Combined effects of cadmium and sulfamethoxazole on Eisenia fetida: Insights into accumulation, subcellular partitioning, biomarkers and toxicological responses.

Cadmium (Cd) and sulfamethoxazole (SMX) frequently coexist in farmlands, yet their synergistic toxicological impacts on terrestrial invertebrates remain unexplored. In this study, earthworms were exposed to artificial soils percolated with Cd (5 mg/kg), SMX (5 mg/kg) or combination of them for 7 days, followed by a 12-day elimination phase in uncontaminated soil. The uptake of Cd and SMX by the earthworms, along with their subcellular distribution, was meticulously analyzed. Additionally, a suite of biomarkers-including superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and weight loss-were evaluated to assess the health status of the earthworms and the toxicological effects of the Cd and SMX mixture. Notably, the cotreatment with Cd and SMX resulted in a significantly higher weight loss in Eisenia fetida (41.25 %) compared to exposure to Cd alone (26.84 %). Moreover, the cotreatment group exhibited substantially higher concentrations of Cd in the total internal body, fraction C (cytosol), and fraction E (tissue fragments and cell membranes) in Eisenia fetida compared to Cd alone counterparts. The combined exposure also significantly elevated the SMX levels in the total body and fraction C compared with the SMX-only treated earthworms. Additionally, Eisenia fetida subjected to the combined treatment showed markedly increased activities of SOD, CAT, and MDA compared to those treated with Cd alone. The effect addition indices (EAIs), ranging from 1.00 to 2.23, unequivocally demonstrated a synergistic effect of the combined treatments. Interestingly, relocating the earthworms to clean soil did not mitigate the observed adverse effects. These findings underscore the increased risk posed by the Cd-SMX complex to terrestrial invertebrates in agricultural areas.

Combined exposure to decabromodiphenyl ether and nano zero-valent iron aggravated oxidative stress and interfered with metabolism in earthworms.

Decabromodiphenyl ether (BDE-209) is a common brominated flame retardant in electronic waste, and nano zero-valent iron (nZVI) is a new material in the field of environmental remediation. Little is known about how BDE-209 and nZVI combined exposure influences soil organisms. During the 28 days study, we determined the effects of single and combined exposures to BDE-209 and nZVI on the oxidative stress and metabolic response of earthworms (Eisenia fetida). On day 7, compared to CK, malondialdehyde (MDA) content increased in most combined exposure groups. To remove MDA and reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities were induced in most combined exposure groups. On day 28, compared to CK, the activities of SOD and CAT were inhibited, while POD activity was significantly induced, indicating that POD plays an important role in scavenging ROS. Combined exposure to BDE-209 and nZVI significantly affected amino acid biosynthesis and metabolism, purine metabolism, and aminoacyl-tRNA biosynthesis pathways, interfered with energy metabolism, and aggravated oxidative stress in earthworms. These findings provide a basis for assessing the ecological impacts of using nZVI to remediate soils contaminated with BDE-209 from electronic waste.

Long-term toxic effects of iron-based metal-organic framework nanopesticides on earthworm-soil microorganism interactions in the soil environment.

With the widespread use of controlled-release nanopesticides in field conditions, the interactions between these nanopesticides and biological systems are complex and highly uncertain. The toxicity of iron-based metal organic frameworks (CF@MIL-101-SL) loaded with chlorfenapyr (CF) to terrestrial invertebrate earthworms in filter paper and soil environments and the potential mechanisms of interactions in the nanopesticide-earthworm-cornfield soil microorganism system were investigated for the first time. The results showed that CF@MIL-101-SL was more poisonous to earthworms in the contact filter paper test than suspension concentrate of CF (CF-SC), and conversely, CF@MIL-101-SL was less poisonous to earthworms in the soil test. In the soil environment, the CF@MIL-101-SL treatment reduced oxidative stress and the inhibition of detoxifying enzymes, and reduced tissue and cellular substructural damage in earthworms compared to the CF-SC treatment. Long-term treatment with CF@MIL-101-SL altered the composition and abundance of microbial communities with degradative functions in the earthworm intestine and soil and affected the soil nitrogen cycle by modulating the composition and abundance of nitrifying and denitrifying bacterial communities in the earthworm intestine and soil, confirming that soil microorganisms play an important role in reducing the toxicity of CF@MIL-101-SL to earthworms. In conclusion, this study provides new insights into the ecological risks of nanopesticides to soil organisms.

