Ecological and physiological effects of PaV1 infection on the Caribbean spiny lobster (Panulirus argus Latreille)

Pathogens can profoundly impact the ecology of the organisms they infect through changes in host behavior that influence demographic processes. For example, juvenile Caribbean spiny lobsters (Panulirus argus Latreille) infected with the PaV1 virus (Panulirus argus Virus 1) are avoided by their normally social conspecifics, which alters local spatial distributions and presumably rates of disease transmission. PaV1 infections are nearly always lethal, but prior to succumbing to the disease, infection may impact other host dynamics (e.g., movement, growth, or survival) that effect transmission of the virus. We used mark-recapture surveys and laboratory studies to determine the impact of PaV1 infection on lobster movement and physiological condition, and in turn, the effect of condition on susceptibility to infection. Significantly more healthy lobsters were recaptured than heavily diseased lobsters in 5-d mark-recapture surveys, indicating either greater emigration or greater mortality of infected lobsters. Results of a laboratory bioassay suggest that lobsters with early-stage infections moved at an equivalent rate to healthy lobsters, however, as infection progressed, lobsters inoculated with PaV1 moved less than healthy lobsters ultimately remaining sedentary. Infected lobsters captured in nature also had significantly lower hemolymph (blood) serum protein values, indicating poor physiological condition. However, poor condition did not predispose lobsters to greater risk of infection because prevalence was not different between starved lobsters and healthy lobsters challenged with PaV1. Although lobsters infected with PaV1 eventually become sedentary and of poor nutritional condition, during the early stages of infection they remain active and are thus capable of dispersing the virus throughout the population.     

Chemotaxis in the Florida spiny lobster, Panulirus argus

The relative importance of input from chemoreceptors on the medial versus the lateral antennular flagellae of the spiny lobster, Panulirus argus, in olfactory orientation is examined. Ablation experiments show that input from the lateral filament, specifically the aesthetase tuft of the lateral filament, is necessary and sufficient to trigger searching behaviour in this organism when stimulated with a gradient of dilute shrimp extract. P. argus locates the odour source by using this lateral filament input in both tropotactic and klinotactic comparisons of odour intensities.     

Hemocyanin-derived phenoloxidase activity in the spiny lobster Panulirus argus (Latreille, 1804)

Hemocyanin and phenoloxidase belong to the type-3 copper protein family, sharing a similar active center whereas performing different roles. In this study, we demonstrated that purified hemocyanin (450 kDa) from the spiny lobster Panulirus argus shows phenoloxidase activity in vitro after treatment with trypsin, chymotrypsin and SDS (0.1% optimal concentration), but it is not activated by sodium perchlorate or isopropanol. The optimal pHs of the SDS-activated hemocyanin were 5.5 and 7.0. Hemocyanin from spiny lobster behaves as a catecholoxidase. Kinetic characterization using dopamine, L-DOPA and catechol shows that dopamine is the most specific substrate. Catechol and dopamine produced substrate inhibition above 16 and 2 mM respectively. Mechanism-based inhibition was also evidenced for the three substrates, being less significant for L-DOPA. SDS-activated phenoloxidase activity is produced by the hexameric hemocyanin. Zymographic analysis demonstrated that incubation of native hemocyanin with trypsin and chymotrypsin, produced bands of 170 and 190 kDa respectively, with intense phenoloxidase activity. Three polypeptide chains of 77, 80 and 89 kDa of hemocyanin monomers were identified by SDS-PAGE. Monomers did not show phenoloxidase activity induced by SDS or partial proteolysis.     

Loss of escape-related giant neurons in a spiny lobster, Panulirus argus

When attacked, many decapod crustaceans perform tailflips, which are triggered by a neural circuit that includes lateral giant interneurons, medial giant interneurons, and fast flexor motor giant neurons (MoGs). Slipper lobsters (Scyllaridae) lack these giant neurons, and it has been hypothesized that behavioral (e.g., digging) and morphological (e.g., flattening and armor) specializations in this group caused the loss of escape-related giant neurons. To test this hypothesis, we examined a species of spiny lobster, Panulirus argus. Spiny lobsters belong to the sister taxon of the scyllarids, but they have a more crayfish-like morphology than scyllarids and were predicted to have escape-related giant neurons. Ventral nerve cords of P. argus were examined using paraffin-embedded sections and cobalt backfills. We found no escape-related giant neurons and no large axon profiles in the dorsal region of the nerve cord of P. argus. Cobalt backfills showed one fewer fast flexor motor neuron than in species with MoGs and none of the fast flexor motor neurons show any of the anatomical specializations of MoGs. This suggests that all palinuran species lack this giant escape circuit, and that the loss of rapid escape behavior preceded, and may have driven, alternative predator avoidance and anti-predator strategies in palinurans.     

