Effect of holding duration on the immune system of western rock lobster, Panulirus cygnus

Live holding of lobsters is a major part of the post-harvest process in lobster fisheries. Following capture, animals are placed in factory tanks and held for varying lengths of time for recovery from stress due to handling, air exposure, disturbance and environmental variations. The aim of the present study was to determine the pattern of response of a range of immune and physiological parameters over varying holding periods and assess their possible application as a measure of recovery from stress of post-harvest procedures and in the determination of an appropriate duration of acclimation prior to live transport. The effect of holding duration on the following immune parameters: total haemocyte counts (THC), haemolymph clotting times, haemolymph pH, haemolymph protein concentration and the differential proportion of haemocytes, was assessed. Lobsters brought to the factory were placed in holding tanks and sampled over a period of up to 10 days. Two studies were conducted, the first on lobsters from different catch groups and the second on lobsters from the same day's catch from the same area. Experiments were conducted on adult animals (10-12 animals/treatment group) with a different group of lobsters being sampled at each time point. Various immune parameters showed alterations with time during holding. A consistent observation was significantly prolonged clotting times following four days of holding. The haemolymph pH showed a strong positive correlation to clotting time and the hyaline cell proportion showed a strong negative correlation with semi-granular cells. Although the levels of THC, clotting time and differential cell counts after one day of holding were similar to those observed in other studies on rested post-harvest lobsters, suggesting that the lobsters had recovered from the acute stress of capture and transport, subsequent alterations in pH, clotting time and differential cell counts indicated other physiological adjustments were still occurring for up to 4 days post capture. Overall the results suggest that though the effects of post-harvest procedures on the immune parameters appear to be resolved after a short duration of holding at low temperatures, no single immune parameter can provide predictable indication of the acclimation process.     

Calcium uptake by juvenile western rock lobster,Panulirus cygnus George,from dietary coralline algae

Foliose coralline algae form a large proportion of the diet of juvenile Panulirus cygnus George at some locations but its rôle in the diet is not clear. In the laboratory, early post-moult rock lobsters fed Corallina cuvieri Lamouroux, labelled with ⁴⁵Ca primarily in the algal skeleton showed significant uptake of calcium compared with lobsters maintained in sea water containing dissolved ⁴⁵Ca. Shortly after ingestion by the rock lobsters the isotopic calcium was distributed through all the soft tissues examined and the exoskeleton. After a 5-day period isotopic calcium levels had fallen in most soft tissues but were not significantly lower in the digestive gland, the mid-gut or the exoskeleton. The data indicate that calcium in the algal skeleton of dietary C. cuvieri is absorbed by the digestive system and may be important in the postmoult re-mineralization of the exoskeleton.     

Osmotic and ionic regulation in the western rock lobster Panulirus longipes (Milne-Edwards)

The western rock lobster, Panulirus longipes (Milne-Edwards) is poikilosmotic over its tolerated salinity range, 25–45 ‰. Blood sodium is accumulated while chloride concentration is reduced. Sodium and chloride vary directly with the external salinity, although maintaining their differences in the same proportions as at normal salinity (36.0 ‰). Calcium is accumulated, ranging from over 150% at salinity 20 ‰ to about 117% at salinity 45 ‰. Potassium concentration is equivalent to the external at normal salinity, but is increased with lowered and decreased with raised salinity. Magnesium is reduced to about one-third that of the external concentration over the salinity range 20–40 ‰, but regulation begins to break down at 45 ‰. Individual ions exhibit, therefore, a range of regulation types, from poikilosmotic to homoiosmotic.Equilibrium for sodium, chloride, and calcium is attained in 10 h at salinities of 25, 30, 40 and 45 ‰ respectively. Rate constants for this exchange are linearly related to salinity differential, and rapid osmotic adjustment is by high permeability, equal in both directions, probably mainly via the gills. Muscle appears to act as a salt pool for sodium, chloride, and potassium but not for magnesium and calcium. Salt-loading causes a slight salt diuresis, the salts being excreted, probably via the gills. Except for calcium, there is no excretion of salt into the gut, but there is evidence of an exchange of chloride with another anion. Magnesium excretion is slow, and in the absence of osmotic stress possibly occurs via the antennal glands. All the ions examined appear to be regulated independently.     

