The digestive system of a cricket pulverizes polyethylene microplastics down to the nanoplastic scale.

Microplastics (MPs; <5 mm) are a growing concern and a poorly understood threat to biota. We used a generalist insect (a cricket; Gryllodes sigillatus) to examine whether individuals would ingest and physically degrade MPs in their food. We fed crickets a range of concentrations (0, 2.5, 5, and 10% w/w) of fluorescent polyethylene MPs mixed into a standard diet and dissected the gut regions to isolate the MPs within. Comparing plastic content and fragment size within gut regions, we sought to identify whether and where crickets can fragment ingested MP particles. Given the digestive tract morphology of this species, we expected that the crickets would both ingest and egest the MPs. We also predicted that the MPs would be fragmented into smaller pieces during this digestive process. We found that G. sigillatus egested much smaller pieces than they ingested, and this fragmentation occurs early in the digestive process of this insect. We found this for both sexes as well as across the range of concentrations of MPs. The degree of plastic breakdown relative to plastic feeding time suggests that the ability to fragment MPs is intrinsic and not altered by how much time crickets have spent eating the plastics. The amount of plastics found in each region of the gut in relation to feeding time also suggests that this size and shape of PE microplastic does not cause any physical blockage in the gut. This lack of evidence for blockage is likely due to plastic breakdown. We found a ∼1000-fold reduction in plastic size occurs during passage through the digestive system, yielding particles very near nanoplastics (NPs; <1 µm), and likely smaller, that are then excreted back into the environment. These findings suggest that generalist insects can act as agents of plastic transformation in their environment if/when encountering MPs.

Quantifying microplastic ingestion, degradation and excretion in insects using fluorescent plastics.

Plastic pollution is a growing threat to our natural environment. Plastic waste/pollution results from high emissions of both macro (>5 mm) and microplastics (MPs; <5 mm) as well as environmental fractioning of macroplastics into MPs. MPs have been shown to have a range of negative impacts on biota. Harmonized methods to accurately measure and count MPs from animal samples are limited, but what methods exist are not ideal for a controlled laboratory environment where plastic ingestion, degradation and elimination can be quantified and related to molecular, physiological and organismal traits. Here, we propose a complete method for isolating and quantifying fluorescent MPs by combining several previously reported approaches into one comprehensive workflow. We combine tissue dissection, organic material digestion, sample filtering and automated imaging techniques to show how fluorescently labelled MPs provided to insects (e.g. in their diet) in a laboratory setting can be isolated, identified and quantified. As a proof of concept, we fed crickets (Gryllodes sigillatus) a diet of 2.5% (w/w) fluorescently labelled plastics and isolated and quantified plastic particles within the gut and frass.

Ingestion of Microplastic Fibres, But Not Microplastic Beads, Impacts Growth Rates in the Tropical House Cricket Gryllodes Sigillatus.

Microplastic is a growing concern as an environmental contaminant as it is ubiquitous in our ecosystems. Microplastics are present in terrestrial environments, yet the majority of studies have focused on the adverse effects of microplastics on aquatic biota. We hypothesized that microplastic ingestion by a terrestrial insect would have localized effects on gut health and nutrient absorption, such that prolonged dietary microplastic exposure would impact growth rate and adult body size. We further hypothesized that plastic form (fibres vs. beads) would influence these effects because of the nature of gut-plastic interactions. Freshly hatched tropical house crickets (Gryllodes sigillatus) were fed a standard diet containing different concentrations of either fluorescent polyethylene microplastic beads (75-105 µm), or untreated polyethylene terephthalate microfibers (< 5 mm) until they died or reached adulthood (approximately 8 weeks). Weight and body length were measured weekly and microplastic ingestion was confirmed through fluorescence microscopy and visual inspection of the frass. While, to our surprise, we found no effect of polyethylene bead ingestion on growth rate or final body size of G. sigillatus, females experienced a reduction in size and weight when fed high concentrations of polyethylene terephthalate microfibers. These results suggest that high concentrations of polyethylene beads of the 100 µm size range can pass through the cricket gut without a substantial negative effect on their growth and development time, but high concentrations of polyethylene terephthalate microfibers cannot. Although we report the negative effects of microplastic ingestion on the growth of G. sigillatus, it remains uncertain what threats microplastics pose to terrestrial insects.

Uptake and Effects of Six Rare Earth Elements (REEs) on Selected Native and Crop Species Growing in Contaminated Soils.

Rare earth elements (REEs) have become increasingly important metals used in modern technology. Processes including mining, oil refining, discarding of obsolete equipment containing REEs, and the use of REE-containing phosphate fertilizers may increase the likelihood of environmental contamination. However, there is a scarcity of information on the toxicity and accumulation of these metals to terrestrial primary producers in contaminated soils. The objective of this work was to assess the phytotoxicity and uptake from contaminated soil of six REEs (chloride forms of praseodymium, neodymium, samarium, terbium, dysprosium, and erbium) on three native plants (Asclepias syriaca L., Desmodium canadense (L.) DC., Panicum virgatum L.) and two crop species (Raphanus sativus L., Solanum lycopersicum L.) in separate dose-response experiments under growth chamber conditions. Limited effects of REEs were found on seed germination and speed of germination. Effects on aboveground and belowground biomass were more pronounced, especially for the three native species, which were always more sensitive than the crop species tested. Inhibition concentrations (IC25 and IC50) causing 25 or 50% reductions in plant biomass respectively, were measured. For the native species, the majority of aboveground biomass IC25s (11 out of 18) fell within 100 to 300 mg REE/kg dry soil. In comparison to the native species, IC25s for the crops were always greater than 400 mg REE/kg, with the majority of results (seven out of 12) falling above 700 mg REE/kg. IC50s were often not detected for the crops. Root biomass of native species was also affected at lower doses than in crops. REE uptake by plants was higher in the belowground parts than in the above-ground plant tissues. Results also revealed that chloride may have contributed to the sensitivity of the native species, Desmodium canadense, one of the most sensitive species studied. Nevertheless, these results demonstrated that phytotoxicity may be a concern in contaminated areas.

