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PubMed1999

Evolution of ITS1 rDNA in the Digenea (Platyhelminthes: trematoda): 3' end sequence conservation and its phylogenetic utility.

A comparison of ribosomal internal transcribed spacer 1 (ITS1) elements of digenetic trematodes (Platyhelminthes) including unidentified digeneans isolated from Cyathura carinata (Crustacea: Isopoda) revealed DNA sequence similarities at more than half of the spacer at its 3' end. Primary sequence similarity was shown to be associated with secondary structure conservation, which suggested that similarity is due to identity by descent and not chance. Using an analysis of apomorphies, the sequence data were shown to produce a distinct phylogenetic signal. This was confirmed by the consistency of results of different tree reconstruction methods such as distance approaches, maximum parsimony, and maximum likelihood. Morphological evidence additionally supported the phylogenetic tree based on ITS1 data and the inferred phylogenetic position of the unidentified digeneans of C. carinata met the expectations from known trematode life-cycle patterns. Although ribosomal ITS1 elements are generally believed to be too variable for phylogenetic analysis above the species or genus level, the overall consistency of the results of this study strongly suggests that this is not the case in digenetic trematodes. Here, 3' end ITS1 sequence data seem to provide a valuable tool for elucidating phylogenetic relationships of a broad range of phylogenetically distinct taxa.

Animals↗
PubMed1999

Epipodite and fat cells as sites of hemoglobin synthesis in the branchiopod crustacean Daphnia magna.

In contrast to the malacostracan crustaceans that use hemocyanin as the oxygen carrier, a number of branchiopod crustaceans, such as the water flea Daphnia magna, utilize hemoglobin (Hb) as the respiratory protein. By means of in situ hybridization (ISH) techniques with subsequent signal amplification using catalyzed reporter deposition, sites of Hb synthesis were localized in Daphnia magna. Based on a previously reported Hb-cDNA sequence, a specific ISH probe was designed and hybridized with the Hb-mRNA in histological sections of adult D. magna. The detection of Hb-mRNA was tissue specific and revealed that Hb is synthesized in fat cells, which play a role in fat and glycogen metabolism, and in epithelial cells of the epipodites, which are involved in osmoregulation. Sites of Hb synthesis have been identified in several invertebrate phyla, including Annelida and Nematoda. However, this is the first example in the class Crustacea, and only the second in the phylum Arthropoda.

Adipocytes↗
PubMed2002

Solid and liquid residues as raw materials for biotechnology.

In the past few decades huge amounts of solid and paste-like wastes of domestic and industrial origin have been deposited on sanitary landfills worldwide. Only a small proportion was incinerated, where incineration plants were available. Since primary resources, such as ores for metal production or crude oil for the production of gasoline, diesel, solvents and plastics, or coal and natural gas as sources for energy or chemicals are not available in unlimited quantities, and because the deposition of residues, wastes and worn-out commodities on sanitary landfills causes pollution of the atmosphere, the soil and the groundwater due to hazardous gaseous emissions and toxic leachates, wastes from households and from industry must be avoided or minimized at an early stage. Whenever waste material can be recycled it must be re-introduced into production processes and the non-recyclable fractions should be used as a fuel for energy recovery. After incineration, the highly toxic dust fractions of ashes and slags resulting from burning the wastes should be deposited on sanitary landfills, while the granulated mineral slag fractions could be used as a substitute for the sand in cement as a construction material. Here we review various processes for the treatment of organic fractions of differently composed wastes to upgrade them to more valuable, re-usable products or at least to recover their energy content. Upgrading processes of organic wastes include composting, biogas fermentation, production of organic acids and solvents, and biopolymer or biosurfactants production. We also include biological purification procedures for the most important components of wastes, such as chitin from the shells of Crustaceae. Typical examples from pilot-scale or full-scale studies are discussed for each process.

Biotechnology↗
PubMed2004

' Candidatus Hepatincola porcellionum' gen. nov., sp. nov., a new, stalk-forming lineage of Rickettsiales colonizing the midgut glands of a terrestrial isopod.