Serum lipoproteins in the spiny lobster, Panulirus interruptus.

1. Most of the lipids in the hemolymph of the spiny lobster, Panulirus interruptus, were associated with a high density lipoprotein (HDL3). The lipid of this lipoprotein was composed of phospholipid (88%), sterol (4%) and triglyceride (3%). 2. In animals fed 14C-labeled triglyceride radioactivity was not seen in the serum until 12 hr after feeding. Most of this serum radioactivity was associated with phosphatidyl choline. 3. Electron micrographs showed that negatively stained high density lipoproteins of the lobster had a polymorphic appearance.     

乌鳢源杀鱼爱德华菌的分离鉴定及药敏特性研究

【背景】江苏省扬州市某乌鳢养殖场发生疾病,给养殖户造成了严重的经济损失。【目的】确定病因并筛选出敏感药物,为乌鳢相关疾病的防治提供参考。【方法】从患病乌鳢体内分离致病菌,并从形态、生理生化特征、16S rRNA和gyrB基因序列及特异性PCR检测等方面对分离菌株进行鉴定,同时开展人工感染试验分析其致病性,通过纸片扩散法进行药敏特性分析。【结果】从患病乌鳢体内分离获得优势菌株SHL,经形态特征、理化特性、16SrRNA和gyrB基因序列及特异性PCR检测鉴定为杀鱼爱德华菌。进一步人工感染试验证实其对乌鳢有较强的致病性,LD50为1.6×105 CFU/g,发病症状与自然发病症状相似。药敏试验结果显示,该菌株对青霉素、氯霉素、四环素等28种抗菌药物高度敏感,对红霉素中度敏感,对苯唑西啉、克拉霉素、万古霉素等6种药物耐药。【结论】引起江苏省扬州市某养殖场的乌鳢体表溃烂及死亡的病原菌为杀鱼爱德华菌,这是我国首次从淡水鱼类中检出致病性杀鱼爱德华菌,表明该菌的感染谱在扩大,需引起水产养殖领域的重视,在养殖过程中可根据药敏实验结果选用合适的国标渔药进行防治。     

Cold-resistant changes in heartbeat of the Japanese spiny lobster

Heartbeat in Panulirus japonicus acclimated to 20°C is often augmented during cooling to 15^oC. Augmented contractions of the heart coincided with increasing amplitude of electrocardiogram. In cold saline, a pericardial hormone serotonin (10⁻⁷ M) increased both the amplitude and duration of the heartbeat while another hormone octopamine (10⁻⁶ M) slightly relieved the cold depression of heart rate despite a smaller increase in beat amplitude. In contrast, the application of the cold saline containing F1 (a FMRFamide-related peptide of pericardial hormones, 10⁻⁹ M) maintained the rate and amplitude of the heartbeat around the control level during cold exposure. This suggests that in the presence of F1, the lobster heart becomes cold resistant clearly. We previously reported that the pericardial organs of spiny lobsters are activated by a small fall in body temperature. The ligamental nerves, extensions of the pericardial organs, terminate in the heart beside the ostia and their ends remain in the isolated hearts. Therefore, the ligamental nerve ends might release their hormones into the ventricle with the fall in temperature even in the isolated hearts.     