Sound production by the western rock lobster Panulirus longipes (Milne Edwards)

Sound characteristics and sound emitting structures of larval, juvenile and adult rock lobsters, Panulirus longipes (Milne Edwards) have been investigated. There is a linear correlation between carapace length and dimensions of the sound producing organ (number and size of plectrum ridges) as well as average duration of a single squeak. The number of pulses per 100 ms squeak, however, decreases asymptotically from $\text{\$}\text{?}$30 in larval forms to about 10 in mature adults. Behavioural changes accompany this process. The energy of a single pulse is distributed equally over the audio frequency range. The same is true for the ‘slow rattle’ sound, produced by mandible-grinding during feeding or moments of extreme agitation. Any predator attacking a rock lobster, is likely to receive injuries from the outstretched, spiny antennae. It is concluded that the antennal sound, accompanying the rock lobsters defence, functions as a warning device aimed primarily at predators, which will learn to associate the sound of rock lobsters with the unpleasant experience of the sharp antennal spines.     

Foraging patterns of juvenile western rock lobsters Panulirus cygnus George

Foraging patterns of juvenile western rock lobster Panulirus cygnus George were investigated in the field by tracking lobsters with electromagnetic tags for up to 3 wk. Most foraging activity begins in response to changes in light levels associated with dusk and not by diurnal changes in water temperature or currents. Similarly, lobsters return from foraging around dawn when light levels start to increase. Foraging activity was constant throughout the night, and males were more active than females. Lobsters usually moved at a rate of ≈ l m · min ⁻¹ at night, although sometimes they travelled up to 18m· min⁻¹ when walking over bare sand. They often lived on one reef but travelled to forage on seagrass beds in front of another reef, sometimes visiting a den on that reef. Some lobsters used several dens on different reefs during the 3 wk of observation. Although lobsters foraged in both of the two main structural varieties of seagrass beds, Amphibolis and Heterozostera with Halophila, they foraged for longer periods in the Heterozostera with Halophila beds. Foraging ranges varied within and between individuals although an individual lobster might forage in the same general area on a few successive nights or even up to several weeks.     

High levels of polyandry,but limited evidence for multiple paternity,in wild populations of the western rock lobster (Panulirus cygnus)

Polyandry, where multiple mating by females results in the temporal and spatial overlap of ejaculates from two or more males, is taxonomically widespread and occurs in varying frequencies within and among species. In decapods (crabs, lobsters, crayfish, and prawns), rates of polyandry are likely to be variable, but the extent to which patterns of multiple paternity reflect multiple mating, and thus are shaped by postmating processes that bias fertilization toward one or a subset of mated males, is unclear. Here, we use microsatellite markers to examine the frequency of multiple mating (the presence of spermatophores from two or more males) and patterns of paternity in wild populations of western rock lobster (Panulirus cygnus). Our data confirm that >45% of females had attached spermatophores arising from at least two males (i.e., confirming polyandry), but we found very limited evidence for multiple paternity; among 24 clutches sampled in this study, only two arose from fertilizations by two or more males. Single inferred paternal genotypes accounted for all remaining progeny genotypes in each clutch, including several instances when the mother had been shown to mate with two or more males. These findings highlight the need for further work to understand whether polyandry is adaptive and to uncover the mechanisms underlying postmating paternity biases in this system.     

