Biotic Ligand Modeling for the Effect of Major Cations on the Uptake of Cadmium in Folsomia candida Exposed in a Sand-Solution Medium.

Biotic ligand modeling (BLM) approaches are already applied to predict the bioavailability and possible risk of metals in surface water, but need further development for soils. The present study investigated the effect of major cations (Ca2+, Mg2+, Na+, K+, and H+) on cadmium bioaccumulation in the springtail Folsomia candida. To avoid the complexity of real soils and enable control of elemental speciation in the exposure medium, the animals were exposed to different cadmium concentrations in an inert quartz sand-solution medium. Accumulation of cadmium in the animals was measured after 7 days exposure at different cation concentrations. Among the cations, only Ca2+ significantly affected the uptake of cadmium in the springtails. Mg2+ also had higher effects compared with other selected cations. Using a BLM approach, the uptake of cadmium in the animals predicted by taking into account both Ca2+ and Mg2+ activities correlated well with the measured values (R2 = 0.68). The final estimated conditional binding constants for cadmium (log KCd-BL), Ca (log KCa-BL), and Mg (log KMg-BL) of 1.06, 2.14, and 1.23 L/mol, respectively, were in agreement with previously reported values. The match between predicted and measured uptake data confirms the applicability and usefulness of the BLM for predicting the bioavailability of cadmium to springtails and opens the way for its application in soil. Environ Toxicol Chem 2024;43:1090-1096. © 2024 SETAC.

Efficiency of plant-based acaricide gels compared to fluvalinate-impregnated strips for control of Varroa destructor in honey bee colonies.

The Varroa mite is one of the most important pests of honey bee colonies. This study compares the efficiency of thymol-based acaricides with the tau-fluvalinate-based commercial treatment to control this mite species. Two experiments were conducted: (1) in the Fall, thymol-based gel was compared with a widely used acaricide, fluvalinate-impregnated strips (Apistan®), and (2) in the Spring, gels containing thymol-only, thymol + menthol + eucalyptus essential oil, and thymol + thyme essential oil were compared with fluvalinate-impregnated strips. In the first experiment, both treatments reduced Varroa mite infestation levels significantly after 70 days of their application compared to the control treatment. The results of the second experiment showed that thymol + thyme essential oil gel, thymol + menthol + eucalyptus essential oil gel, and Apistan strips had always higher efficiency on decreasing mite infestation levels of the selected colonies compared to the control treatment and thymol-only gel during their 7-week application. The results of this study demonstrate that plant-based acaricides for controlling Varroa mites in honey bee colonies have similar efficiency compared to the chemical control methods. Therefore, it is better to use these selected acaricides to reduce mite resistance to chemical control treatments in honey bee colonies.

The Effect of Microbial Diversity and Biomass on Microbial Respiration in Two Soils along the Soil Chronosequence.

Microbial diversity plays an important role in the decomposition of soil organic matter. However, the pattern and drivers of the relationship between microbial diversity and decomposition remain unclear. In this study, we followed the decomposition of organic matter in soils where microbial diversity was experimentally manipulated. To produce a gradient of microbial diversity, we used soil samples at two sites of the same chronosequence after brown coal mining in Sokolov, Czech Republic. Soils were X-ray sterilized and inoculated by two densities of inoculum from both soils and planted with seeds of six local plant species. This created two soils each with four levels of microbial diversity characterized by next-generation sequencing. These eight soils were supplied, or not, by litter of the bushgrass Calamagrostis epigejos, and microbial respiration was measured to assess the rate of decomposition. A strong positive correlation was found between microbial diversity and decomposition of organic matter per gram of carbon in soil, which suggests that microbial diversity supports decomposition if the microbial community is limited by available carbon. In contrast, microbial respiration per gram of soil negatively correlated with bacterial diversity and positively with fungal biomass, suggesting that in the absence of a carbon limitation, decomposition rate is controlled by the amount of fungal biomass. Soils with the addition of grass litter showed a priming effect in the initial stage of decomposition compared to the samples without the addition of litter. Thus, the relationship between microbial diversity and the rate of decomposition may be complex and context dependent.

Methanogenesis in the Digestive Tracts of the Tropical Millipedes Archispirostreptus gigas (Diplopoda, Spirostreptidae) and Epibolus pulchripes (Diplopoda, Pachybolidae).

