Differences in responses to factorial combinations of temperature and salinity by zoeae from two geographically isolated populations of the mud crab Rhithropanopeus harrisii

Ovigerous mud crabs, Rhithropanopeus harrisii, were collected from the Petaluma River (San Francisco Bay Estuarine System, California, USA) and from Sykes Creek (Indian River Lagoon System, Florida, USA) during the summer of 1984 and during February 1985, respectively. Their zoeae were reared in factorial combinations of temperature (20°, 25° or 30°C) and salinity (2, 5, 10, 15, 20, 25, or 30%.). Survival and megalopal dry weight were maximal over a far larger range of temperature-salinity combinations for the Florida population. Absolute values of the two parameters were also greater for this group. Temperature dominated effects on duration of zoeal development in both populations. California zoeae developed more slowly at any of the temperatures tested compared with those from Florida. The pattern of all three indices was markedly different under non-optimal conditions. Putatively adaptive modification of survival, development rate and growth of zoeae is evident in response to prevailing environmental conditions which are, in part, a function of latitudinal position. Even though populations in the Petaluma River, California, are less capable of reaching maximal performance under the prevailing physical regimes than the Florida population, they still can live in habitats where physical conditions exclude competitors and predators.     

Absence of sterol synthesis in larvae of the mud crab Rhithropanopeus harrisii and of the spider crab Libinia emarginata

A dietary requirement for sterols in crustacean larvae was established by the use of isotopic tracer techniques. Larvae of the mud crab Rhithropanopeus harrisii and larvae of the spider crab Libinia emarginata were exposed to acetate-¹⁴C or mevalonate-¹⁴C. Radioanalysis of the fatty acids of each species indicated sufficient incorporation of acetate for lipid synthesis. No radioactivity was detected in the sterols of animals exposed to either acetate-¹⁴C or mevalonate-¹⁴C. It was concluded that R. harrisii and L. emarginata larvae are unable to synthesize sterols from acetate or mevalonate and, therefore, require them in their diet.     

Population-related toxicity responses to two butyltin compounds by zoeae of the mud crab Rhithropanopeus harrisii

We exposed zoeae of the mud crab Rhithropanopeus harrisii to either bis(tri-n-butyltin) oxide (TBT) or di-n-butyltin dichloride (DBT). Experiments were repeated with zoeae from females collected from the Petaluma River, California in June–August 1983 and 1984 or from Sykes Creek, Florida (USA) in February 1985. Using probit analysis, we calculated LC₅₀ values for exposure lasting the duration of zoeal development. Tributyltin was 54 to 65 times more toxic than dibutyltin, the lower value characterizing the response of Florida zoeae. Increases in duration of zoeal development and reduction of dry weights of megalops, both sublethal responses, were dose-dependent for the two populations. However, zoeae from Florida consistently had shorter duration of zoeal development and higher megalopal weights at metamorphosis, indicating less sensitivity to an identical exposure to either organotin compound. The results of these experiments show that dibutyltin, a putative degradation product of tributyltin, is less toxic than the parent compound. In addition, early life-history stages of two populations may have significantly different responses to xenobiotic stress which, in the case of brachyuran larvae, is evident in a differential reduction of survival and growth and an increase in duration of zoeal development.     

Biochemical changes during larval development of the xanthid crab Rhithropanopeus harrisii. III. Isozyme changes during ontogeny

Isozyme changes in the xanthid crab Rhithropanopeus harrisii (Gould) are described for 6 enzyme systems through all four zoeal stages, megalopa stage, and juveniles. There is considerable variation in the number of isozymes and their expression during the ontogeny of R. harrisii.Contribution No. 806 of the Chesapeake Biological Laboratory, Center for Environmental and Estuarine Studies, University of Maryland, USA.     

Biology and ecology of the crab Rhithropanopeus harrisii ssp. tridentatus

Data are presented on the geographic and local distributions as well as on the life cycle of the crab Rhithropanopeus harrisii (Gould) ssp. tridentatus (Maitland). Extensive studies on this subspecies were carried out in its natural habitat (an abandoned arm of the Vistula, Poland), and under laboratory conditions. The effects of some environmental factors on distribution, life cycles, and abundance of the crab are discussed. The results obtained are compared with those reported by other investigators.     

