Lumbricus Terrestris Earthworms Research Articles

Antitumor Activity of Serine Enzyme and Crude Extracts of The Earthworm Lumbricus terrestris Against A549 Human Lung Cancer Cell Line

Background: Numerous disorders have been successfully treated with earthworms, and this has been documented in ancient texts. These invertebrates are a rich natural source of biological and pharmacological compounds with antioxidant and anticancer properties. The present study aims to isolate and partially purify serine protease from the earthworm and evaluate the cytotoxic and antitumor activities of serine protease and crude extracts of the earthworm. Methods: Lumbricus terrestris earthworms were collected and processed to prepare crude extracts, isolate, and partially purify them. The anti-tumour activity's serine enzyme was evaluated by MTT assay and Live/Dead Cell Viability Assay using A549 and NHF cell lines. Results: When the concentration was doubled, it hurt the viability of A549 and NHF cells. The IC50 values for A549cell lines were194.3 and 22.57 μg/ml for crude and serine protease, respectively. Also, the crude extract does not show cytotoxicity against the normal cell line (NHF cells). The IC50 values 391,78.61 mg/ ml, respectively, using Live/Dead Cell Viability Assay, demonstrate that an increased proportion of apoptotic cell deaths in A549 cells after treatment with crude as well as enzyme there is a significant increase in the mean count of dead cells after being treated with enzyme as well as crude the mean count was 172.7 3;127.5 in cells treated with serine enzyme and crude extract, respectively, whereas in the untreated cells, it was 33.1 reflects a significant difference when compared with treated cells with serine enzyme and crud extracts (P<0.001). Conclusion: These findings revealed that Serin enzyme and crude extract of Lumbricus terrestris possesses excellent antitumor activity.

Toxicity of carbon nanofibers in earthworms (Lumbricus terrestris) naturally infected with Monocystis sp.

Although the ecotoxicity of carbon-based nanomaterials (CBNs) is known, the potential effect of carbon nanofibers (CNFs) on edaphic organisms has been insufficiently explored. Thus, we aimed at the ecotoxicity of CNFs (at 10 and 100 mg/kg) in Lumbricus terrestris earthworms naturally infected with Monocystis sp. After 28 days of exposure, treatments did not affect the survival rate. However, we observed a significant loss of body biomass, and Monocystis sp. infection in seminal vesicles was potentiated by exposure to CNFs. Earthworms exposed to CNFs showed a redox imbalance in the seminal vesicle, muscle, and intestine and an alteration in nitric oxide production in these organs. In muscles, we also noticed a significant reduction in AChE activity in earthworms exposed to CNFs. The histopathological analyses revealed the treatments' significant effect on the structures of the different evaluated tissues. Although we did not notice a concentration-response for several of the biomarkers, when taken together and after the application of Integrated Biomarker Response (IBR) and principal component analysis (PCA), we noticed that the response of earthworms to CNFs at 100 mg/kg showed a more significant deviation from the unexposed group. This was mainly determined by inhibiting antioxidant activity in the seminal vesicle, biochemical biomarkers assessed in muscle and intestine, and histomorphometric muscle biomarkers from earthworms exposed to CNFs at 100 mg/kg. Thus, we demonstrate that CNFs increase the parasite load of Monocystis sp. of adult L. terrestris earthworms and induce biochemical and histopathological changes, especially at 100 mg/kg. Our results point to the additional impact these nanomaterials can have on the health of earthworms, signaling the need for greater attention to their disposal and ecotoxicological effects on soil organisms.

Landfill leachate has multiple negative impacts on soil health indicators in Hyrcanian forest, northern Iran

