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Dendrobaena veneta (Rosa, 1886) is widely distributed all over Europe due to its use as compost worm. The specimen presented here was collected in Tiranë district, Albania. Currently, only two species’ complete or nearly complete mitochondrial genome (mitogenome) sequences have been reported in the genus Dendrobaena; D. octaedra (Savigny, 1826) and D. tellermanica Perel, 1966. In this study, the complete mitogenome of D. veneta was sequenced, assembled, and annotated. The mitogenome of D. veneta is a circular DNA molecule, consisting of 15,475 bp with an A + T content of 61.2%. It contains 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and 1 non-coding region (control region). Phylogenetic analysis showed that D. veneta is clustered with the other two Dendrobaena species in the well-supported family Lumbricidae.

Dendrobaena veneta (Rosa, 1886) is widely distributed all over Europe due to its use as compost worm. The specimen presented here was collected in Tiranë district, Albania. Currently, only two species' complete or nearly complete mitochondrial genome (mitogenome) sequences have been reported in the genus Dendrobaena; D. octaedra (Savigny, 1826) and D. tellermanica Perel, 1966. In this study, the complete mitogenome of D. veneta was sequenced, assembled, and annotated. The mitogenome of D. veneta is a circular DNA molecule, consisting of 15,475 bp with an A + T content of 61.2%. It contains 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and 1 non-coding region (control region). Phylogenetic analysis showed that D. veneta is clustered with the other two Dendrobaena species in the well-supported family Lumbricidae. [ABSTRACT FROM AUTHOR]

ABSTRACT The Ethiopian Drylands are rich in their variety of local earthworm species. However, the category of species, and the effect of their performance on additions to plant nutrients have not been adequately evaluated. Thus, local earthworm samples were collected from the three major agroecological zones (highland, midland, and lowland) in Tigray (northern Ethiopia) and classified down to species level. Moreover, a vermicomposting experiment with four treatments (three local earthworm species, Eisenia fetida , and a conventional composting method) and three replications was established. Finally, each bin’s mature compost sample was taken to analyze plant nutrient content. The study results indicated that earthworm species in the highland, midland and lowland agroecological zones were Dendrobaena veneta, Eisenia andrie and Lumbricus rubellus , respectively. The use of these earthworms in the composting process (average of the four earthworm species) yielded higher nutrient content, ranging from 21.9 % for Sodium to 3300 % for Boron, compared to the conventional one. The highest total nitrogen (an increase of 44.4 %) and organic carbon (an increase of 33.4 %) were recorded in the Eisenia fetida and Dendrobaena veneta treated bins, respectively. Composting with Eisenia andrie has resulted in increases in P (96.1 %), K (125 %), Mg (83 %) and all micro-nutrients (between 91 % for Zn and 4400 % for B). Both Eisenia andrie and Lumbricus rubellus species contributed to the increased additions of Sulfur (85.7 %) compared to the control. It can be concluded that the use of local earthworms (particularly Eisenia andrie) in the composting process plays a significant role in plant nutrient addition.

The earthworms’ abundance is usually one of the main indicators of healthy and productive soils. However, agricultural management practices affect the earthworm population and activities. Although there is a lot of information that shows the relation between land use/land cover change and earthworms activity, very little is known about these effects under tea cultivation. Thus the current study was done to determine the effects of long-term tea cultivation on the earthworm’s population and abundance in the tea plantations of Iran to distinguish effects of these practices on soil properties concerning earthworms. Hence, 58 locations of tea cultivations were randomly selected in Guilan and Mazandaran province. Earthworm were sampled by manually excavating and sorting four 30×30 cm pits by 30 cm deep in each location. Earthworms were enumerated in the field and taken to the lab for identification. Once identified, the earthworms will dry in the oven at 60oC for 48h and the dry weight registers. Some physicochemical properties of the mineral soils were determined in the laboratory. The finding indicated that the earthworms were only observed in the two from 58 locations: Bazkiagorab (Lahijan) and Shekarposhteh (Tonekabon). Three species as Perelia kaznakovi, Aporrectodea trapezoides, and Dendrobaena veneta were recorded from Bazkiagorab but only P. kaznakovi was identified in Shekarposhteh. The total population of all identified earthworms was 22 and 3 m-2 in the Bazkiagorab and Shekarposhteh, respectively. Results of the physicochemical analysis showed that 35% and 51% of the soils had a pH less than4.5 and organic carbon less than2%, respectively. Available phosphorus and potassium in 80% and 65% of the soils were less than 25 and 150 mg/kg, respectively. It can be concluded that monoculture and long-term tea cultivations had a negative effect on the earthworm population, in addition, it has strongly acidified the soil. It is recommended that native nitrogen-fixing trees mixed planted with tea, and more attention should be paid to nutrient Best Management Practices in tea plantations.

