Microbial diversity within the water column of a larval rearing system for the ornate rock lobster (Panulirus ornatus)

The ornate tropical rock lobster, Panulirus ornatus has substantial potential as an aquaculture species though disease outbreaks during the animal's extended larval lifecycle are major constraints for success. In order to effectively address such disease-related issues, an improved understanding of the composition and dynamics of the microbial communities in the larval rearing tanks is required. This study used flow cytometry and molecular microbial techniques (clone libraries and denaturing gradient gel electrophoresis (DGGE)) to quantify and characterise the microbial community of the water column in the early stages (developmental stage I-II) of a P. ornatus larval rearing system. DGGE analysis of a 5000 L larval rearing trial demonstrated a dynamic microbial community with distinct changes in the community structure after initial stocking (day I to day 2) and from day 4 to day 5, after which the structure was relatively stable. Flow cytometry analysis of water samples taken over the duration of the trial demonstrated a major increase in bacterial load leading up to and peaking on the first day of the initial larval moult (day 7), before markedly decreasing prior to when > 50% of larvae moulted (day 9). A clone library of a day 10 water sample taken following a mass larval mortality event reflected high microbial diversity confirmed by statistical analysis indices. Sequences retrieved from both clone library and DGGE analyses were dominated by gamma- and alpha-Proteobacteria affiliated organisms with additional sequences affiliated with beta- and epsilon-Proteobacteria, Bacteroidetes, Cytophagales and Chlamydiales groups. Vibrio affiliated species were commonly retrieved in the clone library, though absent from DGGE analysis.

European lobster (Homarus gammarus) larvae under an acidification scenario: addressing biochemical, development and behaviour responses

A acidificação dos oceanos constitui uma problemática global e a realidade de que está, efetivamente, a acontecer não é uma consideração subjetiva. A Acidificação dos oceanos provocada por emissões de dióxido de carbono de origem antropogénica tem vindo a reduzir o pH das águas superficiais do Oceano e projeções preveem a continuidade deste processo. Embora muita investigação tenha sido desenvolvida no âmbito dos invertebrados que calcificam, tais como moluscos e crustáceos, poucos consideraram o estudo de efeitos ao nível sub-celular para avaliar stress oxidativo ou respostas funcionais do metabolismo energético em tais condições, interligando vários níveis de organização biológica. O objetivo do presente estudo foi o de avaliar os efeitos da exposição a diferentes níveis-alvo de pCO2 (controlo: 370 μatm; aumentado: 710 μatm) e de pH (controlo: 8.15; reduzido: 7.85) em parâmetros de crescimento bem como avaliar respostas comportamentais e bioquímicas relacionadas com stress oxidativo e metabolismo energético durante o desenvolvimento larvar de um crustáceo decápode. Este cenário de acidificação está de acordo com os RCPs previstos pelo Painel Intergovernamental para as Alterações Climáticas (IPCC, 2014) para o ano 2100. Para o presente estudo, foi utilizado o crustáceo Homarus gammarus (L.) – sendo uma espécie com elevado valor comercial, que tem vindo a sofrer elevada pressão de pesca em águas Europeias. Fêmeas adultas provenientes da costa Atlântica Oeste Portuguesa foram obtidas de um retalhista local. Após a eclosão em ambiente laboratorial controlado, larvas provenientes da mesma progenitora foram expostas às condições acima descritas desde o momento da eclosão até à chegada ao Estádio III. A sobrevivência individual e ocorrência de ecdise foram avaliados individualmente de 12h em 12h. Réplicas de cada tratamento foram recolhidas em momentos específicos durante o Estádio I (primeiro estádio larvar) e Estádio III (último estádio larvar) para análise morfométrica e de crescimento (peso fresco e comprimento carapaça) e respostas bioquímicas. Os biomarcadores analisados incluíram parâmetros relacionados com stress oxidativo e danos (atividade da enzima superóxido dismutase (SOD), peroxidação lipídica (LPO) e danos no DNA) e metabolismo energético (atividade da cadeia transportadora de eletrões (ETS) e da enzima lactato desidrogenase (LDH) e quantificação de Hidratos de Carbono). Os resultados obtidos sugerem que a sobrevivência diminui e que o período inter-muda é afetado durante a exposição a cenários de acidificação. No que respeita aos parâmetros de crescimento/morfométricos, larvas do cenário de acidificação apresentam uma tendência para crescimento diminuído, menor peso e comprimento de carapaça. As análises bioquímicas realizadas indicam a ocorrência de stress oxidativo sob condições de acidificação. Respostas ao nível do metabolismo energético não variaram significativamente entre tratamentos. Os resultados apontam também para que fases larvares possam possuir um sistema antioxidante ainda em desenvolvimento, tornando-as mais suscetíveis ao stress oxidativo. As fases larvares são uma fase vulnerável e crucial no ciclo de vida das espécies, influenciando o recrutamento e a renovação de stocks. Este estudo contribui para um melhor entendimento sobre a vulnerabilidade desta espécie num cenário de alterações climáticas – Acidificação dos oceanos – ao endereçar os mecanismos envolvidos nas respostas deste crustáceo a este agente causador de stress.