The ontogeny of phototaxis by larvae of the crab Rhithropanopeus harrisii

Phototaxis by each zoeal stage of the crab Rhithropanopeus harrisii (Gould) was quantitatively measured by means of a microscope closed-circuit television system. The megalopa stage is indifferent to light stimulation and was thus not tested. The action spectrum for positive phototaxis is similar at each zoeal stage, having the most pronounced maximum at about 500 nm and smaller maxima at 400 and 280 nm. Responsiveness to various intensities of 500 nm light is also similar at each stage. After dark-adaptation strongest positive phototaxis occurs at intensities between 1.0 and 3.0x10^-4 W/m², with no negative phototaxis to lower intensities. After light-adaptation, the positive response occurs to higher intensities between 20 and 0.01 W/m², with a pronounced negative response to lower intensities. Generally, mean swimming speeds during positive phototaxis do not change with stimulation intensity, although during negative phototaxis mean speeds do vary with intensity. At the highest intensities which initiate a negative response, mean values are significantly greater than values for positive phototaxis. Based on the pattern of phototaxis at each developmental stage, a prediction of events during diurnal vertical migration is possible. The negative response after light-adaptation might also function as a shadow reflex.     

The effect of salinity and cyclic temperature on larval development of the mud-crab Rhithropanopeus harrisii (Brachyura: Xanthidae) reared in the laboratory

Larvae of Rhithropanopeus harrisii (Gould) were reared from hatching to the first or second crab stages in 11 combinations of salinities and cyclic temperatures (5, 20, and 35 S at 20° to 25°C, 25° to 30°C, and 30° to 35°C; 25 S at 20° to 25°C and 30° to 35°C). The larvae survived to the megalops and first crab stages in all salinities and cycles of temperature other than 5 S at 30° to 35°C. The best survival to the megalops (94%) and first crab (90%) stages occurred in 20 S, 20° to 25°C. In all other combinations of salinities and temperatures there was a reduction in survival to the first crab stage. The duration of the larval stages was affected significantly by temperature, whereas the effect of salinity on the mean days from hatching to the first crab stage was not consistent at the different temperature cycles. Development to the first crab stage required the shortest time in 20 S, 30° to 35°C (mean 12.3 days), and the longest time in 5 and 35 S, 20° to 25°C (mean 22.6 days and 21.6 days, respectively). Megalops larvae reared in 35 S at all cycles of temperature, as well as larvae in 20 and 25 S, 30° to 35°C, showed a high percentage of abnormality, with the highest percentage occurring in 35 S, 30° to 35°C. It appears that larval development of R. harrisii is strongly influenced by environmental factors and not solely related to genetic differences.This research was supported by grants from the Nordic Council for Marine Biology and the U.S. Atomic Energy Commission [Grant No. At-(40-1)-4377].Contribution No. 116, Zoological Museum, University of Oslo, Norway.     

Entrainment of the larval release rhythm of the crab Rhithropanopeus harrisii (Brachyura: Xanthidae) by cycles in salinity change

Influences of salinity, Na, K, Ca and Mg on Na–K-ATPase activity in the posterior gills of Carcinus maenas (L.) have been investigated with respect to the role of the enzyme in hyperosmotic regulation. K and Mg ions were obligatory for enzyme activity. The dependence on the K concentration can be seen in a saturation curve of the Michaelis-Menten type. Low concentrations of Ca (0.2–3 mM) in the incubation medium strongly inhibited Na–K-ATPase activity. Activities inhibited by Ca could be reactivated to non-inhibited values by the addition of higher amounts of Mg (25 mM). Activity increased along with the salinity of the sea water used as incubation medium up to about 10 S. Here, maximum activity was observed. Further salinity increases of the incubating sea water were inhibitory. Salinity dependence is assumed to be based on Na dependence of the Na-pump. Comparative investigations of the Na–K-ATPase activity and its affinity to sodium in five species of decapod crustaceans indicated that levels of Na–K-ATPase differed in the posterior gills of stenohaline and euryhaline species. The results obtained confirm previous assumptions of a central role of the branchial Na–K-ATPase in hyperosmotic regulation. Properties of the Na–K-ATPase, such as affinity for substrates or dependence on ionic sea water constituents, are kept constant with respect to salinity changes. Modifications due to salinity only concern enzyme amounts especially in the posterior gills. The finding that the Na-pump is localized in basolateral parts of ion-transporting epithelial cells confirms the aforementioned results.     

