Aquatic toxicity test for enchytraeids

It is the purpose of this report to present a simple method for testing the toxicity of chemical substances by using enchytraeids in an aquatic environment. Up to eight different environmental chemicals were applied to various species, mainly of the genus Enchytraeus. The results were compared with those achieved for D. magna. Significant differences, however, between the LC₅₀ values of the various enchytraeid species and the LC₅₀ values for enchytraeids and daphnids could not be observed. For E. cf. buchholzi the toxicological sensitivity of discrete ontogenetic stages was tested. The Aquatic Enchytraeid Test results were compared with those obtained from the Terrestrial Enchytraeid Test. It was found that in soil a chemical could be 600 times less toxic than in water, although the same species (E. albidus) was used in both environments. Even more pronounced were the discrepancies between the terrestrial and aquatic toxicities when the LC₅₀ values for earthworms and daphnids were compared.     

Freeze tolerance and accumulation of cryoprotectants in the enchytraeid Enchytraeus albidus (Oligochaeta) from Greenland and Europe

The freeze tolerance and accumulation of cryoprotectants was investigated in three geographically different populations of the enchytraeid Enchytraeus albidus (Oligochaeta). E. albidus is widely distributed from the high Arctic to temperate Western Europe. Our results show that E. albidus is freeze tolerant, with freeze tolerance varying extensively between Greenlandic and European populations. Two populations from sub Arctic (Nuuk) and high Arctic Greenland (Zackenberg) survived freezing at −15 °C, whereas only 30% of a German population survived this temperature. When frozen, E. albidus responded by catabolising glycogen to glucose, which likely acted as a cryoprotectant. The average glucose concentrations were similar in the three populations when worms were frozen at −2 °C, approximately 50 μg glucose mg⁻¹ tissue dry weight (DW). At −14 °C the glucose concentrations increased to between 110 and 170 μg mg⁻¹ DW in worms from Greenland. The average glycogen content of worms from Zackenberg and Nuuk were about 300 μg mg⁻¹ DW, but only 230 μg mg⁻¹ DW in worms from Germany showing that not all glycogen was catabolised during the experiment. Nuclear magnetic resonance spectrometry (NMR) was used to screen for other putative cryoprotectants. Proline, glutamine and alanine were up regulated in frozen worms at −2 °C but only in relatively small concentrations suggesting that they were of little significance for freeze survival. The present study confirms earlier reports that freeze tolerant enchytraeids, like other freeze tolerant oligochaete earthworms, accumulate high concentrations of glucose as a primary cryoprotectant.     

Suppression of nematophagous fungi by enchytraeid worms: a field exclosure experiment

The feeding biology of Enchytraeus crypticus and other enchytraeids is poorly understood as is their effect on nematophagous fungi. Because enchytraeids had been associated with nematophagous fungi in the field and had suppressed these fungi in soil microcosms, we tested the hypothesis that exclusion of enchytraeids, largely E. crypticus, would improve establishment of certain nematophagous fungi in field plots. The fungi, Hirsutella rhossiliensis and Monacrosporium gephyropagum, are being studied as potential control agents of plant-parasitic nematodes and were formulated as hyphae in alginate pellets. The pellets were mixed into soil without enchytraeids and placed in cages (PVC pipe, 80 cm³ volume) with fine (20 μm) or coarse (480 μm) mesh; cages were buried 15 cm deep in field plots and then recovered after 6–52 days. When fine mesh was used, enchytraeids were excluded and the fungi increased to large numbers. When coarse mesh was used, enchytraeid numbers in cages increased rapidly and the fungi did poorly. Although mesh also affected other potential fungivores, including collembolans and large dorylaimid nematodes, we suspect that enchytraeids were more important because large numbers were consistently found in cages with coarse mesh soon after the cages were placed in soil. Organisms smaller than enchytraeids (bacteria, fungi, and protozoa) also appeared to be important because the fungi did better in heat-treated soil than in non-heat-treated soil, regardless of mesh size. The rapid increase in enchytraeid numbers in cages with hyphal pellets and coarse mesh was probably caused by movement of enchytraeids toward the pellets with hyphae: increase in enchytraeid numbers was minimal when movement into cages was blocked (or when cages contained pellets without hyphae). Overall, the data were consistent with the hypothesis that enchytraeids, or other meso- or macrofauna, contributed to suppression of nematophagous fungi in our field plots. Received: 22 April 1997 / Accepted: 16 June 1997     

Pygidialporen bei marinen Enchytraeidae (Oligochaeta)

Coelo-rectal pygidial pores were found in the anal area of marine enchytraeids (Lumbricillus lineatus, Marionina spicula, Enchytraeus albidus) from the mud-flats of the Isle of Sylt. The structure and function of these pores have been discussed.     

