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[Chromosomal radiosensitivity as associated with chromatin diminution in cyclops (Crustacea, Copepoda)].

Chromosomal radiosensitivity inferred from the yield of chromosome aberrations (CAs) was for the first time studied in Cyclops (Crustacea, Copepoda) before and after chromatin diminution (CD). A comparison was made for C. kolensis, in which CD denudes somatic embryo cells of the greatest (94%) DNA amount known for multicellular organisms, and C. insignis, which lacks CD. The two species have similar genome sizes, 4.6 and 4.3 pg. respectively. Radiosensitivity of C. kolensis chromosomes proved to be extremely high during prediminution cleavage divisions. This was attributed to membrane damage in granules that contain enzymes (topoisomerases) normally involved in cleavage and ligation of chromosomal DNA during CD.

Anaphase↗

Analysis and management of resistance to chemotherapeutants in salmon lice, Lepeophtheirus salmonis (Copepoda: Caligidae).

In Northern Europe and Canada, the salmon louse, Lepeophtheirus salmonis (Krøyer), seriously affects the marine phase of salmon production. Although the problem is long-standing, the development of sustainable methods of pest management has been unable to keep pace with the intensification of production, leading to large-scale reliance on very few chemotherapeutants. This runs the risk of selecting for genetically determined resistance in target organisms. There are many examples of similar evolutionary adaptations in arthropod pests of arable crops, livestock and human health. Several hundred pest species are now documented as being resistant to one or more chemical classes of insecticides and acaricides. Many of these compounds are identical or closely related to ones currently employed against salmon lice. It is, therefore, opportune to consider what lessons have been learnt from contending with resistance in terrestrial organisms, the implications for sustainable use of chemotherapeutants in aquaculture, and the potential for developing effective resistance management strategies. An EU-funded project named SEARCH (QLK2-CT-2000-00809) has been initiated to explore in more detail the diagnosis, incidence, dynamics and management of resistance to chemotherapeutants in L salmonis.

Animals↗

Molecular cloning of trypsin cDNAs and trypsin gene expression in the salmon louse Lepeophtheirus salmonis (Copepoda: Caligidae).

The salmon louse, Lepeophtheirus salmonis, is a marine ectoparasitic copepod that infects salmonid fishes. We are studying the interactions between this parasite and its salmonid hosts, as it is a common cause of disease in both wild and farmed stocks of salmon. In this paper, we report on the cloning and sequencing of seven trypsin-like enzymes from a cDNA library prepared from whole body preadult female and male L. salmonis. The predicted trypsin activation peptides are 23 or 24 residues in length, considerably longer than previously reported activation peptides of other animals. Differences in the putative signal and activation peptide sequences of the trypsin isoforms suggest that these forms differ in their regulation and function. The calculated molecular weights of the trypsins range from 23.6 to 23.7 kDa. There are eight cysteine residues, which suggest the presence of four disulfide bridges. These trypsins are very similar (>or=46% aa identity) to other crustacean trypsins and insect hypodermins. Using in situ hybridization techniques trypsinogen expression could be identified in all three cell types of the midgut.

Amino Acid Sequence↗

Co-occurrence of Naobranchia lizae (Copepoda) and Metamicrocotyla macracantha (Monogenea), gill parasites of the striped mullet Mugil cephalus.

The copepod Naobranchia lizae (Kroyer, 1863) and the monogenean Metamicrocotyla macracantha (Alexander, 1954; Koratha, 1955) are gill parasites found on the striped mullet (Mugil cephalus) in the Charleston Harbor Estuarine System (South Carolina, USA). Branchial distribution of each species was studied in mono- and bispecific conditions. No preference for the right or left side of the fish host was observed for either species in either condition. Both species exhibited heterogeneous distribution among the gill arches, with a preference for arch I. N. lizae exhibited intraspecific competition and a saturation threshold in both mono- and bispecific conditions. A shift in niche toward the posterior arches was observed for M. macracantha only in bispecific infection. Interspecific competition was detected exclusively on the posterior arches, where M. macracantha seemed out-competed by N. lizae. The data indicated that both neutral and negative interactions played a role in determining the distribution of N. lizae and M. macracantha individuals on the gill arches of M. cephalus.