The midgut glands (hepatopancreas) of terrestrial isopods are densely colonized by hitherto uncultivated bacteria. In the case of the Common Woodlouse, Porcellio scaber (Crustacea: Isopoda), the symbionts represent a novel lineage in the alpha-subdivision of Proteobacteria. Based on comparative sequence analysis of their 16S rRNA genes, their closest (albeit distant) relatives were among the Rickettsiales, which are intracellular symbionts or pathogens of many animals. Transmission electron microscopy and in situ hybridization with fluorescently labeled oligonucleotide probes revealed a homogeneous population of symbionts intimately associated with the endothelium of the hepatopancreas, which apparently interact with the microvilli of the brush border by means of a stalk-like cytoplasmic appendage. Based on isolated phylogenetic position and unique cytological properties, the provisional name ' Candidatus Hepatincola porcellionum' is proposed to classify this new taxon of Rickettsiales colonizing the hepatopancreas of P. scaber.

Alphaproteobacteria↗
PubMed2004

The GC-rich transposon Bytmar1 from the deep-sea hydrothermal crab, Bythograea thermydron, may encode three transposase isoforms from a single ORF.

Mariner-like elements (MLEs) are classII transposons with highly conserved sequence properties and are widespread in the genome of animal species living in continental environments. We describe here the first full-length MLE found in the genome of a marine crustacean species, the deep-sea hydrothermal crab Bythograea thermydron (Crustacea), named Bytmar1. A comparison of its sequence features with those of the MLEs contained in the genomes of continental species reveals several distinctive characteristics. First, Bytmar1 elements contains an ORF that may encode three transposase isoforms 349, 379, and 398 amino acids (aa) in long. The two biggest proteins are due to the presence of a 30- and 49-aa flag, respectively, at the N-terminal end of the 349-aa cardinal MLE transposase. Their GC contents are also significantly higher than those found in continental MLEs. This feature is mainly due to codon usage in the transposase ORF and directly interferes with the curvature propensities of the Bytmar1 nucleic acid sequence. Such an elevated GC content may interfere with the ability of Bytmar 1 to form an excision complex and, in consequence, with its efficiency to transpose. Finally, the origin of these characteristics and their possible consequences on transposition efficiency are discussed.

Amino Acid Sequence↗
PubMed2003

Acute toxicity of methyl-parathion in wetland mesocosms: assessing the influence of aquatic plants using laboratory testing with Hyalella azteca.

Methyl-parathion (MeP) was introduced into constructed wetlands for the purpose of assessing the importance of distance from the source of contamination and the role of emergent vegetation on the acute toxicity to Hyalella azteca (Crustacea: Amphipoda). A vegetated (90% cover: mainly Juncus effuses) and a nonvegetated wetland (each with a water body of 50 x 5.5 x 0.2 m) were each exposed to a simulated MeP storm runoff event. H. azteca was exposed for 48 h in the laboratory to water samples taken from the wetlands at a distance of 5, 10, 20, and 40 m from the pesticide inlet 3 h, 24 h, 96 h, and 10 days following application. Methyl-parathion was detected throughout the nonvegetated wetland, whereas the pesticide was only transported halfway through the vegetated wetland. A repeated-measure three-way analysis of variance (ANOVA) using time, location, and vegetation indicated significantly lower toxicity in the vegetated wetland. Furthermore, the mortality decreased significantly with both increasing distance from the inlet and time (48-h LC50 +/- 95% CI: 9.0 +/- 0.3 microg/L). A significant three-way interaction of time x vegetation x location confirmed higher toxicity at the inlet area of the nonvegetated wetland immediately after contamination. Significant linear regressions of maximum mortality (independent of time) versus distance from the pesticide inlet indicated that 44 m of vegetated and 111 m of nonvegetated wetland would reduce H. azteca mortality to < or = 5%. These results suggest that vegetation contributes to reduced MeP effects in constructed wetlands.

Amphipoda↗
PubMed2006

Toxicity of three polyethoxylated tallowamine surfactant formulations to laboratory and field collected fairy shrimp, Thamnocephalus platyurus.