Responses to acute and chronic desiccation stress in <Emphasis Type="Italic">Enchytraeus</Emphasis> (Oligochaeta: Enchytraeidae)

Enchytraeids are small soil living oligochaete worms with high sensitivity to low soil moisture. The effects of acute and chronic desiccation on survival and reproduction were determined in Enchytraeus albidus and Enchytraeus crypticus. Further, effects of acute drought stress on the water balance physiology and accumulation of osmolytes were investigated in E. albidus. Survival of E. crypticus and E. albidus was significantly influenced by exposure time. Reproduction was much more sensitive to desiccation than survival and was significantly reduced from −0.06 bar, which was surprising because no dehydration or change in the body fluid osmolality of E. albidus occurred until much harsher drought regimes occurred. The body fluid osmolality of E. albidus was relatively high, about 500 mOsm. Congruent with this no water loss or changes in osmotic pressure occurred until equivalent or higher water potential values of the environment were reached. Two osmolytes, glucose and alanine, were up-regulated in drought exposed E. albidus. Even though enchytraeids display moderate physiological protection to rapid changes in soil moisture (by having a high osmotic pressure) in the short term, populations subjected to long-term drought stress can be severely reduced even under moderate drought levels.     

Freeze tolerance and accumulation of cryoprotectants in the enchytraeid Enchytraeus albidus (Oligochaeta) from Greenland and Europe

The freeze tolerance and accumulation of cryoprotectants was investigated in three geographically different populations of the enchytraeid Enchytraeus albidus (Oligochaeta). E. albidus is widely distributed from the high Arctic to temperate Western Europe. Our results show that E. albidus is freeze tolerant, with freeze tolerance varying extensively between Greenlandic and European populations. Two populations from sub Arctic (Nuuk) and high Arctic Greenland (Zackenberg) survived freezing at −15 °C, whereas only 30% of a German population survived this temperature. When frozen, E. albidus responded by catabolising glycogen to glucose, which likely acted as a cryoprotectant. The average glucose concentrations were similar in the three populations when worms were frozen at −2 °C, approximately 50 μg glucose mg⁻¹ tissue dry weight (DW). At −14 °C the glucose concentrations increased to between 110 and 170 μg mg⁻¹ DW in worms from Greenland. The average glycogen content of worms from Zackenberg and Nuuk were about 300 μg mg⁻¹ DW, but only 230 μg mg⁻¹ DW in worms from Germany showing that not all glycogen was catabolised during the experiment. Nuclear magnetic resonance spectrometry (NMR) was used to screen for other putative cryoprotectants. Proline, glutamine and alanine were up regulated in frozen worms at −2 °C but only in relatively small concentrations suggesting that they were of little significance for freeze survival. The present study confirms earlier reports that freeze tolerant enchytraeids, like other freeze tolerant oligochaete earthworms, accumulate high concentrations of glucose as a primary cryoprotectant.     

Tubifex tubifex as a link in food chain transfer of hexachlorobenzene from contaminated sediment to fish

Sediments contaminated with poorly water-soluble organic chemicals pose a risk to aquatic food chains. Sediment-associated chemicals can be accumulated by endobenthic, sediment-ingesting invertebrates. Some tubificid species – or other benthic annelids – serve as food for benthivorous fish, which thereby ingest the sediment-borne chemicals and may accumulate contaminant concentrations far higher than from water exposure only, and transfer them to organisms of higher trophic levels. For measurement of biomagnification, a sediment based food chain was developed and established in the laboratory. The two-step food chain included the sediment-dwelling freshwater oligochaete Tubifex tubifex (Müller) as a representative species of benthic infauna. The three-spined stickleback (Gasterosteus aculeatus, Linné), a small teleost fish which often feeds primarily on benthic invertebrates, served as a model predator. Spiked artificial sediment and reconstituted water as the overlying medium were used. Experiments were performed using ¹⁴C-labelled hexachlorobenzene, a hydrophobic pollutant as a model compound. To examine the influence of benthic prey on the bioaccumulation of the test substance in the predator, fish were exposed to spiked water, spiked sediment, pre-contaminated prey organisms, or combinations of these exposure routes. The results of these experiments indicate that for hexachlorobenzene, the presence of contaminated Tubifex tubifex as a food source in combined exposure leads to significantly higher accumulation in fish than exposure to single pathways.     