Lipid,fatty acid and protein content of late larval to early juvenile stages of the western rock lobster,Panulirus cygnus

Lipid, fatty acid and protein content were determined individually on 7 phyllosomata, 69 clear pueruli, 286 pre-moult pueruli, and 86 juvenile western rock lobster (WRL) collected from four locations between the settlement seasons 2000 to 2006 to evaluate compositional changes during the non-feeding puerulus stage. Only the lipid content, particularly the phospholipids, decreased significantly with development. Protein declined sharply following moult to the juvenile. PL comprised between 86–94% of total lipid in all animals, and declined most between phyllosomata and clear pueruli (238.5 to 121.4 mg g^? 1 DW) (p < 0.001). Triacylglycerols were the only lipid to increase in absolute amounts with development, but declined 53% on average following moult to juvenile. This increase in TAG is likely due to the conversion of phospholipids to triacylglycerols. Monounsaturated fatty acids were the main energy form utilised during benthic development while polyunsaturated fatty acids showed a high degree of sparing. The n-3:n-6 fatty acid ratio of juveniles indicates that they may be approaching critically low levels of stored lipid energy reserves. Both protein, and lipid, declined sharply from the final puerulus phase to the juvenile confirming that a high energetic demand is required to fuel the moulting process.     

Determination of the unknown age at first capture of western rock lobsters (Panulirus cygnus) by random effects model

We propose a method for fitting growth curves to multiple recapture data of lobsters when the age at first capture is unknown. The von Bertalanffy growth curve is used to model the growth. To account for individual variability, the unknown age in logarithmic scale of a lobster at first capture, the individual asymptotic size, and the individual growth coefficient of its carapace length are modeled as random effects with a trivariate normal distribution. Unlike previously suggested models, the present model permits correlation between the growth coefficient and the age at first capture and can be fitted readily using existing software. The error structures between consecutive recaptures of a lobster are assumed to be a first-order autoregressive process with unequally spaced time points. A comparison between this model and the Fabens growth equation is given. The proposed method is a flexible method and can be applied to fit different growth equations when the age at first capture is unknown.     

Modelling deep water habitats to develop a spatially explicit, fine scale understanding of the distribution of the western rock lobster, Panulirus cygnus

Background

The western rock lobster, Panulirus cygnus, is endemic to Western Australia and supports substantial commercial and recreational fisheries. Due to and its wide distribution and the commercial and recreational importance of the species a key component of managing western rock lobster is understanding the ecological processes and interactions that may influence lobster abundance and distribution. Using terrain analyses and distribution models of substrate and benthic biota, we assess the physical drivers that influence the distribution of lobsters at a key fishery site.

Methods and Findings

Using data collected from hydroacoustic and towed video surveys, 20 variables (including geophysical, substrate and biota variables) were developed to predict the distributions of substrate type (three classes of reef, rhodoliths and sand) and dominant biota (kelp, sessile invertebrates and macroalgae) within a 40 km² area about 30 km off the west Australian coast. Lobster presence/absence data were collected within this area using georeferenced pots. These datasets were used to develop a classification tree model for predicting the distribution of the western rock lobster. Interestingly, kelp and reef were not selected as predictors. Instead, the model selected geophysical and geomorphic scalar variables, which emphasise a mix of terrain within limited distances. The model of lobster presence had an adjusted D² of 64 and an 80% correct classification.

Conclusions

Species distribution models indicate that juxtaposition in fine scale terrain is most important to the western rock lobster. While key features like kelp and reef may be important to lobster distribution at a broad scale, it is the fine scale features in terrain that are likely to define its ecological niche. Determining the most appropriate landscape configuration and scale will be essential to refining niche habitats and will aid in selecting appropriate sites for protecting critical lobster habitats.     

中国龙虾微卫星标记的筛选及遗传多样性分析

文章以M13通用引物和重复序列(CT)₁₅、(AT)₁₅引物, 利用PCR法对中国龙虾(Panulirus stimpsoni Hoehuis)部分基因组DNA文库进行筛选。共获得78个微卫星序列, 分别分布于55个阳性重组克隆中, 其中完美型(perfect)共50个, 占64%; 非完美型(imperfect)3个, 占3.8%; 混合完美型(compound perfect)6个, 占7.7%; 混合非完美型(compound imperfect)19个, 占24.5%。根据微卫星序列, 设计并筛选出15对微卫星多态性引物, 对中国龙虾的群体进行了遗传多样性分析。获得3~12个等位基因, 等位基因大小在78~425 bp之间, 基本符合引物设计的理论长度。期望杂合度范围为0.48~0.87, 平均值为0.71, 表明中国龙虾基因组微卫星具有较高的杂合度与遗传多样性。15个微卫星位点的PIC值从0.44到0.84, 平均值为0.60, 说明这些微卫星位点在中国龙虾基因组中包含丰富的遗传信息, 合适用于中国龙虾的各种分子标记及遗传学分析和应用。     