Effects of cadmium and pyrene on earthworm-associated bacterial communities: Unveiling new perspectives for soil pollution management.

Earthworms are considered to be excellent bioindicators of soil pollution. In recent years, there has been increasing interest in examining the effects of soil pollution on earthworm-associated microbiomes, with a particular focus on the gut microbiomes. However, relatively little effort has been invested in comprehensively investigating other microbiomes associated with earthworms and their responses to soil pollution. To fill this gap, we systematically studied the effects of Cd, pyrene, and combined pollution on the bacterial community in different vermicompartments, i.e., burrow wall, gut, and cast, in both epigeic Eisenia fetida and anecic Metaphire guillelmi, using a 2D-terraria incubator and high-throughput sequencing techniques. The results showed that bacterial alpha diversity followed the order of burrow wall > cast > gut, and this did not vary with soil pollution or earthworm ecotypes. Moreover, the dominant phyla in the vermicompartments were similar across different pollution treatments. Principal coordinate analysis (PCoA) revealed that the bacterial communities in different vermicompartments and ecotypes of earthworm were separated from each other, whereas they were grouped together in polluted treatments and unpolluted conditions. These results imply that even in polluted soil, vermicompartment and earthworm ecotypes remain the most significant factors affecting earthworm-associated microbiomes. However, the impacts of soil pollution on the bacterial composition in each vermicompartment were still evident. A comprehensive analysis revealed that the gut bacterial communities are more sensitive to soil contamination than casts and burrow wall in different ecotypes. Additionally, linear discriminant analysis of effect size (LefSe) identified several bacteria in Gemmatimonadota, the Firmicutes phylum in the burrow walls, and Patescibacteria (phyla) in the gut as potential biomarkers for pyrene contamination in soil. This research provides a comprehensive understanding of the effects of soil pollution on earthworm-associated microbiomes, thereby enhancing our understanding of earthworm ecotoxicology and soil pollution management.

Effects of soybean oil exposure on the survival, reproduction, biochemical responses, and gut microbiome of the earthworm Eisenia fetida.

With increasing production of kitchen waste, cooking oil gradually enters the soil, where it can negatively affect soil fauna. In this study, we explored the effects of soybean oil on the survival, growth, reproduction, tissue structure, biochemical responses, mRNA expression, and gut microbiome of earthworms (Eisenia fetida). The median lethal concentration of soybean oil was found to be 15.59%. Earthworm growth and reproduction were significantly inhibited following exposure to a sublethal concentration of soybean oil (1/3 LC50, 5.2%). The activity of the antioxidant enzymes total superoxide dismutase (T-SOD), peroxidase (POD), and catalase (CAT) were affected under soybean oil exposure. The glutathione (GSH) content decreased significantly, whereas that of the lipid peroxide malondialdehyde (MDA) increased significantly after soybean oil exposure. mRNA expression levels of the SOD, metallothionein (MT), lysenin and lysozyme were significantly upregulated. The abundance of Bacteroides species, which are related to mineral oil repair, and Muribaculaceae species, which are related to immune regulation, increased within the earthworm intestine. These results indicate that soybean oil waste is toxic to earthworms. Thus, earthworms deployed defense mechanisms involving antioxidant system and gut microbiota for protection against soybean oil exposure-induced stress.

The influence of ambient temperature and polyphenols from plant leaves on growth and the response to oxidative and nitrosative stress in African nightcrawler earthworm, Eudrilus eugeniae (Kinberg, 1867).