Methanogens represent the final decomposition step in anaerobic degradation of organic matter, occurring in the digestive tracts of various invertebrates. However, factors determining their community structure and activity in distinct gut sections are still debated. In this study, we focused on the tropical millipede species Archispirostreptus gigas (Diplopoda, Spirostreptidae) and Epibolus pulchripes (Diplopoda, Pachybolidae), which release considerable amounts of methane. We aimed to characterize relationships between physicochemical parameters, methane production rates, and methanogen community structure in the two major gut sections, midgut and hindgut. Microsensor measurements revealed that both sections were strictly anoxic, with reducing conditions prevailing in both millipedes. Hydrogen concentration peaked in the anterior hindgut of E. pulchripes. In both species, the intestinal pH was significantly higher in the hindgut than in the midgut. An accumulation of acetate and formate in the gut indicated bacterial fermentation activities in the digestive tracts of both species. Phylogenetic analysis of 16S rRNA genes showed a prevalence of Methanobrevibacter spp. (Methanobacteriales), accompanied by a small fraction of so-far-unclassified "Methanomethylophilaceae" (Methanomassiliicoccales), in both species, which suggests that methanogenesis is mostly hydrogenotrophic. We conclude that anoxic conditions, negative redox potential, and bacterial production of hydrogen and formate promote gut colonization by methanogens. The higher activities of methanogens in the hindgut are explained by the higher pH of this compartment and their association with ciliates, which are restricted to this compartment and present an additional source of methanogenic substrates. IMPORTANCE Methane (CH4) is the second most important atmospheric greenhouse gas after CO2 and is believed to account for 17% of global warming. Methanogens are a diverse group of archaea and can be found in various anoxic habitats, including digestive tracts of plant-feeding animals. Termites, cockroaches, the larvae of scarab beetles, and millipedes are the only arthropods known to host methanogens and emit large amounts of methane. Millipedes are ranked as the third most important detritivores after termites and earthworms, and they are considered keystone species in many terrestrial ecosystems. Both methane-producing and non-methane-emitting species of millipedes have been observed, but what limits their methanogenic potential is not known. In the present study, we show that physicochemical gut conditions and the distribution of symbiotic ciliates are important factors determining CH4 emission in millipedes. We also found close similarities to other methane-emitting arthropods, which might be associated with their similar plant-feeding habits.

Avoidance tests with the oribatid mite Oppia nitens (Acari: Oribatida) in cadmium-spiked natural soils.

Avoidance behavior can be a useful parameter for assessing the ability of organisms to escape from pollutants in their environment. For soil evaluation, a variety of invertebrates is used including the oribatid mite Oppia nitens. Here, we tested the avoidance behavior of O. nitens using a two-chamber test and an escape test with exposures to different cadmium concentrations of up to 800 mg kg-1 dry LUFA 2.2 soil for 2, 4, and 6 days, and up to 7 weeks. With the two-chamber method, the oribatid mites had the choice between clean and polluted soils, whereas they were allowed to escape from a box with polluted soil to clean containers without soil with the escape method. Avoidance of cadmium was observed after 2 days in both tests and the net response of the mites in the two-chamber test increased with increasing cadmium exposure concentrations. Mite responses varied through time, especially with the escape method; with the avoidance behavior becoming more variable and overall non-significant with longer test durations. This is the first study investigating the escape test simultaneously with long-term avoidance of cadmium by O. nitens. This mite species is a promising species for avoidance testing in soil ecotoxicology, but more experiments are needed to evaluate the factors that influence its responses in laboratory tests and the consequences for its distribution in contaminated ecosystems.

The arbuscular mycorrhizal fungus Rhizophagus intraradices and other microbial groups affect plant species in a copper-contaminated post-mining soil.

