Interactions between temperature and salinity during brooding on subsequent zoeal development of the mud crab Rhithropanopeus harrisii

Female mud crabs, Rhithropanopeus harrisii, carrying newly extruded eggs, were collected from the Petaluma River (San Francisco Bay Estuarine System, California, USA) in summer 1985, and exposed to factorial combinations of temperature (20°, 25° or 30°C) and salinity (2, 5, 15, 25, or 32%.). Upon hatching, dry weights of 12 to 15 h-old zoeae were determined. Subgroups of the remaining zoeae were transferred from hatching salinities to the salinities listed above and raised until metamorphosis to megalopa. Low salinities reduced zoeal dry weights by as much as 25%. Temperature played a secondary role in reduction of hatching weight of zoeae. Survival of larvae through zoeal development was best when hatching and rearing salinities were the same; in this case, overall survival increased with temperature. Both duration of zoeal development and megalopal dry weights were strongly influenced by temperature and rearing salinity, with only a small contribution from hatching salinity. The influence of hatching salinity was most obvious at extremes of the range tested. These studies indicate that physical conditions during embryogenesis profoundly influence subsequent larval development. Interpretation of experimental approaches to study ecophysiological adaptations of larval stages should not neglect the role of physical conditions during embryogenesis.     

Effects of the juvenile hormone mimic ZR-512 (Altozar®) on larval development of the mud-crab Rhithropanopeus harrisii at various cyclic temperatures

Effects of the juvenile hormone (JH) mimic hydroprene (Altozar^®: ZR-512), which exhibits high activity against Lepidoptera, were studied on the larval development of the mud-crab Rhithropanopeus harrisii (Gould) (Brachyura: Xanthidae). Larvae reared in 20 S at 3 cycles of temperature of 20° to 25°C, 25° to 30°C and 30° to 35°C, were exposed to 0.01, 0.1 and 0.5 ppm hydroprene from hatching to the first crab stage. Larvae were also exposed to 0.1 and 0.5 ppm hydroprene only from the megalopa stage to the first crab stage. When larvae were treated with hydroprene throughout larval life, survival was significantly reduced with increasing concentrations of the compound at all temperature cycles. Synergistic effect between hydroprene and temperature on survival of zoeal larvae was not observed. On the average there was 11% less survival in the zoeal stages at the 0.01 ppm concentration. of hydroprene than in the control, an additional reduction of 13% occurred at 0.1 ppm, and finally there was a further decrease of 46% at 0.5 ppm hydroprene. Significant decrease in survival in the megalopa stage occurred only in the 0.5 ppm concentration of hydroprene at the lowest temperature cycle when larvae were exposed to the compound from hatching. When larvae were treated with hydroprene only within the megalopa stage, a significant reduction in survival was not observed. First-stage zoeae were the most sensitive of the larval stages to hydroprene. Duration of zoeal development was significantly delayed at 0.5 ppm hydroprene at the two lower temperature cycles, whereas in the megalopa stage the delay began at the 0.1 ppm level at all 3 temperature cycles when larvae were exposed to hydroprene from hatching. A significant delay was also observed at 0.1 ppm hydroprene at the two lower cycles when larvae were exposed to hydroprene only in the megalopa stage; at 30° to 35°C a significant delay was observed only at the 0.5 ppm level. The results show that metamorphosis to the first crab stage was not inhibited at the 0.5 ppm level of hydroprene or lower. Reduction in survival and increase in duration of larval development were presumably related to stress conditions caused by hydroprene. The results also suggest an interaction between temperature and hydroprene on survival of megalopa larvae and duration of larval development.     

Patterns of larval and early juvenile growth in a semiterrestrial crab, Armases angustipes (Decapoda: Sesarmidae): comparison with a congener with abbreviated development