Animals↗

Three new species of Enterocola van Beneden, 1860 (Crustacea: Copepoda: Cyclopoida) from ascidian hosts.

Three new species of Enterocola are described: E. dicaudatus n. sp., from the ascidian host Polysyncraton rostrum Monniot & Monniot collected in Tanzania, E. monnioti n. sp., from the ascidian host Polyclinum constellatum Savigny collected in Bahrain, and E. parapterophorus n. sp., from the ascidian host Didemnum granulatum Tokioka taken in Djibouti. In all three species, the mandibles are interpreted as being represented by a pair of setulose seta-like elements that are largely concealed beneath similar paired setulose structures, the labral palps, carried on the labrum. The paired elements interpreted as mandibles are transversely orientated and lie in a groove, the anterior margin of which forms a defined ridge ornamented with spinules.

Animals↗

A new genus and species of nicothoid copepod (Crustacea: Copepoda: Siphonostomatoida) parasitic on the mysid Siriella okadai Ii from off Japan.

Neomysidion rahotsu, a new genus and species of the siphonostomatoid family Nicothoidae Dana, 1849, is described from the marsupium of the mysid Siriella okadai Ii in the shallow waters of the Seto Inland Sea of Japan. This is the first record of nicothoids parasitic on mysids in Asian waters. The new genus is closely related to the three other mysid-infesting nicothoid genera, Aspidoecia Giard & Bonnier, 1889, Mysidion Hansen, 1897 and Hansenulus Heron & Damkaer, 1986, but is distinguishable by: the trunk of adult females being remarkably transformed at maturity; the male cephalosome covered with numerous rounded knobs posterodorsally; the absence of caudal rami in the female; three-segmented antennules in both sexes; two-segmented antenna in the male; remarkable sexual dimorphism in the maxillae; the presence of a pair of spinulate sclerites between the maxillae in both sexes; three- segmented maxillipeds in both sexes; the absence of legs 1 to 5 in the female; the presence of two pairs of legs in the male; and egg-sacs laid free.

Anatomy, Comparative↗

Lamproglena hepseti n. sp. (Copepoda: Lernaeidae), a gill parasite of the African pike Hepsetus odoe (Bloch) from the Okavango River and Delta, Botswana.

During surveys of the biodiversity of fish parasites in the Okavango River and Delta, Botswana, specimens of Lamproglena von Nordmann, 1832 were found associated with the African pike Hepsetus odoe (Bloch). This Lamproglena species distinctly differs from all known species based on morphological features, in particular the cephalothorax and the maxilliped; it is described as L. hepseti n. sp. and is specific to its host, the African pike.

Animals↗

A new species of Prohatschekia Nunes-Ruivo, 1954 (Copepoda: Hatschekiidae) parasitic on Scorpaena elongata (Cadenat) off Algeria.

A new species of parasitic copepod, Prohatschekia mediterranea n. sp. (Siphonostomatoida: Hatschekiidae), is described from a scorpaenid fish, Scorpaena elongata, collected off Algeria. This is the seventh species of Prohatschekia Nunes-Ruivo, 1954 to be described and the first record of the genus from the Mediterranean Sea. The new species is most closely related to P. cremouxi Nunes-Ruivo, 1954, known from a congeneric host collected in Senegal. A key is provided to distinguish the new species from other members of the genus.

Algeria↗

Effects of salinity and temperature on the expression of enzymatic biomarkers in Eurytemora affinis (Calanoida, Copepoda).