BACKGROUND AND AIM: Arbuscular mycorrhizal fungi (AMF) have an important role in plant-microbe interactions. But, there are few studies in which the combined effect of AMF with a stress factor, such as the presence of a metal, on plant species were assessed. This study investigated the effect of arbuscular mycorrhizal (AM) fungus Rhizophagus intraradices and other soil microbial groups in the presence of copper on three plant species in a microcosm experiment. METHODS: Two grass species Poa compressa and Festuca rubra and one herb species Centaurea jacea were selected as model plants in a pot-design test in which soils were artificially contaminated with copper. Treatments were bacteria (control), saprophytic fungi, protists, and a combined treatment of saprophytic fungi and protists, all in the presence or absence of the AM fungal species. After sixty days, plants were harvested and the biomass of grass and herb species and microbial respiration were measured. RESULTS: The results showed almost equal above- and belowground plant biomass and microbial respiration in the treatments in the presence or absence of R. intraradices. The herb species C. jecea responded significantly to the soil inoculation with AM fungus, while grass species showed inconsistent patterns. Significant effect of AMF and copper and their interactions was observed on plant biomass when comparing contaminated vs. non-contaminated soils. CONCLUSION: Strong effect of AMF on the biomass of herb species and slight changes in plant growth with the presence of this fungal species in copper-spiked test soils indicates the importance of mycorrhizal fungi compared to other soil microorganisms in our experimental microcosms.

Comparison Among Test Substrates in Metal Uptake and Toxicity to Folsomia candida and Hordeum vulgare.

The main aim of this short review was to assess the effect of test medium on the bioavailability of metals to the soil invertebrate Folsomia candida and the barley plant Hordeum vulgare. Solution-only exposures and sand-solution media were suitable media with control survival of > 80%. Comparing toxicity and accumulation data, LC50 and/or EC50 values as well as internal concentrations of cadmium (Cd) and copper (Cu) were similar in the tests with different porewater composition for springtails and barley plants. Similar results for toxicity and bioaccumulation of Cd and Cu using different test substrates, suggest the importance of physiological handling of the effects by the organisms rather than the influence of test medium composition.

The effect of the earthworm Lumbricus rubellus on the bioavailability of cadmium and lead to the springtail Folsomia candida in metal-polluted field soils.

The bioavailability of metals can be influenced not only by soil properties but also by other species living at polluted sites. However, in laboratory experiments, usually only one test species is used to estimate bioavailability. In this study, a two-species approach was applied to assess the impact of the earthworm Lumbricus rubellus on the bioavailability of cadmium and lead to the springtail Folsomia candida using natural soils from a gradient of metal pollution. Earthworms were kept in half of the soil replicates for 4 weeks. Subsequently, the uptake and elimination kinetics of cadmium and lead in F. candida exposed for 21 days to the soils was determined. Earthworm activity affected soil properties but did not significantly affect metal uptake rate constants in springtails. The slightly higher uptake due to the presence of earthworms, which was consistent in all tested soils and for both metals, suggests that further research is needed on the role of species interactions in affecting metal bioavailability in soil.

The effect of major cations on the toxicity of cadmium to Folsomia candida in a sand-solution medium analyzed by biotic ligand modeling.

The aim of this study was to assess the effect of major cations (Ca2+, Mg2+, Na+, K+, and H+) on cadmium toxicity to the springtail Folsomia candida. Survival of the animals was determined after seven days exposure to different cadmium concentrations in an inert sand-solution medium, in different experimental setups with modification of the cation concentrations. Among the cations tested, Ca2+ and Mg2+ had protective effects on the toxicity of cadmium to the springtails while Na+, K+, and H+ showed less competition with free cadmium ions for binding to the uptake sites of the collembolans. Toxicity predicted with a biotic ligand model agreed well with the observed values. Calculated conditional binding constants and the fraction of biotic ligands occupied by cadmium to show 50% effects were similar to values reported in the literature. The results emphasize the important role of solution chemistry in determining metal toxicity to soil invertebrates.

Sorption and pH determine the long-term partitioning of cadmium in natural soils.

The bioavailability of metals in soil is a dynamic process. For a proper extrapolation to the field of laboratory studies on fate and effects, it is important to understand the dynamics of metal bioavailability and the way it is influenced by soil properties. The aim of this study was to assess the parallel (concurrent) effect of pH and aging time on the partitioning of cadmium in natural LUFA 2.2 soil. Cadmium nitrate-spiked pH-amended LUFA 2.2 soils were incubated under laboratory conditions for up to 30 weeks. Measured pHpw was lower after 3 weeks and decreased only slightly toward the end of the test. Cadmium concentrations in the pore water increased with time for all soil pH levels, while they decreased with increasing pH. Freundlich kf values ranged between 4.26 and 934 L kg(-1) (n = 0.79 to 1.36) and were highest at the highest pH tested (pH = 6.5). Multiple linear regression analysis, based on a soil ligand modeling approach, resulted in affinity constants of 2.61 for Ca(2+) (log KCa-SL) and 5.05 for H(+) (log KH-SL) for their binding to the active sites on the soil surface. The results showed that pH and aging time are two important factors which together affect cadmium partitioning and mobility in spiked natural soils.

