Optimizing the lyophilization of Lumbricus terrestris erythrocruorin.

Haemorrhagic shock is a leading cause of death worldwide. Blood transfusions can be used to treat patients suffering severe blood loss but donated red blood cells (RBCs) have several limitations that limit their availability and use. To solve the problems associated with donated RBCs, several acellular haemoglobin-based oxygen carriers (HBOCs) have been developed to restore the most important function of blood: oxygen transport. One promising HBOC is the naturally extracellular haemoglobin (i.e. erythrocruorin) of Lumbricus terrestris (LtEc). The goal of this study was to maximise the portability of LtEc by lyophilising it and then testing its stability at elevated temperatures. To prevent oxidation, several cryoprotectants were screened to determine the optimum formulation for lyophilisation that could minimise oxidation of the haem iron and maximise recovery. Furthermore, samples were also deoxygenated prior to storage to decrease auto-oxidation, while resuspension in a solution containing ascorbic acid was shown to partially reduce LtEc that had oxidised during storage (e.g. from 42% Fe3+ to 11% Fe3+). Analysis of the oxygen equilibria and size of the resuspended LtEc showed that the lyophilisation, storage, and resuspension processes did not affect the oxygen transport properties or the structure of the LtEc, even after 6 months of storage at 40 °C. Altogether, these efforts have yielded a shelf-stable LtEc powder that can be stored for long periods at high temperatures, but future animal studies will be necessary to prove that the resuspended product is a safe and effective oxygen transporter in vivo.

Trophic Position of the White Worm (Enchytraeus albidus) in the Context of Digestive Enzyme Genes Revealed by Transcriptomics Analysis.

To assess the impact of Enchytraeidae (potworms) on the functioning of the decomposer system, knowledge of the feeding preferences of enchytraeid species is required. Different food preferences can be explained by variations in enzymatic activities among different enchytraeid species, as there are no significant differences in the morphology or anatomy of their alimentary tracts. However, it is crucial to distinguish between the contribution of microbial enzymes and the animal's digestive capacity. Here, we computationally analyzed the endogenous digestive enzyme genes in Enchytraeus albidus. The analysis was based on RNA-Seq of COI-monohaplotype culture (PL-A strain) specimens, utilizing transcriptome profiling to determine the trophic position of the species. We also corroborated the results obtained using transcriptomics data from genetically heterogeneous freeze-tolerant strains. Our results revealed that E. albidus expresses a wide range of glycosidases, including GH9 cellulases and a specific digestive SH3b-domain-containing i-type lysozyme, previously described in the earthworm Eisenia andrei. Therefore, E. albidus combines traits of both primary decomposers (primary saprophytophages) and secondary decomposers (sapro-microphytophages/microbivores) and can be defined as an intermediate decomposer. Based on assemblies of publicly available RNA-Seq reads, we found close homologs for these cellulases and i-type lysozymes in various clitellate taxa, including Crassiclitellata and Enchytraeidae.

Assessing dietary specialization to inform the conservation of the fairy pitta (Pitta nympha), an endangered vermivore.

Quantifying the diet of endangered species is crucial for conservation, especially for diet specialists, which can be more susceptible to environmental changes. The vulnerable fairy pitta (Pitta nympha) is considered a specialist that primarily feeds its nestlings with earthworms. However, there have been few studies of the nestling diet provisioned by parents, and no assessments of earthworm proportion in the diet of adults. Our study aimed to fill these gaps, shedding light on crucial factors for conservation. Combining new observations with existing literature, we confirmed a consistent dominance of earthworms in the nestling diet, regardless of rainfall, nestling age, and time of day. We extrapolated the total earthworm consumption during a breeding event, accounting for potential variation in the availability of earthworms and their prevalence in the adult diet. We used literature-based earthworm densities in pitta habitats and our estimates of family earthworm consumption to calculate the habitat area that could provide a pitta family with the number of earthworms consumed during a breeding event. The predictions matched observed pitta home range sizes when assumed that the adult diet is comprised of approximately 70% earthworms. The results highlight the importance of earthworm-rich habitats for conservation planning of the fairy pitta. To mitigate the effects of habitat destruction, we discuss conservation practices that may involve enhancing earthworm abundance in natural habitats and providing vegetation cover for foraging pittas in adjacent anthropogenic habitats rich in earthworms. To guide conservation efforts effectively, future studies should investigate whether previously reported breeding in developed plantation habitats is due to high earthworm abundance there. Future studies should also quantify correlations between local earthworm densities, home range size, and the breeding success of the fairy pitta.

