative FaMeT expression using PCR and fragment analysis. Each tissue type expressed all three isoforms of putative FaMeT irrespective of sex or moult stage. Protein domain analysis revealed the presence of a deduced casein kinase II phosphorylation site present only in the long isoform of putative FaMeT.

Gene expression profiling of cuticular proteins across the moult cycle of the crab Portunus pelagicus

Background: Crustaceans represent an attractive model to study biomineralization and cuticle matrix formation, as these events are precisely timed to occur at certain stages of the moult cycle. Moulting, the process by which crustaceans shed their exoskeleton, involves the partial breakdown of the old exoskeleton and the synthesis of a new cuticle. This cuticle is subdivided into layers, some of which become calcified while others remain uncalcified. The cuticle matrix consists of many different proteins that confer the physical properties, such as pliability, of the exoskeleton. Results: We have used a custom cDNA microarray chip, developed for the blue swimmer crab Portunus pelagicus, to generate expression profiles of genes involved in exoskeletal formation across the moult cycle. A total of 21 distinct moult-cycle related differentially expressed transcripts representing crustacean cuticular proteins were isolated. Of these, 13 contained copies of the cuticle_1 domain previously isolated from calcified regions of the crustacean exoskeleton, four transcripts contained a chitin_bind_4 domain (RR consensus sequence) associated with both the calcified and un-calcified cuticle of crustaceans, and four transcripts contained an unannotated domain (PfamB_109992) previously isolated from C. pagurus. Additionally, cryptocyanin, a hemolymph protein involved in cuticle synthesis and structural integrity, also displays differential expression related to the moult cycle. Moult stage-specific expression analysis of these transcripts revealed that differential gene expression occurs both among transcripts containing the same domain and among transcripts containing different domains. Conclusion: The large variety of genes associated with cuticle formation, and their differential expression across the crustacean moult cycle, point to the complexity of the processes associated with cuticle formation and hardening. This study provides a molecular entry path into the investigation of the gene networks associated with cuticle formation.

Additional records of the rock shrimp, Sicyonia australiensis Hanamura & Wadley, 1998 (Decapoda, Sicyoniidae) from eastern Australia, western central Pacific

This paper reports on the collection of S. australiensis from the continental shelf off southern Queensland, easter Australia, in the western Central Pacific, documenting for the first time the occurrence of the species outside of eastern Bass Strait.

Aspects of reproduction and diet of the Australian endemic skate Dipturus polyommata (Ogilby) (Elasmobranchii: Rajidae), by-catch of a commercial prawn trawl fishery

The Australian endemic skate Dipturus polyommata collected from by-catch of a benthic prawn fishery off southern Queensland was examined to provide information on reproduction and diet. Morphological relationships of total length (LT) to disc width and LT to mass were estimated. Size at birth was estimated at c. 100-110 mm and size at first feeding at c. 105-110 mm LT. Size at 50% maturity (LT50 and 95% CI) was 321 (305-332) and 300 (285-306) mm LT for females and males, respectively. Size at first maturity corresponded to 87.7% of observed maximum size in females (366 mm LT) and 87.5% in males (343 mm L T). Two females, representing 18.2% of mature females sampled in the austral winter were each carrying two egg cases. Descriptions of egg cases are given. Diet described by the index of relative importance as a percentage (%IRI) was predominantly crustacean based with carid shrimps (53.64%) and penaeoid prawns (23.30%) the most significant prey groups. Teleosts (11.72%), gammarid amphipods (5.31%) and mysids (4.72%) were also important to the diet of the species, while a further six prey groups made only a minor contribution to diet (1.31%). An ontogenetic change was evident between the diets of immature and mature skates. Immature animals fed more extensively on carids and amphipods and mature animals on penaeoids, teleosts and mysids.

Limb-loss in pond-reared blue swimmer crabs Portunus pelagicus (L.): Effect on growth in an indoor shedding system

Limb-loss in crustaceans can reduce moult increment and delay or advance the timing of moulting, both aspects that are likely to impact upon soft-shell crab production. Pond-reared blue swimmer crabs Portunus pelagicus were harvested and maintained in a crab shedding system. The wet weight, carapace width (CW) and the occurrence of limb-loss were assessed before stocking in the shedding system and after each of the next three moults. Many of the crabs were initially missing one or two limbs and these did not grow as much as the crabs that were intact at the start of the trial. Despite its strong correlation with wet weight, CW changes proved to be misleading. Limb-loss reduced the %CW increment but not the per cent weight increment (where the later is calculated from the actual pre-moult weight). Pre-moult weight explained much of the variation in post-moult weight, with crabs moulting to approximately double their weight. Limb-loss reduced 'growth' and production from the pond because it reduced pre-moult weight but limb-loss did not alter the weight change on shedding a given weight of crabs, although some of that change now included regeneration of limbs. One can hypothesize that much of the size variation seen in pond-reared crabs may be due to accumulated effects of repeated limb-loss, rather than genetic variation.