In order to establish effective enzymatic biomarkers that could provide in situ early warning of contaminant exposure in estuarine ecosystems, the potential effects of the principal abiotic factors (temperature and salinity) were investigated on common biomarkers, the acetylcholinesterase (AChE) and the glutathione S-transferase (GST) in Eurytemora affinis. Short term salinity stress effects simulated during an experimental tide indicated that enzymatic activities of this species are characterized by maximum expression related to an optimal salinity range (between 5 and 15 psu). Moreover, longer time exposure to various salinity tanks confirmed the effects of this factor on both AChE and GST activities. Therefore, optimal AChE activity was measured at 10 psu, while optimal GST activity was measured at 5 psu. Furthermore, significant effects of temperature were also recorded, particularly for AChE expression (slight effects were measured on GST expression) with an optimal condition at 11 degrees C. These experiments indicated a more pronounced effect of salinity over temperature especially on the AChE expression and confirmed the need to standardize sampling procedures in relation with environmental parameters for biomonitoring studies based on enzymatic analyses.

Acetylcholinesterase↗

Molecular characterization and classification of a clip domain containing peptidase from the ectoparasite Lepeophtheirus salmonis (Copepoda, Crustacea).

Clip domain containing serine peptidases (CSPs) include one or more N-terminal clip domain(s) and a C-terminal serine peptidase domain that shares traits with both chymotrypsin and trypsin. CSPs are found in arthropods and are involved in embryonic patterning, immune responses and blood clotting. Among crustaceans only one CSP, which activates prophenoloxidase in crayfish, have previously been reported. We here present LsCSP1, the first CSP found in copepods. LsCSP1 is expressed in the subcuticular tissue and the transcription appears to be upregulated during development. In conjunction with previous studies of CSPs, this study suggests that LsCSP1 may play a role in the immune responses of L. salmonis. Phylogenetic and structural analyses indicate that the CSPs and catalytically inactive CSP homologs (CSPHs) constitute a monophyletic lineage.

Amino Acid Sequence↗

Organisation of trypsin genes in the salmon louse (Lepeophtheirus salmonis, Crustacea, copepoda) genome.

Trypsins constitute a subclass of the S1A family of serine peptidases found in all groups of animal and some bacteria. At present, no information about the genomic organisation of trypsins is available for copepods. The only data of copepod trypsins indicate several different trypsins in the marine parasitic copepod Lepeophtheirus salmonis. In the present study, 31.7 kbp of genomic DNA surrounding the previously described LsTryp1-5 sequences was sequenced. The sequenced regions contain nine full-length and three partial trypsin genes. A conservative estimate based on PCR analysis and genomic sequence indicated at least 22 different trypsin genes in L. salmonis, of which 18 are most similar to the previously described LsTryp1 and -2 cDNA sequences. Four of these genes are putative pseudogenes. In addition, a putative mariner like transposase gene was identified. The genomic sequences suggest that the L. salmonis trypsin genes reside within one or more gene clusters. Three different LsTryp intron exon structures were identified, and all three are different from the intron exon organisation previously reported for other S1A peptidases. This implies several intron loss and gain events in the evolution of the L. salmonis trypsin genes.

Animals↗

Genetic characterization of the mitochondrial DNA from Lepeophtheirus salmonis (Crustacea; Copepoda). A new gene organization revealed.

The mitochondrial DNA (mtDNA) from the salmon louse, Lepeophtheirus salmonis, is 15445 bp. It includes the genes coding for cytochrome B (Cyt B), ATPase subunit 6 and 8 (A6 and A8), NADH dehydrogenase subunits 1-6 and 4L (ND1, ND2, ND3, ND4, ND4L, ND5 and ND6), cytochrome c oxidase subunits I-III (COI, COII and COIII), two rRNA genes (12S rRNA and 16S rRNA) and 22 tRNAs. Two copies of tRNA-Lys are present in the mtDNA of L. salmonis, while tRNA-Cys was not identified. Both DNA strands contain coding regions in the salmon louse, in contrast to the other copepod characterized Tigriopus japonicus, but only a few genes overlap. In vertebrates, ND4 and ND4L are transcribed as one bicistronic mRNA, and are therefore localized together. The same organization is also found in crustaceans, with the exceptions of T. japonicus, Neocalanus cristatus and L. salmonis that deviate from this pattern. Another exception of the L. salmonis mtDNA is that A6 and A8 do not overlap, but are separated by several genes. The protein-coding genes have a bias towards AT-rich codons. The mitochondrial gene order in L. salmonis differs significantly from the copepods T. japonicus, Eucalanus bungii, N. cristatus and the other 13 crustaceans previously characterized. Furthermore, the mitochondrial rRNA genes are encoded on opposite strands in L. salmonis. This has not been found in any other arthropods, but has been reported in two starfish species. In a phylogenetic analysis, using an alignment of mitochondrial protein sequences, L. salmonis groups together with T. japonicus, being distant relatives to the other crustaceans.