Effect of conventional and biodegradable microplastics on earthworms during vermicomposting process.

The potential effect of microplastics is an increasingly growing environmental issue. However, very little is known regarding the impact of microplastics on the vermicomposting process. The present study explored the effect of non-biodegradable (low density polyethylene; LDPE) and biodegradable (polybutylene succinate-co-adipate; PBSA) microplastics on earthworm Eisenia fetida during vermicomposting of cow dung. For this, earthworms were exposed to different concentrations (0, 0.5, 1 and 2%) of LDPE and PBSA of 2 mm size. The cow dung supported the growth and hatchlings of earthworms, and the toxicity effect of both LDPE and PBSA microplastics on Eisenia fetida was analyzed. Microplastics decreased the body weight of earthworms and there was no impact on hatchlings. The body weight of earthworm decreased from 0 to 60th day by 18.18% in 0.5% of LDPE treatment, 5.42% in 1% of LDPE, 20.58% in 2% of LDPE, 19.99% in 0.5% of PBSA, 15.09% in 1% of PBSA and 16.36% in 2% of PBSA. The physico-chemical parameters [pH (8.55-8.66), electrical conductivity (0.93-1.02 (S/m), organic matter (77.6-75.8%), total nitrogen (3.95-4.25 mg/kg) and total phosphorus (1.16-1.22 mg/kg)] do not show much significant changes with varying microplastics concentrations. Results of SEM and FTIR-ATR analysis observed the surface damage of earthworms, morphological and biochemical changes at higher concentrations of both LDPE and PBSA. The findings of the present study contribute to a better understanding of microplastics in vermicomposting system.

Soft skeletons transmit force with variable gearing.

A hydrostatic skeleton allows a soft body to transmit muscular force via internal pressure. A human's tongue, an octopus' arm and a nematode's body illustrate the pervasive presence of hydrostatic skeletons among animals, which has inspired the design of soft engineered actuators. However, there is a need for a theoretical basis for understanding how hydrostatic skeletons apply mechanical work. We therefore modeled the shape change and mechanics of natural and engineered hydrostatic skeletons to determine their mechanical advantage (MA) and displacement advantage (DA). These models apply to a variety of biological structures, but we explicitly consider the tube feet of a sea star and the body segments of an earthworm, and contrast them with a hydraulic press and a McKibben actuator. A helical winding of stiff, elastic fibers around these soft actuators plays a critical role in their mechanics by maintaining a cylindrical shape, distributing forces throughout the structure and storing elastic energy. In contrast to a single-joint lever system, soft hydrostats exhibit variable gearing with changes in MA generated by deformation in the skeleton. We found that this gearing is affected by the transmission efficiency of mechanical work (MA×DA) or, equivalently, the ratio of output to input work. The transmission efficiency changes with the capacity to store elastic energy within helically wrapped fibers or associated musculature. This modeling offers a conceptual basis for understanding the relationship between the morphology of hydrostatic skeletons and their mechanical performance.

Congener-specific fate and impact of microcystins in the soil-earthworm system.