Biotic ligand modeling approach: Synthesis of the effect of major cations on the toxicity of metals to soil and aquatic organisms.

The biotic ligand model (BLM) approach is used to assess metal toxicity, taking into account the competition of other cations with the free metal ions for binding to the biotic ligand sites of aquatic and soil organisms. The bioavailable fraction of metals, represented by the free metal ion, is a better measure than the total concentration for assessing their potential risk to the environment. Because BLMs are relating toxicity to the fraction of biotic ligands occupied by the metal, they can be useful for investigating factors affecting metal bioaccumulation and toxicity. In the present review, the effects of major cations on the toxicity of metals to soil and aquatic organisms were comprehensively studied by performing a meta-analysis of BLM literature data. Interactions at the binding sites were shown to be species- and metal-specific. The main factors affecting the relationships between toxicity and conditional binding constants for metal binding at the biotic ligand appeared to be Ca(2+) , Mg(2+) , and protons. Other important characteristics of the exposure medium, such as levels of dissolved organic carbon and concentrations of other cations, should also be considered to obtain a proper assessment of metal toxicity to soil and aquatic organisms.

The relationship between metal toxicity and biotic ligand binding affinities in aquatic and soil organisms: a review.

The biotic ligand model (BLM) is a theoretical, potentially mechanistic approach to assess metal bioavailability in soil and aquatic systems. In a BLM, toxicity is linked to the fraction of biotic ligand occupied, which in turn, depends on the various components of the solution, including activity of the metal. Bioavailability is a key factor in determining toxicity and uptake of metals in organisms. In this study, the present status of BLM development for soil and aquatic organisms is summarized. For all species and all metals, toxicity was correlated with the conditional biotic ligand binding constants. For almost all organisms, values for Ag, Cu, and Cd were higher than those for Zn and Ni. The constants derived for aquatic systems seem to be equally valid for soil organisms, but in the case of soils, bioavailability from the soil solution is greatly influenced by the presence of the soil solid phase.

Uptake and elimination kinetics of metals in soil invertebrates: a review.

Uptake and elimination kinetics of metals in soil invertebrates are a function of both soil and organism properties. This study critically reviewed metal toxicokinetics in soil invertebrates and its potential use for assessing bioavailability. Uptake and elimination rate constants of different metals are summarized. Invertebrates have different strategies for essential and non-essential metals. As a consequence, different types of models must be applied to describe metal uptake and elimination kinetics. We discuss model parameters for each metal separately and show how they are influenced by exposure concentrations and by physiological properties of the organisms. Soil pH, cation exchange capacity, clay and organic matter content significantly affect uptake rates of non-essential metals in soil invertebrates. For essential metals, kinetics is hardly influenced by soil properties, but rather prone to physiological regulation mechanisms of the organisms. Our analysis illustrates that toxicokinetics can be a valuable measurement to assess bioavailability of soil-bound metals.

A combined toxicokinetics and toxicodynamics approach to assess the effect of porewater composition on cadmium bioavailability to Folsomia candida.

The aim of the present study was to improve our understanding of cadmium bioavailability by linking toxicokinetics and toxicodynamics. The springtail Folsomia candida was exposed to different cadmium concentrations in solutions embedded in inert quartz sand. Survival and cadmium uptake in the animals were followed for 21 d. After 10 d, some animals were transferred to clean medium to assess cadmium elimination. Using a first-order one-compartment model, an overall uptake rate constant (k1) of 0.18 L kg(animal)(-1) d(-1) and an elimination rate constant (k(2-TK)) of 0.02 d(-1) were calculated. Survival decreased with time, resulting in an estimated final median lethal concentration (LC50) of 0.51 mM. A lethal body concentration (LBC) of 4.6 µmol Cd g(-1) dry body weight was estimated by multiplying the final LC50 by the bioconcentration factor (k1/k(2-TK)). The LC50(animal) values based on internal cadmium concentrations were between 3.56 µmol Cd g(-1) and 9.91 µmol Cd g(-1) dry body weight, with an overall value of 7.9 µmol Cd g(-1) dry body weight (95% confidence interval [CI]: 3.8-12.0 µmol Cd g(-1) dry body wt). Because the 95% CI of the LC50(animal) included the LBC, there was good agreement of cadmium toxicokinetics and toxicodynamics.