Cerium chloride heptahydrate (CeCl3 · 7H2O) induces muscle paralysis in the generalist herbivore, Melanoplus sanguinipes (Fabricius) (Orthoptera: Acrididae), fed contaminated plant tissues.

Of increasing economic importance are the rare earth elements (REEs). Pollution from mining and processing activity is expected to rise with industrial demand. Plants are known to accumulate REEs, although levels vary with species and soil content. However, the effect on wildlife of ingesting REE contaminated vegetation is not well understood. Here we examined the effect of consuming vegetation with elevated levels of cerium on the generalist grasshopper, Melanoplus sanguinipes (Fabricius). Adults excreted a substantial portion of ingested contamination. However, after only four-days of feeding, accumulation in the body occurred at all doses and paralysis of appendages resulted at the highest doses. Short-term toxicity studies may underestimate the impact of ingesting REE contamination. Metals tend to be low in toxicity; however, their persistence in the environment may be better represented by exposure over longer portions of the life cycle.

Rare earth elements (REEs): effects on germination and growth of selected crop and native plant species.

The phytotoxicity of rare earth elements (REEs) is still poorly understood. The exposure-response relationships of three native Canadian plant species (common milkweed, Asclepias syriaca L., showy ticktrefoil, Desmodium canadense (L.) DC. and switchgrass, Panicum virgatum L.) and two commonly used crop species (radish, Raphanus sativus L., and tomato, Solanum lycopersicum L.) to the REEs lanthanum (La), yttrium (Y) and cerium (Ce) were tested. In separate experiments, seven to eight doses of each element were added to the soil prior to sowing seeds. Effects of REE dose on germination were established through measures of total percent germination and speed of germination; effects on growth were established through determination of above ground biomass. Ce was also tested at two pH levels and plant tissue analysis was conducted on pooled samples. Effects on germination were mostly observed with Ce at low pH. However, effects on growth were more pronounced, with detectable inhibition concentrations causing 10% and 25% reductions in biomass for the two native forb species (A. syriaca and D. canadense) with all REEs and on all species tested with Ce in both soil pH treatments. Concentration of Ce in aboveground biomass was lower than root Ce content, and followed the dose-response trend. From values measured in natural soils around the world, our results continue to support the notion that REEs are of limited toxicity and not considered extremely hazardous to the environment. However, in areas where REE contamination is likely, the slow accumulation of these elements in the environment could become problematic.

Influence of soil organic matter on the sensitivity of selected wild and crop species to common herbicides.

Current regulatory protocols for assessing herbicide effects on plants rely heavily on the use of crops grown under controlled greenhouse conditions to indicate risks to wild vegetation. Guideline test protocols call for low levels of soil nutrients, approximately 3% organic matter (OM), to test the worst-case scenario for non-target species growing in poor soils. However, species sensitivity to herbicides may be affected by growing conditions, especially soil nutrient levels. The purpose of this study was to assess the impact of high and low soil OM content on the sensitivity of plants to several common agricultural herbicides (glyphosate, chlorimuron ethyl and dicamba). Ten plant species (wild and crop) with high (nitrophilous) or low (non-nitrophilous) affinity for nitrogen were grown under greenhouse conditions in soil with two levels of OM (3 and 9%) and were exposed to seven doses of the three herbicides in concurrent experiments. Results showed that most species were more sensitive to glyphosate under high OM conditions while chlorimuron ethyl and dicamba results were mixed with respect to OM levels. Overall, 15 species-herbicide combinations were more sensitive in high OM soil, while 11 were more sensitive in low OM soil. No clear pattern in sensitivity emerged among nitrophilous and non-nitrophilous species. Several species showed a difference in response at herbicide levels expected to reach non-target habitats adjacent to crop fields through drift (5% commonly and 25% occasionally). In terms of regulatory testing, guidelines may need alteration to allow testing with nutrient levels that more accurately reflect natural environments.

Effects of chlorimuron ethyl on terrestrial and wetland plants: Levels of, and time to recovery following sublethal exposure.

Current pesticide registration guidelines call for short-term testing of plants; long-term effects on vegetative parts and reproduction remain untested. The aims of our study were to determine level of recovery and recovery times for plants exposed to the sulfonylurea herbicide chlorimuron ethyl using data collected from single species, dose-response greenhouse experiments. The nine terrestrial and eight wetland species tested showed variable levels of recovery and recovery timeframes. Many species (six terrestrial and five wetland) were vegetatively stunted at sublethal doses and were reproductively impaired. Full recovery did not occur at all doses and maximum recovery times varied from 3 to 15 weeks in this controlled environment. In a complex community, affected species may be displaced by tolerant species, through interspecific competition, before they fully recover. It is plausible that individual populations could be diminished or eliminated through reduced seedbank inputs (annuals and perennials) and asexual reproduction (perennials).