Earthworms are one of the organisms that may be affected by climate change. Finding ways to help them deal with this problem is, therefore, important and necessary. The objective of this experiment was to understand the influence of ambient temperature and polyphenols from mulberry (Morus alba L.), almond (Terminalia catappa L.) and cassava (Manihot esculenta (L.) Crantz) leaves on growth, ferric reducing antioxidant power (FRAP), malondialdehyde (MDA), hydrogen peroxide (H2O2) and nitric oxide (NO) concentration of the African night crawler, Eudrilus eugeniae (Kinberg, 1867) earthworm. The earthworms were cultured in 2 different conditions of ambient temperature, and in 4 types of substrate i.e. dairy cow faeces (BS), dairy cow faeces + mulberry leaves (BS + MA), almond leaves (BS + TC), and cassava leaves (BS + ME), respectively. At week 2 of the experiment, body weight, FRAP, MDA, H2O2 and NO were measured in the earthworms. It was found that the body weight gain (BWG) of the earthworms cultured in BS at cyclical temperature (26 + 1oC - 34 + 1oC - 26 + 1oC, CyT) was higher than the constant temperature (26 ± 1 °C, CoT) (P < 0.05). FRAP of earthworms cultured in BS + TC was higher than in other groups (P < 0.05). MDA of earthworms cultured at CyT was higher than ambient temperature at CoT (P < 0.05). At CyT, the MDA of earthworms cultured in BS + MA was higher than that of those cultured in BS, BS + TC and BS + ME (P < 0.05). NO of earthworms at CoT was higher than at CyT(P < 0.05). At CoT, the NO of earthworms cultured in BS + TC was lower than that of those cultured in BS + MA and BS + ME (P < 0.05). H2O2 of earthworms at CoT was higher than those at CyT (P < 0.05). The level of H2O2 of the earthworms cultured in BS + ME at CoT was higher than at CyT (P < 0.05). In addition, the H2O2 of earthworms cultured in both ambient temperatures and cultured in BS + MA was higher than the other groups (P < 0.05). These phenomena indicated that low and high ambient temperatures induced nitrosative and oxidative stress in earthworms, respectively. Mulberry leaves are toxic to earthworms. On the other hand, almond leaves could reduce nitrosative stress in earthworms. While at the CoT, cassava leaves induced the production of H2O2 in the earthworms.

An earthworm-like modular soft robot for locomotion in multi-terrain environments.

Robotic locomotion in subterranean environments is still unsolved, and it requires innovative designs and strategies to overcome the challenges of burrowing and moving in unstructured conditions with high pressure and friction at depths of a few centimeters. Inspired by antagonistic muscle contractions and constant volume coelomic chambers observed in earthworms, we designed and developed a modular soft robot based on a peristaltic soft actuator (PSA). The PSA demonstrates two active configurations from a neutral state by switching the input source between positive and negative pressure. PSA generates a longitudinal force for axial penetration and a radial force for anchorage, through bidirectional deformation of the central bellows-like structure, which demonstrates its versatility and ease of control. The performance of PSA depends on the amount and type of fluid confined in an elastomer chamber, generating different forces and displacements. The assembled robot with five PSA modules enabled to perform peristaltic locomotion in different media. The role of friction was also investigated during experimental locomotion tests by attaching passive scales like earthworm setae to the ventral side of the robot. This study proposes a new method for developing a peristaltic earthworm-like soft robot and provides a better understanding of locomotion in different environments.

Predicting the bioremediation potential of earthworms of different ecotypes through a multi-biomarker approach.