Overview,opportunities and outlook for Australian spiny lobster fisheries

Australia’s lobster fisheries are relatively small in volume (9500t) compared with global production (289,000t), but are the country’s most valuable in terms of both overall production and value of export (2014 Gross Value of Production of $610 million AUD). Further, they support commercial, recreational and indigenous fishers along most of the continent’s coastline. Here we review similarities and key differences between these lobster fisheries, based on biological characteristics, fishery data collection, assessment and management methods, and supply chain considerations. A diverse range of palinurid lobsters occur in Australia, but only three genera, distributed across eight different management jurisdictions, support significant fisheries. Catches of western rock lobster Panulirus cygnus dominate landings (61%), followed by southern rock lobster Jasus edwardsii, tropical lobster Panulirus ornatus and the eastern rock lobster Sagmariasus verreauxi. Large-scale environmental influences such as climate change are impacting on these fisheries in similar or different ways forcing new management and raising the need for greater resilience in current supply chains. Although these are separate fisheries, the integrated nature of the dominant Chinese export markets suggests potentially important economic and market-related interactions. Our overview highlights the critical role of continued monitoring of recruitment pulses, in combination with robust harvest strategies, to ensure that harvests respond adequately and fisheries achieve biological and economic sustainability. Approaches that also include socio-cultural considerations (triple bottom line) are important given many fisheries include indigenous Australians. Our integrated analysis of Australian lobster fisheries highlights differences and similarities with spiny lobster fisheries worldwide and lessons from opportunities, including adapting to new free trade agreements, enhancing the reputation of wild lobsters as a whole, sharing expertise, and better alignment of supply and demand.     

Organization of the stomatogastric ganglion of the spiny lobster

Summary The Stomatogastric ganglion ofPanulirus interruptus contains about 30 neurons, and controls the movements of the lobster's stomach. When experimentally isolated, the ganglion continues to generate complex rhythmic patterns of activity in its motor neurons which are similar to those seen in intact animals.In this paper, we describe the synaptic organization of a group of six neurons which drive the stomach's lateral teeth (Figs. 2, 6). This group includes four motor neurons and two interneurons, all but one of which were recorded and stimulated with intracellular microelectrodes.One pair of synergistic motor neurons, LGN and MGN, are electrotonically coupled and reciprocally inhibitory (Figs. 9, 12). A second pair of synergistic motor neurons, the LPGNs, are antagonists of LGN and MGN. The LPGNs are electrotonically coupled (Fig. 14), and are both inhibited by LGN and MGN (Figs. 8, 11). The LPGNs inhibit MGN (Fig. 15) but not LGN. One of the two interneurons in the ganglion, Int 1, reciprocally inhibits both LGN and MGN (Figs. 10, 13). The other interneuron, Int 2, excites Int 1 and inhibits the LPGNs (Fig. 16). The synaptic connections observed in the ganglion are reflected in the spontaneous activity recorded from the isolated ganglion and from intact animals.From the known synaptic organization and observations on the physiological properties of each of the neurons, we have formulated some hypotheses about the pattern-generating mechanism. We found no evidence that any of the neurons are endogenous bursters.We thank D. Kennedy, Eve Marder, and D. Russell for criticizing early drafts of these papers, Nina Pollack and Betty Jorgensen for expert technical assistance, Diane Newsome, SanDee Newcomb, and Pattie Macpherson for typing the many drafts. The authors' research is supported by grant number NS-09322 from N.I.H. and by the Alfred P. Sloan Foundation. B. M. is an NINDS-NIH postdoctoral fellow.     

Polymorphic tetranucleotide microsatellite markers in the Caribbean spiny lobster,Panulirus argus

Ten tetranucleotide microsatellite loci are described for the Caribbean spiny lobster Panulirus argus. Loci were polymorphic (4–15 alleles per locus) and exhibited high levels of expected (0.553–0.921) and observed heterozygosity (0.469–0.906) from samples caught off Belize and Puerto Rico coasts. No significant departure from Hardy–Weinberg equilibrium conditions were observed for any locus. All microsatellite loci should be useful for assessing population discrimination for this valuable marine animal currently subjected to excessive fishing efforts.     

Organization of the stomatogastric ganglion of the spiny lobster

Summary Three direct synaptic connections occur between neurons in the gastric and pyloric systems of the stomatogastric ganglion ofPanulirus interruptus. Two synapses are inhibitory, and one is electrical. This electrical synapse is both excitatory and inhibitory at different times. These synapses, and others within each system, let the two systems interact under some conditions. The synapses also form multisynaptic pathways which modulate the firing of many neurons in both systems. The consequences of these multisynaptic pathways are described and discussed.I thank Allen I. Selverston, Karen Sigvardt, Eve Marder, David Russell and Mary Chamberlin for criticizing a draft of this paper, Forrest Gompf and Doug Tissdale for technical support, and Nina Pollack and Betty Jorgensen for laboratory assistance. The research was supported by USPHS grant NS-12295 to BM and USPHS grant NS-09322 to Alien I. Selverston. BM was a USPHS NIH Postdoctoral Fellow in A.I. Selverston's laboratory during part of this research and is now a Research Fellow of the Alfred P. Sloan Foundation.     