The storage of municipal solid wastes in unengineered landfills poses a severe threat to soil functions and health. Wastes seriously threaten human health and the terrestrial ecosystem, especially due to heavy metals. There is a general knowledge gap about the long-term impacts of storage wastes on the soil health indicators which are effective on soil functions. This investigation focuses on the examination of landfill leachate on soil health indicators from different years in the Hyrcanian forest region in northern Iran. For this purpose, soil sampling was done in the summer of 2012 and 2022 (from three depths of 0–10, 10–20, and 20–30 cm and on a surface of 30 cm × 30 cm). Soil samples were randomly collected from a polluted forest used as waste storage and a nearby unpolluted protected forest. In addition to the general soil physical, chemical and biological parameters, the amounts of cadmium (Cd) and lead (Pb) in the soil were also measured. Simultaneously with soil sampling, earthworms (from a depth of 0–30 cm) were collected and identified. Also, the concentration of Cd and Pb in the earthworm's biomass were measured in the laboratory. We found that unpolluted sites had maximum values of N, K, P, and Ca than the polluted sites. In addition, a decrease of soil aggregates stability, nutrient contents, microbial and enzyme activities, and also fauna and microflora abundance were found in the polluted sites in the period 2012–2022. Soil Cd and Pb contents were more in the polluted site in 2022 compared to the unpolluted site. Lumbricus rubellus and Lumbricus terrestris earthworms had significantly higher population in the polluted sites and higher accumulation of Cd and Pb in biomass. According to our results, soil health decreased in the order unpolluted site 2022 > unpolluted site 2012 > polluted site 2012 > polluted site 2022, which corresponds with the reduction of soil health during the release of landfill leachate. This investigation contributes to understand landfill pollution derived from leachate and its effects on soil physical, chemical and biological parameters to help managing landfill leachate. Therefore, the main issue is choosing a landfill system that minimizes the risk of pollution, installing a leachate collection system and constructing a landfill with engineering principles that can reduce the effects of urban waste pollution on soil health. We emphasize that landfilling is dangerous for the environment, so the government should implement sanitary landfilling to prevent further contamination of surface and underground waters, as well as soil in the precious Hyrcanian forest.

Monitoring the effects of pesticide pellets to address farmers' concerns on soil fauna, specifically earthworms

AbstractSlugs are a major pest in arable agriculture that are typically controlled by using pesticide‐impregnated pellets (molluscicides). We received multiple requests from farmers in 2018 for research into the impact of slug pellets on earthworms because they had been told that ferric‐based pellets were ‘worm‐friendly’. We measured soil fauna feeding activities within Lumbricus terrestris middens on a replicated field plot experiment comparing the recommended product rates of a metaldehyde (4%) based or ferric phosphate EDDS (3%) based slug pellets. There was a significant decrease in feeding activity in the presence of either type of slug pellet, particularly in the top 25 mm of the soil indicating movement away from the pesticide. This could be explained by the significantly lower abundance (numbers and biomass) of earthworms in the topsoil. Four months after ferric phosphate‐based slug pellet application, there was no significant difference between the control and treated plots in earthworm abundance, indicating the effects were temporary. In the laboratory, individual Lumbricus terrestris earthworms consumed up to 4 slug pellets over 14 days and no significant differences in biomass were measured. A burrowing fitness assay indicated that earthworms exposed to ferric phosphate‐based slug pellets burrowed to a greater depth. This suggests an avoidance behaviour response, which could also explain the field results. A consultation with farmers (n = 68) rated this information service as useful and indicated widespread interest in minimizing the effects of pesticides on non‐target soil fauna.

Potential interactive effects between invasive Lumbricus terrestris earthworms and the invasive plant Alliaria petiolata on a native plant Podophyllum peltatum in northeastern Ohio, USA.

We test whether the invasive earthworm Lumbricus terrestris and leaf litter of the invasive herbaceous plant Alliaria petiolata interact to influence the native plant, Podophyllum peltatum, using both observational field data and a multi-year experiment. We hypothesized invader interactive effects on the native plant might result from either changes in allelochemical distribution in the soil or nutrient availability mediated by the invasive earthworm pulling leaf litter down into the soil. Within the field data we found that Alliaria petiolata presence and higher soil nitrogen correlated with reduced Podophyllum peltatum cover, and no evidence for an invader–invader interaction. Within the factorial experiment, we found a super-additive effect of the two invaders on plant biomass only when activated carbon was present. In the absence of activated carbon, there were no differences in Podophyllum peltatum biomass across treatments. In the presence of activated carbon, Podophyllum peltatum biomass was significantly reduced by the presence of both Lumbricus terrestris and Alliaria petiolata leaf litter. The absence of an effect of Alliaria petiolata leaves without activated carbon, combined with a failure to detect arbuscular mycorrhizal colonization, suggests that indirect effects of allelochemicals on arbuscular mycorrhizal fungi were not the primary driver of treatment responses. Rather direct nutrient availability might influence a potential interaction between these invaders. Leaf nitrogen content was higher and leaf CO2 concentration was lower in the presence of Lumbricus terrestris, but treatment did not influence maximum photosynthetic rate. While the field data do not suggest a negative interaction between these invaders, the experiment suggests that such an interaction is possible with greater environmental stress, such as increasing nitrogen deposition. Further, even plants with rapid physiological responses to increased nitrogen availability may have other physiological limits on growth that prevent them from compensating from the harm caused by multiple invaders.

