Microplastic-stressor responses are rarely synergistic in freshwater fishes: A meta-analysis.

Microplastic exposure can cause a range of negative effects on the biochemistry, condition and ecology of freshwater fishes depending on aspects of the exposure and the exposed fish. However, fishes are typically exposed to microplastics and additional multiple stressors simultaneously, for which the combined effects are poorly understood and may have important management consequences. Additive effects are those where the combined effect is equal to the sum, antagonistic where combined effects are less than the sum and for synergistic effects the combined effect is greater to the sum of the individual effects. Here, we performed a meta-analysis of studies recording freshwater fish responses to microplastic-stressor exposures to test if interactions were primarily non-additive (synergistic or antagonistic), and factors impacting the net response. Individual responses were classified (antagonistic/additive/synergistic) and the fit of net responses to a null additive model determined for 838 responses (36 studies) split by categorical variables for the microplastic exposure (environmental relevance, interacting stressor, microplastic morphology and response category measured), as well as the exposed fish (lifestage, ecology and family). Most responses were classified as antagonistic (48 %) and additive (34 %), with synergistic effects least frequent (17 %). Net responses fitted null additive models for all levels of interacting stressor, fish family and microplastic morphology. In contrast, net antagonism was present for biochemical responses, embryo lifestages, environmentally relevant microplastic exposures and fish with benthopelagic ecology, while synergism was identified for fishes with demersal ecology. While substantial knowledge gaps remain and are discussed, the data thus far suggest microplastic-stressor responses in freshwater fishes are rarely synergistic and, therefore, addressing either or both stressors will likely result in positive management and biological outcomes.

Low microplastic loads in riverine European eel (Anguilla anguilla) from southwest England during their marine-freshwater transition.

The microplastic loads in elvers of the critically endangered European eel Anguilla anguilla, sampled in the lower reaches of three English rivers, were very low (incidence: 3.3%, mean ± s.d.: 0.03 ± 0.18 particles) and did not vary with body length or between rivers. Particles were mostly black, polyolefins, fibres and fragments of size 101-200 µm. Current levels indicate a low contamination pressure locally and, consequently, management efforts might prioritise mitigating the effects of other stressors affecting the species.

Parasite infection but not chronic microplastic exposure reduces the feeding rate in a freshwater fish.

Microplastics (plastics <5 mm) are an environmental contaminant that can negatively impact the behaviour and physiology of aquatic biota. Although parasite infection can also alter the behaviour and physiology of their hosts, few studies have investigated how microplastic and parasite exposure interact to affect hosts. Accordingly, an interaction experiment tested how exposure to environmentally relevant microplastic concentrations and the trophically transmitted parasite Pomphorhynchus tereticollis affected the parasite load, condition metrics and feeding rate of the freshwater fish final host chub Squalius cephalus. Microplastic exposure was predicted to increase infection susceptibility, resulting in increased parasite loads, whereas parasite and microplastic exposure were expected to synergistically and negatively impact condition indices and feeding rates. Following chronic (≈170 day) dietary microplastic exposure, fish were exposed to a given number of gammarids (4/8/12/16/20), with half of the fish presented with parasite infected individuals, before a comparative functional response experiment tested differences in feeding rates on different live prey densities. Contrary to predictions, dietary microplastic exposure did not affect parasite abundance at different levels of parasite exposure, specific growth rate was the only condition index that was lower for exposed but unexposed fish, with no single or interactive effects of microplastic exposure detected. However, parasite infected fish had significantly lower feeding rates than unexposed fish in the functional response experiment, with exposed but unexposed fish also showing an intermediate decrease in feeding rates. Thus, the effects of parasitism on individuals were considerably stronger than microplastic exposure, with no evidence of interactive effects. Impacts of environmentally relevant microplastic levels might thus be relatively minor versus other stressors, with their interactive effects difficult to predict based on their single effects.

Distinct microplastic patterns in the sediment and biota of an urban stream.