Microcystins (MCs) have a significant influence on aquatic ecosystems, but little is known about their terrestrial fate and impact. Here, we investigated the fate of two MCs (MC-LR and MC-RR) in the soil-earthworm system, with consideration of their congener-specific impact on earthworm health, soil bacteria, and soil metabolome. Although MCs had little acute lethal effect on earthworms, they caused obvious growth inhibition and setae rupture. Relative to MC-RR, MC-LR exhibited higher bioaccumulation and the resulting dermal lesions and deformation of longitudinal muscles. While the incorporation of both MCs into soils stimulated pathogenic bacteria and depressed oxidative stress tolerant bacteria, the response among soil nitrification and glutathione metabolism differed between the two congeners. The dissipation kinetics of MCs obeyed the first-order model. Earthworms stimulated soil N-cycling enzyme activities, increased the abundance of MC-degrading bacteria, and promoted bacterial metabolic functions related to glutathione metabolism, xenobiotics biodegradation, and metabolism of amino acids that comprise MCs, which accelerated the dissipation of MC-LR and MC-RR by 227% and 82%, respectively. These results provide evidence of significant congener differences in the terrestrial fate and impact of MCs, which will enable a better understanding of their role in mediating soil functions and ecosystem services.

Predicting the bioavailability of polycyclic aromatic hydrocarbons in rhizosphere soil using a new novel in situ solid-phase microextraction technique.

In situ measurement of the bioavailability of organic pollutants in soil is crucial for understanding their environmental behavior and assessing health risks. Due to the high heterogeneity of soil, microscale determination is crucial for achieving high accuracy, but few methods are available. In this study, microsized probes coated with polydimethylsiloxane (PDMS) were used to measure the bioavailability of polycyclic aromatic hydrocarbons (PAHs) in soil in situ. The concentrations of PAHs enriched by the PDMS-coated probes correlated well with the results of bioassays using earthworms (R2 = 0.92-0.99) and ryegrass roots (R2 = 0.92-0.99). Compared with other chemical extraction methods, such as n-butanol extraction, the proposed method has advantages such as in situ operation, microvolume analysis, and negligible interference to the soil environment. In the soil rhizosphere zone, PAHs bioavailability decreased in the following order: rhizosphere > near-rhizosphere > far-rhizosphere. The bioavailability of PAHs in soil amended with biochar was also successfully characterized by the proposed method. Thus, this study developed an in situ and microscale method to predict the bioavailability of organic pollutants in contaminated soils and provides new insight into migration and transformation processes in rhizosphere soil.

Plant growth-promoting bacteria isolated from earthworms enhance spinach growth and its phytoremediation potential in metal-contaminated soils / Las bacterias promotoras del crecimiento vegetal aisladas de lombrices de tierra mejoran el crecimiento de las espinacas y su potencial de fitorremediación en suelos contaminados con metales

The aim of this study was to evaluate the impact of metal-tolerant plant growth-promoting bacteria (PGPB) isolated from the chloragogenous tissue of Aporrectodea molleri, which represents a unique habitat. Our objectives were to investigate their effects on the growth of Spinacia oleracea under heavy metal stress and assess their potential for enhancing phytoremediation capabilities. The experiment was conducted in an alkaline soil contaminated with 7 mg kg-1 of cadmium, 100 mg kg-1 of nickel, 150 mg kg-1 of copper, 300 mg kg-1 of Zinc, and mg kg-1 of 600 Manganese. The results showed that heavy metal stress considerably diminished root (42.8%) and shoot length (60.1%), biomass (80%), chlorophyll content (41%), soil alkaline (45%), and acid (51%) phosphatases (42%) and urease (42%). However, soil inoculation with bacterial isolates remarkably improved plant growth. Soil bioaugmentation increased spinach growth (up to 74.5% for root length, up to 106.3% for shoot length, and up to 5.5 folds for fresh biomass) while significantly increasing soil enzyme activity and NPK content. Multivariate data analysis indicated that soil inoculation with Bacillus circulans TC7 promoted plant growth while limiting metal bioaccumulation, whereas Pseudomonas sp. TC33 and Bacillus subtilis TC34 increased metal bioaccumulation in spinach tissues while minimizing their toxicity. Our study confirms that earthworms are a reservoir of multi-beneficial bacteria that can effectively improve phytoremediation efficiency and mitigate the toxic effects of heavy metals on plant growth. Further studies are needed to investigate the long-term effects and feasibility of using these isolates as a consortium in field applications.(AU)

Annelid adult cell type diversity and their pluripotent cellular origins.

