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.     

Behavioral response to hydrostatic pressure in larvae of two species of xanthid crabs

Responses of pelagic larvae of two species of xanthid crabs to manipulations of hydrostatic pressure were examined and compared. One species, Rhithropanopeus harrisii (Gould), is a temperate estuarine species, while the other, Leptodius floridanus (Gibbes), inhabits shallow water and reefs in tropical and subtropical regions. All zoeal stages of R. harrisii detected pressure stimuli presented in increments as small as 0.025 atm. Both Stage I and II zoeae of L. floridanus responded to pressure stimuli presented in increments of 0.1 atm. Stage I responded when pressure was changed by the smallest increment tested, 0.025 atm, but Stage II did not. L. floridanus zoeal Stages III and IV, however, did not change vertical position relative to control larvae when subjected to pressure stimuli. The capacity of R. harrisii zoeae to respond to changes in hydrostatic pressure may be related to their retention in the estuarine environment. In L. floridanus, a non-estuarine species, the pressure response is important only in the first zoeal stage and may function promarily in dispersal.     

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.     

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.     

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.     

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.     

A comparative analysis of sperm storage and mating strategies in five species of brachyuran crabs

Variation in female sperm storage is explained, in part, by the amount of sperm transferred at mating. Laboratory mating experiments were conducted on Eurypanopeus depressus and Rhithropanopeus harrisii from the Chesapeake Bay and Pachygrapsus transversus from Florida, while mated pairs of Uca beebei and U. terpsichores were collected from mudflats in Panama. All experiments and collections were conducted during the summer of 2006 and 2007. More sperm was transferred to larger than smaller females, and by species with long copulation durations (R. harrisii and E. depressus). These two species live in cryptic habitats, have high sperm/egg ratios, and likely store sperm across multiple broods. In contrast, P. transversus and U. beebei mate conspicuously, have short copulations, transfer fewer sperm, and have low sperm/egg ratios. Comparisons of sperm transfer across different mating strategies and habitats provide a better understanding of female sperm storage in the Brachyura.     

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.     

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.     

Comparative vulnerability to predators,and induced defense responses,of eastern oysters Crassostrea virginica and non-native Crassostrea ariakensis oysters in Chesapeake Bay

Management agencies are considering introducing the Suminoe oyster Crassostrea ariakensis into Chesapeake Bay, USA. It is unknown if the growth of feral populations of this non-native oyster would be regulated by the same predators that once controlled the abundance of the native eastern oyster C. virginica. In laboratory studies, we compared the relative susceptibility of juvenile diploids (shell height < 25 mm) of both oyster species to invertebrate predators of eastern oyster juveniles. Predators included four species of mud crabs [Rhithropanopeus harrisii (carapace width 7–11 mm), Eurypanopeus depressus (6–21 mm), Dyspanopeus sayi (8–20 mm), and Panopeus herbstii (9–29 mm)], the blue crab Callinectes sapidus (35–65 mm), and two sizes of polyclad flatworms (Stylochus ellipticus and possibly Euplana gracilis; planar area ≲5 mm² and ∼14 to 88 mm²). All four species of mud crab and the blue crab preyed significantly (ANOVA, P ≤ 0.05) more on C. ariakensis than on C. virginica, but predation by flatworms of both sizes did not differ significantly between oyster species. The greater susceptibility of C. ariakensis to crab predation was likely due to its shell compression strength being 64% lower than that of C. virginica (P = 0.005). To test for predator-induced enhancement of shell strength, we held oysters of both species for 54 days in the presence of, but protected from, C. sapidus and R. harrisii. Crabs were fed congeneric oysters twice weekly within each aquarium. Compared to controls, shell strength of C. virginica exposed to R. harrisii increased significantly (P < 0.043), as did shell strength of both oyster species exposed to C. sapidus (P < 0.01). Despite the changes in shell strength by both oyster species in the presence of C. sapidus, the shell of C. ariakensis remained 57% weaker than C. virginica. We conclude that, because C. ariakensis exposed to predators continued to have a weaker shell relative to C. virginica, the natural suite of crab and flatworm predators in Chesapeake Bay will likely serve to control the abundance of feral C. ariakensis. We caution that the situation in the natural environment may be sufficiently different in some locations that C. ariakensis may be able to compensate for its greater vulnerability to crab predation and hence become a nuisance species.     

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.     

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.     

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.     

Biochemical and energy changes during embryogenesis in the rock scallopCrassadoma gigantea

The biochemical composition of eggs spawned from rock scallopsCrassadoma gigantea (Gray) collected from Clayoquot Sound, Vancouver Island (British Columbia, Canada) in the winter of 1987 and the summer of 1988, differed significantly only in lipid and soluble-ash contents. Major macronutrients were catabolized simultaneously and linearly with time of embryogenesis. Total egg energy expended during development was 11.8 kJ g^–1 derived 46.7, 9.8 and 43.5%, from lipid, carbohydrate, and protein, respectively. Glucose in the egg was the dominant source of carbohydrate energy. Deposition of shell was linear with time of embryogenesis and the formation of 44.8% dry wt of shell in the early larval stage, referred to as the D-larva, consumed 64.4% of the total energy expended. Energy required for shell formation was 17.55 kJ g^–1. The RNA:DNA ratio declined exponentially from 18.6 in the egg to 2.21 in the D-larva, which was consistent with that of adult bivalves. Many changes in fatty acid composition were linear with time of embryogenesis, and the percentage ofn3- andn6-higher unsaturated fatty acids to total polyethylenic acids exhibited significant declining and increasing trends, respectively. The linear rate of catabolism of eicosapentaenoic acid, 20:5n3, and the constant level of docosahexaenoic acid, 22:6n3, illustrated an energy and a structural role, respectively, for these dominant acids in the embryonic development of the rock scallop.     