Urban freshwaters, their sediments and resident biota are often highly susceptible to microplastic contamination from catchment-specific sources. Water velocity and spatiotemporal dynamics within the system can impact microplastic loads, while biological features may additionally impact levels within freshwater biota. Here, we investigated the spatiotemporal variations in microplastic loads collected from sediment, macroinvertebrate and fish samples from an urban watercourse (Bourne Stream) in Dorset, southwest England. Sediment particles were mostly fragments of colours (especially orange and purple) whereas microplastics in both macroinvertebrates and fishes were blue/green and fibres. Across all sample types, the dominant particle size class was ≤100 µm. Median (M) and range (R) of microplastic loads within each sample type were sediment: M = 0.06, R = 0-0.36 particles g-1; macroinvertebrates: M = 0, R = 0-4 particles per batch; and fishes: M = 1, R = 0-6 particles per individual. Sediment loads varied spatially, with the highest load in the most upstream site, whereas biotic loads did not vary across space and time. Macroinvertebrate batch loadings varied between taxa and feeding guild, with counts significantly higher in annelids but lower in herbivores. Fish counts were higher in species with true, differentiated stomachs, but with the effects of species, feeding guild and body size being non-significant. Within sites, mean microplastic loads did not correlate between sediment, macroinvertebrate and fish samples. These results suggest that sediment freshwater microplastic loadings may vary spatially but that these trends are not reflected by, or correlated to, those in the biota where ingestion varies with biological traits. Assessments of freshwater microplastic contamination must therefore consider sampling spatiotemporally and across different biotic communities to fully understand the scale of contamination, and to subsequently undertake effective mitigation steps.

Microplastic loads within riverine fishes and macroinvertebrates are not predictable from ecological or morphological characteristics.

Microplastics are a relatively new but important form of freshwater contamination that can be ingested by a range of different species, with particle counts thought to be predictable from species ecology and morphology. Here, we report levels of microplastics in a 26 µm-5 mm size range within the macroinvertebrate and fish community of a lowland river (Dorset Stour, SW England), and test the hypothesis that counts are predictable from characteristics such as feeding guild, body length and trophic position. Macroinvertebrates (n = 257, 12 taxa) and fish (n = 418, 9 species) were collected from distinct river reaches by kick sampling and rod and line angling, respectively. Batches of whole macroinvertebrates and individual fish gastrointestinal tracts were digested with 30% hydrogen peroxide before microplastic screening and FTIR polymer confirmation on a particle subset. Particles were found in 40% of pooled macroinvertebrate batches (taxa incidences: 14-75%) and 39% of fishes (species incidences: 29-47%). Dominant particle feature categories were ≤100 µm, blue/green, fragments and fibres identified as various polyolefins. Although particle counts in macroinvertebrates were highest in Ephemeroptera (mean of 0.74 particles per individual), the relationships between particle loads, batch number and guild were all non-significant. In fishes, particle counts were not significantly related to species, stomach structure, feeding guild or body length, with spatial differences also not apparent across the catchment. Individual fish particle counts were similarly not significantly associated with their trophic positions (calculated from bulk δ15N values for a subset of fishes) and parasite load of Pomphorhynchus tereticollis. Correlations between fish and macroinvertebrate particle counts within specific river reaches were also not significant. In entirety, these results indicated although loadings of microplastic particles were relatively consistent within the two communities, they were not predictable from any of their ecological or morphological characteristics.

Effects of Copper and the Insecticide Cypermethrin on a Soil Ciliate (Protozoa: Ciliophora) Community.

Ciliated protozoa play important ecological roles in soils, yet few studies have investigated the effect of soil pollution on them. We determined the effect of copper (Cu) and cypermethrin on a soil ciliate community under microcosm conditions. Soils were treated with Cu or cypermethrin and the abundance and species richness of ciliates determined 15 days later. Cu treatment increased soil ciliates abundance at the highest concentration (960 mg kg-1), as did cypermethrin at a treatment of 160 mg kg-1. No negative effect on ciliate abundance was found for either substance due to increased numbers of tolerant species, particularly Homalogastra setosa and Chilodonella uncinata in the case of Cu and Colpoda stenii and Colpoda inflata for cypermethrin treatments. However, several species were absent at high treatment levels. Notably, Halteria grandinella was not found in Cu treatments above 240 mg kg-1, whilst Oxytricha setigera was not found in cypermethrin treatments above 160 mg kg-1. For Homalogastra setosa, there was an initial positive response to cypermethrin, but abundance then decreased at a treatment of 320 mg kg-1, and treatment at 640 mg kg-1 eradicated the species from the microcosms. Accordingly, both substances affected the structure of the soil ciliate community at high concentrations.