Polyethoxylated tallowamine (POEA) is a non-ionic surfactant used in herbicide formulations to increase the efficacy of active ingredients. POEA promotes penetration of herbicide active ingredients into plant cuticles, and in animal species is known to cause alterations in respiratory surfaces. POEA use has increased recently with the advent of "Roundup-Ready" crops; however, its potential effects on aquatic invertebrates are relatively unknown. The aquatic macroinvertebrate Thamnocephalus platyurus (Crustacea, Anostraca) was used to assess the acute toxicity of POEA. Three formulations of POEA consisting of a 5:1, 10:1, and 15:1 average oxide:tallowamine were used in this study. All POEA formulations were found to be extremely toxic to T. platyurus with 48-h LC50 concentrations as low as 2.01 microg/L for 15:1. POEA toxicity increased as the tallowamine chain length was reduced, whereas the oxide chain length appeared to only slightly increase toxicity. Based on these results, POEA has the potential to adversely affect aquatic organisms in areas in which it is used.

Animals↗
PubMed1998

Effects of copper and zinc on embryos and larvae of the horseshoe crab, limulus polyphemus

We investigated the short-term and long-term effects of copper and zinc on horseshoe crab (Limulus polyphemus) embryos and first instar larvae from two estuaries in New Jersey, Delaware Bay and Sandy Hook Bay. Animals were exposed to nominal metal concentrations from 0.01 to 1,000 mg/L for 24 h, 48 h, 72 h, or continuously until the endpoint for the study (successful molting or death) was reached. Larvae showed greater tolerance of Cu and Zn than embryos. Under most treatment regimes, embryos and larvae were able to survive and molt in the presence of up to 100 mg/L Cu or Zn. Unexpectedly, horseshoe crabs from the more heavily polluted site (Sandy Hook Bay) were more susceptible to heavy metals than animals from a less polluted site (lower Delaware Bay). Horseshoe crab embryos and larvae were highly resistant to heavy metals in comparison to marine crustacea. The ability of Limulus embryos and larvae to survive in the presence of heavy metals implies the potential for these contaminants to be passed on to shorebirds and other predators.

Journal Article↗
PubMed1999

Polychlorinated biphenyl- and mercury-associated alterations on benthic invertebrate community structure in a contaminated salt marsh in southeast Georgia.

The community structure of a benthic macroinvertebrate assemblage in a contaminated salt marsh was evaluated as part of an ecological characterization of a former chloralkali production facility in Georgia. Sample locations were chosen based on a gradient of the primary contaminants of concern, total mercury and polychlorinated biphenyls (PCBs), primarily Aroclor 1268. Sediment concentrations of Aroclor 1268 ranged from 2.3 to 150 mg/kg dry weight, while mercury concentrations ranged from 15 to 170 mg/kg dry weight in the study area. Mercury and PCBs were determined to be co-located in the sediments. Total organic carbon composition of the sediments was negatively associated with PCB and mercury concentrations. A total of 29 benthic taxa was identified in 49 samples; replicate samples were taken at each of five sampling locations. Mean infaunal density across all sampling locations was estimated at approximately 61,000 to 234,000 organisms m(-2). Overall, polychaetes comprised 57% of the infaunal community with Manayunkia aestuarina as the dominant species. Oligochaetes, nematodes, crustacea, insects, and gastropods comprised 23.0, 18.0, 1.0, 0.7, and 0.2% of the overall benthic community, respectively. Density estimates of individual species between sampling locations showed no consistent patterns in response to pollutants. However, an analysis of higher taxonomic levels revealed some general trends. In uncontaminated areas, the benthic community was dominated by nematodes and oligochaetes, whereas moderate to highly contaminated areas were dominated by polychaetes and a smaller percentage of oligochaetes and nematodes. A trophic analysis of the same data set revealed that the community shifted from an evenly distributed percentage of surface and subsurface feeders in the uncontaminated areas to a community dominated by surface feeders in the more contaminated locations. Carnivores comprised from 0.13 to 0.90% of the trophic structure, with the percentage of carnivores generally decreasing with increasing contamination. Mercury and PCBs were bioaccumulating in representative marsh benthic invertebrates, presenting a potential source of contaminants to marsh consumers. Tissue PCB and tissue mercury concentrations were positively related to sediment PCB and mercury concentrations, respectively. A standard 14-day toxicity test using the amphipod Leptocheirus plumulosus showed no acute toxicity across the sampling locations.