To feed humans on a future Mars settlement, a sustainable closed agricultural ecosystem is a necessity. On Mars, both the faeces of astronauts as well as any plant residues or other organic waste needs to be (re)used to fertilise the present regolith. The activity of earthworms may play a crucial role in this ecosystem as they break down and recycle the dead organic matter. The contribution of worms to Mars regolith forming is yet an unexplored territory. The first goal of our research was to investigate whether earthworms (Caligonella genus and Dendrobaena veneta) can survive in Mars soil simulant. The second goal was to investigate whether earthworm activity on Mars soil simulant can stimulate the growth of crops, in our case Rucola. The third goal was if earthworm activity can enhance the effect of pig slurry on the growth of Rucola. In a 75-day greenhouse experiment, we sowed Rucola in Mars soil simulant as well as in silver sand as an Earth control, amended with pig slurry, plant residues, and earthworms. During the experimental period, we observed worm activity. At the end of the experiment, the worms had propagated both in the Mars soil simulant and Earth control. However, we found no significant effect of worm activity on plant biomass production. This was probably due to the relative short duration of the experiment, being one life cycle of Rucola. Adding pig slurry stimulated plant growth significantly as expected, especially for the Mars soil simulant.

The earthworms' abundance is usually one of the main indicators of healthy and productive soils. However, agricultural management practices affect the earthworm population and activities. Although there is a lot of information that shows the relation between land use/land cover change and earthworms activity, very little is known about these effects under tea cultivation. Thus the current study was done to determine the effects of long-term tea cultivation on the earthworm's population and abundance in the tea plantations of Iran to distinguish effects of these practices on soil properties concerning earthworms. Hence, 58 locations of tea cultivations were randomly selected in Guilan and Mazandaran province. Earthworm were sampled by manually excavating and sorting four 30x30 cm pits by 30 cm deep in each location. Earthworms were enumerated in the field and taken to the lab for identification. Once identified, the earthworms will dry in the oven at 60°C for 48h and the dry weight registers. Some physicochemical properties of the mineral soils were determined in the laboratory. The finding indicated that the earthworms were only observed in the two from 58 locations: Bazkiagorab (Lahijan) and Shekarposhteh (Tonekabon). Three species as Perelia kaznakovi, Aporrectodea trapezoides, and Dendrobaena veneta were recorded from Bazkiagorab but only P. kaznakovi was identified in Shekarposhteh. The total population of all identified earthworms was 22 and 3 m-2 in the Bazkiagorab and Shekarposhteh, respectively. Results of the physicochemical analysis showed that 35% and 51% of the soils had a pH less than 4.5 and organic carbon less than 2%, respectively. Available phosphorus and potassium in 80% and 65% of the soils were less than 25 and 150 mg/kg, respectively. It can be concluded that monoculture and long-term tea cultivations had a negative effect on the earthworm population, in addition, it has strongly acidified the soil. It is recommended that native nitrogen-fixing trees mixed planted with tea, and more attention should be paid to nutrient Best Management Practices in tea plantations. [ABSTRACT FROM AUTHOR]