Size selectivity of diamond and square mesh cod ends for rose shrimp (Parapenaeus longirostris) and Norway lobster (Nephrops norvegicus) off the Portuguese south coast

The effects of an increase in cod end mesh size from 55 to 60 and 70 mm and a change of mesh configuration from diamond to square mesh on the size selectivity for rose shrimp Parapenaeus longirostris and Norway lobster Nephrops norvegicus captured off the Portuguese south coast were evaluated. The results were analysed taking into account between-haul variation in selectivity, and indicate a significant increase in L-50 for rose shrimp with an increase in mesh size or with the use of a square mesh cod end, while for Norway lobster only mesh configuration was found to affect this parameter. Two other important external variables were identified; the trawling depth and the cod end catch, which influence between-haul variation, by increasing the selection range for rose shrimp and Norway lobster, respectively. The results obtained suggest that an increase in the current minimum mesh size of 55 mm would be advisable for rose shrimp in order to respect the minimum landing size of 24 mm carapace length presently established for this species. Moreover, trawling for rose shrimp should be avoided at depths above 200 m, in order to avoid catches consisting almost exclusively of juveniles. Such an increase in mesh size would have a minor impact in terms of losses of individuals above the minimum landing size for Norway lobster and would contribute to reducing the amount of discards in this fishery. (C) 2002 Elsevier Science B.V. All rights reserved.

Innate Immunity in Lobsters: Partial Purification and Characterization of a Panulirus cygnus Anti-A Lectin

A lectin detected in haemolymph from the Australian spiny lobster Panulirus cygnus agglutinated human ABO Group A cells to a higher titre than Group O or B. The lectin also agglutinated rat and sheep erythrocytes, with reactivity with rat erythrocytes strongly enhanced by treatment with the proteolytic enzyme papain, an observation consistent with reactivity via a glycolipid. The lectin, purified by affinity chromatography on fixed rat-erythrocyte stroma, was inhibited equally by N-acetylglucosamine and N-acetylgalactosamine. Comparison of data from gel filtration of haemolymph (behaving as a 1,800,000 Da macromolecule), and polyacrylamide gel electrophoresis of purified lectin (a single 67,000 Da band), suggested that in haemolymph the lecin was a multimer. The purified anti-A lectin autoprecipitated unless the storage solution contained chaotropic inhibitors (125 mmol/L sucrose: 500 mmol/L urea). The properties of this anti-A lectin and other similar lectins are consistent with a role in innate immunity in these invertebrates.

A quantitative metric to identify critical elements within seafood supply networks

A theoretical basis is required for comparing key features and critical elements in wild fisheries and aquaculture supply chains under a changing climate. Here we develop a new quantitative metric that is analogous to indices used to analyse food-webs and identify key species. The Supply Chain Index (SCI) identifies critical elements as those elements with large throughput rates, as well as greater connectivity. The sum of the scores for a supply chain provides a single metric that roughly captures both the resilience and connectedness of a supply chain. Standardised scores can facilitate cross-comparisons both under current conditions as well as under a changing climate. Identification of key elements along the supply chain may assist in informing adaptation strategies to reduce anticipated future risks posed by climate change. The SCI also provides information on the relative stability of different supply chains based on whether there is a fairly even spread in the individual scores of the top few key elements, compared with a more critical dependence on a few key individual supply chain elements. We use as a case study the Australian southern rock lobster Jasus edwardsii fishery, which is challenged by a number of climate change drivers such as impacts on recruitment and growth due to changes in large-scale and local oceanographic features. The SCI identifies airports, processors and Chinese consumers as the key elements in the lobster supply chain that merit attention to enhance stability and potentially enable growth. We also apply the index to an additional four real-world Australian commercial fishery and two aquaculture industry supply chains to highlight the utility of a systematic method for describing supply chains. Overall, our simple methodological approach to empirically-based supply chain research provides an objective method for comparing the resilience of supply chains and highlighting components that may be critical.