Effect of the insect growth regulator diflubenzuron (Dimilin®) on the uptake of glucose and N-acetylglucosamine into the cuticle of crab larvae

Incorporation of ³H-labelled glucose and ³H-labelled N-acetylglucosamine (NAGA) — both precursors to chitin —into the cuticle of Rhithropanopeus harrisii (Gould) larvae (Crustacea: Brachyura) has been examined at different stages of the moult cycle in control larvae as well as in larvae treated with the insect growth regulator diflubenzuron (Dimilin^®). As far as the control larvae were concerned, the incorporation of both precursors was high at the postmoult stage when endocuticle was secreted. NAGA appeared to be a more specific precursor of cuticular material than glucose during the premoult stage when exocuticle was produced. Incorporation of both precursors was low immediately before ecdysis and during the intermoult stage when secretion of the cuticle is complete. The results show that incorporation of glucose into chitin was greatly inhibited by the pollutant during the postmoult stage when endocuticle is produced, while incorporation of NAGA was reduced to a lesser extent at this stage. Diflubenzuron treatment markedly affected the incorporation of both NAGA and glucose in the premoult stage during secretion of exocuticle.     

Heterogeneity in the photoprotective capacity of three Antarctic diatoms during short-term changes in salinity and temperature

The Antarctic marine ecosystem changes seasonally, forming a temporal continuum of specialised niche habitats including open ocean, sea ice and meltwater environments. The ability for phytoplankton to acclimate rapidly to the changed conditions of these environments depends on the species’ physiology and photosynthetic plasticity and may ultimately determine their long-term ecological niche adaptation. This study investigated the photophysiological plasticity and rapid acclimation response of three Antarctic diatoms—Fragilariopsis cylindrus, Pseudo-nitzschia subcurvata and Chaetoceros sp.—to a selected range of temperatures and salinities representative of the sea ice, meltwater and pelagic habitats in the Antarctic. Fragilariopsis cylindrus displayed physiological traits typical of adaptation to the sea ice environment. Equally, this species showed photosynthetic plasticity, acclimating to the range of environmental conditions, explaining the prevalence of this species in all Antarctic habitats. Pseudo-nitzschia subcurvata displayed a preference for the meltwater environment, but unlike F. cylindrus, photoprotective capacity was low and regulated via changes in PSII antenna size. Chaetoceros sp. had high plasticity in non-photochemical quenching, suggesting adaptation to variable light conditions experienced in the wind-mixed pelagic environment. While only capturing short-term responses, this study highlights the diversity in photoprotective capacity that exists amongst three dominant Antarctic diatom species and provides insight into links between ecological niche adaptation and species’ distribution.     

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.     

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.     

Patch use as an indicator of habitat preference,predation risk,and competition

Summary A technique for using patch giving up densities to investigate habitat preferences, predation risk, and interspecific competitive relationships is theoretically analyzed and empirically investigated. Giving up densities, the density of resources within a patch at which an individual ceases foraging, provide considerably more information than simply the amount of resources harvested. The giving up density of a forager, which is behaving optimally, should correspond to a harvest rate that just balances the metabolic costs of foraging, the predation cost of foraging, and the missed opportunity cost of not engaging in alternative activities. In addition, changes in giving up densities in response to climatic factors, predation risk, and missed opportunities can be used to test the model and to examine the consistency of the foragers' behavior. The technique was applied to a community of four Arizonan granivorous rodents (Perognathus amplus, Dipodomys merriami, Ammospermophilus harrisii, and Spermophilus tereticaudus). Aluminum trays filled with 3 grams of millet seeds mixed into 3 liters of sifted soil provided resource patches. The seeds remaining following a night or day of foraging were used to determine the giving up density, and footprints in the sifted sand indicated the identity of the forager. Giving up densities consistently differed in response to forager species, microhabitat (bush versus open), data, and station. The data also provide useful information regarding the relative foraging efficiencies and microhabitat preferences of the coexisting rodent species.     

Biosynthesis of trimethylamine oxide in calanoid copepods. seasonal changes in trimethylamine monooxygenase activity

Live copepods, Calanus finmarchicus (Gunnerus) and C. hyperboreus (Krøyer), exposed to dissolved ¹⁴C-labeled trimethylamine (TMA) in seawater, oxidized TMA to trimethylamine oxide (TMAO), which accumulated in the organisms. The amount of TMAO synthesized was dependent on the time of exposure to TMA and the concentration of TMA in the seawater. It was inferred that copepods can produce TMAO by oxidation of TMA found in their plant food. Choline and methionine did not appear to be of importance as precursors of TMAO. There were large seasonal changes in TMA monooxygenase activity of both copepods. The activity was high in spring, and decreased through summer and autumn to a winter low in October to March. The changes in TMAO content of C. finmarchicus were, in comparison, small (this aspect was not tested for C. hyperboreus).