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.     

Laboratory study of effects of temperature and salinity on survival and larval development of a population of Rhithropanopeus harrisii from the Mondego River estuary,Portugal

Rhithropanopeus harrisii (Gould) is an introduced species in the estuary of the Mondego River (Portugal): it was first recorded from the Iberian Peninsula in 1989. The larval development of this population was studied under laboratory conditions at different temperatures and salinities, and showed a larval development pattern very similar to that reported for American estuarine populations of this species. Larval development was negatively correlated with temperature. Time to megalopa varied between 7 and 35 d; the first crab (C₁) was reached after a maximum of 11 to 43 d. Larval development was optimum at 25°C and 15S. Larval survival was maximum at 10, 15 and 20S at all three temperatures studied (20, 25 and 30°C). The percentage of abnormal megalopae increased with increasing salinity to a maximum (100%) at 30S; incidence of abnormality was not affected by temperature.     

Behavioral responses of larvae of the crab Rhithropanopeus harrisii (Brachyura: Xanthidae) during diel vertical migration

Larval Rhithropanopeus harrisii (Gould) show nocturnal vertical migration. Larval behavioral responses to different rates of increase and decrease in light intensity were measured in an apparatus with a natural angular light distribution. A central objective was to establish whether phototaxis actually participates in vertical migration. At sunset the level of light adaptation controlled the readiness of the larvae to migrate, while an ascent was initiated by a preductable relative decrease in intensity (e.g. 4.0x10^-3s^-1). Rates of relative decrease around sunset would evoke continuous upward swimming. Gravity was the orienting cue and there was no change in swimming speed during the ascent. At sunrise, the larval descent was initiated by exposure to an absolute light intensity of about 0.23 log unit above the lower visual threshold. Light served as an orienting cue, as larvae descended by a negative phototaxis. Thus, phototaxis is not a laboratory artifact and does participate in vertical migration. A consideration of behavioral responses of other crustacean zooplankton indicates there is considerable variation in the initiating and directing cues for vertical movements. The variety of behavioral responses of R. harrisii suggests that a synthesis of hypotheses about migration may provide the proper basis for explaining the mechanisms underlying diel vertical migration.     

Interactive effects of salinity,temperature and polycyclic aromatic hydrocarbons on the survival and development rate of larvae of the mud crabRhithropanopeus harrisii

Zoeae of the mud crabRhithropanopeus harrisii (Gould) were exposed continuously throughout larval development to factorial combinations of salinity, temperature and specific aromatic hydrocarbon concentrations. Salinities and temperatures were 5, 15, or 25 and 20°, 25°, or 30°C, respectively. Either phenanthrene or naphthalene was tested separately at respective concentrations of 0, 100, 150 or 200 ppb and 0, 125, 250 or 500 ppb. Phenanthrene was much more toxic than naphthalene. Naphthalene was not acutely toxic at any physical factor combination-naphthalene concentration tested. Both compounds caused the highest mortality at low salinities. The time course of mortality due to phenanthrene exposure showed that ecdysis between the first and second zoeal stage was the most sensitive period for the larvae exposed to aqueous hydrocarbons. Phenanthrene-exposed larvae had a decreased development rate, but the naphthalene-exposed larvae developed faster than the controls.     

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.     