B- and T-cell epitopes of tropomyosin, the major shrimp allergen

Seafood allergy is often encountered on ingestion of crustaceans such as shrimp, lobster, crab, and crayfish (1). On eating cooked shrimp, sensitive individuals experience a wide spectrum of reactions ranging from abdominal discomfort to anaphylaxis. The presence of cross-reacting heat-stable allergens in crustacean food was first recognized by Hoffman et al. (2) and Lehrer et al. (3). Subsequently, the major allergen was isolated and characterized from the shrimp species Paneaus indicus (Pen i 1) (4) and I? aztecm (Pen a 1) (5). Pen i 1 (originally designated Sa-TI) and Pen a 1, with mol. mass of 34 and 36 kDa, respectively, contain 301 and 312 amino-acid residues with a predominance of gluta- mate/glutamine and asparatate/asparagine.

Stochastic growth models for analyzing crustacean data

The contemporary methodology for growth models of organisms is based on continuous trajectories and thus it hinders us from modelling stepwise growth in crustacean populations. Growth models for fish are normally assumed to follow a continuous function, but a different type of model is needed for crustacean growth. Crustaceans must moult in order for them to grow. The growth of crustaceans is a discontinuous process due to the periodical shedding of the exoskeleton in moulting. The stepwise growth of crustaceans through the moulting process makes the growth estimation more complex. Stochastic approaches can be used to model discontinuous growth or what are commonly known as "jumps" (Figure 1). However, in stochastic growth model we need to ensure that the stochastic growth model results in only positive jumps. In view of this, we will introduce a subordinator that is a special case of a Levy process. A subordinator is a non-decreasing Levy process, that will assist in modelling crustacean growth for better understanding of the individual variability and stochasticity in moulting periods and increments. We develop the estimation methods for parameter estimation and illustrate them with the help of a dataset from laboratory experiments. The motivational dataset is from the ornate rock lobster, Panulirus ornatus, which can be found between Australia and Papua New Guinea. Due to the presence of sex effects on the growth (Munday et al., 2004), we estimate the growth parameters separately for each sex. Since all hard parts are shed too often, the exact age determination of a lobster can be challenging. However, the growth parameters for the aforementioned moult processes from tank data being able to estimate through: (i) inter-moult periods, and (ii) moult increment. We will attempt to derive a joint density, which is made up of two functions: one for moult increments and the other for time intervals between moults. We claim these functions are conditionally independent given pre-moult length and the inter-moult periods. The variables moult increments and inter-moult periods are said to be independent because of the Markov property or conditional probability. Hence, the parameters in each function can be estimated separately. Subsequently, we integrate both of the functions through a Monte Carlo method. We can therefore obtain a population mean for crustacean growth (e. g. red curve in Figure 1). [GRAPHICS]

Estimation of growth parameters from multiple-recapture data

This article develops a method for analysis of growth data with multiple recaptures when the initial ages for all individuals are unknown. The existing approaches either impute the initial ages or model them as random effects. Assumptions about the initial age are not verifiable because all the initial ages are unknown. We present an alternative approach that treats all the lengths including the length at first capture as correlated repeated measures for each individual. Optimal estimating equations are developed using the generalized estimating equations approach that only requires the first two moment assumptions. Explicit expressions for estimation of both mean growth parameters and variance components are given to minimize the computational complexity. Simulation studies indicate that the proposed method works well. Two real data sets are analyzed for illustration, one from whelks (Dicathais aegaota) and the other from southern rock lobster (Jasus edwardsii) in South Australia.

Estimating equations for parameters in stochastic growth models from tag-recapture data

James (1991, Biometrics 47, 1519-1530) constructed unbiased estimating functions for estimating the two parameters in the von Bertalanffy growth curve from tag-recapture data. This paper provides unbiased estimating functions for a class of growth models that incorporate stochastic components and explanatory variables. a simulation study using seasonal growth models indicates that the proposed method works well while the least-squares methods that are commonly used in the literature may produce substantially biased estimates. The proposed model and method are also applied to real data from tagged rack lobsters to assess the possible seasonal effect on growth.