Predicting potential impacts of climate change on the geographical distribution of enchytraeids: a meta-analysis approach

The expectation that atmospheric warming will be most pronounced at higher latitudes means that Arctic and montane systems, with predominantly organic soils, will be particularly influenced by climate change. One group of soil fauna, the enchytraeids, is commonly the major soil faunal component in specific biomes, frequently exceeding above‐ground fauna in biomass terms. These organisms have a crucial role in carbon turnover in organic rich soils and seem particularly sensitive to temperature changes. In order to predict the impacts of climate change on this important group of soil organisms we reviewed data from 44 published papers using a combination of conventional statistical techniques and meta‐analysis. We focused on the effects of abiotic factors on total numbers of enchytraeids (a total of 611 observations) and, more specifically, concentrated on total numbers, vertical distribution and age groupings of the well‐studied species Cognettia sphagnetorum (228 observations). The results highlight the importance of climatic factors, together with vegetation and soil type in determining global enchytraeid distribution; in particular, cold and wet environments with mild summers are consistently linked to greater densities of enchytraeids. Based on the upper temperature distribution limits reported in the literature, and identified from our meta‐analyses, we also examined the probable future geographical limits of enchytraeid distribution in response to predicted global temperature changes using the HadCM3 model climate output for the period between 2010 and 2100. Based on the existing data we identify that a maximum mean annual temperature threshold of 16 °C could be a critical limit for present distribution of field populations, above which their presence would decline markedly, with certain key species, such as C. sphagnetorum, being totally lost from specific regions. We discuss the potential implications for carbon turnover in these organic soils where these organisms currently dominate and, consequently, their future role as C sink/source in response to climate change.     

Earthworm and enchytraeid activity under different arable farming systems,as exemplified by biogenic structures

A study was conducted in order to compare soil faunal activity in four experimental farming systems using different tillage, chemical input and crop rotation practices: A conventional system with deep-ploughing (CT), an integrated system with reduced tillage and minimum chemical input (IN), a system with reduced tillage and high chemical input (RT) and a system with minimum tillage and high chemical input (MT). In nine experimental fields with two sampling points each, earthworms were sampled and biogenic structures were identified and counted in topsoil profiles (0–14 cm depth). Components of these profiles were identified by morphological features. Quantitative analyses of these morphological features provided information about soil compaction, earthworm and enchytraeid activity and distribution of roots and crop residues in the soil matrix. The dominant species in the earthworm community was the endogeic Aporrectodea rosea. Earthworm densities were unexpectedly lowest under reduced tillage (6 specimens per m²), and highest under deep-ploughing (67 specimen per m²), the reverse effect being observed with enchytraeid worms, as ascertained by deposition of their faecal pellets in topsoil profiles. Strong very fine granular structure (STVFGR) was most frequent in the integrated farming system (IN). We concluded that in the studied site embracing four farming systems, enchytraeids play an important role in creating a stable soil structure and porosity at the low level of earthworm densities found in the integrated system (IN). This revised version was published online in June 2006 with corrections to the Cover Date.     

Trace Element (Pb,Cd, and As) Contamination in the Sediments and Organisms in Zhalong Wetland,Northeastern China

This study examined Tessier's fractions of Pb, Cd, and As in the sediments and their potential toxic effects on four aquatic animals, i.e., Enchytraeus of Lumbricidae, Cipangopaludina chinensis of Viviparidae, Cybister japonicus Sharp of Dytiscidae, and Perccottus glenii Dybowski of Odontobutidae in Zhalong Wetland, Northeastern China. Results show that Cd poses a high risk to the biota in Zhalong Wetland. The geo-accumulation indices indicate that the region only has background concentrations of As, although it is slightly polluted by Pb and is seriously polluted by Cd. The exchangeable fraction receives minor contributions from these metals: 0.8% for Pb, 2.8% for Cd, and 4% for As. Three reed organs and four aquatic animal groups contain detectable concentrations of Pb, Cd, and As, with the following order of increasing concentrations: stem < rhizome < root < Viviparidae < Odontobutidae < Dytiscidae < Lumbricidae. Relatively low bioaccumulation factor (BAF) of As occurred in four aquatic organisms, whereas significantly higher BAFs of Pb and Cd were found in the earthworm than other aquatic organisms (P < 0.05). The earthworms (Enchytraeus of Lumbricidae) were recommended to be used as a suitable indicator for Pb, Cd, and As toxic risk level in this region.     