Olive oil industry is economically important in Mediterranean countries. Disposal of olive mill waste (OMW) presents an environmental concern in those countries due to its high salinity and its high level of polyphenols. In order to reuse OMW, those properties have to change either through the filtration process and addition of adsorbents or by composting. One of the most important organisms in composting of organic wastes is earthworms. However, data on the effects of OMW on earthworms are scarce. The main aim of our study was to investigate whether OMW contaminated soil (OMW CS) causes adverse effects on molecular and organism level in epigeic earthworm Dendrobaena veneta and on microbiological activity. Changes of measured biochemical biomarkers (AChE, CAT, GST, lipids, MDA) varied depending on the quantity of added OMW CS and the exposure duration. Oxidative stress occurred after 7 days of exposure, while in most cases enzyme activity recovered after 28 days. At the highest ratio of contaminated soil (50%), reproduction was completely inhibited. The second aim was to investigate the impact of earthworms on phenol degradation and microbial activity, indicating an important role in the bioremediation of contaminated soils. Our results show that above a certain quantity an OMW CS has an adverse effect on earthworms, while the impact of earthworms on soil microbial activity was positive but transient. Yet, as the results also imply that earthworms have an impact on phenol degradation, they can be used to help remediation of OMW CS and its subsequent usage in agriculture. However, the quantity of OMW CS that can be safely added should be determined first. [ABSTRACT FROM AUTHOR]

To feed humans on a future Mars settlement, a sustainable closed agricultural ecosystem is a necessity. On Mars, both the faeces of astronauts as well as any plant residues or other organic waste needs to be (re) used to fertilise the present regolith. The activity of earthworms may play a crucial role in this ecosystem as they break down and recycle the dead organic matter. The contribution of worms to Mars regolith forming is yet an unexplored territory. The first goal of our research was to investigate whether earthworms (Caligonella genus and Dendrobaena veneta) can survive in Mars soil simulant. The second goal was to investigate whether earthworm activity on Mars soil simulant can stimulate the growth of crops, in our case Rucola. The third goal was if earthworm activity can enhance the effect of pig slurry on the growth of Rucola. In a 75-day greenhouse experiment, we sowed Rucola in Mars soil simulant as well as in silver sand as an Earth control, amended with pig slurry, plant residues, and earthworms. During the experimental period, we observed worm activity. At the end of the experiment, the worms had propagated both in the Mars soil simulant and Earth control. However, we found no significant effect of worm activity on plant biomass production. This was probably due to the relative short duration of the experiment, being one life cycle of Rucola. Adding pig slurry stimulated plant growth significantly as expected, especially for the Mars soil simulant. [ABSTRACT FROM AUTHOR]

Earthworms shape the biological and physicochemical qualities of the soil they choose to reside in, but our understanding of the specific chemicals that attract or repel a particular species of earthworm remains incomplete. Current research indicates that some species feed on and are attracted to fungi, such as Geotrichum candidum. In the present study, as part of our continuing effort to characterize mechanisms of earthworm chemosensation, we tested whether ethyl hexanoate and ethyl pentanoate, two compounds produced by G. candidum, are appetitive to the European nightcrawler (Dendrobaena veneta). In a soil T-maze, both of these compounds significantly repelled individual earthworms in a dosage-dependent manner, this result ran counter to our initial hypothesis. D. veneta also avoided ethyl hexanoate and ethyl pentanoate in an assay we specifically developed to test an earthworms aversion to chemical stimuli in soil. In both of these assays, ethyl hexanoate was aversive at lower concentrations than ethyl pentanoate. These findings further clarify our understanding of the chemical cues that trigger the decision of D. veneta to select a particular soil-environment, and emphasize that different earthworm species may react very differently to commonly encountered chemical stimuli. [ABSTRACT FROM AUTHOR]