Morphology and Phylogeny of the Ciliate Psilotricha silvicola n. sp. (Alveolata, Ciliophora) from Woodland Soils in the United Kingdom.

The genus Psilotricha was established by Stein in 1859, with P. acuminata as the type species within the family Oxytrichidae. This species lacked a full description until it was re-discovered in 2001, showing that its morphological and morphogenetic characters confirmed the inclusion in the family Oxytrichidae. Since then, the genus Psilotricha has had a convoluted taxonomy despite the morphological evidence available. In this paper, we describe a new Psilotricha species, Psilotricha silvicola n. sp., from woodland soils in Southern England (United Kingdom). The morphology was investigated in live and protargol-impregnated specimens. Our findings show that P. silvicola n. sp. shares morphological characteristics with P. acuminata, including the distinctive cell shape and the long and sparse cirri. Phylogenetic analysis of the 18S rRNA gene places this new species within the family Oxytrichidae, nested apart from the family Psilotrichidae (which includes the genera Urospinula, Psilotrichides and Hemiholosticha), in a clade containing species of the family Oxytrichidae. Furthermore, the morphology of another Psilotricha species, P. viridis, found in a freshwater pond in the same woodland area, is also here described, bringing additional insight into the taxonomy of the genus. Our findings provide further evidence for inclusion of the genus Psilotricha within the oxytrichids.

Assessment of biotransfer and bioaccumulation of cadmium, lead and zinc from fly ash amended soil in mustard-aphid-beetle food chain.

The present study investigates the extent of biotransfer and bioaccumulation of cadmium (Cd), lead (Pb) and zinc (Zn) from fly ash amended soil in mustard (Brassica juncea)-aphid (Lipaphis erysimi)-beetle (Coccinella septempunctata) food chain and its subsequent implications for the beetle. The soil was amended with fly ash at the rates of 0, 5, 10, 20 and 40% (w/w). Our results showed that the uptake of Cd, Pb and Zn from soil to mustard root increased with the increase in fly ash application rates, but their root to shoot translocation was relatively restricted. Increase in chlorophyll content and dry mass of mustard plant on treatments ≥20% even at elevated accumulation of Cd (1.67mgkg-1), Pb (18.25mgkg-1) and Zn (74.45mgkg-1 dry weight) in its shoot showed relatively higher tolerance of selected mustard cultivar to heavy metal stress. The transfer coefficient (TC1) of Cd from mustard shoot to aphid was always >1, indicating that Cd biomagnified in aphids at second trophic level. But, there was no biomagnification of Cd in adult beetles at third trophic level. Zinc accumulation was 2.06 to 2.40 times more in aphids than their corresponding host shoots and 1.26-1.35 times more in adult beetles than their prey (aphids) on which they fed. Lead was only metal whose TC was <1 at both second and third trophic levels. The elimination of Cd via honeydew of aphids was most efficient as the ratio of metal in honeydew to aphid (ranging from 0.21 to 0.26) was higher than the Pb (0.16 to 0.20) and Zn (0.07 to 0.09). The statistically consistent (p>0.05) biomass and predation rate of predatory beetles indicated that all levels of soil amendments with fly ash did not have any lethal or sub-lethal effects on beetles.

Effect of cadmium accumulation on mineral nutrient levels in vegetable crops: potential implications for human health.

Consumption of vegetables is often the predominant route whereby humans are exposed to the toxic metal Cd. Health impacts arising from Cd consumption may be influenced by changes in the mineral nutrient content of vegetables, which may occur when plants are exposed to Cd. Here, we subjected model root (carrot) and leaf (lettuce) vegetables to soil Cd concentrations of 0.3, 1.5, 3.3, and 9.6 µg g(-1) for 10 weeks to investigate the effect of Cd exposure on Cd accumulation, growth performance, and mineral nutrient homeostasis. The findings demonstrated that Cd accumulation in lettuce (20.1-71.5 µg g(-1)) was higher than that in carrot (3.2-27.5 µg g(-1)), and accumulation exceeded the maximum permissible Cd concentration in vegetables when soil contained more than 3.3 µg g(-1) of Cd. There was a marked hormetic effect on carrot growth at a soil Cd concentration of 3.3 µg g(-1), but increasing the Cd concentration to 9.6 µg g(-1) caused decreased growth in both crops. Additionally, in most cases, there was a positive correlation between Cd and the mineral nutrient content of vegetables, which was due to physiological changes in the plants causing increased uptake and/or translocation. This may suggest a general mechanism whereby the plant compensated for disrupted mineral nutrient metabolism by increasing nutrient supply to its tissues. Increased nutrient levels could potentially offset some risks posed to humans by increased Cd levels in crops, and we therefore suggest that changes in mineral nutrient levels should be included more widely in the risk assessment of potentially toxic metal contamination. Graphical abstract The Cd concentration (µg g-1 in dry matter) in the root, shoot and translocation factor (TF) of Cd from root to shoot in the carrot and lettuce, and the percentage of root Cd to the gross Cd contents (%) in carrot (C) and lettuce (D) exposed to soil Cd (0 (control), 1, 3, and 9 µg g-1) for 70 days. Values are means ± SD (n = 5).