Pattern generation in the lobster (Panulirus) stomatogastric ganglion

1. Results from the companion paper were incorporated into a physiologically realistic computer model of the three principal cell types (PD/AB, LP, PY) of the pyloric network in the stomatogastric ganglion. Parameters for the model were mostly calculated (sometimes estimated) from experimental data rather than fitting the model to observed output patterns. 2. The initial run was successful in predicting several features of the pyloric pattern: the observed gap between PD and LP bursts, the appropriate sequence of the activity periods (PD, LP, PY), and a substantial PY burst not properly simulated by an earlier model. 3. The major discrepancy between model and observed patterns was the too-early occurrence of the PY burst, which resulted in a much shortened LP burst. Motivated by this discrepancy, additional investigations were made of PY properties. A hyperpolarization-enabled depolorization-activated hyperpolarizing conductance change was discovered which may make an important contribution to the late phase of PY activity in the normal burst cycle. Addition of this effect to the model brought its predictions more in line with observed patterns. 4. Other discrepancies between model and observation were instructive and are discussed. The findings force a substantial revision in previously held ideas on pattern production in the pyloric system. More weight must be given to functional properties of individual neurons and less to properties arising purely from network interactions. This shift in emphasis may be necessary in more complicated systems as well. 5. An example has been provided of the value quantitative modeling can be to network physiology. Only through rigorous quantitative testing can qualitative theories of how the nervous system operates be substantiated.     

Indices of nutritional state in the western rock lobster,Panulirus longipes (Milne Edwards). II. Gastric fluid constituents

The concentrations of gastric fluid protein (GFPC) and of non-protein amino acids of Panulirus longipes (Milne Edwards) are both considerably reduced during starvation. Because of its simplicity and reliability of measurement, GFPC was selected for further testing. A reduction in GFPC with time was a negative exponential function so that a measurement of the GFPC has a maximum sensitivity during early starvation.Food odours, stress (as induced by anoxia), and time of day did not affect the GFPC; increased temperature enhanced the effects of starvation, but did not alter the GFPC in fed rock lobsters. The GFPC fell sharply after ecdysis, but returned to normal levels by the end of stage C₃ in well-fed animals. In starved animals there was a small post-moult increase but the concentrations remained low. Nutritional history prior to ecdysis affected the GFPC in the subsequent stadium.Sampling of gastric fluid in the field was shown to be feasible, and the technique was facilitated by use of a serum protein refractometer. In general, the GFPC supported field observations on the state of the animals as well as estimates of growth by modal shifts and marking-recapture experiments; there were, however, some discrepancies.It is concluded that GFPC is a reliable index of nutritional state, but must not be used uncritically. The wide range of values for wild populations, the effects of handling, and possible seasonal influences all need to be taken into account. Moult-staging in the field is essential. A combination of GFPC and blood volume estimations may provide the best simple means of estimating nutritional state in decapod Crustacea.     

Pattern generation in the lobster (Panulirus) stomatogastric ganglion

There are a number of perspectives gained from a quantitative analysis of the pyloric system which may be applicable to other simple pattern generators: 1. The system is organized around a dominant, endogenously-bursting neuron group, and its properties are tailored to that dominance. In particular, synaptic strengths and firing frequencies of that group appear just sufficient to suppress postsynaptic "follower" cells if the latter are not too highly excited. 2. Repetitive firing properties of follower neurons are such as to facilitate their switch-like mode of activity. This includes pacemaker response nonlinearities, rebound properties, and "burstiness" properties. 3. Proper sequencing of follower cells may be controlled by particular synaptic strengths and time-courses, feedback on the oscillator cells, and functional cellular properties of follower neurons (e.g., rebound; see also next paper). All such properties interact and must be tuned to each other for proper patterns to result.     