Earthworm coelomic fluid (CF) is known as a rich source of various bioactive compounds with promising anticancer features. However, it has been demonstrated that CF affects functionality of both, cancer and normal cells. This non-selective activity causes a major problem for medical application of CF.In this study, we present the anticancer activity of the active protein-carbohydrate fraction (AF) isolated from thermally treated CF of earthworm Dendrobaena veneta. The in vitro effect of the AF was examined in human colon model including normal human colon epithelium (CCD 841 CoTr) and human colon adenocarcinoma (HT-29 and LS180) cell lines. We investigated the impact of AF on cell viability neutral red and lactate dehydrogenase assays, morphology May-Grünwald-Giemsa staining assay proliferation MTT tetrazolium salt and BrdU incorporation assays as well as cell cycle progression propidium iodide/RNase staining and the activity of human 20S proteasome the hydrolysis of AMC from a Suc-LLVY-AMC peptide substrate. Additionally, the influence of AF on apoptosis was examined in HT-29 cells by Annexin V/PI, Hoechst 33342 staining and active caspase-3 assays.Our investigation demonstrated that AF at the tested concentration range does not affect the viability and morphology of CCD 841 CoTr cells. Simultaneously, AF inhibits human 20S proteasome activity as well as significantly decreases mitochondrial metabolism, disturbs cell cycle and induces apoptosis via activation of procaspase-3 in HT-29 cancer cells. Obtained results demonstrate the antiproliferative and proapoptotic activity of AF that can be useful in developing therapeutic strategies to treat human colon cancer.

Due to a need for sustainability in agriculture, waste products ought to be utilized in the most appropriate way. A study was undertaken relating to the vermicomposting of Cameroon sheep dung (CSD) by the earthworm Dendrobaena veneta. Processing of this waste was investigated using unadulterated CSD and in a 1:1 mixture with unpalatable (waste) hay (CSDH). Results demonstrated that these materials were actively processed by D. veneta with vermicomposts obtained which can be characterized by a higher amount of total nitrogen, phosphorus and potassium (average, respectively, 17.0, 10.5, 13.2 g kg−1 d.m.), as well as lower total carbon and magnesium content (respectively, 340 and 3.2 g kg−1 d.m.), compared with the initial waste material. No significant differences were found between CSD and CSDH vermicomposts with respect to chemistry. Levels of selected trace elements (average: Cu 17.5–18.8, Cr 5.7–5.8, Pb 13.5–14.4, Ni < 3, Cd < 0.4 mg kg−1 d.m.) in both vermicomposts did not exclude their application to agricultural soil as a fertilizer. [ABSTRACT FROM AUTHOR]

It is known that earthworm coelomic fluid (CF) can affect not only cancer but also normal cells. The study demonstrated that the CF of the earthworm Dendrobaena veneta exhibited cytotoxicity against A549 lung cancer cells but did not toward the bronchial epithelial cell line BEAS‐2B. The selective effect on the tumor cells was achieved after a short‐term CF heat pre‐treatment at 70 °C. The cytotoxic effect of the CF was time‐ and concentration‐dependent. The CF noticeably decreased the viability and affected the morphology of the A549 cells. Scanning electron microscopy revealed a different degree of destruction of the nucleus and cytoplasm of A549 cells. As determined by atomic force microscopy, the cell surface roughness increased while the cell stiffness was reduced upon the CF treatment. A twofold increase in the caspase 3, 4, 5, and 10 levels was observed in the A549 cells after the incubation with the CF. The results obtained by flow cytometry using Annexin V confirmed the proapoptotic effect of the earthworm CF on A549 lung cancer cells. The D. veneta CF and active fraction obtained with cytotoxicity toward A549 lung cancer is an interesting and promising preparation for further biological, chemical, and biomedical research. [ABSTRACT FROM AUTHOR]

Dendrobaena veneta is one of the most widely used compost worms now distributed all over Europe and has been described under several synonymous names. Besides the typical stripy red specimens, another form bearing pinkish pigmentation also exists, mostly found in natural habitats in the East Mediterranean. Here we provide the first integrative taxonomic analysis of D. veneta, including some closely related species. Both the morphological and molecular results highly support the resurrection of D. succinta, a former synonym of D. veneta. In addition, we describe a peculiar new species, D. karacadagi sp. n. from Anatolia, and prove the highly homoplasious character of the cross section of the longitudinal musculature. [ABSTRACT FROM AUTHOR]