Many annelids can regenerate missing body parts or reproduce asexually, generating all cell types in adult stages. However, the putative adult stem cell populations involved in these processes, and the diversity of cell types generated by them, are still unknown. To address this, we recover 75,218 single cell transcriptomes of the highly regenerative and asexually-reproducing annelid Pristina leidyi. Our results uncover a rich cell type diversity including annelid specific types as well as novel types. Moreover, we characterise transcription factors and gene networks that are expressed specifically in these populations. Finally, we uncover a broadly abundant cluster of putative stem cells with a pluripotent signature. This population expresses well-known stem cell markers such as vasa, piwi and nanos homologues, but also shows heterogeneous expression of differentiated cell markers and their transcription factors. We find conserved expression of pluripotency regulators, including multiple chromatin remodelling and epigenetic factors, in piwi+ cells. Finally, lineage reconstruction analyses reveal computational differentiation trajectories from piwi+ cells to diverse adult types. Our data reveal the cell type diversity of adult annelids by single cell transcriptomics and suggest that a piwi+ cell population with a pluripotent stem cell signature is associated with adult cell type differentiation.

Community Structure and Water Quality Assessment of Benthic Macroinvertebrates in Hongze Lake.

This study investigated the species, density, biomass and physicochemical factors of benthic macroinvertebrates in Hongze Lake from 2016 to 2020. Redundancy analysis (RDA) was used to analyze the relationship between physicochemical parameters and the community structure of macroinvertebrates. Macroinvertebrate-based indices were used to evaluate the water quality conditions in Hongze Lake. The results showed that a total of 50 benthic species (10 annelids, 21 arthropods and 19 mollusks) were collected. The community structure of benthic macroinvertebrates varied in time and space. The dominant species were Limnodrilus hoffmeisteri (L.hoffmeisteri), Corbicula fluminea (C.fluminea), Nephtys oligobranchia (N.oligobranchia). In 2016, arthropods such as Grandidierella sp. were the dominant species of benthos in Hongze Lake while annelids and mollusks dominated from 2017 to 2020, such as L.hoffmeisteri, N.oligobranchia, C.fluminea. The benthic fauna of Chengzi Lake and Lihewa District were relatively abundant and showed slight variation, while the benthic macroinvertebrates of the Crossing the water area were few and varied greatly. RDA showed that changes in benthic macroinvertebrate structure were significantly correlated with dissolved oxygen (DO), Pondus Hydrogenii (pH) and transparency (SD). The Shannon Wiener, Pielou, and Margalef indices indicate that Hongze Lake is currently in a moderately polluted state. Future studies should focus on the combined effects of various physicochemical indicators and other environmental factors on benthic communities.

Size-dependent vector effects of microplastics on bioaccumulation of hydrophobic organic contaminants in earthworm: A dual-dosing study.

The potential of microplastics to act as a vector for anthropogenic contaminants is of rising concern. However, directly quantitatively determining the vector effects of microplastics has been rarely studied. Here, we present a dual-dosing method that simulates the chemical bioaccumulation from soil and microplastics simultaneously, wherein unlabeled hydrophobic organic contaminants (HOCs) were spiked in the soil and their respective isotope-labeled reference compounds were spiked on the polyethylene microplastics. The comparison of the bioavailability, i.e., the freely dissolved concentration in soil porewater and bioaccumulation by earthworm, between the unlabeled and isotope-labeled HOCs was carried out. Relatively higher level of bioavailability of the isotope-labeled HOCs was observed compared to the unlabeled HOCs, which may be attributed to the irreversible desorption of HOCs from soil particles. The average relative fractions of bioaccumulated isotope-labeled HOCs in the soil treated with 1 % microplastics ranged from 6.9 % to 46.4 %, which were higher than those in the soil treated with 0.1 % microplastics. Treatments with the smallest microplastic particles were observed to have the highest relative fractions of bioaccumulated isotope-labeled HOCs, with the exception of phenanthrene, suggesting greater vector effects of smaller microplastic particles. Biodynamic model analysis indicated that the contribution of dermal uptake to the bioaccumulation of isotope-labeled HOCs was higher than that for unlabeled HOCs. This proposed method can be used as a tool to assess the prospective vector effects of microplastics in complex environmental conditions and would enhance the comprehensive understanding of the microplastic vector effects for HOC bioaccumulation.