Structure and function of spiny lobster ligamental nerve plexuses: Evidence for synthesis,storage, and secretion of biogenic amines

Three pairs of ligaments support not only the heart of spiny lobsters but also ligamental nerve plexuses, the complex terminal aborizations of segmental nerves. Segmental nerves 1–4 project from the thoracic ganglia into the pericardial cavity and ultimately ramify along the strands of the anterior, medial, and posterior ligaments. In each branch, a core of large axons sends fibers to terminate in a surrounding cortex of fine and varicose secretory processes. Electron micrographs reveal at least five distinct populations of granule-filled neuronal profiles, many with vesicles clustered at membrane thickenings adjacent to the epineural sheath. The ligamental nerve plexuses synthesize and accumulate octopamine, dopamine, 5-HT, and acetylcholine. Octopamine and 5-HT are predominant, comprising 33% and 65%, respectively, of the synthetic activity devoted to the four amines. Thus, the anatomy, ultrastructure, and neurochemistry of the ligamental nerve plexuses establishes their homology with the pericardial organs of other Crustacea. Octopamine and 5-HT are released by a Ca⁺⁺-dependent mechanism upon electrical stimulation of preterminal nerve trunks, and, in vivo, would be swept immediately through ostia into the heart. These observations, when considered with known effects of octopamine and 5-HT on crustacean cardiac activity, neuromuscular transmission, muscle tension, and cyclic AMP metabolism provide a strong case for hormonal actions at target sites throughout the animal. Segmental nerve processes in the dorsal nerve trunk ramify into a plexus around the dorsal nerve apparatus, a small muscular bulb that lies recessed in the cardiac surface. The dorsal nerve, carrying excitatory and inhibitory input to the cardiac ganglion directly through the bulb's hollow interior. The apparatus synthesizes and contains acetylcholine and the three amines mentioned above. In situ, it may beat rhythmically out of phase with the heart.     

Ecto-ATPase/phosphatase activity in the olfactory sensilla of the spiny lobster,Panulirus argus: localization and characterization

Summary Electrophysiological studies have shown that the olfactory organ (antennule) of the spiny lobster, Panulirus argus, has chemoreceptors that are selectively excited by adenine nucleotides in seawater. Biochemical studies have revealed that these same nucleotides can be rapidly dephosphorylated by ectoenzymes associated with the olfactory sensilla (aesthetascs). In this study the distribution of ecto-ATPase/phosphatase activity within aesthetascs was determined cytochemically and the nature of the adenine-nucleotide dephosphorylating activity was dissected biochemically. Cytochemically, the distribution of ATP-dephosphorylating activity was similar to that shown previously for AMP and -glycerol phosphate; i.e., cerium phosphate reaction product was specifically localized to the transitional zone where the sensory dendrites develop cilia and branch to form the outer dendritic segments. Unlike the dephosphorylation of AMP and -glycerol phosphate, Mg²⁺ or Ca²⁺ was required for ecto-ATPase/phosphatase activity. Biochemical measures of both AMP-and ATP-dephosphorylating activity within aesthetascs corroborated the cytochemical evidence that these activities are localized to the transitional zone. A major portion of the AMP dephosphorylation (about 67%) derives from nonspecific alkaline phosphatase activity that is insensitive to levamisole and L-bromotetramisole. In contrast, nonspecific phosphatase activity accounted for a much smaller part of the ATP dephosphorylation (about 15%). Ectoenzymatic activity in the transitional zone may be an important means of removing excitatory/inhibitory nucleotides from this region.Abbreviations ADP Adenosine 5-diphosphate - AMP adenosine 5-monophosphate - AMPCP , -methylene ADP - ASW artificial seawater - ATP adenosine 5-triphosphate - -GP -glycerol phosphate - EM electron microscopy