Cold tolerance and dehydration in Enchytraeidae from Svalbard

When cooled in contact with moisture, eight species of arctic Enchytraeidae from Svalbard were killed by freezing within minutes or hours at −3 and −5 °C; an exception was Enchytraeus kincaidi which survived for up to 2 days. When the temperature approached 0 °C the enchytraeids apparently tried to escape from the moist soil. The supercooling capacity of the enchytraeids was relatively low, with mean supercooling points of −5 to −8 °C. In contrast, specimens of several species were extracted from soil cores that had been frozen in their intact state at −15 °C for up to 71 days. Compared to freezing in a moist environment, higher survival rates were obtained during cooling at freezing temperatures in dry soil. Survival was recorded in species kept at −3 °C for up to 35 days, and in some species kept at −6 °C for up to 17 days. Slow warming greatly increased survival rates at −6 °C . The results strongly suggest that arctic enchytraeids avoid freezing by dehydration at subzero temperatures. In agreement with this, weight losses of up to ca. 42% of fresh weight were recorded in Mesenchytraeus spp. and of up to 55% in Enchytraeus kincaidi at water vapour pressures above ice at −3 to −6 °C. All specimens survived dehydration under these conditions. Accepted: 12 December 1997     

Succession of fungi and fauna during decomposition of needles in a small area of Scots pine litter

During micromorphological investigations on Scots pine litter, several decomposition stages have been recognized on fallen pine needles, each being associated with the activity of animal and microbial organisms, both. To well-known fungal successions that have been so far described by mycologists we must add succession of animal groups such as nematodes, amoebae, enchytraeids, sciarid larvae, oribatid mites and earthworms. A bacterial development was observed in the L₂ layer, following penetration by microfauna (nematodes, amoebae). After that stage pine needles were actively tunnelled by enchytraeids, sciarid larvae and oribatid mites and at the same time were nibbled on by epigeic earthworms (L₂ and F₁ layers). When the fine root system of pine developed through accumulated old needles (F₁ layer), mycorrhizal fungi penetrated the needles and seemed to impede any further bacterial development. Pine foliar tissues were progressively incorporated into the fecal material of earthworms and other members of the soil fauna. A more realistic scheme was suggested for plant litter decomposition in moder humus.     

Roles of carbohydrate reserves for local adaptation to low temperatures in the freeze tolerant oligochaete Enchytraeus albidus

Geographic variation in cold tolerance and associated physiological adaptations were investigated in the freeze tolerant enchytraeid Enchytraeus albidus (Oligochaeta). Specimens from Svalbard, Greenland (Nuuk), Iceland (Hólar and Mossfellsbær) and continental Europe [Norway (Bergen), Sweden (Kullen) and Germany] were reared in the laboratory in a common-garden experiment. The aim was to test for variations in minimum lethal temperature, freeze duration tolerance, carbohydrate reserves and metabolic rate among the populations. Cold tolerance was related to environmental temperature of the respective location. Populations from the coldest climatic regions were able to tolerate freezing down to at least ?15 °C and endured being frozen at ?5 °C for 27–48 days, respectively. Populations from milder climates had a lower freeze duration tolerance (about ?9 °C) and endured ?5 °C for a shorter period (between 9 and 16 days). Glucose accumulation and glycogen reserves varied significantly between populations, but was not related directly to cold tolerance. Metabolic rate varied significantly between populations, but was not significantly related to cold tolerance. The metabolic rates at ?2 °C of frozen and unfrozen worms from Germany and Svalbard were tested. The metabolic depression due to freezing of these populations was relatively small (<50 %), suggesting that the large carbohydrate accumulations may also be important as fuel during long-term freezing at moderately low temperatures. Differences in metabolic depression may partly explain the difference in cold tolerance of these two populations, however, the mechanisms behind local adaptation to low winter temperatures in these enchytraeid populations seem more complex than earlier studies have indicated.     

Phylogeny and origins of Enchytraeidae

Phylogenetic analyses of Achaetinae (Enchytraeidae: Oligochaeta) and other bisetate enchytraeids indicate that the achaetines include the earliest species of Enchytraeidae but that Achaetinae is not monophyletic. The earliest species of bisetate enchytraeid now extant are restricted to South America, Africa, and India. As this part of the enchytraeid lineage is ancestral to other enchytraeid taxa, it is suggested that Enchytraeidae may have arisen in South America or a contiguous Southern land mass.Less than 50% of the 11 genera of Enchytraeidae considered are supported by the results of these analyses as evolutionary or phylogenetic groups. Five are substantiated as monophyletic: Achaeta, Lumbricillus, Fridericia, Randidrilus, and Enchytronia. In addition to most of the achaetine genera, Marionina is shown to be in great need of revision. Its recognition is a continuing source of confusion to taxonomic resolution of Enchytraeidae.     

