A pheromone bouquet controls the reproductive behaviour of the male shore crab, Carcinus maenas

The reproduction of many brachyuran crustaceans involves the formation of mating pairs often around the time of the female moult with attraction of a sexual partner and mating behaviour controlled by sex pheromones. In shore crabs, Carcinus maenas, females produce sex pheromones that are released in the urine. High Performance Liquid Chromatography analysis (HPLC) of female urine shows that the pheromone, identified as the nucleotide uridine diphosphate (UDP), elutes as an unresolved peak with structurally related nucleotides. We examined female urine samples over the moult cycle and detected UDP as well as uridine triphosphate (UTP). Bioassays were conducted to establish the possibility of a blend of nucleotides forming a sex pheromone bouquet in C. maenas. Whilst UDP induced the male mate guarding behaviour (cradling), a mixture of the two nucleotides at a ratio of 4:1 UDP:UTP elicited an even stronger mating response than either UDP or UTP individually. The urine concentration and composition of these nucleotides changes over the moult period pre and post ecdysis, providing evidence that a pheromone bouquet composition is not always constant. The change of the bouquet is related to the physiological state of the sender, here the moult cycle. Our study unravels the functionality of reaction-specific molecules in a pheromone bouquet. Whilst UDP is the mating signal, UTP acts as an attractant and combined they maximise the reproductive response. The use of bouquets provides species-specificity, potentially enabling reproductive isolation of sympatric species, and contains valuable information on the physiological state of the sender.

First mass spectrometric report of cryptocyanin, a moulting protein from the mud crab Scylla serrata (Forskål, 1775) (Decapoda: Brachyura: Portunidae) in India

The mud crab Scylla serrata (Forskål, 1775) is a highly abundant and economically important species throughout coastal India. Cryptocyanin plays an important role during the moult cycle of brachyuran crabs, and the presence of cryptocyanin and haemocyanin in oocytes, embryos, and zoeas of various species suggests that cryptocyanin is available from the very early stage of cuticle formation in development. Cryptocyanin is highly similar to haemocyanin in structure, but it is a copper-free enzyme that lacks an oxygen-binding capacity. Cryptocyanin has furthermore been reported as an important factor in immunity in crustaceans. The precise molecular weight of the cryptocyanin protein from gill tissues of S. serrata (79.11 kDa) is reported with the help of the matrix-assisted laser desorption/ionization mass spectrophotometry (MALDI-MS) technique.

Biphasic moult cycle of the parasitic isopod Norileca indica (H. Milne Edwards, 1840) (Isopoda: Cymothoidea): stage-wise characterisation and haemolymph ecdysteroids titre

Norileca indica (H. Milne Edwards, 1840) is a protandrous hermaphroditic and obligatory parasitic isopod of scombrid fishes. The posterior and anterior biphasic moult cycles in the adult phases (male, transitional, and female) of this species are characterised based on the microscopic changes reflected in the uropod exopodite and maxillule, respectively. Biphasic parturial moult as well as the moult removing the old brood pouch characteristic of ovigerous females was recorded. Assay of haemolymph ecdysteroids titre with respect to different biphasic moult cycle stages appears to be first report of its kind among aquatic and/or parasitic isopods. The ecdysteroids titre showed significant fluctuation in relation to the moult cycle stages, while the parasite was at a biphasically intermoult stage, titre was estimated to be 13.64 ± 2.09 ng ml–1. By the onset of the premoult at the posterior body region, titre showed a gradual increase and reached a maximum level (328.54 ± 70.17 ng ml–1; P = 0.0060) when the parasite was at late premoult at the posterior body part and mid premoult stage at the anterior body part. There was a subsequent sharp decline (92.8 ± 12.95 ng ml–1; P = 0.002) soon after the ecdysis of the posterior part by the time premoult changes in the anterior part advanced as late premoult and about to moult. The level of ecdysteroids titre was 55.56 ± 14.47 ng ml–1 when the parasite completed anterior ecdysis and reached postmoult. Data on the monthly occurrence of biphasic moult cycles revealed that a mean average of 23.26% of the adult population undergoes premoult/ecdysis/postmoult activities throughout the year. The rate of moulting showed a decline (16.12 ± 0.60%) during the monsoon months (June-August) when compared to pre-summer/summer (February-May) (24.83 ± 2.42%) and post-monsoon/winter (September-January) (26.28 ± 1.75; P = 0.005).

Expression patterns of MIH, EcR2, and RXR3 in the moult cycle of the red swamp crayfish, Procambarus clarkii (Girard, 1852) (Decapoda, Cambaridae)

In arthropods, the moult-inhibiting hormone (MIH), the ecdysone receptor (EcR), and the retinoid X receptor (RXR) are key regulators in moulting. In the present study, the full-length cDNAs of the MIH, EcR2, and RXR3 genes from the red swamp crayfish, Procambarus clarkii (Girard, 1852) (denoted as PcMIH, PcEcR2, and PcRXR3) were cloned. Tissue-specific and moult stage-specific mRNA expression patterns of these genes were detected by real-time quantitative polymerase chain reaction. PcMIH was detected only in the eyestalk, whereas PcEcR2 and PcRXR3 mRNA were expressed in all tissues tested. The highest levels of PcEcR2 and PcRXR3 were detected in the gill and hepatopancreas. Expression of PcMIH mRNA in the eyestalk increased from postmoult to peak in intermoult and then decreased in premoult. Expression of PcEcR2 mRNA in the eyestalk, hepatopancreas, and muscle increased from postmoult to peak in early premoult and then decreased. However, expression of PcEcR2 mRNA in the gill increased from postmoult to reach a maximum in intermoult and then decreased in premoult. Expression of PcRXR3 mRNA also fluctuated in the eyestalk, hepatopancreas, muscle, and gill, with a decrease from postmoult to late premoult. Expression of PcEcR2 and PcRXR3 mRNA increased relative to the control in the hepatopancreas and gill after unilateral and bilateral eyestalk ablation, which suggested that PcMIH can inhibit their mRNA expression. Double-stranded RNA-mediated RNA interference of PcRXR3 caused different changes in mRNA expression of these genes in different tissues and resulted in decreased expression of PcEcR2 mRNA, which suggested a collaborative relationship between PcEcR2 and PcRXR3.

Modelling growth and reproduction of Antarctic krill, Euphausia superba, based on temperature, food and resource allocation amongst life history functions

Estimates of productivity of Antarctic krill, Euphausia superba, are dependent on accurate models of growth and reproduction. Incorrect growth models, specifically those giving unrealistically high production, could lead to over-exploitation of the krill population if those models are used in setting catch limits. Here we review available approaches to modelling productivity and note that existing models do not account for the interactions between growth and reproduction and variable environmental conditions. We develop a new energetics moult-cycle (EMC) model which combines energetics and the constraints on growth of the moult-cycle. This model flexibly accounts for regional, inter- and intra-annual variation in temperature, food supply, and day length. The EMC model provides results consistent with the general expectations for krill growth in length and mass, including having thin krill, as well as providing insights into the effects that increasing temperature may have on growth and reproduction. We recommend that this new model be incorporated into assessments of catch limits for Antarctic krill.