Exploring the potential of earthworm gut bacteria for plastic degradation.

The use of plastic mulch films in agriculture leads to the inevitable accumulation of plastic debris in soils. Here, we explored the potential of earthworm gut-inhabiting bacterial strains (Mycobacterium vanbaalenii (MV), Rhodococcus jostii (RJ), Streptomyces fulvissimus (SF), Bacillus simplex (BS), and Sporosarcina globispora (SG) to degrade plastic films (⌀ = 15 mm) made from commonly used polymers: low-density polyethylene film (LDPE-f), polylactic acid (PLA-f), polybutylene adipate terephthalate film (PBAT-f), and a commercial biodegradable mulch film, Bionov-B® (composed of Mater-Bi, a feedstock with PBAT, PLA and other chemical compounds). A 180-day experiment was conducted at room temperature (x̄ =19.4 °C) for different strain-plastic combinations under a low carbon media (0.1× tryptic soy broth). Results showed that the tested strain-plastic combinations did not facilitate the degradation of LDPE-f (treated with RJ and SF), PBAT-f (treated with BS and SG), and Bionov-B (treated with BS, MV, and SG). However, incubating PLA-f with SF triggered a reduction in the molecular weights and an increase in crystallinity. Therefore, we used PLA-f as model plastic to study the influence of temperature ("room temperature" & "30 °C"), carbon source ("carbon-free" & "low carbon supply"), and strain interactions ("single strains" & "strain mixtures") on PLA degradation. SF and SF + RJ treatments significantly fostered PLA degradation under 30 °C in a low-carbon media. PLA-f did not show any degradation in carbon-free media treatments. The competition between different strains in the same system likely hindered the performance of PLA-degrading strains. A positive correlation between the final pH of culture media and PLA-f weight loss was observed, which might reflect the pH-dependent hydrolysis mechanism of PLA. Our results situate SF and its co-culture with RJ strains as possible accelerators of PLA degradation in temperatures below PLA glass transition temperature (Tg). Further studies are needed to test the bioremediation feasibility in soils.

[Effects of Vermicomposting on Compost Quality and Heavy Metals：A Meta-analysis].

In order to comprehensively evaluate the effects of vermicomposting on compost quality and the conversion of heavy metals under different control conditions, 109 studies were reviewed. The effects of earthworm species, pre-compost time, ventilation methods, initial C/N, initial pH, and initial moisture of the raw materials on compost quality and the heavy metal toxicity were quantitatively discussed during the vermicomposting process through Meta-analysis. The results showed that the six subgroups of factors all showed obvious influences on the compost quality and heavy metal toxicity. After vermicomposting, the contents of NO3--N (116.2%), TN (29.1%), TP (31.2%), and TK (15.0%) were significantly increased, whereas NH4+-N (-14.8%) and C/N (-36.3%) were significantly decreased. Meanwhile, the total amount of Cu and Cr of the final compost and their bioavailability were significantly reduced. Considering the influences of grouping factors on compost quality and heavy metals, it is recommended to adjust the initial moisture of pile materials to 70%-80%, C/N to 30-85, and pH to 6-7 and to conduct pre-composting for 0-15 d; additionally, vermicomposting should be naturally placed when the composting is aimed at promoting the compost quality. If the main purpose is to weaken the perniciousness of heavy metals in the raw material, it is recommended to adjust the initial moisture of the material to 50%-60%, C/N to less than 30, and pH to 7-8; to conduct no pre-compost; regularly turn the piles; and use the earthworm Eudrilus eugeniae for vermicomposting.