Toxicodynamics of copper and cadmium in Folsomia candida exposed to simulated soil solutions.

To improve our understanding of metal bioavailability to soil-living invertebrates, the effect of porewater composition on the toxicodynamics of copper and cadmium in Folsomia candida (Collembola) was investigated. Assuming that porewater is the main exposure route, F. candida was exposed to simulated soil solutions of different composition. Toxicity of copper was slightly lower in a calcium-only solution than in a multication solution. With increasing copper concentrations from 0.005 mM to 1.37 mM, internal copper concentrations similarly increased in both exposure solutions, suggesting that a single cation nutrient solution is suitable for testing F. candida. In the second experiment, animals were exposed for 7 d to copper and cadmium in simplified soil solutions with different calcium (0.2 mM, 0.8 mM, 3.2 mM, 12.8 mM) and pH (5.0, 6.0, 7.0) levels. The median lethal concentration (LC50) values decreased with time in both the calcium and pH series. A hormetic-type effect was observed for copper in the second test, as well as in the calcium-only solution in the first experiment. Because of stronger hormesis, LC50s for copper were higher at lower calcium concentrations. For cadmium, LC50 values were higher at higher calcium concentrations, suggesting competition of calcium with the free cadmium ion. Toxicity of cadmium increased with decreasing pH, while copper was more toxic at intermediate pH. The results show that a toxicodynamics approach can help to improve the interpretation of metal toxicity to soil invertebrates, taking into account soil solution properties.

The influence of calcium and pH on the uptake and toxicity of copper in Folsomia candida exposed to simplified soil solutions.

The aim of the present study was to investigate the influence of Ca and pH on the uptake and effects of Cu in Folsomia candida (Collembola). Assuming that soil pore water is the main route of exposure, F. candida were exposed for seven days to Cu in simplified soil solutions at different Ca concentrations and different pH levels. A hormetic-type effect was seen for the effect of Cu on F. candida survival. Toxicity of Cu was slightly decreased and Cu uptake increased at the highest Ca concentrations. Cu toxicity and uptake were not significantly affected by pH. Conditional binding constants for Cu(2+), Ca(2+) and H(+), calculated with a Langmuir isotherm, were used to relate Cu toxicity to the fraction of occupied binding sites (BL). The estimated 50% effect level (fCu-BL50) was 0.14 when all data were combined. To verify the Langmuir parameters, binding constants calculated based on internal Cu concentrations were used to estimate predicted effects and compared with the measured values. A good correlation between predicted and measured survival indicated that the principles of a biotic ligand model may be applicable to explain Cu toxicity to F. candida in simplified soil solutions.

Using a toxicokinetics approach to explain the effect of soil pH on cadmium bioavailability to Folsomia candida.

The aim of this study was to improve our understanding of metal bioavailability in soil by linking the biotic ligand approach with toxicokinetics modelling. We determined cadmium bioaccumulation kinetics in Folsomia candida (Collembola) as a function of soil pH. Animals were exposed for 21 days to LUFA 2.2 soil at 5 or 20 µg Cd g(-1) dry soil followed by 21 days elimination in clean soil. Internal cadmium concentrations were modelled using a first-order one-compartment model, relating uptake rate constants (k1) to total soil, water or 0.01 M CaCl2 extractable and porewater concentrations. Based on total soil concentrations, k1 was independent of soil pH while it strongly increased with increasing pH based on porewater concentrations explaining the reduced competition of H(+) ions making cadmium more bioavailable in pore water at high pH. This shows that the principles of biotic ligand modelling are applicable to predict cadmium accumulation kinetics in soil-living invertebrates.

Influence of Ca and pH on the uptake and effects of Cd in Folsomia candida exposed to simplified soil solutions.