Evaluation of partial replacement of live and fresh feeds with a formulated diet and the influence of weaning Panulirus ornatus phyllosomata onto a formulated diet during early ontogeny

We have evaluated the potential of a formulated diet as a replacement for live and fresh feeds for 7-day post-hatch Panulirus ornatus phyllosomata and also investigated the effect of conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a 100% formulated diet. In the first trial, the highest survival (>55%) was consistently shown by phyllosomata fed a diet consisting of a 50% combination of Artemia nauplii and 50% Greenshell mussel, followed by phyllosomata fed 50% Artemia nauplii and 50% formulated diet and, thirdly, by those receiving 100% Artemia nauplii. The second trial assessed the replacement of on-grown Artemia with proportions of formulated diet and Greenshell mussel that differed from those used in trial 1. Phyllosomata fed a 75% combination of formulated diet and 25% on-grown Artemia and 50% on-grown Artemia and 50% Greenshell mussel consistently showed the highest survival (>75%). Combinations of Greenshell mussel and formulated diet resulted in significantly (P < 0.05) reduced survival. In trial 3, phyllosomata were conditioned for 14, 18 or 21 days on Artemia nauplii prior to weaning onto a 100% formulated diet, which resulted in survival rates that were negatively related to the duration of feeding Artemia nauplii. In the final trial, phyllosomata were conditioned for 14 days on live on-grown Artemia prior to weaning onto one of three formulated diets (one diet with 44% CP and two diets with 50%). Phyllosomata fed a 44% CP diet consistently showed the highest survival (>35%) among all treatments, while those fed a 50%-squid CP diet showed a significant (P < 0.05) increase in mortality at day 24. The results of these trials demonstrate that hatcheries can potentially replace 75% of live on-grown Artemia with a formulated diet 7 days after hatch. The poor performance associated with feeding combinations of Greenshell mussel and formulated diet, and 100% formulated diet as well as conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a formulated diet highlights the importance of providing Artemia to stimulate feeding.

Ghost fishing in the pot fishery for blue swimmer crabs Portunus pelagicus in Queensland, Australia

Blue swimmer crabs (Portunus pelagicus) are an economically important crab caught in baited traps throughout the Indo-west Pacific and Mediterranean. In Australia they are traditionally caught using rigid wire traps (approximate to pots) but there has been a recent increase in the use of collapsible pots constructed from polyethylene trawl mesh. Two experiments were conducted in Moreton Bay, Queensland, to determine the ghost fishing potential of lost crab pots on both target and bycatch species and to evaluate the differences between traditional and contemporary pot designs. A lost contemporary, collapsible trawl mesh pot will catch between 3 and 223 R pelagicus per year after the bait has been exhausted, while a traditional wire mesh pot would catch 11-74 crabs peryear. As most fishers now use the collapsible trawl mesh pots, ghost fishing mortality could be as high as 111,811-670,866 crabs per year. Bycatch retention was also higher in contemporary designs. Periods of strong winds appeared to increase the ghost fishing potential of lost pots. The use of escape gaps, larger mesh sizes and construction options that allow for the deterioration of entrance funnels to minimise ghost fishing are recommended to reduce environmental impacts.

Stocking density and artificial habitat influence stock structure and yield from intensive nursery systems for mud crabs Scylla serrata (Forsskål 1775)

Intensive nursery systems are designed to culture mud crab postlarvae through a critical phase in preparation for stocking into growout systems. This study investigated the influence of stocking density and provision of artificial habitat on the yield of a cage culture system. For each of three batches of postlarvae, survival, growth and claw loss were assessed after each of three nursery phases ending at crab instars C1/C2, C4/C5 and C7/C8. Survival through the first phase was highly variable among batches with a maximum survival of 80% from megalops to a mean crab instar of 1.5. Stocking density between 625 and 2300 m-2 did not influence survival or growth in this first phase. Stocking densities tested in phases 2 and 3 were 62.5, 125 and 250 m -2. At the end of phases 2 and 3, there were five instar stages present, representing a more than 20-fold size disparity within the populations. Survival became increasingly density-sensitive following the first phase, with higher densities resulting in significantly lower survival (phase 2: 63% vs. 79%; phase 3: 57% vs. 64%). The addition of artificial habitat in the form of pleated netting significantly improved survival at all densities. The mean instar attained by the end of phase 2 was significantly larger at a lower stocking density and without artificial habitat. No significant effect of density or habitat on harvest size was detected in phase 3. The highest incidence of claw loss was 36% but was reduced by lowering stocking densities and addition of habitat. For intensive commercial production, yield can be significantly increased by addition of a simple net structure but rapidly decreases the longer crablets remain in the nursery.

Patterns in the distribution and abundance of female eastern king prawns, Melicertus plebejus (Hess, 1865), capable of spawning and reproductive potential in waters off

Linear mixed models were used to test the null hypothesis that there were no differences between seasons and locations in the reproductive potential of female eastern king prawns, Melicertus plebejus along the east coast of Australia. Three samples were collected in each season between autumn 1991 and winter 1992 (inclusive). Females capable of spawning were found at all locations but proportions were greater in lower than higher latitudes. Females capable of spawning were not found at the southern (highest latitude) most location in all seasons. There was a significant interaction in reproductive potential between seasons and locations suggesting that patterns among seasons differed between locations and vice versa. Reproductive potential was greatest amongst the northern (lower latitudes) most locations and was greatest in autumn at these locations. Seasonal patterns were less pronounced further south (higher latitudes). The length composition of females in catches differed between locations with more larger prawns found in samples from northern locations. The challenge that remains is to quantify the oceanic sources of larvae that contribute to recruitment in each nursery area and the estuarine sources of juveniles that contribute adults back to the effective spawning stock. Maintaining the effective spawning stock and important nursery areas are crucial to the sustainability of this resource.