Effects of pyroligneous acid on acute, chronic, and cyto-genotoxicity to earthworms (Eisenia fetida).

The pyroligneous acid (PA), or wood vinegar, is a byproduct of wood carbonization during the slow pyrolysis process. PA is recognized globally as a safe compound for agriculture due to its various beneficial properties, such as antioxidant, antibacterial, antifungal, and termiticidal properties. However, the impact of different PA concentrations on beneficial soil organisms, such as earthworms has not been investigated. The present study aims to understand the effects of different PA concentrations on earthworm Eisenia fetida. The earthworms were exposed to nine different concentrations of PA in soils, including their control. The acute toxicity assay was performed after 14 days of exposure, and the chronic toxicity assay was performed up to 8 weeks after exposure. The results from the acute toxicity assay demonstrated no significant effect on earthworm mortality. The chronic toxicity assay showed that lower PA concentrations (0.01-0.2% of weight/weight PA in soil) promoted cocoon and juvenile production in soils, whereas higher PA concentrations (0.5 and 1%) had a negative effect. These findings highlight the potential of PA to enhance soil fertility at lower concentrations, up to 0.2%, by stimulating worm activity and subsequent manure production. The outcomes of this study have significant implications for the careful management of PA concentrations within agricultural operations.

Variation and trade-offs in life history traits of the protist parasite Monocystis perplexa (Apicomplexa) in its earthworm host Amynthas agrestis.

The life history of a parasite describes its partitioning of assimilated resources into growth, reproduction, and transmission effort, and its precise timing of developmental events. The life cycle, in contrast, charts the sequence of morphological stages from feeding to the transmission forms. Phenotypic plasticity in life history traits can reveal how parasites confront variable environments within hosts. Within the protist phylum Apicomplexa major clades include the malaria parasites, coccidians, and most diverse, the gregarines (with likely millions of species). Studies on life history variation of gregarines are rare. Therefore, life history traits were examined for the gregarine Monocystis perplexa in its host, the invasive earthworm Amynthas agrestis at three sites in northern Vermont, United States of America. An important value of this system is the short life-span of the hosts, with only seven months from hatching to mass mortality; we were thus able to examine life history variation during the entire life cycle of both host and parasite. Earthworms were collected (N = 968 over 33 sample periods during one host season), then parasites of all life stages were counted, and sexual and transmission stages measured, for each earthworm. All traits varied substantially among individual earthworm hosts and across the sites. Across sites, timing of first appearance of infected earthworms, date when transmission stage (oocysts packed within gametocysts) appeared, date when number of both feeding (trophic) cells and gametocysts were at maximum, and date when 100% of earthworms were infected differed from 2-8 weeks, surprising variation for a short season available for parasite development. The maximal size of mating cells varied among hosts and across sites and this is reflected in the number of oocysts produced by the gametocyst. A negative trade-off was observed for the number of oocysts and their size. Several patterns were striking: (1) Prevalence reached 100% at all sites by mid season, only one to three weeks after parasites first appeared in the earthworms. (2) The number of parasites per host was large, reaching 300 × 103 cells in some hosts, and such high numbers were present even when parasites first appeared in the host. (3) At one site, few infected earthworms produced any oocysts. (4) The transmission rate to reach such high density of parasites in hosts needed to be very high for a microbe, from >0.33% to >34.3% across the three sites. Monocystis was one of the first protist parasites to have its life cycle described (early 19th century), but these results suggest the long-accepted life cycle of Monocystis could be incomplete, such that the parasites may be transmitted vertically (within the earthworm's eggs) as well as horizontally (leading to 100% prevalence) and merogony (asexual replication) could be present, not recognized for Monocystis, leading to high parasitemia even very early in the host's season.