Parasitological and molecular study of Toxocara spp. in Lumbericus terrestricus earthworms

Soil-associated invertebrates which are coprophagic are capable of acting as paratenic hosts for Toxocara species larvae eggs. The aim of the present study was to assess the role of Lumbricus terrestris earthworms as potential intermediate host for Toxocara species. A total of 240 earthworms were collected from the public places in four areas of Karaj, Alborz Province, Iran. The subjects of the study were digested to detect Toxocara species using microscopy and molecular analyses. Toxocara larvae and larval eggs were recovered from 1.7% (4/240) of the sampled earthworms. The recovery rate in Shah'Abbasi and Talaghani regions in the studied areas was higher than that of the other regions. In the mentioned area, 8.3% (4/48) of earthworms were revealed to be positive for Toxocara species by microscopic observation and only one was confirmed to be positive for Toxocara canis by polymerase chain reaction (PCR). This is the first molecular report of Toxocara canis infective larvae in Lumbricus terrestris earthworms that could be demonstrated in common environment.

Plastic Bag Derived-Microplastics as a Vector for Metal Exposure in Terrestrial Invertebrates.

Microplastics are widespread contaminants in terrestrial environments but comparatively little is known about interactions between microplastics and common terrestrial contaminants such as zinc (Zn). In adsorption experiments fragmented HDPE bags c. one mm2 in size showed similar sorption characteristics to soil. However, when present in combination with soil, concentrations of adsorbed Zn on a per mass basis were over an order of magnitude lower on microplastics. Desorption of the Zn was minimal from both microplastics and soil in synthetic soil solution (0.01 M CaCl2), but in synthetic earthworm guts desorption was higher from microplastics (40-60%) than soil (2-15%), suggesting microplastics could increase Zn bioavailability. Individual Lumbricus terrestris earthworms exposed for 28 days in mesocosms of 260 g moist soil containing 0.35 wt % of Zn-bearing microplastic (236-4505 mg kg-1) ingested the microplastics, but there was no evidence of Zn accumulation, mortality, or weight change. Digestion of the earthworms showed that they did not retain microplastics in their gut. These findings indicate that microplastics could act as vectors to increase metal exposure in earthworms, but that the associated risk is unlikely to be significant for essential metals such as Zn that are well regulated by metabolic processes.

Milled cereal straw accelerates earthworm (Lumbricus terrestris) growth more than selected organic amendments

Earthworms benefit agriculture by providing several ecosystem services. Therefore, strategies to increase earthworm abundance and activity in agricultural soils should be identified, and encouraged. Lumbricus terrestris earthworms primarily feed on organic inputs to soils but it is not known which organic amendments are the most effective for increasing earthworm populations. We conducted earthworm surveys in the field and carried out experiments in single-earthworm microcosms to determine the optimum food source for increasing earthworm biomass using a selection of crop residues and organic wastes available to agriculture. We found that although farmyard manure increased earthworm populations more than cereal straw in the field, straw increased earthworm biomass more than manures when milled and applied to microcosms. Earthworm growth rates were positively correlated with the calorific value of the amendment and straw had a much higher calorific value than farmyard manure, greenwaste compost, or anaerobic digestate. Reducing the particle size of straw by milling to <3mm made the energy in the straw more accessible to earthworms. The benefits and barriers to applying milled straw to arable soils in the field are discussed.

Changes in the composition and physiological and biochemical properties of fungi during passage through the digestive tract of earthworms

It was established that the biomass of fungal mycelium decreased by 30-50% during passage through the in- testine of the Aporrectodea caliginosa and Lumbricus terrestris earthworms, while its content in empty intes- tines was 40-60% less than in the soil. It was found that the amount of mycelium increases again in three-day-old coprolites due to the rapid growth of the species. It was demonstrated that the physiological activity of fungi (estimated according to the time of the appearance of colonies on the medium and probability of propagation) is lower in the intestine content, digestive tract, and fresh excrement of the worms than in the soil. It was noted that the activity and diversity of organic substrates (utilized by fungi), as well as proteolytic activity is lower in fungal isolates from the intestine than from the soil. It was registered that the death of a part of the fungi occurs in the worm intestine, while the physiological state changes in the animals withstanding the effect of the digestive medium.