In a semiterrestrial and estuarine tropical crab, Armases angustipes Dana (Grapsoidea: Sesarmidae), changes in biomass (measured as dry mass, W; carbon, C; nitrogen, N; and hydrogen, H; per individual) and relative elemental composition (C, N, H, in percent of W; C:N mass ratio) were studied during development from an early egg stage through hatching, the complete larval phase, metamorphosis and the first juvenile crab stage (CI). In the megalopa and CI, growth was measured also within the moulting cycle, and biomass and elemental composition were determined in cast exuviae. From an early egg stage to the freshly hatched larva, A. angustipes lost about 20% of W, 29% of C, 5% of N and 32% of H. Proportionally higher losses in C than in N were reflected also in a significantly decreasing C:N mass ratio (from 5.02 to 3.74). These results indicate that lipids mobilised from yolk reserves represented the principal metabolic substrate for embryonic energy production, while proteins were catabolised at a much lower rate. The present data of growth and exuviation are compared with previously published data from a congener, A. miersii Rathbun, which has an abbreviated and facultatively lecithotrophic mode of larval development (with three instead of four zoeal stages; stages I and II in principle independent of food). When growth is measured as an increase in the final (premoult) biomass of successive developmental stages, both species show an exponential pattern. Within the moulting cycles of the megalopa and the first juvenile, both species show parabola-shaped growth curves, with a rapid biomass increase in postmoult and intermoult stages, and losses in the premoult phase. Thus, the two Armases species show, in general, similar patterns of larval and early juvenile growth. However, the initial size of eggs and larvae is about four times larger in A. miersii, and its biomass remains higher throughout the period of larval and early juvenile development. A. angustipes is able to partially make up for this difference, as it has an additional zoeal stage, and its megalopa and CI stages show higher relative biomass increments (in percent of initial values). Due to this compensatory growth pattern, A. angustipes reaches in its CI stage about half the biomass of a juvenile A. miersii. When exuvial losses of megalopae and juveniles are compared between these two species, A. miersii shows higher biomass losses per individual (corresponding with its larger size), but lower relative losses (C, N, H, in percent of late premoult body mass or in percent of previously achieved growth increments). Differences in larval and early juvenile growth and in the exuvial losses of megalopae and juveniles of these two congeners are discussed in relation to their differential ecology, life history and reproductive strategy.     

Long-term effects of water-soluble fractions of Kuwait crude oil on the larval and juvenile development of the mud crab Eurypanopeus depressus

Larvae of the mud crab Eurypanopeus depressus (Smith) were reared in various concentrations of the water-soluble fraction of Kuwait crude oil. The 48-h TL_m (median tolerance limit) for Zoea Stage I was approximately 10 ppm total dissolved hydrocarbons and that for Zoea Stage II approximately 17 ppm. Chronic toxicity of more dilute solutions (4.3 and 8.7 ppm) was assessed independently for each larval stage and for subsequent developmental stages through Crab Stage 5. In the group continuously exposed to oil from hatching, there was differential mortality relative to controls in every larval stage and increased duration of intermolt periods was observed at every stage through Crab Stage 5. Mortality in groups not exposed until larvae had reached Zoea Stages III or IV was not greater than controls, suggesting that toxicity to advanced larval stages may be related to accumulation of toxic compounds by the larvae. Neither concentration (4.3 or 8.7 ppm) of crude oil caused increased mortality among juvenile crab stages regardless of the time of initial exposure. Increased occurrence of an extra and morphologically abnormal megalopa stage was associated with exposure to the crude oil. This has not been reported before.Communicated by I. Morris, West Boothbay Harbor     

A long-term study of the effects of water-soluble fractions of No. 2 fuel oil on the survival,development rate,and growth of the mud crab Rhithropanopeus harrisii

Mud crabs, Rhithropanopeus harrisii (Gould), were exposed continuously for 6 months after hatching to water-soluble fractions (WSF) of No. 2 fuel oil. Survival, growth and development rate were monitored during this time. The zoeal stages were the most sensitive to fuel oil. A 20% WSF (0.36 ppm total naphthalenes, 1.26 ppm total hydrocarbons) was acutely toxic to these stages. Of the zoeal stages, the first stage appeared to be the most sensitive. The combined duration of the 4 zoeal stages was significantly increased by increasing WSF exposure concentrations. The megalopa and crab stages were not particularly sensitive to continued petroleum hydrocarbon exposure, particularly when compared to zoeal stages. However, mean duration of the megalopa and first crab stages was significantly affected by oil exposure. Individuals which survived the highest exposure concentrations as larvae appeared to grow larger during the crab stages, so that at the end of 6 months comparably staged crabs were equal to or larger than both control crabs and those exposed to low WSF concentrations. Stage distributions at the end of 6 months showed no differences due to WSF exposure. Sex ratios, which could be determined at the end of 6 months, were approximately 1, indicating no sex-related differential sensitivity to WSF exposure, at least as larvae or juveniles. The data indicate that these crabs possess considerable ability to recover from the effects of chronic sublethal exposure to petroleum hydrocarbons. The most deleterious effects of oil pollution on this species may be due to its impact on larval recruitment into the adult population.     