An improved Fabens method for estimation of growth parameters in the vonBertalanffy model with individual asymptotes

In the analysis of tagging data, it has been found that the least-squares method, based on the increment function known as the Fabens method, produces biased estimates because individual variability in growth is not allowed for. This paper modifies the Fabens method to account for individual variability in the length asymptote. Significance tests using t-statistics or log-likelihood ratio statistics may be applied to show the level of individual variability. Simulation results indicate that the modified method reduces the biases in the estimates to negligible proportions. Tagging data from tiger prawns (Penaeus esculentus and Penaeus semisulcatus) and rock lobster (Panulirus ornatus) are analysed as an illustration.

The fishery and reproductive biology of Barking crayfish, Linuparus trigonus (Von Siebold, 1824) along Queenslands east coast

This paper describes the fishery and reproductive biology for Linuparus trigonus obtained from trawl fishermen operating off Queensland’s east coast, Australia. The smallest mature female lobster measured 59.8 mm CL, however, 50% maturity was reached between 80 and 85 mm CL. Brood fecundity (BF) was size dependent and ranged between 19,287 and 100,671 eggs in 32 females from 59.8 to 104.3 mm CL. The relationship was best described by the power equation BF = 0.1107*CL to the power of 2.9241 (r to the power of 2 = 0:74). Egg size ranged from 0.96 to 1.12 mm in diameter (mean = 1:02 (+or-) 0:01 mm). Egg weight and size were independent of lobster size. Length frequencies displayed multi-modal distributions.The percentage of female to male lobsters was relatively stable for small size classes (30 to 70 mm CL; 50.0 to 63.6% females), but female proportions rose markedly between 75 and 90 mm (72.2 to 85.4%) suggesting that at the onset of sexual maturity female growth rates are reduced. In size classes greater than 95 mm, males were numerically dominant. A description of the L. trigonus fishery in Queensland is also detailed.

Predicting survival of western rock lobsters Panulirus cygnus using discriminant analysis of hemolymph parameters taken immediately following simulated handling treatments

Instances of morbidity amongst rock lobsters (Panulirus cygnus) arriving at factories in Western Australia (WA) have been attributed to stress during post-harvest handling. This study used discriminant analysis to determine whether physiological correlates of stress following a period of simulated post-harvest handling had any validity as predictors of future rejection or morbidity of western rock lobsters. Groups of 230 western rock lobsters were stored for 6 h in five environments (submerged/flowing sea water, submerged/re-circulating sea water, humid air, flowing sea water spray, and re-circulated sea water spray). The experiment was conducted in late spring (ambient sea water 22°C), and repeated again in early autumn (ambient sea water 26°C). After 6 h treatment, each lobster was graded for acceptability for live export, numbered, and its hemolymph was sampled. The samples were analysed for a number of physiological and health status parameters. The lobsters were then stored for a week in tanks in the live lobster factory to record mortality. The mortality of lobsters in the factory was associated with earlier deviations in hemolymph parameters as they emerged from the storage treatments. Discriminant analysis (DA) of the hemolymph assays enabled the fate of 80-90% of the lobsters to be correctly categorised within each experiment. However, functions derived from one experiment were less accurate at predicting mortality when applied to the other experiments. One of the reasons for this was the higher mortality and the more severe patho-physiological changes observed in lobsters stored in humid air or sprays at the higher temperature. The analysis identified lactate accumulation during emersion and associated physiological and hemocyte-related effects as a major correlate of mortality. Reducing these deviations, for example by submerged transport, is expected to ensure high levels of survival. None of the indicators tested predicted mortality with total accuracy. The simplest and most accurate means of comparing emersed treatments was to count the mortality afterwards.

Advances in the culture of lobsters.

Commercial aquaculture of marine lobsters is an attractive proposition, as most species are high value with established market demand, and fishery production is static or diminishing. Nevertheless, achievement of commercial success will necessitate resolution of technical difficulties associated with on-growing of aggressive species (clawed lobsters) or with rearing the larvae, which for spiny and slipper lobsters is generally a painstaking and protracted process. Notwithstanding these technical challenges, increasing market demand for the product is driving a substantial research and development effort around the world to develop commercial lobster farming technology. This chapter reports on the status of that effort, the successes and obstacles.

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.