Due to a need for sustainability in agriculture, waste products ought to be utilized in the most appropriate way. A study was undertaken relating to the vermicomposting of Cameroon sheep dung (CSD) by the earthworm Dendrobaena veneta. Processing of this waste was investigated using unadulterated CSD and in a 1:1 mixture with unpalatable (waste) hay (CSDH). Results demonstrated that these materials were actively processed by D. veneta with vermicomposts obtained which can be characterized by a higher amount of total nitrogen, phosphorus and potassium (average, respectively, 17.0, 10.5, 13.2 g kg−1 d.m.), as well as lower total carbon and magnesium content (respectively, 340 and 3.2 g kg−1 d.m.), compared with the initial waste material. No significant differences were found between CSD and CSDH vermicomposts with respect to chemistry. Levels of selected trace elements (average: Cu 17.5–18.8, Cr 5.7–5.8, Pb 13.5–14.4, Ni < 3, Cd < 0.4 mg kg−1 d.m.) in both vermicomposts did not exclude their application to agricultural soil as a fertilizer.

Dendrobaena veneta (Rosa, 1886) is widely distributed all over Europe due to its use as compost worm. The specimen presented here was collected in Tiranë district, Albania. Currently, only two species' complete or nearly complete mitochondrial genome (mitogenome) sequences have been reported in the genus Dendrobaena ; D. octaedra (Savigny, 1826) and D. tellermanica Perel, 1966. In this study, the complete mitogenome of D. veneta was sequenced, assembled, and annotated. The mitogenome of D. veneta is a circular DNA molecule, consisting of 15,475 bp with an A + T content of 61.2%. It contains 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and 1 non-coding region (control region). Phylogenetic analysis showed that D. veneta is clustered with the other two Dendrobaena species in the well-supported family Lumbricidae., Competing Interests: The authors report there are no competing interests to declare., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Earthworms are exposed to herbicides both through their skin and digestive system. Herbicides can influence earthworms' survival, physiology and reproduction. However, there is a lack of data on herbicide effects on earthworms as they are often regarded as low or non-toxic. The aim of our study was to investigate whether widely used commercial formulations of glyphosate (GLF), tembotrione (TBT) and nicosulfuron (NCS) each applied at three environmentally relevant concentrations have adverse effects on various biomarkers and reproduction in epigeic earthworm Dendrobaena veneta. The activities of measured biomarkers varied depending on the herbicide used and the exposure duration and suggest that oxidative stress plays an important role in the toxicity of tested herbicides. Namely, GLF caused an acetylcholinesterase (AChE) activity induction after seven days, and NCS after 28 days, while TBT caused an inhibition up to 47% (6.6 μg kg dw soil −1 ) after seven days. Only TBT caused a significant change (H2 = 13.96, p = 0.002) to catalase (CAT) after seven days of exposure. Malondialdehyde concentrations (MDA) were increased all the time after NCS exposure, but only after seven days in GLF and 28 days in TBT treatments, respectively. The tested herbicides did not have a significant effect on reproduction success, expect of NCS which increased the number of juveniles (p < 0.05). [ABSTRACT FROM AUTHOR]