Earth worm proportions change as they grow

![Figure][1] Juvenile and adult Lumbricus terrestris earthworms. Photo credit: William Kier. Most animals' proportions change as they grow. Human babies are born with massive heads relative to their bodies that scale down as they grow and many young animals look as if they need to grow

Environmental controls on the production of calcium carbonate by earthworms

Lumbricus terrestris earthworms produce calcium carbonate (CaCO3) granules with unknown physiological function. To investigate carbon sequestration potential, the influence of temperature and CO2 concentration ([CO2]) on CaCO3 production was investigated using three soils, five temperatures (3–20 °C) and four atmospheric [CO2] (439–3793 ppm). Granule production rates differed between soils, but could not be related to any soil characteristics measured. Production rates increased with temperature, probably because of higher metabolic rate, and with soil CO2 concentration. Implications for carbon sequestration are discussed. CaCO3 production in earthworms is probably related to pH regulation of blood and tissue fluid in the high CO2 environment of the soil.

Soil pH governs production rate of calcium carbonate secreted by the earthworm Lumbricus terrestris

Lumbricus terrestris earthworms exposed to 11 soils of contrasting properties produced, on average, 0.8 ± 0.1 mgCaCO3 earthworm−1 day−1 in the form of granules up to 2 mm in diameter. Production rate increased with soil pH (r2 = 0.68, p < 0.01). Earthworms could be a significant source of calcite in soils.

The Effects of PBDE‐99 on Lysosomal Membrane Stability in Coelomocytes of the Earthworm Lumbricus terrestris Using the Neutral Red Retention Test

Flame retardants, such as polybrominated diphenyl ethers (PBDEs) have recently been found to have toxic side effects due to their bioaccumulative nature. Their widespread use on materials such as electronics, furniture, and plastics has led to contamination of natural environments and exposure to various living organisms, including humans. Lysosomal membrane stability is used as a biomarker of pollutant stressors and is measured using the Neurtral Red Retention test. Neutral Red Retention time measures the length of time it takes red dye to escape from the lysosome into the cytosol, indicating cellular damage. Lumbricus terrestris earthworms were exposed to a medium concentration of PBDE‐99 (50 ng/g dry soil) and a high concentration (250 ng/g dry soil) for one week. After exposure, coelomocytes (gut cells) were extracted using sonication, techniques. Data indicated that treatment with PBDEs resulted in a significant increase in the percentage of cells exhibiting red cytosol per minute. The percent increase in red cells/min. increased from 0.21 to 0.53 from the control to the high group. Decreased lysosomal membrane stability in the high concentration group suggested that PBDEs impaired cellular activity in the earthworm model, potentially making the cells susceptible to degradation by their own hydrolytic enzymes.

Impacts of earthworms on soil nutrients and plant growth in soybean and maize agroecosystems

The objective of this experiment was to determine the effects of earthworms on soil N pools and plant growth in soybean and maize agroecosystems. The species and number of individuals in earthworm communities were manipulated in plot-scale field enclosures (2.4 m × 1.2 m) by first reducing earthworm populations within enclosures with carbaryl pesticide, and then adding earthworm treatments to the enclosures. Soybean was grown in the enclosures in the first year and stover maize in the second year. The success of earthworm manipulations in field enclosures was affected by climate conditions and available food resources. The endogeic earthworm species Aporrectodea caliginosa was dominant in all enclosures, while introduced anecic Lumbricus terrestris earthworms had poor survival. In the first season, when climate conditions were favourable for earthworm survival and growth, there was a significant ( P < 0.05) linear increase in soil mineral-N and microbial biomass N concentrations in the 0–15 cm depth of enclosures with more earthworms. Similarly, soybean grain and grain-N yield was significantly ( P < 0.05) greater in enclosures with the largest earthworm populations than the control which had no earthworms added. In the second season, when climate conditions were less favourable, there was no effect of earthworms on soil N pools or maize plants, probably due to poor survival of introduced earthworms.

Effects of norepinephrine and epinephrine on resting membrane potential in body wall muscle cells of Lumbricus terrestris earthworm.

Norepinephrine and to a lesser extent epinephrine increased the resting membrane potential of earthworm body wall muscle cells. Ouabain, phentolamine, propranolol, and replacement of Ca(2+) with Mg(2+) in the incubation medium abolished this effect. External 3'5'-cAMP in high concentration, dibutyryl cAMP, and dibutyryl cGMP did not induced hyperpolarization of muscle cell membranes. It was concluded that norepinephrine and epinephrine increased the resting membrane potential in earthworm body wall muscle cells via activation of Na(+), K(+) pump. This process involves Ca(2+) and sarcolemmal adrenergic structures, similar to alpha-adrenoceptors in vertebrate, but not the cyclic nucleotide systems.