The responses of CO2 emission to nitrogen application and earthworm addition in the soybean cropland.

The effects of nitrogen application or earthworms on soil respiration in the Huang-Huai-Hai Plain of China have received increasing attention. However, the response of soil carbon dioxide (CO2) emission to nitrogen application and earthworm addition is still unclear. A field experiment with nitrogen application frequency and earthworm addition was conducted in the Huang-Huai-Hai Plain. Results showed nitrogen application frequency had a significant effect on soil respiration, but neither earthworms nor their interaction with nitrogen application frequency were significant. Low-frequency nitrogen application (NL) significantly increased soil respiration by 25%, while high-frequency nitrogen application (NH), earthworm addition (E), earthworm and high-frequency nitrogen application (E*NH), and earthworm and low-frequency nitrogen application (E*NL) also increased soil respiration by 21%, 21%, 12%, and 11%, respectively. The main reason for the rise in soil respiration was alterations in the bacterial richness and keystone taxa (Myxococcales). The NH resulted in higher soil nitrogen levels compared to NL, but NL had the highest bacterial richness. The abundance of Corynebacteriales and Gammaproteobacteria were positively connected with the CO2 emissions, while Myxococcales, Thermoleophilia, and Verrucomicrobia were negatively correlated. Our findings indicate the ecological importance of bacterial communities in regulating the carbon cycle in the Huang-Huai-Hai Plain.

Evaluating drained water quality in a pilot worm-sludge treatment reed bed planted with Arundo donnas in the Mediterranean climate.

This study evaluated the impact of incorporating earthworms (Eisenia fetida) on the drained water quality from a sludge treatment reed bed. The experiment encompassed four setups of treatment beds in two replicates: planted with Arundo donax and addition of earthworms, planted without earthworms, unplanted with earthworms, and treatment bed without plants nor earthworms as control. The units were fed every two weeks with mixed sewage sludge, a blend of primary and secondary sludge over 24 cycles. The mixed sewage sludge had mean dry and volatile solid contents of 24.71 g.DS.L-1 (± 13.67) and 19.14 g.VS.L-1 (± 10.29) resulting a sludge loading rate of 43.59 kg.DS.m-2.year-1 (± 14.49). The inclusion of earthworms in the planted unit reduced release masses of total suspended solids, chemical oxygen demand, nitrate and phosphorous by 43, 45, 75 and 45 % compared to the planted unit. Plant biomass production increased by 43 % with the earthworm presence. The removal efficiency of the units improved after a ramp-up phase (after six months feeding) of which the concentration of TSS, COD and Escherichia coli met limits for water reuse while nitrogen components and phosphorous surpassed the limits. The planted unit with earthworms removed 99 and 99 % of TSS and COD, respectively. Overall, water loss namely through evapotranspiration and earthworm hydration need, positively correlated with pollutant concentration, and earthworm-planted unit had 46 % higher water loss compared to control unit.

Microbial community structure and diversity attached to the periphyton in different urban aquatic habitats.

