Effects of Microphallus turgidus (Trematoda: Microphallidae) on the predation, behavior, and swimming stamina of the grass shrimp Palaemonetes pugio

The effect of the trematode Microphallus turgidus on its second intermediate host, the grass shrimp, Palaemonetes pugio, was tested. To do so, we measured the susceptibility of infected and uninfected shrimp to predation by the mummichog, Fundulus heteroclitus. Shrimp behavior was compared in the presence and absence of a fish predator, and the swimming stamina and backthrust escape responses of infected and uninfected shrimp were measured. Infected shrimp were more likely to be eaten by a predator than uninfected shrimp, had lower swimming stamina, and spent more time swimming and less time motionless in the presence of a predator. There was no difference between backthrust distances traveled in response to a stimulus by either infected or uninfected shrimp. Thus, M. turgidus may increase the predation of P. pugio in the wild, possibly by affecting the swimming stamina and predator avoidance responses of the shrimp.     

Ultrastructure of the eyes of the grass shrimp,Palaemonetes pugio

Summary The cone cells and corneagenous cells possess extensive networks of smooth tubular endoplasmic reticulum that may be involved in optical reflectance and light-adaptational responses, respectively. The extracellular basal lamina of the basement membrane is confluent with glial cell capillary walls and may prove to be a viaduct for the transmission of hemolymph-borne substances to the retina or of retinal degradation products to the hemolymph. In addition to dense pigment granules, the distal pigment cells are shown for the first time to contain migratory reflecting platelets that are usually polymorphic in light-adapted eyes but are rectangular in dark-adapted eyes. In the latter these plates become aligned against the crystalline cones and presumably contribute to the reflection superposition optics of the grass shrimp. Dark-adapted retinular cells possess well-developed perirhabdomal cisternae, oblong or ovoid mitochondria, generally vesicular rough endoplasmic reticulum, and occasional, spherical, calcium-like intrarhabdomal inclusions. Light-adapted retinular cells possess poorly developed perirhabdomal cisternae, lamelliform rough endoplasmic reticulum, and condensed mitochondria frequently associated with lipid droplets and pigment granules. The cytoplasmic boundaries of the reflecting pigment cells expand into the extracellular spaces between individual ommatidial retinular cells during dark adaptation and recede to the interommatidial extracellular spaces during light adaptation. Cytoplasmic microfilament bundles found only at the bases of partially light-adapted rhabdomeric microvilli may be involved in microvillar shortening.     

Effects of cyclic hypoxia on gene expression and reproduction in a grass shrimp, Palaemonetes pugio

Cyclic changes in dissolved oxygen occur naturally in shallow estuarine systems, yet little is known about the adaptations and responses of estuarine organisms to cyclic hypoxia. Here we examine the responses of Palaemonetes pugio, a species of grass shrimp, to cyclic hypoxia (1.5-8 mg/l dissolved oxygen; 4.20-22.42 kPa) at both the molecular and organismal levels. We measured alterations in gene expression in hepatopancreas tissue of female grass shrimp using custom cDNA macroarrays. After short-term (3-d) exposure to cyclic hypoxia, mitochondrial manganese superoxide dismutase (MnSOD) was upregulated and 70-kd heat shock proteins (HSP70) were downregulated. After 7-d exposure, nuclear genes encoding mitochondrial proteins (ribosomal protein S2, ATP synthase, very-long-chain specific acyl-CoA dehydrogenase [VLCAD]) were downregulated, whereas mitochondrial phosphoenol pyruvate carboxykinase (PEP Cbk) was upregulated. After 14 d, vitellogenin and apolipoprotein A1 were upregulated. Taken together, these changes suggest a shift in metabolism toward gluconeogenesis and lipid export. Long-term (77-d) exposure to hypoxia showed that profiles of gene expression returned to pre-exposure levels. These molecular responses differ markedly from those induced by chronic hypoxia. At the organismal level, cyclic hypoxia reduces the number of broods and eggs a female can produce. Demographic analysis showed a lower estimated rate of population growth in grass shrimp exposed to both continuous and short-term cyclic hypoxia, suggesting population-level impacts on grass shrimp.     