Population dynamics and functional roles of Enchytraeidae (Oligochaeta) in hardwood forest and agricultural ecosystems

Population dynamics of enchytraeids are described for 2 montane forested watersheds in southwestern North Carolina (Coweeta) and an agricultural site under conventional (CT) and no-tillage (NT) management in the northeastern Georgia piedmont (Horseshoe Bend, HSB). Given that much of the taxonomy, ecology and community structure of enchytraeids is poorly known, our objective was to identify key indicators of enchytraeid community structure which could be used, in this case, to better understand their role in soil structure formation. Although population densities of enchytraeids were higher in the forested (Coweeta) than in the arable soils (HSB), the average ash free dry weight per enchytraeid at HSB was nearly double that found at Coweeta. Based on these measurements and an estimate of their gut transit time, we calculated that the enchytraeids at HSB transported 2180 g of soil per m² per year compared to 443 and 393 g m^-2 yr^-1 for watershed 18 and 27, respectively at Coweeta. We therefore hypothesize that enchytraeids have a larger influence on soil structure in agricultural fields than in forested areas, in spite of lower population densities. The ash free dry weight and ash wt. per enchytraeid may qualify as key indicator parameters of enchytraeid community structure which helps to understand their functional role in ecosystems, though more studies are called for.     

Coprophilous streptomycetes and fungi—Food sources for enchytraeid worms (Enchytraeidae)

Food selection experiments demonstrated thatEnchytraeus crypticus (Oligochaeta, Enchytraeidae) was attracted byStreptomyces species and microscopic fungi in vermiculture substrates and in the gut content ofEisenia andrei earthworms. Consumption of spores and/or mycelia of attractive strains influenced markedly the proliferation ofE. crypticus. There was a 74-fold increase in the numbers of enchytraeids fed on the mixture ofAspergillus flavus andVerticillium tenerum mycelia or on mycelium of one strain ofStreptomyces in reproduction tests. Lower rates of increase ofE. crypticus (50-fold or less) were observed in variants whereV. tenerum or mixtures of fungi and streptomycetes were offered as food. We showed a potential importance of microbial populations in vermicultures and indicated that their regulation may provide a way to increase the productivity of such systems.     

Biological studies upon Enchytraeus variatus Bouguenec & Giani 1987 in breeding cultures

The life history of an enchytraeid worm, Enchytraeus variatus, was studied under laboratory conditions at 18–22 °C. This species can reproduce simultaneously by asexual (architomy) or sexual reproduction. The number of ova per cocoon varies from 5 to 20 (x = 10.9). The generation period (from cocoon to next cocoon) varies from 14 to 39 days (x = 26.1) according to the period of the year. The number of generations per year is between 7.3 and 26.1 (x = 14). A mature worm can lay between 23.7 and 25.8 cocoons during its life (254 days as maximum observed) at a mean rate of 0.12 cocoon worm^–1 day^–1. Experimental cultures were carried out to determine the structure, density and biomass of the populations. A maximal density of 1 396 314 worms was recorded after 85 days of culture. Net production reached 21.48 g m^–2 day^–1 after 26 days in a culture initiated from cocoons.     

Ultrastructure of the nephrostome in Enchytraeus albidus (Annelida, Clitellata)

 The nephrostome of Enchytraeus albidus exhibits a longitudinal slit-like opening on the dorsal side of a bulbous organ which is mainly composed of three cells, one flame cell situated centrally and endowed with a ciliary flame, and two cells lying superficially, called the mantle cell and the accessory mantle cell. The mantle cell occupies the ventral side of the organ extending on both sides up to the opening to constitute its immediate border on the frontal and lateral sides. Here it forms a kind of crest which is heavily subdivided into many protrusions and infoldings endowed with long cilia which exclusively spread into the coelomic cavity. The accessory mantle cell borders the narrow posterior slit of the opening, forming the roof of the initial canal which is devoid of cilia. From its anterior region a projection arises extending above the opening. The flame cell forms a groove from which the ciliary flame arises which extends into the canal. At its posterior region it replaces the accessory mantle cell displacing it onto the dorsal surface of the organ. It is argued that the mantle cell and the accessory mantle cell have presumbly originated from coelothelial cells. Thus the metanephridium may be a composite organ developing from a nephridioblastic and a coeloblastic source. A discussion of results in other annelid species indicates that metanephridia in the Annelida may have evolved more than once. Accepted: 13 October 1997