Periphyton is a complex community composed of diverse prokaryotes and eukaryotes; understanding the characteristics of microbial communities within periphyton becomes crucial for biogeochemical cycles and energy dynamics of aquatic ecosystems. To further elucidate the community characteristics of periphyton across varied aquatic habitats, including unpolluted ecologically restored lakes, aquaculture ponds, and areas adjacent to domestic and industrial wastewater treatment plant outfalls, we explored the composition and diversity of prokaryotic and eukaryotic communities in periphyton by employing Illumina MiSeq sequencing. Our findings indicated that the prokaryotic communities were predominantly composed of Proteobacteria (40.92%), Bacteroidota (21.01%), and Cyanobacteria (10.12%), whereas the eukaryotic communities were primarily characterized by the dominance of Bacillariophyta (24.09%), Chlorophyta (20.83%), and Annelida (15.31%). Notably, Flavobacterium emerged as a widely distributed genus among the prokaryotic community. Unclassified_Tobrilidae exhibited higher abundance in unpolluted ecologically restored lakes. Chaetogaster and Nais were enriched in aquaculture ponds and domestic wastewater treatment plant outfall area, respectively, while Surirella and Gomphonema dominated industrial sewage treatment plant outfall area. The alpha diversity of eukaryotes was higher in unpolluted ecologically restored lakes. pH and nitrogen content ( NO 2 - - N , NO 3 - - N , and TN) significantly explained the variations for prokaryotic and eukaryotic community structures, respectively. Eukaryotic communities exhibited a more pronounced response to habitat variations compared to prokaryotic communities. Moreover, the association networks revealed an intensive positive correlation between dominant Bacillariophyta and Bacteroidota. This study provided useful data for identifying keystone species and understanding their ecological functions.

Lipid-related ion suppression on the herbicide atrazine in earthworm samples in ToF-SIMS and matrix-assisted laser desorption ionization mass spectrometry imaging and the role of gas-phase basicity.

In mass spectrometry imaging (MSI), ion suppression can lead to a misinterpretation of results. Particularly phospholipids, most of which exhibit high gas-phase basicity (GB), are known to suppress the detection of metabolites and drugs. This study was initiated by the observation that the signal of an herbicide, i.e., atrazine, was suppressed in MSI investigations of earthworm tissue sections. Herbicide accumulation in earthworms was investigated by time-of-flight secondary ion mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Additionally, earthworm tissue sections without accumulation of atrazine but with a homogeneous spray deposition of the herbicide were analyzed to highlight region-specific ion suppression. Furthermore, the relationship of signal intensity and GB in binary mixtures of lipids, amino acids, and atrazine was investigated in both MSI techniques. The GB of atrazine was determined experimentally through a linear plot of the obtained intensity ratios of the binary amino acid mixtures, as well as theoretically. The GBs values for atrazine of 896 and 906 kJ/mol in ToF-SIMS and 933 and 987 kJ/mol in MALDI-MSI were determined experimentally and that of 913 kJ/mol by quantum mechanical calculations. Compared with the GB of a major lipid component, phosphatidylcholine (GBPC = 1044.7 kJ/mol), atrazine's experimentally and computationally determined GBs in this work are significantly lower, making it prone to ion suppression in biological samples containing polar lipids.

Biochemical and molecular-level effects of co-exposure to chlorpyrifos and lambda-cyhalothrin on the earthworm (Eisenia fetida).

Farmland soil organisms frequently encounter pesticide mixtures presented in their living environment. However, the underlying toxic mechanisms employed by soil animals to cope with such combined pollution have yet to be explored. This investigation aimed to reveal the changes in cellular and mRNA levels under chlorpyrifos (CPF) and lambda-cyhalothrin (LCT) co-exposures in earthworms (Eisenia fetida). Results exhibited that the combination of CPF and LCT triggered an acute synergistic influence on the animals. Most exposures resulted in significant alterations in the activities of total superoxide dismutase (T-SOD), copper/zinc superoxide dismutase (Cu/Zn-SOD), caspase 3, and carboxylesterase (CarE) compared to the basal level. Moreover, when exposed to chemical mixtures, the transcription levels of four genes [heat shock protein 70 (hsp70), gst, sod, and calreticulin (crt)] also displayed more pronounced changes compared with their individual exposures. These changes in determined parameters indicated the occurrence of oxidative stress, cell death, detoxification dysfunction, and endoplasmic reticulum damage after co-exposure to CPF and LCT in E. fetida. The comprehensive examination of mixture toxicities of CPF and LCT at different endpoints would help to understand the overall toxicity they cause to soil invertebrates. The augmented deleterious effect of these pesticides in a mixture suggested that mixture toxicity assessment was necessary for the safety evaluation and application of pesticide mixtures.