Compensatory partitioning of carbon budgets by the grass shrimp (Palaemonetes pugio) and implications for predator prey relationships

Carbon utilization and allocation were examined between two populations of shrimp (Palaemonetes pugio) to determine the possible effects of living in an area of high anthropogenic impact. Carbon assimilation has been studied in P. pugio, but no study has looked at how assimilation might be influenced by contaminants. Anthropogenic effects on carbon assimilation in grass shrimp represent a major unmeasured impact on the carbon budget of multi-cellular organisms in estuaries and near shore environments. The influence of anthropogenic contamination on carbon assimilation has implications for predicting the environmental impact of contaminants, for models of estuarine function, and trophic transfer from the dominant macroscopic detritus processor to species of direct economic importance. Shrimp budgets were compared between two populations, one from a highly polluted marsh creek system in Northern New Jersey, and one from a clean reference site in Southern New Jersey. All components of the carbon budgets were measured directly including carbon allocated to reproduction. Carbon lost to respiration was lower in shrimp from the polluted system allowing them to have increased reproductive output. This is examined in the context of previous studies that show lowered predation by a piscine predator at the polluted site, resulting in a trophic cascade and changes in ecosystem function due to anthropogenic impacts.     

Endocrine-mediated effects of UV-A irradiation on grass shrimp (Palaemonetes pugio) reproduction

Although much is known regarding photoperiodic effects on crustacean egg production, the effects of ultraviolet (UV) light on reproduction has not been investigated. Likewise, little is known concerning the interaction between UV and xenobiotic exposure on crustacean reproductive cycles. In this study, male and female grass shrimp, Palaemonetes pugio, were exposed to sublethal concentrations of endosulfan (200 ng/l and 400 ng/l ES) under both white fluorescent (WF) and UV-A (315-400 nm) light conditions for 50 days in laboratory bioassays. Female endocrine (vitellogenin, ecdysteroids, and cholesterol), reproductive (percent gravid, clutch size), and embryo (days to hatch, hatching success, and hatching survival) responses were assessed. UV-exposure alone caused a significant (>4-fold) increase in total Palaemonetes pugio female egg production over the course of 50 days. Exposure to ES and UV significantly lowered the percentage of gravid females relative to UV controls, whereas ES-exposed shrimp under WF lighting did not exhibit these trends. Although higher vitellogenin concentrations and lower ecdysteroid titers were correlated with increased female egg production, cholesterol titers only exhibited a dose-dependent change when exposed to ES. Embryos from females exposed to UV had significantly lower ecdysteroid titers and shorter hatching times but there were no differences in embryo vitellogenin concentrations, hatching success, or hatching survival. These results indicate that UV-A exposure has a pronounced effect on grass shrimp (Palaemonetes pugio) reproduction and is likely mediated through 5-hydroxytrptamine (5-HT)-related neuroendocrine pathways. The implications for decapod aquaculture and evaluating chronic contaminant effects are discussed.     

Prevalence,geographic distribution,and fitness effects of Microphallus turgidus (trematoda: Microphallidae) in grass shrimp (Palaemonetes spp.) from coastal Georgia

Grass shrimp, Palaemonetes pugio Holthuis and P. vilgaris (Say), were collected at 11 localities along the coast of Georgia and surveyed for digenetic trematode metacercariae. The effect of trematode infection on grass shrimp fitness was also examined. Microphallus turgidas (Leigh) was the only trematode observed. The prevalence of metacercarial cysts of this parasite in P. pugio (75%) was higher than in P. vulgaris (24%), as were the mean intensity, abundance, and population density (no. cysts/cm host body length) of the parasite. Infected shrimp were found at every collection locality and parasite prevalence and density were greatest in P. pugio from higher salinity localities (> or = 20 parts per thousand [ppt]). There was no relation between host body size and parasite density in P. vulgaris, and parasite density increased with host body size in P. pugio, suggesting that the parasite does not affect host survival. There was no relation between parasite density and shrimp egg mass, but nonovigerous female P. pugio were more heavily infected than ovigerous ones. In addition, 1 metacercaria of M. turgidus in each of 2 specimens of P. vulgaris was parasitized by the haplosporidian Urosporidium crescens De Turk. This represents a new shrimp host record for this hyperparasite.     