Earthworms are attracting the attention of bioremediation research because of their short-term impact on pollutant fate. However, earthworm-assisted bioremediation largely depends on the earthworm sensitivity to target pollutants and its metabolic capacity to break down contaminants. The most studied species in soil bioremediation has been Eisenia fetida, which inhabits the soil surface feeding on decomposing organic residues. Therefore, its bioremediation potential may be limited to organic matter-rich topsoil. We compared the detoxification potential against organophosphate (OP) pesticides of three earthworm species representative of the main ecotypes: epigeic, anecic, and endogeic. Selected biomarkers of pesticide detoxification (esterases, cytochrome P450-dependent monooxygenase, and glutathione S-transferase) and oxidative homeostasis (total antioxidant capacity, glutathione levels, and glutathione reductase [GR] and catalase activities) were measured in the muscle wall and gastrointestinal tract of E. fetida (epigeic), Lumbricus terrestris (anecic) and Aporrectodea caliginosa (endogeic). Our results show that L. terrestris was the most suitable species to bioremediate OP-contaminated soil for the following reasons: 1) Gut carboxylesterase (CbE) activity of L. terrestris was higher than that of E. fetida, whereas muscle CbE activity was more sensitivity to OP inhibition than that of E. fetida, which means a high capacity to inactivate the toxic oxon metabolites of OPs. 2) Muscle and gut phosphotriesterase activities were significantly higher in L. terrestris than in the other species. 3) Enzymatic (catalase and GR) and molecular mechanisms of free radical inactivation (glutathione) were 3- to 4-fold higher in L. terrestris concerning E. fetida and A. caliginosa, which reveals a higher potential to keep the cellular oxidative homeostasis against reactive metabolites formed during OP metabolism. Together with biological and ecological traits, these toxicological traits suggest L. terrestris a better candidate for soil bioremediation than epigeic earthworms.

Bioresponses of earthworm-microbiota symbionts to polychlorinated biphenyls in the presence of nano zero valent iron in soil.

Both earthworms and nano zero-valent iron (nZVI) have been recently regarded as important approaches for in-situ remediation of polychlorinated biphenyls (PCBs) in soil. However, the combined action of earthworms and nZVI toward PCBs, and the biological responses of earthworm-microbiota symbionts to nZVI-PCBs co-exposure in soil remediation systems remain unclear. In this study, a 28-d exposure with different levels of polychlorinated biphenyls (PCBs) and nZVI was applied to earthworm Eisenia fetida in an agricultural soil. Both physiological responses of earthworms and their surrounding microbiota in gut and soil were examined. Kinetic modelling parameters showed a doubled PCB accumulation in earthworms with the presence of nZVI. Meanwhile, nZVI-PCBs coexposure synergistically stimulated the activities of superoxide dismutase (SOD) and catalase (CAT), along with the elevated levels of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) in earthworms. Based on integrated metabolomic and 16S rRNA analysis, it was found that earthworms provided certain metabolites, e.g., S-(2-hydroxyethyl)glutathione, 16-hydroxypalmitic acid, and formamide, beneficial to PCB-degrading microbiota (Novosphingobium and Achromobacter) in the intestine. Our findings of nZVI-enhanced PCB bioaccumulation and the defense mechanism afforded by the earthworm-microbiota symbionts toward PCB-nZVI exposure show the promise of combining earthworms with nZVI for the remediation of PCBs-contaminated soil.

Novel Venetin-1 nanoparticle from earthworm coelomic fluid as a promising agent for the treatment of non-small cell lung cancer.

The present research shows the antitumor activity of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworms against A549 cancer cells. The investigations are a continuation of experiments on the antitumor activity of coelomic fluid obtained from this species. The Venetin-1 nanoparticle was obtained after thermal treatment of the coelomic fluid, separation from coelomocytes, filtration, and lyophilization. The preparation showed a selective effect on cancer cells, whereas normal cells were unaffected. Venetin-1 was effective against the lung cancer cells at doses of 31.3 and 62.5 µg/ml, and the results were imaged using light microscopy and scanning electron microscopy (SEM). The cells died mainly via the apoptosis pathway. Necrotic cells appeared sporadically in the microscopic view. SEM imaging revealed complete destruction of the A549 cells after the incubation with Venetin-1. The atomic force microscopy (AFM) analyses showed changes in the topography, peak force error images, and Young's modulus (elasticity) of the A549 cells after the incubation with Venetin-1. The transmission electron cryomicroscopy (Cryo-TEM) analysis indicated a polymeric nature of the analyzed preparation. The samples of Venetin-1 showed a very homogeneous size profile with the microparticle size of approximately 58.23 nm. A significant decrease in Venetin-1 binding to sphingomyelin was observed. Venetin-1 lost its pore-forming activity or deactivation of the pore-forming activity occurred. This confirms the absence of hemolytic capacity of Venetin-1 towards red blood cells. The conducted analyses show the suitability of the obtained complex for biomedical research. The next step will consist in analyses of the effect of Venetin-1 on the immune system in mice.