Behavioral thermoregulation in the spiny lobster Panulirus Argus (latreille)

Ten spiny lobsters (Panulirus argus) were allowed to thermoregulate individually for 3-day periods in an electronic thermoregulatory shuttlebox which allowed them to control water temperatures (and thereby their own body temperatures) by their movements. The range of preferred (voluntarily occupied) temperatures was 25–35°C (mean 29.9°C; mode 30.0°C; median 30.0°C; midpoint 30.0°C; S_k (skewness, Pearson's coefficient) –0.04; s.e.m. 0.19°C; S.D. 2.32°C). The final thermal preferendum (by the gravitation method) in this species is 30°C.     

Changes in lipid and fatty acid composition of late larval and puerulus stages of the spiny lobster (Panulirus cygnus) across the continental shelf of Western Australia

The feeding of the late larval stages of the spiny lobster, Panulirus cygnus, and the energy reserves used by the non-feeding post-larvae for crossing the continental shelf of Western Australia were examined through their lipid composition. Lipid was a significant component of the biomass of all larval and post-larval samples (range 63-213 mg g(-1) of dry biomass). The fatty acid profiles of late stage larvae (8-9) suggest that they were feeding on salps and small crustaceans, such as euphausids, from oligotrophic pelagic communities where herbivorous and microbial grazing is an important basis to the food web. There was a marked decrease in lipid content of post-larvae progressively across the continental shelf, and this corresponded closely with decreasing dry mass, suggesting that post-larvae were using lipid as an energy source during cross-shelf movement at a rate of 1.6 J km(-1). This is considerably lower than for other spiny lobster species, suggesting that the post-larvae of the western rock lobster may use physical processes as well as active swimming for onshore transport. This may help to explain the large inter-annual variability in the post-larval settlement of this species, which is closely related to differences in weather patterns capable of greatly varying onshore advection processes.     

Water uptake at ecdysis in the western rock lobster

Water ingestion at ecdysis by the western rock lobster. Panulirus longipes (Milne Edwards) was investigated using the reference markers ⁵¹Cr-EDTA and ⁵⁸Co-EDTA. Two possible mechanisms controlling water absorption were examined: first, changes in osmolarity of blood and muscle and secondly, the effects of extracts of central nervous system.Water ingestion was 16.071 ± 2.365 mlkg^?1h^?1 during swelling just before ecdysis (stage D₄(S)) and 23.099 ± 1.238 mlkg^?1h^?1 during stage A. There was no significant absorption in the foregut or hindgut and the digestive gland appeared to be the site of major absorption. Total water ingested during stages D₄(S) and A was 13.7% of the proecdysis weight. Calculating total water uptake by wet weight differences plus wet weight of exuviae gave a value that was too high and instead weight increases were calculated from a carapace length-weight formula. Allowing for postecdysis increase in weight the net increase at ecdysis was 18.4–21.4% which was 4.7–7.7% more than the water ingested. It was concluded from this that water enters the body at ecdysis both by ingestion and by absorption through the external surface. It is suggested that water ingestion provides the main source of swelling of the cephalothorax in stage D₄(S) and after ecdysis both ingested water and external absorption enables the flaccid abdomen and appendages to swell rapidly.Statistically significant differences were found in the concentrations of total cations and chloride in leg muscle during the transition from stage C₄ to late D₄ but the trends were not consistent and probably have no functional significance. There were no changes in the concentration of osmotically active organic constituents. The freezing-point depression of the blood in stage D₄ was significantly higher than that in stage C₄(P < 0.02) but the mean difference was only 1.8%. It was concluded that osmoticchanges were unlikely to be an important mechanism of water uptake.Water-soluble extract (WSE) and acetone-soluble extract (ASE) of brains and first ventral ganglia were without significant effect when compared together with controls. There was a barely significant decrease, however, in water in the proventriculus of WSE-treated animals compared with that of controls (P < 0.05). and further investigation on the effects of such extracts on water uptake at ecdysis is warranted.     