Animals↗
PubMed2001

Visual interneurons of the crab Chasmagnathus studied by intracellular recordings in vivo.

Comparative physiology of visual systems has become an important field of investigation. However, despite the fact that Crustacea represents a major phylogenetic group, research on the physiology of vision of these animals is scant and almost limited to the crayfish. We developed a preparation to study in vivo the visual nervous system of a semiterrestrial crab through intracellular recordings. The response to a pulse of light was investigated in 206 interneurons from 38 animals. Seventy-eight of these neurons could be classified by functional criteria as sustaining cells, dimming cells, nonspiking hyperpolarizing cells and nonspiking depolarizing cells. Quantitative description is provided for the first two groups and qualitative description is given for the last two. The remaining neurons presented a broad range of different types of phasic responses to light. Although semiterrestrial crabs are behaviorally more reactive to visual stimuli than the crayfish, the general physiological properties of identified lamina and medullary neurons of Chasmagnathus resemble those of the crayfish. The results described here represent the first attempt to study the visual system of crabs with intracellular recordings and constitute the beginning of a project aimed to investigate the neuronal functions underlying behavioral responses elicited by visual stimuli.

Animals↗
PubMed2002

Origin and evolution of arthropod hemocyanins and related proteins.

Arthropod hemocyanins are large, multimeric, (n x 6) copper-containing proteins that deliver oxygen in the haemolymph of many chelicerate, crustacean, myriapod, and also possibly some insect species. The arthropod hemocyanins belong to a large protein superfamily that also includes the arthropod phenoloxidases, certain crustacean and insect storage proteins (pseudo-hemocyanins and hexamerins), and the insect hexamerin receptors. Here I summarise the present knowledge of the origin, functional adaptations, and evolution of these proteins. Arthropod and mollusc hemocyanins are, if at all, only distantly related. As early as in the arthropod stem line, the hemocyanins emerged from a phenoloxidase-like enzyme. The evolution of distinct hemocyanin subunits, as well as the formation of multi-hexamers occurred independently within the arthropod subphyla. Hemocyanin subunit evolution is strikingly different in the Chelicerata, Myriapoda and Crustacea. Hemocyanins individually gave rise to two distinct copper-less storage proteins, the insect hexamerins and the crustacean pseudo-hemocyanins (cryptocyanins). The receptor responsible for the uptake of hexamerin by the larval fat body of the insects emerged from a hexamerin-precursor. Molecular phylogenetic analyses show a close relationship of the crustacean and insect proteins, providing strong support for a pancrustacean taxon, while structural data suggest a myriapod-chelicerate clade.

Animals↗
PubMed2002

Photopigment gene expression and rhabdom formation in the crayfish (Procambarus clarkii).

This study examines the expression of the photopigment gene in the developing retina of the freshwater crayfish Procambarus clarkii(Crustacea, Malacostraca, Decapoda). Both sense and anti-sense RNA probes were used for in situ hybridization (ISH) of whole embryos collected at various stages during development. A characteristic of retinal development is the formation of screening pigment in the retinular cells of the retinal ommatidia. This pigmentation is seen as a band that begins at the lateral side of the retinal field and progresses medially. At hatching the retina is approximately 50% pigmented. ISH of whole embryos shows that expression of the photopigment gene by the retinular cells correlates with the extent of the screening pigment band in the retina and with the presence of rhabdoms within the ommatidia. Sections taken through embryos after being hybridized indicate that staining is localized in the cytoplasm of the retinular cells and in the axonal region below the basement membrane. No staining reaction was seen in the rhabdoms of older ommatidia. ISH staining was also seen at the anterior midline of the protocerebrum where extraretinal photoreceptors have been reported. The data presented here show a close correlation of opsin expression within the retinular cells of the ommatidia and the formation of the very early rhabdoms, similar to Drosophila. The results will be discussed in relation to recent studies in Drosophila that suggest rhodopsin plays a role in effecting the organization of the terminal web-like cytoskeleton at the base of the developing rhabdom microvilli.

Animals↗
PubMed2004

Incidence of diapause varies among populations of Daphnia pulicaria.