Larval growth in the estuarine crab Chasmagnathus granulata: the importance of salinity experienced during embryonic development, and the initial larval biomass

The importance of salinity experienced during embryonic development and initial larval biomass on larval growth was studied in the South American estuarine crab Chasmagnathus granulata. Ovigerous females were maintained at three salinities (15, 20, and 32‰) from egg laying to hatching of zoea l. Larvae from all treatments were reared under constant conditions of photoperiod (12∶12), temperature (18°C), and salinity (first instar at 20‰, subsequent instars at 32‰). Biomass was measured as dry weight, carbon, and nitrogen content per individual at egg laying, hatching of zoea l, premoult zoea l, and zoea 4, and in 8-day-old megalopa. From hatching to premoult zoea 4, biomass was higher for larvae from prehatching salinities of 15 and 32‰. There was a significant positive correlation between biomass at hatching and at premoult zoea l and zoea 4. Accumulated biomass during zoeal stages tended to be higher for larvae from broods with higher biomass at hatching, although this trend was not always significant. Zoea 4 either directly metamorphosed to megalopa or moulted to zoea 5, following, respectively, a short or long developmental pathway. The proportion of zoea 4 that followed the long pathway was negatively correlated with biomass of zoeal stages. Biomass at hatching was correlated with biomass of megalopae developed through the short pathway, although it was not correlated with the accumulated biomass at this stage. Megalopae developed through the long pathway (i.e. metamorphosed from zoeae 5) had higher biomass than those from the short pathway. The present results suggest that prehatching salinity and initial egg and larval biomass can be very important for larval growth. Published online: 9 August 2002     

Effects of the juvenile hormone mimic ZR-515 (Altosid®) on larval development of the mud-crab Rhithropanopeus harrisii in various salinities and cyclic temperatures

Effects of 0.01, 0.1 and 1.0 ppm methoprene (Altosid^®: ZR-515), a juvenile hormone (JH) mimic which shows high activity against some economically important insect pests, especially Diptera, were tested on larvae of the mud-crab Rhithropanopeus harrisii (Gould) (Brachyura: Xanthidae) from hatching to the first crab stage under optimum and stress conditions of a number of salinities and cyclic temperatures. There was a significant reduction in survival of zoeal larvae with increasing concentrations of methoprene in nearly all combinations of salinity and temperature. On the average there was 9% less survival in the 0.01 ppm concentration of methoprene than in the control, and in the 0.1 ppm concentration the survival was further reduced by another 16%. At 1.0 ppm methoprene no larvae survived beyond the first zoeal stage under optimum conditions or under stressful combinations of salinity and temperature. Except at 0.2 ppm in 27.5% S, survival of the megalopa was not significantly reduced in 0.01 or 0.1 ppm methoprene in any salinity or temperature, although the percentage of abnormal megalopa increased under stress conditions. The first zoeal stage was the most sensitive of the larval stages to methoprene as well as to salinity and temperature stress. The duration of zoeal development was significantly lengthened with an increase in concentration of methoprene under nearly all conditions of salinity and temperature. The JH mimic had, however, no significant effect on the duration of megalopa development. A significant synergism between methoprene, salinity and temperature was not observed. It can be concluded from the results that methoprene does not inhibit metamophosis of R. harrisii larvae at the 0.1 ppm level or lower. Reduction in survival of zoeal stages and increased duration of zoeal development with increasing concentrations of methoprene are presumably related to stress.     