Animals↗

Epizootiological studies of Amblyospora camposi (Microsporidia: Amblyosporidae) in Culex renatoi (Diptera: Culicidae) and Paracyclops fimbriatus fimbriatus (Copepoda: Cyclopidae) in a bromeliad habitat.

The epizootiology of Amblyospora camposi was studied in a natural population of Culex renatoi, a bromeliad-inhabiting mosquito, and its intermediate host, Paracyclops fimbriatus fimbriatus, over a 2-year period. Twenty Eryngium cabrerae plants were sampled monthly from January 2003 to January 2005 and the prevalence of A. camposi in P.f. fimbriatus and Cx. renatoi populations was determined. The monthly prevalence rates of meiospore infections in Cx. renatoi larvae never exceeded 5.5% and was detected in 50% of the monthly samples. Meiospores were available in plants over the course of the study at a mean concentration of 2 x 10(4) meiospores/ml. Within each plant the parasite was maintained by horizontal transmission. P.f. fimbriatus with vegetative stages and mature spores were found regularly in bromeliads suggesting efficient meiospore infectivity to field copepod populations. The mean concentration of spores from copepods found in plants was 8 x 10(2) spores/ml. Infections in copepods were detected in 54% of the monthly samples with a prevalence rate ranging from 0.55 to 17.4% and an overall average of 5.1%. Vegetative stages in fourth instar mosquito larvae (probably derived from the horizontal pathway via spores formed in copepods) were detected in 12.5% of the monthly samples with an overall prevalence rate of 1.1%. Infections in female and male adults were detected in 20.8% of the monthly samples with an overall average of 4.1% and 6.8%, respectively.

Amblyospora↗

The effects of power station entrainment passage on three species of marine planktonic crustacean, Acartia tonsa (Copepoda), Crangon crangon (Decapoda) and Homarus gammarus (Decapoda).

Experiments have been undertaken exposing larval common shrimp (Crangon crangon) and lobster (Homarus gammarus) and adult copepods (Acartia tonsa) to the key stresses of entrainment within power-station cooling-water systems. The apparatus has enabled the testing of mechanical, thermal, chlorine and realistic pressure effects both alone and in combination, the range of stressors spanning the standard conditions found within a temperate coastal direct-cooled power station. Mechanical stresses affected only lobster larvae, pressure changes affected only the Acartia adults. Residual chlorine caused significant mortality of Acartia and shrimp larvae, but had no effect on lobster larvae even at 1 ppm. The temperature increment significantly affected all three species, with a synergistic effect on chlorine sensitivity in the shrimp larvae, but only temperatures higher than would be experienced in a normally-operating power station affected the copepods. The majority of individuals of each species would survive passage through a power-station system under normal conditions. It is notable that, within the species tested, generalizations from the responses of one species to those of another are not valid.

Animals↗

Analysis of 686 expressed sequence tags (ESTs) from the intertidal harpacticoid copepod Tigriopus japonicus (Crustacea, Copepoda).

The intertidal harpacticoid copepod Tigriopus japonicus is an important species in the study of marine pollution. To facilitate molecular biomonitoring using T. japonicus, we constructed a T. japonicus unidirectional cDNA library using lambdaZAP expression vector, excised to pBluescript vector with the aid of helper phage, and analyzed 686 randomly picked expressed sequence tags (ESTs) from this species. From the 686 ESTs sequenced, we found several functional genes such as vitellin, kinases and potential detoxification-related genes. We are now preparing a T. japonicus cDNA chip for molecular ecotoxicological studies. In this paper, we discuss the potential use of T. japonicus ESTs and their importance in ecotoxicology.

Animals↗