Dormancy is a common way in which organisms survive environmental conditions that would be lethal to the active individual. However, while dormant, individuals forego reproduction. Hence theory suggests an optimal time in which to enter dormancy, depending on risks associated with both remaining active and entering dormancy. When these relative risks differ among habitats, dormancy strategies are predicted to vary as well. For freshwater zooplankton, it has been suggested that sensitivity to the cues that initiate dormancy should be selected against when females have the opportunity to remain in the water column year round. We tested this prediction with 12 populations of lake-dwelling Daphnia pulicaria (Crustacea: Cladocera). Differences among lakes in basin morphometry, predators and resources create a gradient of risk for Daphnia in the water column. Some populations persist in high numbers year round while others are abundant only in spring. We used this difference in persistence ability as an estimate of risk in the water-column. For 3 years of field sampling we found consistent differences among the lake populations in the incidence of dormancy. In some populations, only a small fraction of females switched to producing dormant eggs each year whereas in others the majority of eggs produced in the late spring were dormant. In general, populations that experienced predictably low abundances in the active form exhibited higher incidence of dormancy than did populations that persisted in high abundance year round, but there were exceptions. Our results confirm that the incidence of dormancy varies considerably among populations in a fashion consistent with general theory, but suggest that persistence in the water column is not the sole predictor of the diapause strategy found in any particular lake.

Adaptation, Physiological↗
PubMedDate not supplied

Seafood allergy and allergens: a review.

Seafoods are composed of diverse sea organisms and humans are allergic to many of them. Tropomyosin is a major allergen in many shellfish, especially crustacea and mollusks. Interestingly, tropomyosin has also been identified as an important allergen in other invertebrates including dust mites and cockroaches, and it has been proposed by some to be an invertebrate pan allergen. Different regions of shrimp tropomyosin bind IgE; 5 major IgE-binding regions have been identified in shrimp tropomyosin containing 8 epitopes. Mutations of these shrimp allergenic epitopes can reduce seafood allergenicity; methods utilizing such mutations will provide safer vaccines for more effective treatment of seafood-allergic patients, and in the future less-allergenic seafood products for consumption.

Allergens↗
PubMed2004

PCR-based approach for sequencing mitochondrial genomes of decapod crustaceans, with a practical example from kuruma prawn (Marsupenaeus japonicus).

An approach for sequencing the entire mitochondrial genomes (mitogenomes) of decapod crustaceans using 79 newly designed and 7 published polymerase chain reaction (PCR) primers is described. The approach comprises the following steps: (1) the entire mitogenome is amplified in 2 or 3 long PCRs; (2) the 86 primers are used in different combinations to amplify contiguous, overlapping short segments of the entire mitogenome with the diluted long PCR products as templates; (3) direct cycle sequencing is conducted using the short PCR products. This strategy allows a more rapid determination of decapod mitogenomic sequences than a traditional method using cloned mitochondrial DNA and primer walking strategy. As a practical example, the mitogenomic sequence for a kuruma prawn Marsupenaeus japonicus (Crustacea: Decapoda), was determined using the PCR-based approach.

Animals↗
PubMed2005

Toxicity of organic compounds to marine invertebrate embryos and larvae: a comparison between the sea urchin embryogenesis bioassay and alternative test species.