Changes of respiration and biomass of spider crab (Hyas araneus) larvae during starvation

The influence of starvation on respiration (R), dry weight (W), carbon (C), nitrogen (N), hydrogen (H), and energy content (E; calculated fromC) of spider crab (Hyas araneus L.) larvae was studied in the laboratory. In all larval stages (zoea I and II, megalopa)W increased during postmoult, independent of food, and decreased subsequently. The final reduction inW after continued starvation increased from stage to stage (9, 13, and 20% respectively), but it was always much lower than the decrease inC (44 to 52%),N (42 to 46%),H (50 to 58%), andE (53 to 62%). Individual (R) and weight-specific respiration rates (QO ₂) were reduced by 83 to 88%. The time-dependence of these reductions in metabolism and biomass as well as the rates of change in all parameters studied were described with non-linear regression models and differential equations, respectively. Rates and total amounts of energy lost during starvation were independently calculated fromC andR values, and similar results were mostly obtained. Only in the megalopa stage was there a conspicuous difference between the two estimates: higher losses were calculated fromR. This shows that further (biochemical) data are required for a more complete understanding of the energetics of this stage. Estimates of total protein (fromN) and lipid (fromC) suggest that both constituents serve as metabolic substrates during starvation, but most of the energy originates from the breakdown of protein.Supported by the Deutsche Forschungsgemeinschaft (An 145/1-1)     

Effects on larval crabs of exposure to algal toxin via ingestion of heterotrophic prey

We tested whether ingesting toxic algae by heterotrophic prey affected their nutritional value to crab larval predators, using toxic algal strains that are either ingested directly by larval crabs or rejected by them. Ingestion of toxic strains of the dinoflagellates Alexandrium andersoni and A. fundyense by the rotifer Brachionus plicatilis was confirmed. Rotifers having ingested either algal type for five days were fed to freshly hatched larvae of three crab species, with larval survival and stage durations determined. For both algal/rotifer treatments in all three crab species, larvae fed algae directly died during the first zoeal stage, while those fed rotifers that had been fed either algal strain survived to the experiment’s end (zoeal stage 3). Survival was lower, and stage duration longer, for larvae fed rotifers cultured on toxic algae when compared to those fed non-toxic algae. The role of toxic algae in the planktonic food web may be influenced by its direct or indirect ingestion by larval crabs.     

Effects of the insect growth regulator Dimilin® (TH 6040) on larval development of two estuarine crabs

Effects of Dimilin^® (TH 6040), an insect growth regulator which interferes with the formation of the insect cuticle, were studied on the larval development of Rhithropanopeus harrisii (Gould) and Sesarma reticulatum (Say) (Crustacea: Brachyura). When larvae were exposed to 0.5 (R. harrisii only), 1, 3, 5, 7, and 10 ppb Dimilin from hatching to the first crab stage, survival in both species decreased in relation to increased concentrations of Dimilin. Survival of R. harrisii larvae wa significantly lower at 1 ppb and higher levels compared with control experiments, and in S. reticulatum a significant decrease in survival began at the 3 ppb level. At 10 ppb Dimilin, no larvae survived to the megalopa stage in either of the two species. The results indicate that early stage larvae of R. harrisii are more sensitive to Dimilin than those of S. reticulatum. When R. harrisii larvae were treated with 10 ppb Dimilin during the intermolt period of each of the 4 zoeal stages, nearly all larvae died during molting to the succeeding stage. First zoeal larvae of R. harrisii exposed to 10 ppb Dimilin at various days during the intermolt period were more sensitive to the compound late than early in the period. It is suggested that Dimilin also may interfere with the formation of the cuticle in crab larvae.     

Biochemical changes during larval development of the xanthid crab Rhithropanopeus harrisii. I. Protein,total lipid,alkaline phosphatase,and glutamic oxaloacetic transaminase

Protein, total lipid, and the activities of the enzymes alkaline phosphatase (AP) and glutamic oxaloacetic transaminase (GOT) were measured daily during larval development of the xanthid crab Rhithropanopeus harrisii (Gould). Percent increase in fresh weight was greatest on the first day after each molt. Protein and total lipid per larva increased during the second, third, and fourth zoeal stages, with the most marked increase during the third. When calculated as percent fresh weight, marked accumulation of both protein and total lipid occurred during the third zoeal stage. GOT activity remained stable during the third zoeal stage, but increased significantly during the fourth zoeal stage. AP showed a 4-day cycle of activity apparently related to the molt cycle. It is suggested that increased protein and total lipid content during the third zoeal stage indicate nutritional requirements prior to metamorphosis. Biochemical change may be a more relevant indicator of growth during larval development than either size increase or molt frequency when assessing the value of various diets in the culture of crabs.     