Comparative genetic stock structure in three species of commercially exploited Indo-Malay Carangidae (Teleosteii,Perciformes)

We examine genetic structuring in three commercially important species of the teleost family Carangidae from Malaysian waters: yellowtail scad Atule mate, bigeye scad Selar crumenophthalmus and yellowstripe scad Selaroides leptolepis, from the Indo-Malay Archipelago. In view of their distribution across contrasting habitats, we tested the hypothesis that pelagic species display less genetic divergence compared with demersal species, due to their potential to undertake long-distance migrations in oceanic waters. To evaluate population genetic structure, we sequenced two mitochondrial (mt)DNA [650 bp of cytochrome oxidase I (coI), 450 bp of control region (CR)] and one nuclear gene (910 bp of rag1) in each species. One hundred and eighty samples from four geographical regions within the Indo-Malay Archipelago including a population of yellowtail from Kuwait were examined. Findings revealed that the extent of genetic structuring among populations in the semi-pelagic and pelagic, yellowtail and bigeye were lower than demersal yellowstripe, consistent with the hypothesis that pelagic species display less genetic divergence compared with demersal species. The yellowtail phylogeny identified three distinct clades with bootstrap values of 86%–99% in mtDNA and 63%–67% in rag1. However, in bigeye, three clades were also observed from mtDNA data while only one clade was identified in rag1 dataset. In yellowstripe, the mtDNA tree was split into three closely related clades and two clades in rag1 tree with bootstraps value of 73%–99% and 56% respectively. However, no geographic structure appears in both mtDNA and rag1 datasets. Hierarchical molecular variance analysis (AMOVA), pair wise F_ST comparisons and the nearest-neighbour statistic (S_nn) showed significant genetic differences among Kuwait and Indo-Malay yellowtail. Within the Indo-Malay Archipelago itself, two distinct mitochondrial lineages were detected in yellowtail suggesting potential cryptic species. Findings suggests varying degrees of genetic structuring, key information relevant to management of exploited stocks, though more rapidly evolving genetic markers should be used in future to better delimit the nature and dynamics of putative stock boundaries.     

Fussy feeders: phyllosoma larvae of the Western Rocklobster (Panulirus cygnus) demonstrate prey preference

The Western Rocklobster (Panulirus cygnus) is the most valuable single species fishery in Australia and the largest single country spiny lobster fishery in the world. In recent years a well-known relationship between oceanographic conditions and lobster recruitment has become uncoupled, with significantly lower recruitment than expected, generating interest in the factors influencing survival and development of the planktonic larval stages. The nutritional requirements and wild prey of the planktotrophic larval stage (phyllosoma) of P. cygnus were previously unknown, hampering both management and aquaculture efforts for this species. Ship-board feeding trials of wild-caught mid-late stage P. cygnus phyllosoma in the eastern Indian Ocean, off the coast of Western Australia, were conducted in July 2010 and August-September 2011. In a series of experiments, phyllosoma were fed single and mixed species diets of relatively abundant potential prey items (chaetognaths, salps, and krill). Chaetognaths were consumed in 2-8 times higher numbers than the other prey, and the rate of consumption of chaetognaths increased with increasing concentration of prey. The highly variable lipid content of the phyllosoma, and the fatty acid profiles of the phyllosoma and chaetognaths, indicated they were from an oligotrophic oceanic food chain where food resources for macrozooplankton were likely to be constrained. Phyllosoma fed chaetognaths over 6 days showed significant changes in some fatty acids and tended to accumulate lipid, indicating an improvement in overall nutritional condition. The discovery of a preferred prey for P. cygnus will provide a basis for future oceanographic, management and aquaculture research for this economically and ecologically valuable species.