This study investigated the toxic effects of the insecticides lindane and chlorpyrifos, the herbicide diuron, the organometallic antifoulant tributyltin (TBT), and the surfactant sodium dodecyl sulfate (SDS) on the early life stages of Paracentrotus lividus (Echinodermata, Euechinoidea), Ciona intestinalis (Chordata, Ascidiacea), Maja squinado and Palaemon serratus (Arthropoda, Crustacea) in laboratory acute toxicity tests. The assays studied embryogenesis success from fertilized egg to normal larvae in P. lividus (48 h incubation at 20 degrees C) and C. intestinalis (24 h incubation at 20 degrees C), and larval mortality at 24 and 48 h in M. squinado and P. serratus. For P. lividus, the median effective concentrations (EC50) reducing percentages of normal larvae by 50% were: 350 microg l(-1) for chlorpyrifos, 5500 microg l(-1) for diuron, 4277 microg l(-1) for SDS, and 0.309 microg l(-1) for TBT. For C. intestinalis, the EC50 values affecting embryogenesis success were 5666 microg l(-1) for chlorpyrifos, 24,397 microg (l-1) for diuron, 4412 microg l(-1) for lindane, 5145 microg I(-1) for SDS, and 7.1 microg l(-1) for TBT. The median lethal concentrations (LC50) for M. squinado larval survival were 0.84 microg l(-1) (24 h) and 0.79 microg l(-1) (48 h) for chlorpyrifos, 2.23 microg(l(-1) (24 h) and 2.18 microg l(-1) (48 h) for lindane, and 687 microg l(-1) (48 h) for SDS. For P. serratus the LC50 values obtained were 0.35 microg l(-1) (24 h) and 0.22 microg l(-1) (48 h) for chlorpyrifos, 3011 microg l(-1) (24 h) and 3044 microg l(-1) (48 h) for diuron, 5.20 microg l(-1) (24 h) and 5.59 microg l(-1) (48 h) for lindane, and 22.30 microg l(-1) (24 h) and 17.52 microg l(-1) (48 h) for TBT. Decapod larvae, as expected, were markedly more sensitive to the insecticides than sea urchins and ascidians, and SDS was the least toxic compound tested for these organisms. Lowest observed effect concentrations (LOEC) of TBT for sea urchin and ascidian embryos, chlorpyrifos and lindane for crustacean larvae, and SDS, were similar to those found in many coastal areas indicating that there would be a risk to invertebrate embryos and larvae from exposure in the field to these pollutants.

Animals↗
PubMed2005

Water quality assessment by ecotoxicological and limnological methods in water supplies, southeast Brazil.

The quality of the water supplied to Araraquara city (São Paulo State, Brazil) was analyzed at four points: two reservoirs from which water is pumped and the water incoming and exiting a drinking-water treatment plant. Physical and chemical analyses of water were performed, as well as acute and chronic toxicity tests of the water from all four points, using Ceriodaphnia silvestrii (Cladocera, Crustacea) as test-organism. The study was carried out monthly in the period between December 2000 and November 2001. Water from the reservoirs caused neither acute nor chronic toxicity to test-organisms. On the contrary, it promoted an increase in the fecundity of cladocerans compared to control, suggesting greater availability of food. However, the water from exiting the treatment plant did cause chronic toxicity to C. silvestrii, regarding both fecundity and mortality. The toxicity was probably caused by residual chlorine, perhaps not completely removed by aeration, or by chlorine disinfection by-products (trihalomethanes) in the treated water, considering the level of organic matter present in the reservoir water.

Animals↗
PubMed2006

Ecotoxicological assessment of effluents in the Basque country (Northern Spain) by acute and chronic toxicity tests using Daphnia magna straus.

Acute pass/fail, multi-concentration tests, and 3-brood chronic toxicity tests with Daphnia magna Straus (Cladocera, Crustacea) were used to characterise industrial and municipal effluents from various sources. The effluents that "passed" the pass/fail tests had 48-h EC50 values >100% and reproduction No Observed Effect Concentration (NOECs) > or =100%, except for one effluent that had a reproduction NOEC of 31.6%. The acute multi-concentration toxicity tests allowed a rapid classification of effluents from Very Toxic (48-h EC50 < 25%), to Non-Toxic (48-h EC50 >100%). The acute-to-chronic ratio (ACR: 48-h EC50 divided by the NOEC for reproduction) in the studied effluents ranged from 5 to about 100. From these results, we propose a step-wise protocol for assessing effluent toxicity. First, effluent is evaluated by means of simple and rapid pass/fail acute toxicity tests, to discriminate Non-Toxic from potentially Toxic effluents, thus facilitating the establishment of priority actions. Second, 48-h ECx is estimated to classify effluents on a toxicity scale from Non-Toxic to Very Toxic. Third, chronic multi-concentration tests are used to calculate reproduction NOECs. These parameters combined with data on effluent chemical composition, chemical and hydrological characteristics of receiving waters, and biological quality criteria can be jointly used for more rational regulatory practices and risk assessment of effluents.

Animals↗