Larval settlement and metamorphosis in the sand crab Emerita talpoida (Crustacea: Decapoda: Anomura)

The effects of various potential cues on the survival and time to metamorphosis of larvae of the sand crab Emerita talpoida (Say) were examined. Zoeal duration, which ranged from 25 to 43 d, was not correlated with subsequent megalopal survival but had a weak, negative correlation with the duration of the megalopal stage. Neither food, sand nor water from containers with adults directly affected megalopal survival or duration. In a second experiment, the presence of sand both increased the probability of survival and shortened the duration of late zoeal stages. This is the first experimental evidence for delayed zoeal metamorphosis in a decapod. Settlement of E. talpoida appears to occur primarily during the zoeal phase, not at the megalopal stage.     

Facultative lecithotrophy during larval development of the burrowing shrimp Callianassa tyrrhena (Decapoda: Callianassidae)

Survival, developmental and consumption rate (Artemia nauplii ingested per day) as well as predation efficiency (ingested per available Artemia nauplii) were studied during the larval development of the shallow-water burrowing thalassinid Callianassa tyrrhena (Petagna, 1792), which exhibits an abbreviated type of development with only two zoeal stages and a megalopa. The larvae, hatched from berried females from S. Euboikos Bay (Aegean Sea, Greece), were reared at 10 temperature–food density combinations (19 and 24 °C; 0, 2, 4, 8 and 16 Artemia nauplii d⁻¹). Enhanced starvation resistance was evident: 92 and 58% of starved zoeas I molted to zoea II, while metamorphosis to megalopa was achieved by 76 and 42% of the hatched zoeas at 19 and 24 °C, respectively. The duration of both zoeal stages was affected by temperature, food density and their interaction. Nevertheless, starvation showed different effects at the two temperatures: compared to the fed shrimp, the starved zoeae exhibited accelerated development at 19 °C (8.4 d) but delayed metamorphosis at 24 °C (5.9 d). On the other hand, both zoeal stages were able to consume food at an increased rate as food density and temperature increased. Predation efficiency also increased with temperature, but never exceeded 0.6. Facultative lecithotrophy, more pronounced during the first zoeal stage of C.tyrrhena, can be regarded as an adaptation of a species whose larvae can respond physiologically to the different temperature–food density combinations encountered in the wide geographical range of their natural habitat. Received: 28 February 1998 / Accepted: 21 October 1998     

Digestive enzymes in the ontogenetic stages of the southern king crab, Lithodes santolla

The early ontogenetic stages of the sub-Antarctic king crab Lithodes santolla were analyzed for the presence and activities of a set of important digestive enzymes. The eggs and non-feeding larvae (zoea I-III, megalopa) showed high activities of esterases, phosphatases, and exopeptidases indicating the enzymatic ability to utilize endogeneous yolk reserves. SDS-PAGE showed a continuous decrease of proteins or proteids in the range of 59–81 kDa during ontogenetic development from the eggs through the zoeal stages to the first juvenile crab stage, CI. This reduction reflects the degradation of storage compounds during lecithotrophic larval development. Activities of the endopeptidases, trypsin and chymotrypsin, were low in eggs and larvae but increased significantly in the first juvenile crab stage. These enzymes typically facilitate the first steps of proteolysis in the extra-cellular spaces of the midgut gland and in the stomach. Their scarcity indicates that the larvae of L. santolla are physiologically not prepared to digest external food. This ability seems to appear first in the CI stage. Extracts of juvenile midgut glands and the gastric fluids of adults showed high activities of a variety of digestive enzymes including phosphatases, carbohydrases, as well as endo- and exopeptidases. High activities of digestive enzymes in adults may compensate for scarce food supply and rate-limiting low temperatures in the predominantly sub-Antarctic habitats of L. santolla.     