Toxicokinetics and toxicodynamics of copper and cadmium in the soil invertebrate Enchytraeus crypticus (Oligochaeta).

The aim of this study was to evaluate the toxicokinetics-toxicodynamics (TKTD) of Cu and Cd in the soil model organism Enchytraeus crypticus, and assess the development of internal effect concentrations over time. Animals were exposed in LUFA 2.2 soil spiked with increasing concentrations of Cu and Cd. Survival, reproduction and internal metal concentrations in the animals were evaluated at different points in time over a period of 21 days. Internal concentrations increased with time, for Cu reaching a steady state after c. 10 days, except for the highest test concentration, and for Cd continuing to increase after 21 days. Applying a one-compartment model to all data together, estimated uptake and elimination rate constants for Cu and Cd were 0.08 and 0.45 kg soil/kg organism/day and 0.4 and 0.04 per day, respectively. Median lethal concentrations, based on total soil concentrations, decreased with time for Cu and did not reach a steady state level, but they did not change with time for Cd. The LC50inter (based on internal concentrations) was 75 mg Cu/kg body DW and > 800 mg Cd/kg body weight. Animals were able to regulate Cu internal concentrations, keeping them low, while for Cd internal concentrations continued to increase showing lack of regulation and also the importance of exposure time. This study highlights the advantages of using a TKTD approach to understand the relation between organism survival and internal Cu or Cd concentrations over time.

Ovary micromorphology and oogenesis in a rhyacodriline oligochaete (Clitellata: Naididae, Rhyacodrilinae).

The main goal of the article is to describe the ovary organization and oogenesis in Peristodrilus montanus, an aquatic oligochaete of the subfamily Rhyacodrilinae. The presented analysis will not only enrich the knowledge about how eggs are formed but, because of the suggested conservatism of ovary organization in clitellate annelids, can contribute to disentangling the complex phylogenetic relationships of the rhyacodrilines within Naididae. The paired, conically shaped ovaries are located in segment XI. They are composed of a dozen or so syncytial germ-line cysts, which are associated with somatic cells. Each germ cell in a cyst has one intercellular bridge that joins it to a central and anuclear cytoplasmic mass, the cytophore. This pattern of cyst organization is typical for all clitellates that have been studied to date. Initially, the germ cells in a cyst undergo a synchronous development, however, there is no synchrony between cysts, and therefore there is a developmental gradient (oogonia, pre-diplotene germ cells, germ cells in diplotene) of oogenesis along the long ovary axis. The cysts are composed of a maximum of 32 cells. Cysts with cells in diplotene detach from the ovaries and the extraovarian phase of oogenesis begins. The developmental synchrony is lost, one cell (an oocyte) per cyst starts to gather cell components and yolk and grows considerably. The remaining cells grow to some extent and function as nurse cells. Like in other microdriles, P. montanus oocytes are rich in yolk; other features of oogenesis are also similar to those that are known from other microdrile taxa. The system of ovary organization found in the studied species is broadly similar to the corresponding features known from Naidinae and Phreodrilidae and, to some extent, in Enchytraeidae. However, this system is different from the one that is known in Tubificinae, Limnodriloidinae and Branchiurinae.

Lateral bending and buckling aids biological and robotic earthworm anchoring and locomotion.

Earthworms (Lumbricus terrestris) are characterized by soft, highly flexible and extensible bodies, and are capable of locomoting in most terrestrial environments. Previous studies of earthworm movement focused on the use of retrograde peristaltic gaits in which controlled contraction of longitudinal and circular muscles results in waves of shortening/thickening and thinning/lengthening of the hydrostatic skeleton. These waves can propel the animal across ground as well as into soil. However, worms benefit from axial body bends during locomotion. Such lateral bending and buckling dynamics can aid locomotor function via hooking/anchoring (to provide propulsion), modify travel orientation (to avoid obstacles and generate turns) and even generate snake-like undulatory locomotion in environments where peristaltic locomotion results in poor performance. To the best of our knowledge, lateral bending and buckling of an earthworm's body has not yet been systematically investigated. In this study, we observed that within confined environments, worms use lateral bending and buckling to anchor their body to the walls of their burrows and tip (anterior end) bending to search the environment. This locomotion strategy improved the performance of our soft-bodied robophysical model of the earthworm both in a confined (in an acrylic tube) and above-ground heterogeneous environment (rigid pegs), where present peristaltic robots are relatively limited in terradynamic capabilities. In summary, lateral bending and buckling facilitates the mobility of earthworm locomotion in diverse terrain and can play an important role in the creation of low cost soft robotic devices capable of traversing a variety of environments.