Antioxidants and total oxyradical scavenging capacity during grass shrimp, Palaemonetes pugio, embryogenesis

During embryogenesis in grass shrimp the capacity to scavenge oxyradicals increased as measured by the Total Oxyradical Scavenging Capacity (TOSC) assay. The increase in TOSC during embryogenesis was associated with increasing concentrations of a number of antioxidants, including coenzyme Q (ubiquinone), alpha-tocopherol and reduced glutathione. Glutathione concentrations ranged from 0.004 to 0.005 nmol/embryo in early embryo stages and reached concentrations between 0.16 to 0.23 nmol/embryo in late embryo stages. Ascorbate remained essentially constant (0.16-0.20 nmol/embryo) throughout embryogenesis and may provide the preponderance of TOSC during early embryo development. Carotenoids were associated with yolk lipovitellin and these antioxidants decreased as yolk was absorbed during embryogenesis. Astaxanthin and beta-carotene were identified in embryos with astaxanthin always the principal carotenoid. In early embryo stages there are maternally derived antioxidants but as embryogenesis proceeds there is an assembly of a complex antioxidant system by newly formed cells and tissues.     

Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio

The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations.     

Some factors affecting size distribution and density of grass shrimp (Palaemonetes pugio) populations in two New Jersey estuaries

Mature Palaemonetes pugio, from a polluted estuary, Piles Creek (PC) are larger than those in a more pristine estuary in Tuckerton (TK). Possible causes for the differences in size-structure could be differences in environmental factors at the two creeks, differential competition at the two sites, inherent factors causing a greater growth rate in the PC population, differences in reproductive timing, or differential predation at the two sites. Lab microcosm studies were used to examine the possibility of inherent population differences or environmental conditions (including differences in salinity) causing faster growth in PC. There was no evidence that PC shrimp grew faster than TK shrimp, nor that PC conditions fostered greater growth compared to TK. Salinity adjustments in PC and TK tanks also had no effect on growth. Therefore, it appears that genetic and environmental factors did not play a significant role in greater shrimp growth in PC. In the field, data on relative abundances of Fundulus heteroclitus and P. pugio, and size-frequency distributions of P. pugio were collected from the two estuaries. Gravid females and recruits of young shrimp physically appeared at approximately the same time within both systems, eliminating earlier reproduction as a cause of the size discrepancy. There were three times as many shrimp in PC than in TK (eliminating the possibility of increased population density and competition at TK as a cause) and three times as many F. heteroclitus present in TK than in PC. Since it has already been established that the Fundulus at PC are smaller than at TK and that they are poor predators, differences in Fundulus predation would appear to be an important factor in determining the number and size-frequency of the grass shrimp.     

Prevalence of the bopyrid isopod Probopyrus pandalicola in the grass shrimp, Palaemonetes pugio, in four tidal creeks on the South Carolina-Georgia coast

The grass shrimp, Palaemonetes pugio, is an important food source for many invertebrate and fish species, including several of commercial importance. The bopyrid Probopyrus pandalicola prevents reproduction in P. pugio by sexual sterilization. The purpose of our research was to determine bopyrid prevalence in grass shrimp over the course of a year. Shrimp were collected from 2 estuarine systems in South Carolina and 2 estuarine systems in Georgia and examined for parasite presence, sex, and gravidity. Site-specific monthly prevalence ranged from 0 to 6.3%. Country Club Creek had the maximum mean +/- SE prevalence of 3.1 +/- 0.3%, and Harbour Town had the minimum of 1.3 +/- 0.3%. Maximum prevalence was concurrent with peak gravidity for Moon River; thus, at this site the negative effect of this parasite on reproductive output may be greater. Reduced egg production may affect grass shrimp abundance and ultimately the recruitment success of its predators.     

Rosette glands in the gills of the grass shrimp,Palaemonetes pugio. I. Comparative morphology,cyclical activity,and innervation

Two types of exocrine rosette glands (called type A and type B), located in the gill axes of the grass shrimp Palaemonetes pugio, are described. The type A glands are embedded within the longitudinal median septum of the gill axes, whereas the type B glands typically project into the efferent hemolymph channels of the gill axes. Although both glands have certain common characteristics (i.e., a variable number of radially arranged secretory cells, a central intercalary cell, and a canal cell that forms the cuticular ductule leading to the branchial surface), they differ in the following respects. The type B gland is innervated, but the type A gland is not; axonal processes, containing both granular (ca. 900–1300 Å) and agranular (ca. 450–640 Å) vesicles, occur at a juncture between adjacent secretory cells and the central cell of the type B gland. The secretory cells of type A and type B glands differ in their synthetic potential and membrane specializations. These differences are more pronounced in well-developed, mature glands, most frequently encountered in larger (24–28 mm, total length) grass shrimp, than in the underdeveloped, immature glands that are most abundant in smaller (14–18 mm, total length) grass shrimp. Thus, in mature glands, the secretory cells of the type A rosette glands are characterized by extensive RER, abundant Golgi, and numerous secretory granules, whereas the secretory cells of the type B gland are characterized by extensively infolded and interdigitated basal plasmalemmas and by the presence of numerous mitochondria. In general, both types of glands exhibit increased secretory activity soon after ecdysis. The central and canal cells in both glands seem to have a role in the modification of the secreted materials. The possible functions assigned to the type A gland and the type B gland include phenol-oxidase secretion and osmoregulation, respectively.     