Accumulation and loss of biomass inLiocarcinus holsatus larvae during growth and exuviation

Liocarcinus holsatus (Fabricius) larvae, of females originating from the Elbe Estuary, FRG, were reared in the laboratory at constant 15°C in May 1988. For each larval stage, developmental time was measured by individual cultures (Zoea I: 6.7±0.7d; Zoea II: 5.0±0.6d; Zoea III: 4.8±0.7 d; Zoea IV: 5.3±0.6d; Zoea V: 6.1±1.1d; Megalopa: 10.45±0.7d). During the entire period of development, dry weight (W), carbon (C), nitrogen (N), and hydrogen (H) were measured daily (Zoea I to V) or every second day (Megalopa). The energy content (E) was estimated from C. Biomass and energy (per individual) increased in each larval stage as a parabolic function of age and is described by power functions. C, H, and E exhibit a higher percentage gain (relative to initial values at the time of hatching) than W and N. It is suggested that proportionally more lipid than protein is accumulated during larval development. Cyclical changes in the relative biomass (% W) correspond to the larval moult cycle, indicating a rapid uptake of water and minerals immediately after hatching and a later increase in tissue growth. Changes in the C:N ratio suggest that during the first period more lipid than protein is accumulated. These patterns of growth and elemental composition are compared with literature data and a high degree of similarity in the growth characteristics of decapod larvae is seen. In addition W, C, N, and H values as well as E were measured for the exuviae of Zoea I to V and Megalopa. The percentage loss of growth rate by exuviae for each larval instar were higher in W (12 to 16%) and C (8 to 12%), and varied between 5 and 10% for N, H, and E.     

Larval development of Ocypode quadrata (Brachyura: Crustacea) under laboratory conditions

The larvae of Ocypode quadrata (Fabricius) have been reared in the laboratory, from hatching to megalopa stage, at 35 S, 25°C. The five zoeal stages and the megalopa are described, including functional appendages of each stage. On the basis of morphological characteristics, the first zoeal and megalopa stages of O. quadrata can be distinguished from similar stages of closely related Ocypodinae. At 25°C, the megalopa appeared in a minimum of 34 days following hatching.     

Dissolved histamine: a potential habitat marker promoting settlement and metamorphosis in sea urchin larvae

Many species of marine invertebrate larvae settle and metamorphose in response to chemicals produced by organisms associated with the adult habitat, and histamine is a cue for larvae of the sea urchin Holopneustes purpurascens. This study investigated the effect of histamine on larval metamorphosis of six sea urchin species. Histamine induced metamorphosis in larvae of three lecithotrophic species (H. purpurascens, Holopneustes inflatus and Heliocidaris erythrogramma) and in one planktotrophic species (Centrostephanus rodgersii). Direct comparisons of metamorphic rates of lecithotrophic and planktotrophic larvae in assays cannot be made due to different proportions of larvae being competent. Histamine (10 μM) induced metamorphosis in 95% of larvae of H. purpurascens and H. inflatus after 1 h, while the coralline alga Amphiroa anceps induced metamorphosis in 40–50% of these larvae. Histamine (10 μM) and A. anceps induced 40 and 80% metamorphosis, respectively, in the larvae of H. erythrogramma after 24 h. Histamine (10 μM) and the coralline alga Corallina sp. induced 30 and 70% metamorphosis, respectively, in the larvae of C. rodgersii after 24 h. No metamorphosis of any larval species occurred in seawater controls. Larvae of two planktotrophic species (Tripneustes gratilla and Heliocidaris tuberculata) did not metamorphose in response to histamine. Seagrasses, the host plants of H. inflatus, induced rapid metamorphosis in larvae of the two Holopneustes species, and several algae induced metamorphosis in C. rodgersii larvae. Histamine leaching from algae and seagrasses may act as a habitat marker and metamorphic cue for larvae of several ecologically important sea urchin species.     

Larval development of the red crab Pleuroncodes monodon (Decapoda Anomura: Galatheidae) under laboratory conditions

Larvae of Pleuroncodes monodon (Milne-Edwards, 1837), a red crab of commercial importance in South America, were reared in the laboratory at 2 different temperatures (15° and 20°C), from hatching up to the last larval stage. The 5 typical stages, with their corresponding functional appendages, are described and figured. The main characteristics useful in differentiating larvae of P. monodon from those of the other Chilean species of Galatheidae and its northern congener P. planipes are discussed. Data on duration of each larval stage, length of moulting intervals and mortality at the 2 test temperatures are also given.This study was financially supported by the Chilean Ministry of Agriculture and by the National Commission for Scientific and Technological Research (CONICYT).