Identification and expression of adenosine deaminases acting on tRNA (ADAT) during early tail regeneration of the earthworm.

BACKGROUND: RNA editing is a widespread phenomenon in all metazoans. One of the common RNA editing event is the chemical conversion of adenosine to inosine (A-to-I) catalyzed by adenosine deaminases acting on tRNA (ADAT). During D. melanogaster development, the ADAT1 transcript was found to localize mainly to the central nervous system including brain and ventral nerve cord during brain development. Although an earthworm adenosine deaminases acting on mRNA (ADAR) has been identified and its possible implication in earthworm regeneration has been investigated, there is little accumulated information on ADAT and tRNA editing in the annelid including terrestrial earthworms. OBJECTIVE: This study aimed to investigate the molecular characteristics and the expression pattern of earthworm ADAT during tail regeneration to understand its physiological significance. METHODS: Nucleotide sequence of Ean-ADAT was retrieved from the genome assembly of Eisenia andrei via Basic Local Alignment Search Tool (BLAST). The genome assembly of Eisenia andrei was downloaded from National Genomics Data Center ( http://bigd.big.ac.cn/gwh/ ). The alignment and phylogenetic relationship of the core deaminase domains of ADATs and ADARs were analyzed. Its temporal expression during early tail regeneration was measured using real-time PCR. RESULTS: The open reading frame of Ean-ADAT consists of 1719 nucleotides encoding 573 amino acids. Domain analysis indicates that Ean-ADAT has a deaminase domain composed of 498 amino acids and a predicted nuclear localization signal at the N-terminal. Its subcellular localization was predicted to be nuclear. The core deaminase region of Ean-ADAT encompasses the three active-site motifs, including zinc-chelating residues and a glutamate residue for catalytic activity. In addition, Ean-ADAT shares highly conserved RNA recognition region flanking the third cysteine of the deaminase motif with other ADAT1s even from the yeast. Multiple sequence alignment and phylogenetic analysis indicate that Ean-ADAT shows greater similarity to vertebrate ADARs than to yeast Tad1p. Ean-ADAT mRNA expression began to remarkably decrease before 12 h post-amputation, showing a tendency to gradual decrease until 7 dpa and then it slightly rebounded at 10 dpa. CONCLUSIONS: Our results demonstrate that Ean-ADAT belongs to a class of ADAT1s and support the hypothesis of a common evolutionary origin for ADARs and ADATs. The temporal expression of Ean-ADAT could suggest that its activity is unrelated to the molecular mechanisms of dedifferentiation.

Assessing single effects of sugarcane pesticides fipronil and 2,4-D on plants and soil organisms.

The continuous growth in global population since the beginning of the 20th century result in the necessity of food and energy provision favoring the intensive use of agricultural products such as pesticides. Although pesticides are important to prevent losses in the conventional chemically based agriculture, they frequently present side effects, which goes against agricultural production. The use of pesticides cause direct and indirect effects to soil organisms unbalancing essential soil processes (e.g. primary production, organic matter decomposition, nutrient cycling). Under tropical conditions, very little is known regarding the effects of pesticides to terrestrial organisms. Hence, the aim of the present study was to assess the ecotoxicological effects of the herbicide DMA® 806 BR (active ingredient: 2,4-D) and the insecticide Regent® 800 WG (active ingredient: fipronil), on terrestrial plant species (the dicot Raphanus sativus var. acanthioformis and the monocot Allium cepa), and soil invertebrates (the collembolan Folsomia candida and the enchytraeid Enchytraeus crypticus), using natural (NS) and artificial soils (TAS). For both pesticides, negative effects on non-target species were observed at concentrations lower than the doses recommended to prevent pests in sugarcane fields. For both soils, the dicot species was the most affected by the herbicide (R. sativus > A. cepa > F. candida > E. crypticus) and the collembolan species was the most affected by the insecticide (F. candida > E. crypticus = R. sativus = A. cepa). Although the order of the organisms' sensitivity for both pesticides was the same in both soils, results showed that the extent of the effects was soil dependent. Considering the ecologically relevant concentrations tested, and their severe effects to non-target organisms, it may be concluded that the use of fipronil and 2,4-D under recommended conditions may pose a risk to the terrestrial environment.