Earthworms may be subjected to mechanical/chemical stimuli and/or sub-lethal predator attacks leading to the extrusion of celomocytes and/or loss of body parts; thus, regeneration of cells, tissues and organs has adaptive value. The aim of present study on the lumbricid earthworm Dendrobaena veneta was to determine the interactive effects of celomocytes and the brain on the regeneration of either system after experimental depletion or extirpation, and to assess the effects of such treatments on reproductive performance. Decerebration was achieved either by amputating the first six anterior segments, or by surgery; celomocyte depletion was achieved by a standard electro-stimulation procedure. Celomocytes (amebocyte and eleocytes, respectively) were counted by hemocytometry, and riboflavin content in celomocyte lysates measured by spectrofluorimetry. The main findings were: (i) D. veneta regenerated anatomically intact brain, including neurosecretory cells, within 10 - 18 weeks after its removal plus celomocyte depletion (i.e. dual treatment); (ii) amoebocyte counts recovered to control levels by 10 weeks after extrusion treatment alone, but were still lower (60 %) than in controls at 18 weeks after dual treatment; eleocyte recovery after electro-stimulation alone was slow, reaching control levels only after 18 weeks, and was further retarded (31 % of controls at 18 weeks) by brain extirpation; (iii) riboflavin content was lower than controls only in the dual-treatment worms at 5 weeks; riboflavin content relative to eleocyte numbers was initially higher than controls in both treatment groups; this index was restored to control levels by 18 weeks in the electro-stimulation only treatment whilst recovery was somewhat retarded in the brain-extirpated group; (iv) celomocyte depletion treatment alone slightly impaired reproductive output, whilst brain removal had pronounced and protracted inhibitory effects. The observations engender the hypothesis that brain-derived neurosecretions and immune-competent celomocytes act in tandem to modulate neural regeneration and reproduction.

Dissolved organic carbon (DOC) has recently been proposed as an indicator of compost stability. We assessed the earthworms’ effect on DOC content and composition during composting, and linked compost stability to greenhouse gas emissions and feeding ratio. Earthworms reduced total DOC content, indicating larger stability of vermicompost than of thermophilic compost. The concentrations of humic acid and fulvic acid were reduced by earthworms, whereas there was no significant effect on hydrophobic neutrals and hydrophilics. The humic acid fraction was depleted more quickly than the other compounds, indicating humic acid degradation during composting. The optimum feeding ratio decreased DOC content compared to the high feeding ratio. The lowest N 2 O emissions were also observed at the optimum feeding ratio. Our study confirmed the use of DOC content and composition as an indicator of compost stability and suggested that feeding ratio should be considered when assessing the earthworms’ effect on stabilisation and greenhouse gas emissions. [ABSTRACT FROM AUTHOR]

Selenium (Se) is an essential micronutrient for humans, animals, and certain lower plants, but at higher concentrations Se becomes toxic to organisms. The boundary between the Se beneficial effect and its toxicity is narrow and depends on its chemical form, applied concentration, and other environmentally regulating factors. Due to the potential risk of toxicity in higher concentration, the aim of this study was to estimate the impact of increased concentrations of different forms of Se on the response of the wheat-soil-earthworm system. Soil, earthworms, and wheat grains were exposed to the Se in form of selenite and selenate in concentrations of 0.01, 0.1, and 1 mg kg−1. As an indicator of oxidative stress in wheat, lipid peroxidation levels (LPO) and total H2O2 content were determined, while antioxidative response was determined by catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR) activities. The biomarker responses in earthworms were determined by acetylcholinesterase (AChE), carboxylesterase (CES), and antioxidative enzymes (CAT and glutathione S-transferase) activities. Selenite and selenate increased Se content in the wheat and earthworms, while selenate application was more efficient, indicating higher bioaccumulation of this Se form. Both Se forms did not cause significant changes in the LPO level and H2O2 content, while GPX activities were elevated in all treatments, suggesting that oxidative stress was not induced in wheat. In earthworms, Se significantly reduced activities of AChE and CAT at some concentrations, while CES activity was increased at all concentrations applied. This study showed significant impact of Se on measured biochemical responses in wheat and earthworms, indicating the disruption of homeostasis. Obtained results can serve as basis for further studies on Se effects and will help in including different aspects necessary for understanding of Se impact on different components of soil ecosystems. [ABSTRACT FROM AUTHOR]

