Oligochaetes and eutrophication; an experiment over four years in outdoor mesocosms

Eight experimental ditch mesocosms were used to study the effect of eutrophication over four years. The experimental ditches had a sand or clay bottom. The ditches were treated with additions of phosphorus, phosphorus and nitrogen, or without additions (controls). Oligochaetes were sampled by deploying trays with substratum for colonization over twenty weeks. Both the important variables phosphorus, nitrogen and oxygen as well as the oligochaete species and numbers are presented. The effects of nutrient additions on phosphorus, nitrogen and oxygen concentrations were described together with changes in oligochaete species composition and numbers. The results were further analyzed by redundancy analysis (RDA). In the clay-lined ditches nutrient addition coincided with fluctuation in oxygen concentration. The higher the nutrient addition levels the longer the period of oxygen depletion became. During oxygen depletion the number of oligochaetes was strongly reduced or even became zero. The low nutrient status of the sandy bed in the sand-lined ditches slowed down the rate of colonization. Only a few tubificids were collected. Eutrophication effects were only observed at the highest nutrient addition level. Considerable variation is attributed to stochastic factors in the sand-lined ditches. Whether oligochaete species were present was related to the length of the colonization period. The substratum composition and food together with oxygen regime decided whether they become more or less abundant in ditches. Large-scale mesocosm experiments require time to develop. Only after the first colonization period variables of species presences and abundances can be employed to detect changes associated with eutrophication. Oligochaetes can be used to measure colonization as well as eutrophication processes.     

Contributions of adult oligochaete emigration and immigration in a dynamic soft-sediment community

It is well known that adult dispersal is common in soft bottom intertidal and shallow subtidal communities. We here report on the first study that attempts to quantify the effects of both immigration and emigration on patches of soft sediment communities. Some species show adaptive emigration from the seabed, although dispersal direction, distance, and colonization success are probably strongly dependent on hydrodynamics, morphological adaptations to dispersal, and the ability to select appropriate target microsites. The naid oligochaete Paranais litoralis is a numerically dominant benthic species in southern New England and New York mud flats and tends to reproduce mainly or exclusively by means of budding of new individuals. When population density is high and resources in short supply, budding frequency is reduced, worms grow longer, and may emigrate from the sediment. We quantified emigration by means of a conical trap and quantified immigration with sediment dishes. We followed emigration/immigration during the typical late spring population explosion and crash cycle of worms within the sediment, which is driven by a seasonal cycle of provision and exhaustion of organic detrital food supply. Emigration was proportionally maximal either at or after the population peak, consistent with a response to food shortage. Over a span of ca. 50 m, we found no net movement in either direction along a transect, nor was emigration or immigration correlated with local density in the sediment. Nevertheless, both emigration and immigration were important in our 2004 sampling, and immigration especially had an important impact on population densities. We do not know the relative capture efficiencies of the emigration and immigration apparatus, so more needs to be done to understand the impacts of dispersal in this and other systems.     

Glutamine synthetase gene expression during the regeneration of the annelid Enchytraeus japonensis

Enchytraeus japonensis is a highly regenerative oligochaete annelid that can regenerate a complete individual from a small body fragment in 4–5 days. In our previous study, we performed complementary deoxyribonucleic acid subtraction cloning to isolate genes that are upregulated during E. japonensis regeneration and identified glutamine synthetase (gs) as one of the most abundantly expressed genes during this process. In the present study, we show that the full-length sequence of E. japonensis glutamine synthetase (EjGS), which is the first reported annelid glutamine synthetase, is highly similar to other known class II glutamine synthetases. EjGS shows a 61–71% overall amino acid sequence identity with its counterparts in various other animal species, including Drosophila and mouse. We performed detailed expression analysis by in situ hybridization and reveal that strong gs expression occurs in the blastemal regions of regenerating E. japonensis soon after amputation. gs expression was detectable at the cell layer covering the wound and was found to persist in the epidermal cells during the formation and elongation of the blastema. Furthermore, in the elongated blastema, gs expression was detectable also in the presumptive regions of the brain, ventral nerve cord, and stomodeum. In the fully formed intact head, gs expression was also evident in the prostomium, brain, the anterior end of the ventral nerve cord, the epithelium of buccal and pharyngeal cavities, the pharyngeal pad, and in the esophageal appendages. In intact E. japonensis tails, gs expression was found in the growth zone in actively growing worms but not in full-grown individuals. In the nonblastemal regions of regenerating fragments and in intact worms, gs expression was also detected in the nephridia, chloragocytes, gut epithelium, epidermis, spermatids, and oocytes. These results suggest that EjGS may play roles in regeneration, nerve function, cell proliferation, nitrogenous waste excretion, macromolecule synthesis, and gametogenesis.     

Expression pattern of annelid <Emphasis Type="Italic">Zic</Emphasis> in embryonic development of the oligochaete <Emphasis Type="Italic">Tubifex tubifex</Emphasis>

Embryonic expression of a Zic homologue (Ttu-Zic) was examined in the oligochaete annelid Tubifex tubifex. The body plan of T. tubifex is characterized by obvious segmentation in the ectoderm and mesoderm. Ttu-Zic expression is detected in the mesodermal germ band and a subset of micromere descendants. Ttu-Zic is transiently expressed in primary m-blast cells (i.e., founder cells of mesodermal segments) as early as the time of their birth from M teloblasts. During its development, each mesodermal segment experiences two additional phases of Ttu-Zic expression. Ttu-Zic expression in micromere descendants is seen on the anterior surfaces of embryos undergoing teloblastogenesis; subsequently, these cells proliferate to form bilateral clusters, which then become internalized. Finally, clusters of Ttu-Zic-expressing cells are found in the center of the prostomium, corresponding to the cerebral ganglion. The Ttu-Zic expression profile in the early embryogenesis of T. tubifex may be homologous to those of evolutionarily distant animals.     