Microorganization of ovaries and oogenesis of Haplotaxis sp. (Clitellata: Haplotaxidae).

Ovaries of Haplotaxis sp. were studied in active and nonactive states, that is, in a sexually mature specimen and in specimens outside of the reproductive period. Two pairs of ovaries were found in segments XI and XII. Especially in the nonactive state, they were in close contact with copulatory glands. Each ovary was composed of germ cells interconnected with syncytial cysts, which were enveloped by a layer of somatic cells. Within cysts each germ cell had one ring canal connecting it to the common anuclear cytoplasmic mass called a cytophore. During oogenesis clustering germ cells differentiated into nurse cells and oocytes; thus, the oogenesis was recognized as meroistic. Vitellogenic oocytes were detached from the ovaries and continued yolk absorption within the body cavity. Because recent studies have shown the variety of ovaries and germ line cyst organization in clitellates and suggest their evolutionary conservatism at the family or subfamily level, the data presented here can be valid in understanding the phylogenetic relationships among Clitellata. In this context, ovaries found in Haplotaxis sp. resembled those of the "Tubifex" type. "Tubifex" ovaries are characteristic for numerous microdrile taxa (tubificines, limnodriloidines, propappids, lumbriculids, and leech-like branchiobdellids) and can be regarded as the primary character for these Clitellata in which germ-line cysts are formed during early oogenesis. As the family Haplotaxidae is currently considered to be paraphyletic and the species studied here belongs to Haplotaxidae sensu stricto, our results support the close relationship of Haplotaxidae sensu stricto to the clade consisting of Lumbriculidae, Branchiobdellida, and Hirudinida, in which lumbriculids are sister to the latter two.

Oxidative stress in benthic oligochaete worm, Tubifex tubifex induced by sublethal exposure to a cationic surfactant cetylpyridinium chloride and an anionic surfactant sodium dodecyl sulfate.

The present study was assessed to determine the in vivo toxic effects of a cationic surfactant, cetylpyridinium chloride (CPC), and an anionic surfactant, sodium dodecyl sulfate (SDS) in terms of oxidative stress biomarkers in benthic oligochaete worm Tubifex tubifex for 14 days. The investigation demonstrated that sublethal concentrations of CPC (0.0213, and 0.0639 mg L-1) and SDS (1.094 and 3.092 mg L-1)induced paramount alterations in the oxidative stress enzymes in Tubifex tubifex. Superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH), and glutathione peroxidase (GPx) exhibited an initial notable increase in their activities in the surfactants exposed worms at 1d and 7d of exposure period followed by consequential reduction at 14d exposure period with respect to control, while catalase (CAT) and malondialdehyde (MDA) activities markedly incremented gradually throughout the exposure periods. Through the construction of the correlation matrix and integrated biomarker response (IBR), the effects of CPC and SDS on Tubifex tubifex were distinguished. These results indicate that exposure to these cationic and anionic surfactants modulates the levels of oxidative stress enzymes in Tubifex tubifex.

Wormcasts produced by three earthworm species (Alma millsoni, Eudrilus eugeniae and Libyodrilus violaceus) exposed to a glyphosate-based herbicide reduce growth, fruit yield and quality of tomato (Lycopersicon esculentum).