The selected chemical composition (dry matter, ash, total protein, and crude fat) of the integumentary muscles of Dendrobaena veneta were determined, plus the dry matter (DM) percentage content of 17 amino acids and the profile (%) of fatty acids. Results were compared with a more fully studied earthworm, Eisenia fetida . In addition, the composition of exogenous amino acids was compared to the WHO standard for pork, beef, and chicken eggs. Both earthworm species were grown on the same kitchen waste, and protein composition was analyzed using the same methods. Studies indicated that the muscle of D. veneta was characterized by a high level of protein (76.82% DM). A similar content of exogenous amino acids was observed in the protein of both earthworms, but for phenylalanine and isoleucine, slightly higher levels were recorded for E. fetida . More histidine, lysine, threonine, isoleucine, and arginine were found in earthworms compared with chicken egg white. Fatty acids play an essential role in balancing human or animal feed and their content determines the dietary and nutritional value of the food. Both earthworm species contained the appropriate content of saturated and unsaturated acids. In D. veneta , a higher content of arachidonic acid was found, and in E. fetida , lauric, tridecanoic, and palmitic acids were present. Future issues of food security may force us to seriously consider earthworm protein for indirect or even direct human consumption.

Abstract: In the present study, lysozyme-like activity against Micrococcus luteus was detected in the coelomic fluid, the extract from coelomocytes, intestine and in the homogenates from cocoons of Dendrobaena veneta. Four hours after immunization with Escherichia coli, the lysozyme activity in the coelomic fluid increased about three times and in the extract of coelomocytes – four times, in comparison to the control. In three cases: of the coelomic fluid, the homogenates from cocoons and the extract from coelomocytes, the antibody against HEWL (hen egg white lysozyme) recognized only one protein with a molecular mass of about 14.4kDa. In the coelomic fluid, apart from the protein with molecular mass of 14.4kDa the antibody directed against human lysozyme recognized an additional protein of 22kDa. Using the bioautography technique after electrophoretic resolution of native proteins in acidic polyacrylamide gels, two lytic zones of M. luteus were observed in the case of the coelomic fluid and three after the analysis of the extract of coelomocytes and the egg homogenates. The results indicated the existence of several forms of lysozyme with a different electric charge in the analyzed D. veneta samples. The highest lysozyme activity in the intestine of D. veneta was observed in the midgut. The antibody directed against human lysozyme indicated a strong positive signal in epidermal and midgut cells of earthworm. [Copyright &y& Elsevier]

Abstract: This study aimed to evaluate short-term earthworm-induced changes in the availability of metals applied to soil directly (metal-spiked) or via an organic matrix (sludge-amended). A laboratory experiment was performed using destructive sampling of microcosms filled with agricultural soil. A concentration gradient of industrial sludge contaminated predominantly with Cr, Cu, Ni, and Zn, and a soil freshly spiked with the same metal concentrations were applied on top of the soil columns. Individuals of Dendrobaena veneta (mimicking a realistic density of 500 earthworms per m2) were introduced in half of the replicates of each treatment. Total and 0.01M CaCl2 extractable metal concentrations were measured in soil after 0, 3, 6 and 12 weeks and metal concentrations in earthworms and percolates were measured after 3, 6, and 12 weeks. Earthworm activity did not affect metal availability of any treatment over time, but Ni and Cu concentrations in D. veneta were higher at the highest treatment levels. Earthworm Zn concentrations were similar in all treatments while Cr concentrations increased with increasing soil total metal content only for sludge treatments. Existing relationships of earthworm metal concentrations with total metal content in soil, taken from the literature, were not able to predict the metal levels measured in D. veneta. Results demonstrated that although over 12 weeks earthworm activity did not affect metal availability in soil, their burrowing activities did influence the metal concentrations of percolates over time. [Copyright &y& Elsevier]