Some new species of Haplotaxidae (Oligochaeta) from Guinea and remarks on the history of the family

Three new species of Haplotaxidae collected by Dr. A. Villiers in a cave near Kindia, Guinea, are described. One species belongs to the genus Metataxis Righi, 1985 (M. carnivorus sp. nov.) which is distributed in Western Africa and Eastern America. Another new species is similar to Lycodrilus kraepelini Michaelsen, 1914 incertae sedis: a new genus Villiersia is proposed for both species (V. guanivora sp. nov., V. kraepelini nov. comb.).A third species, of which only immature specimens are available, is tentatively attributed to the genus Haplotaxis: it is remarkable because of its peculiar setae and commissural vessels (H. villiersi sp. nov.).The three species appear to feed on bat guano and, occasionally, on aquatic arthropods. Metataxis, Tiguassu and Villiersia belong to the section of Haplotaxidae endowed with paired sperm sacs and ovisacs.The relations of Haplotaxidae with other families are discussed and it is suggested that the passage from the haplotaxid condition to that of sparganophilids and more advanced lumbriciforms is connected to a change of reproductive strategy and to an enhanced metabolism.     

The effects of FMRFamide, serotonin, and acetylcholine on the isolated crop-gizzard of the earthworm, Lumbricus terrestris

We examined the effects of FMRFamide, serotonin, and acetylcholine on the isolated crop-gizzard of the earthworm, Lumbricus terrestris. FMRFamide caused a decrease in contraction amplitude with a threshold between 10⁻⁹ and 10⁻⁸ M and a biphasic change in contraction rate. The contraction rate increased with a threshold between 10⁻⁸ and 10⁻⁷ M and decreased with a threshold between 10⁻⁶ and 10⁻⁵ M. Serotonin decreased the contraction rate with a threshold between 10⁻⁸ and 10⁻⁷ M and the amplitude with a threshold between 10⁻⁸ and 10⁻⁷ M. Acetylcholine increased the contraction rate with a threshold between 10⁻⁸ and 10⁻⁷ M and caused a biphasic change in contraction amplitude. Amplitude rose with a threshold between 10⁻⁹ and 10⁻⁸ M and decreased with a threshold between 10⁻⁶ and 10⁻⁵ M. These results suggest that all three neurotransmitters may play a role in controlling the movement of the digestive tract in L. terrestris.     

Cell dynamics during cocoon secretion in the aquatic leech, Theromyzon tessulatum (Annelida: Clitellata: Glossiphoniidae)

One distinguishing feature of clitellate annelids is the presence of specialized segments comprising the clitellum, whose primary function is to secrete a cocoon. Using histological analyses, we have documented cell types (I-V) and cellular processes associated with cocoon secretion in the aquatic leech, Theromyzon tessulatum. Our data indicate that the bulk of the cocoon's biomass arises from precursor cells of a single type that hypertrophy and proliferate ∼1 week prior to egg laying, and then differentiate into either of two cell types (i.e., Type II or Type III) depending on their position within the clitellum. Type II cells are concentrated along the lateral edges and venter of the clitellum and secrete alcian blue-staining granules that form opercula (i.e., glue-like material that seals both cocoon ends), while Type III cells populate the dorsal midline and secrete azocarmine-staining granules that build the cocoon wall. Both cell types occupy spaces between deep muscle layers and extend long-neck tubules to the surface epithelium as they fill with granules a few days prior to egg laying. Other cell types appear to make minor contributions to the cocoon (e.g., Type I, Type IV) or have supporting or signaling roles (e.g., Type V). Our observations suggest that post-translational modification (i.e., glycosylation) of the same core protein(s) distinguishes the granules of Type II/III cells, and that the default state of the Type II/III precursor may be evolutionarily linked to secretory cells in basal polychaetes.     

Regeneration and asexual reproduction share common molecular changes: upregulation of a neural glycoepitope during morphallaxis in Lumbriculus

Neural morphallaxis is a regenerative process characterized by wide-spread anatomical and physiological changes in an adult nervous system. During segmental regeneration of the annelid worm, Lumbriculus variegatus, neural morphallaxis involved a reorganization of sensory, interneuronal, and motor systems as posterior fragments gained a more anterior body position. A monoclonal antibody, Lan 3-2, which labels a neural glyco-domain in the leech, was reactive in Lumbriculus. In the worm, this antibody labeled neural structures, particularly axonal tracts and giant fiber pathways of the central nervous system. A 60kDa protein, possessing a lumbriculid mannose-rich glycoepitope, was upregulated during neural morphallaxis, peaking in its expression at 3 weeks post-amputation. Peak upregulation of the Lan 3-2 epitope, or the protein possessing it, corresponded to the time of major neurobehavioral plasticity during regeneration. Analyses of asexually reproducing animals also revealed induction of the Lan 3-2 epitope. In this developmental context, Lan 3-2 epitope upregulation was also confined to segments expressing both changes in positional identity and neurobehavioral plasticity, but these molecular and behavioral changes occurred prior to body fragmentation. These results suggest that the lumbriculid Lan 3-2 glycoepitope and proteins that bear them have been co-opted for neural morphallactic programs, induced both in anticipation of reproductive fragmentation and in compensation for injury-induced fragmentation.