Biochemical changes during larval development of the xanthid crab Rhithro panopeus harrisii. II. Nucleic acids

Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) contents were measured daily during the zoeal development of the crab Rhithropanopeus harrisii (Gould). DNA concentration (per unit protein) decreased as larvae increased in size. Total DNA content per larva showed an abrupt increase at the second molt and increased steadily during the third and fourth zoeal stages. Ratios of fresh weight to DNA were highest during the first two zoeal stages, dropped sharply at the second molt, then increased during the third and fourth zoeal stages. RNA:DNA ratios showed cyclical activity apparently related to the molt cycle. Assuming that fresh weight: DNA ratio reflects cell size and that DNA content reflects cell number, growth during the first two stages is due primarily to increase in cell size and during the last two stages to a combination of increase in cell size and number. Assuming further that RNA:DNA ratios reflect protein-synthesis activity, such activity in these larvae follows a rhythm related to the molt cycle.     

Effect of temperature and salinity on larval development of southern king crab (Lithodes antarcticus)

Larvae of Lithodes antarcticus Jacquinot were reared in October, 1981 from hatching to the glaucothoe stage at 16 temperature/salinity combinations (5.5°; 7.5°; 9.5° and 13.5°C; 26, 29, 32 and 35 S) to determine optimal environmental conditions for larval development. The highest survival percentage was obtained in the culture at 7.5°C and diminished according to temperature increase or decrease. High temperature cultures significantly shorten the larval life duration, but produce large mortalities. At 5.5°C mortality occurred almost exclusively during the moult to glaucothoe stage. Higher survival percentages were obtained as salinity was increased. In the lowest salinity culture (26 S) no zoea reached the post-larvae stage at culture temperatures. The best T/S combination was obtained at 7.5°C and 35 S, with a survival percentage of 29%. The shortest zoeal developments were obtained at 32 S in all culture temperatures. Salinity also affects larvae coloration: there is a pigment concentration on erythrophores, which causes a color decrease.     

Action of eyestalks on the ovary in Rhithropanopeus harrisii and Crangon crangon (Crustacea: Decapoda)

The annual cycle of ovarian activity and eyestalk effects on the ovary in different phases of its activity were examined in the crab Rhithropanopeus harrisii Gould ssp. tridentatus Maitland, and the sand shrimp Crangon crangon L. In both species, the annual ovarian cycle consists of two phases of vitellogenesis and the resting phase. In R. harrisii, the eyestalks exert an inhibitory effect on vitellogenesis during the phase of vitellogenesis outside the breeding season, and are necessary for the process of oviposition during the breeding season. In C. crangon, ovary-inhibiting hormone is present in the eyestalks outside the breeding season, and has not been detected during this season. The resting phase of the ovary after the breeding season is caused by the ovary-inhibitiing hormone. Females of C. crangon are a suitable material for testing ovary-inhibiting hormone activity, while female R. harrisii are not.     

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.     

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.     

Ultrastructural study of the exoskeleton of the estuarine crab Rhithropanopeus harrisii: Effect of the insect growth regulator Dimilin® (diflubenzuron) on the formation of the larval cuticle

Ultrastructure of larval cuticle during the molt cycle of the estuarine crab Rhitropanopeus harrisii (Gould) (Crustacea: Brachyura) was studied in control larvae as well as in larvae exposed to 10 ppb of the insect growth regulator Dimilin^® (diflubenzuron). First zoeal larvae were used as test organisms. It has earlier been shown that 10 ppb Dimilin is lethal to zoeal larvae of R. harrisii, and nearly all exposed larvae died during molting to the next stage (Christiansen et al., 1978). Distinct differences in structure of the cuticle were found between the two groups of larvae. Both endocuticle and exocuticle appear to be deformed in Dimilin-treated larvae, whereas formation of epicuticle did not seem to be affected. The results indicate that Dimilin probably inhibits chitin synthesis in crab larvae as shown earlier by several authors for insect larvae.     

Interaction of temperature and salinity on oxygen consumption of the estuarine crab Panopeus herbstii

Marine Biology - The combined effects of temperature and salinity on the rate of oxygen consuption by the estuarine crab Panopeus herbstii Milne-Edwards (Crustacea: Decapoda: Xanthidae) were...