Abstract: The new bacterial strain with antimycobacterial activity has been isolated from the midgut of Dendrobaena veneta (Annelida). Biochemical and molecular characterization of isolates from 18 individuals identified all as Raoultella ornithinolytica genus with 99% similarity. The bacterium is a possible symbiont of the earthworm D. veneta. The isolated microorganism has shown the activity against four strains of fast-growing mycobacteria: Mycobacterium butiricum, Mycobacterium jucho, Mycobacterium smegmatis and Mycobacterium phlei. The multiplication of the gut bacterium on plates with Sauton medium containing mycobacteria has caused a lytic effect. After the incubation of the cell free extract prepared from the gut bacterium with four strains of mycobacteria in liquid Sauton medium, the cells of all tested strains were deformed and divided to small oval forms and sometimes created long filaments. The effect was observed by the use of light, transmission and scanning microscopy. Viability of all examined species of mycobacteria was significantly decreased. The antimycobacterial effect was probably the result of the antibiotic action produced by the gut bacterium of the earthworm. The application of ultrafiltration procedure allowed to demonstrate that antimicrobial substance with strong antimycobacterial activity from bacterial culture supernatant, is a protein with the molecular mass above 100kDa. [ABSTRACT FROM AUTHOR]

Abstract: Earthworms irritated naturally (e.g. by predators) or experimentally extrude coelomocyte-containing coelomic fluid through the dorsal pores of the body wall. In the present study, the earthworms, Dendrobaena veneta, experimentally depleted of free-floating coelomocytes by multiple electric shocks (1min, 4.5V) remained fully vital and coelomocyte depletion was followed by the extensive cell replenishment, which was more efficient in the case of amoebocytes than autofluorescent eleocytes/chloragocytes, quantified by flow cytometry. Immunohistochemical procedure with antibodies against human Ki-67 proliferation antigens revealed proliferating cells on cytospin preparations of coelomocytes extruded by electric shock. Quantification of proliferating cells in the suspension of extruded coelomocytes was performed by flow cytometry on FL-2 profiles of propidium iodide-stained samples; riboflavin-derived autofluorescence of eleocytes/chloragocytes was lost during detergent treatment. As expected, the percentage of coelomocytes proliferating in coelomic fluid was increased during restoration of coelomocyte number after experimental depletion. The method described here may be very useful for investigations of antigen-driven proliferation of earthworm coelomocytes. [Copyright &y& Elsevier]

Abstract: Three-day dermal exposure of Dendrobaena veneta to metal ions differentially disrupts the immunocompetence/pathogen balance. Zn does not accumulate in the earthworm body, Cu accumulation is temperature-independent while Cd accumulation is stronger at 22°C than at 10°C. During in vitro incubation with metal ions at 22°C, growth of coelom-derived bacteria is enhanced by Zn, but significantly or almost completely inhibited by Cu or Cd. In contrast, under in vivo conditions at 22°C, bacterial load is decreased only after Cd exposure, but increased after Zn and Cu exposures. At 10°C bacteria growth is almost completely inhibited in all groups except Cu-treated animals. Coelomocyte number is unaffected in animals exposed to Zn, but significantly decreased after exposure to Cd (at 22°C) and Cu (at 22°C and 10°C) with concomitant changes of amoebocyte-to-eleocyte ratio in favour of amoebocytes. Metal exposure up-regulates expression of metallothioneins in coelomocytes, mainly amoebocytes. [Copyright &y& Elsevier]

Abstract: Microscope techniques, light microscope (LM), transmission electron microscope (TEM), scanning electron microscope (SEM) were employed to describe and classify coelomocytes of the oligochaete Dendrobaena veneta. Three main cell types were distinguished in the coelomic fluid: eleocytes, amoebocytes and granulocytes. Eleocytes are large, oval cells containing characteristic granules called chloragosomes. Amoebocytes are most numerous coelomocytes and have been divided into two types (I and II). Both amoebocytes of the types I and II often form aggregations of a few to about a dozen cells. Granulocytes are oval cells with spherical nuclei and cytoplasm containing polymorphic, electron dense granules. Contrary to the amoebocytes, the granulocytes do not form aggregations. Morphology and ultrastructure of coelomocytes are presented on micrographs: similarities and differences are compared to coelomocytes of related species. [Copyright &y& Elsevier]