The present study sought to quantify the components of a biotic ligand model (BLM) for the effects of Cd on Folsomia candida (Collembola). Assuming that soil porewater is the main route of exposure and to exclude the effects of soil particles on metal availability, animals were exposed for 7 d to different Cd concentrations between 0.1 mM and 100 mM in simplified soil solutions at different Ca concentrations (0.2 mM, 0.8 mM, 3.2 mM, and 12.8 mM) or at different pH (5.0, 6.0, and 7.0). Higher Ca concentrations decreased the toxicity of Cd (adult survival) in test solutions, whereas toxicity was slightly lower at pH 7 and 6 than at pH 5, suggesting a mitigating effect of Ca and to a lesser extent pH on Cd toxicity to F. candida. Internal Cd concentrations in the animals increased with increasing exposure level but were significantly reduced by increasing Ca concentrations and were not significantly affected by pH. By using Langmuir isotherms, binding constants for Cd, Ca, and protons and the fraction of binding sites occupied by Cd were calculated and used to predict effects of Cd on survival. Predicted toxicity showed a good agreement with measured responses when Ca and pH were used as separate factors or combined together. The present study shows indications of protective effects of Ca but less of protons on the toxicity and uptake of Cd in F. candida on exposure to simplified soil solutions, which can be described using the principles of a biotic ligand model.

Dynamic bioavailability of copper in soil estimated by uptake and elimination kinetics in the springtail Folsomia candida.

This study aimed to assess the bioavailability of copper in soil, by measuring its uptake kinetics into a representative soil invertebrate, the collembolan Folsomia candida. The animals were exposed to 25 or 100 µg Cu g(-1) dry LUFA 2.2 soil at nominal pH(CaCl2) 4.5, 5.5, or 6.5 during 14 days after which they were transferred to clean soil for 14 days elimination. Uptake and elimination rate constants were calculated based on total and extractable soil concentrations and porewater concentrations using one-compartment first-order kinetics modelling. Copper was present in the animals at a basal physiological level of 40-90 µg g(-1)dry weight, on top of this uptake and elimination kinetics were observed. Uptake rates constants varied between 0.02 and 0.17 g(soil) g(animal)(-1) day(-1), being higher at lower exposure level, but did not differ significantly between different soil pH levels. Elimination rate constants ranged between 0.04 and 0.20 day(-1) and were negligible (k(2) < 0.001 day(-1)) at pH 4.5 and 6.5. Multiple linear regressions showed that the pH effect on copper uptake was only significant when taking into account cation exchange capacity, or calcium and dissolved organic carbon levels in the pore water. Copper concentrations in the animals however, never were higher than 185 µg g(-1) dry weight, independent of exposure level and pH, suggesting homeostatic regulation. These results show that the chemical composition of the pore water does affect bioavailability of copper in soil, but that copper uptake in collembolans is dominated by homeostatic regulation rather than by soil properties like pH.

Regular dorsal dimples and damaged mites of Varroa destructor in some Iranian honey bees (Apis mellifera).

The frequency of damaged Varroa destructor Anderson and Trueman (Mesostigmata: Varroidae) found on the bottom board of hives of the honey bee, Apis mellifera L. (Hymenoptera: Apidae) has been used as an indicator of the degree of tolerance or resistance of honey bee colonies against mites. However, it is not clear that this measure is adequate. These injuries should be separated from regular dorsal dimples that have a developmental origin. To investigate damage to Varroa mites and regular dorsal dimples, 32 honey bee (A. mellifera) colonies were selected from four Iranian provinces: Isfahan, Markazi, Qazvin, and Tehran. These colonies were part of the National Honey bee Breeding Program that resulted in province-specific races. In April, Varroa mites were collected from heavily infested colonies and used to infest the 32 experimental colonies. In August, 20 of these colonies were selected (five colonies from each province). Adult bees from these colonies were placed in cages and after introducing mites, damaged mites were collected from each cage every day. The average percentage of injured mites ranged from 0.6 to 3.0% in four provinces. The results did not show any statistical differences between the colonies within provinces for injuries to mites, but there were some differences among province-specific lines. Two kinds of injuries to the mites were observed: injuries to legs and pedipalps, and injuries to other parts of the body. There were also some regular dorsal dimples on dorsal idiosoma of the mites that were placed in categories separate from mites damaged by bees. This type of classification helps identifying damage to mites and comparing them with developmental origin symptoms, and may provide criteria for selecting bees tolerant or resistant to this mite.