It remains unknown if casts produced by earthworms exposed to a glyphosate-based herbicide (GBH) will retain their agricultural benefit. This study investigated the agricultural importance of surface casts produced by three earthworm species (Alma millsoni, Eudrilus eugeniae and Libyodrilus violaceus) exposed to a GBH on growth, fruit yield and quality of tomato (Lycopersicon esculentum). We sprayed 60 buckets (i.e 20 buckets/earthworm species) containing 20 adult earthworms of each species with 115.49 ml/m2 of Roundup® Alphée (Exposed) while another 60 buckets with earthworm species were sprayed with water (Control). Surface casts produced by the earthworms were collected for 1st, 2nd, 4th, 6th and 8th week post herbicide application. Tomato planting experiment on soil treated with the casts, NPK fertilizer and normal soil were grouped into 32 treatments. Tomato growth performance, yield and quality were evaluated with standard methods. Only the tomatoes planted with the casts of the exposed earthworms were unable to set fruit. There was no significant difference (p < 0.05) in yields of tomatoes planted with the control casts of E. eugeniae and A. millsoni compared to those planted with fertilizer. Vitamin C and ß-carotene contents increased in fruits of tomatoes planted with control cast of A. millsoni while reductions were recorded in the fruits of tomatoes planted with the control casts of E. eugeniae and L. violaceus relative to those planted with fertilizer. Surface casts of earthworms exposed to GBH could not enhance tomato growth while casts produced by unexposed earthworms greatly improved the performance of tomato plant.

The toxicity of silver nanomaterials (NM 300K) is reduced when combined with N-Acetylcysteine: Hazard assessment on Enchytraeus crypticus.

The widespread production and use of silver nanomaterials (AgNMs) in consumer and medical products have been raising environmental concerns. Once in the environment, the soil is one of the major sinks of AgNMs due to e.g. sewage sludge applications, and invertebrates are directly exposed. In this study, we investigate the potential of N-acetylcysteine (NAC) to reduce the toxic effects of Ag NM300 K (and AgNO3) on the soil invertebrate Enchytraeus crypticus. Ag NM300 K induces mortality, reproduction impairment, and avoidance. The addition of NAC to the soil showed a remarkable reduction in the toxicity of Ag, indicating that NAC can act as a detoxifying agent for terrestrial organisms exposed to Ag materials. That the reduction in toxicity likely is caused by thiol groups, was confirmed by GSH and GSSH studies. Identifying the mechanisms and hence alternatives that allow the recovery of contaminated soils is an important mitigation measure to promote environmental safety and reduce the associated risks to human health. Further, it may inform on strategies to implement in safe-by-design industry development.

Subcellular partitioning of cadmium and lead in Eisenia fetida and their effects to sperm count, morphology and apoptosis.

Earthworms and their biomarkers are considered good indicators for assessing the effects of toxic chemicals. Therefore, in this study, we exposed Eisenia fetida to lethal and sub-lethal concentrations of Cd and Pb nitrate in artificial soil for 14 and 28 days to evaluate the impact on subcellular partitioning, lethal toxicity (LC50), growth, sperm count, morphology and apoptosis (using TUNEL assay). The soluble internal pools of both metals were good predictors of the responses of biomarkers. We found sperm deformation, TUNEL positive sperms and weight loss positively and sperm count negatively correlated with the concentrations of Cd and Pb in the total internal and cytosolic fraction (p < 0.01) and to a lesser extent with Pb concentrations in the granular fraction (p < 0.05). Fourteen days LC50 for Cd and Pb were 2169 ±â€¯322 and 6387 ±â€¯904 µg/g, respectively. Cadmium and Pb caused a significant depression in sperm count after 14 (Cd: up to 46.9%; Pb: up to 36.24%) and 28 (Cd: up to 72.47%; Pb: up to 43.12%) days of exposure relative to the control (p < 0.05). Cadmium induced higher abnormality in sperm heads than Pb. For both metals, TUNEL positive sperms significantly increased after 14 (Cd: up to 14.17%; Pb: up to 16.33%) and 28 (Cd: up to 16.33%; Pb: up to 11.67%) days of exposure compared with the control (p < 0.05). The findings of this study, illustrate the importance of considering sperm parameters as a rapid, easy and sensitive biomarker for the evaluation of metal toxicity.