Purification of vitellin from grass shrimp Palaemonetes pugio, generation of monoclonal antibodies, and validation for the detection of lipovitellin in Crustacea

Much effort has been put into developing vitellogenin antibodies against a wide variety of aquatic vertebrate species to study potential estrogen or anti-estrogen endocrine disrupters. Little work has been done on endocrine disruption in aquatic invertebrates. Although some antibodies have been produced against blue crab and penaeid shrimp lipovitellin, they have only poor cross-reactivity with the important estuarine grass shrimp, Palaemonetes pugio. Vitellin was purified from eggs, monoclonal antibodies were produced using standard techniques, and hybridoma supernatants were screened by ELISA. Western blots were done using extracts from male and female grass shrimp to verify specificity of the monoclonal antibodies. Two low molecular mass bands in the range of 68-85 kD and two high molecular mass bands in the range of 190-221 kD were found. In addition to grass shrimp, several other crustacean species were screened and cross-reactivity found, including blue crab (Callinectes sapidus), mud crab (Rhithropanopeus harrisii), red swamp crayfish (Procambarus clarkii ) and Daphnia magna. To further investigate the use of the antibody, we performed a chronic 6-week pyrene exposure study. We found that vitellin was upregulated in females after 6 weeks and that this may be a protective measure against lipophilic xenobiotics.     

Behavioral thermoregulation in the grass shrimp Palaemonetes vulgaris (Say).

Grass shrimp (Palaemonetes vulgaris) tested in a horizontal linear gradient trough exhibited a modal final thermal preferendum of 27 degrees C, and voluntarily occupied a 24-31 degrees C range.     

A new species of Pleistophora (Microsporida: Pleistophoridae) parasitic in the shrimp Palaemonetes pugio

Pleistophora sp. Sprague, 1970, in the muscle of Palaemonetes pugio was studied. Formalin fixed spores were ellipsoidal, 2.5–3.3 × 1.4–2.0 μm, and with large anterior and posterior clear areas representing the polaroplast and posterior vacuole. This species, after comparison with four other species of Pleistophora in decapod crustacea, was found to be new. It is named Pleistophora lintoni n. sp.     

Ultrastructure of an exocrine dermal gland in the gills of the grass shrimp,Palaemonetes pugio: Occurrence of transitory ciliary axonemes associated with the sloughing and reformation of the ductule

Exocrine dermal glands, comparable to the class 3 glandular units of insects, are found in the gills of the grass shrimp, Palaemonetes pugio. The dermal glands are composed of three cells: secretory cell, hillock cell and canal cell. Originating as a complex invagination of the apical cytoplasm of the granular secretory cell, a duct ascends through the hillock and canal cells to the cuticular surface. The duct is divisible into four regions: the secretory apparatus in the granular secretory cell, the locular complex, the hillock region within the hillock cell and the canal within the canal cell. A tubular ductule is contained within the latter two regions. As the ductule ascends to the cuticular surface, its constitution gradually changes from one of a fibrous material to one which possesses layers of epicuticle. During the proecdysial period, the ductule is extruded into the ecdysial space and this is followed by the secretion of a new ductule. Temporary ciliary structures, located near the secretory apparatus of the secretory cell, are associated with the extrusion and reformation of the ductule. Characterized only by a basal body and rootlets throughout most of the intermolt cycle, the ciliary organelles give rise to temporary axonemic processes which ascend through the ductule toward the ecdysial space at the onset of proecdysis. Subsequently, the old ductule is sloughed off and a new ductule is reformed around the ciliary axonemes. Following this reformation, the ciliary axonemes degenerate. The function of cytoplasmic processes, derived from the apical cytoplasm of the secretory cell, is also discussed.