The transfer and fate of Pb from sewage sludge amended soil in a multi-trophic food chain: a comparison with the labile elements Cd and Zn.

The contamination of agroecosystems due to the presence of trace elements in commonly used agricultural materials is a serious issue. The most contaminated material is usually sewage sludge, and the sustainable use of this material within agriculture is a major concern. This study addresses a key issue in this respect, the fate of trace metals applied to soil in food chains. The work particularly addresses the transfer of Pb, which is an understudied element in this respect, and compares the transfer of Pb with two of the most labile metals, Cd and Zn. The transfer of these elements was determined from sludge-amended soils in a food chain consisting of Indian mustard (Brassica juncea), the mustard aphid (Lipaphis erysimi) and a predatory beetle (Coccinella septempunctata). The soil was amended with sludge at rates of 0, 5, 10 and 20 % (w/w). Results showed that Cd was readily transferred through the food chain until the predator trophic level. Zn was the most readily transferred element in the lower trophic levels, but transfer to aphids was effectively restricted by the plant regulating shoot concentration. Pb had the lowest level of transfer from soil to shoot and exhibited particular retention in the roots. Nevertheless, Pb concentrations were significantly increased by sludge amendment in aphids, and Pb was increasingly transferred to ladybirds as levels increased. The potential for Pb to cause secondary toxicity to organisms in higher trophic levels may have therefore been underestimated.

Assimilation of Cd and Cu by the carnivorous plant Sarracenia leucophylla raf. fed contaminated prey.

Many species of carnivorous plants have become endangered through exposure to multiple risks such as habitat loss, illegal poaching, and pollution. A potential threat to these plants posed by pollution stems from the contamination of their invertebrate prey with trace metals. This study examined the potential for prey to act as sources of the trace metals Cd and Cu for the pitcher plant Sarracenia leucophylla. Cd- and Cu-contaminated Diptera larvae were fed to S. leucophylla plants in separate experiments. The results demonstrated that Cd and Cu were readily transferred to the shoots of S. leucophylla in a dose-dependent manner. While the assimilation of Cu decreased with treatment level, the assimilation of Cd did not. Some assimilated Cu appeared to be translocated to the roots, but Cd was strongly retained in the shoots, where it was related to a reduction in shoot biomass. This suggested that on exposure to Cd-contaminated prey, the plants either experienced phytotoxicity or there was disruption of nutrient acquisition from the prey. Accumulation of Cu was not related to any sign of phytotoxicity.

Differential uptake, partitioning and transfer of Cd and Zn in the soil-pea plant-aphid system.

The biomagnification of trace metals during transfer from contaminated soil to higher trophic levels may potentially result in the exposure of predatory arthropods to toxic concentrations of these elements. This study examined the transfer of Cd and Zn in a soil-plant-arthropod system grown in series of field plots that had received two annual applications of municipal biosolids with elevated levels of Cd and Zn. Results showed that biosolids amendmentsignificantly increased the concentration of Cd in the soil and the shoots of pea plants and the concentration of Zn in the soil, pea roots, shoots, and pods. In addition, the ratio of Cd to Zn concentration showed that Zn was preferentially transferred compared to Cd through all parts of the system. As a consequence, Zn was biomagnified by the system whereas Cd was biominimized. Cd and Zn are considered to exhibit similar behaviors in biological systems. However, the Cd/Zn ratios demonstrated that in this system, Cd is much less labile in the root-shoot-pod and shoot-aphid pathways than Zn.

Contrasting behaviour of cadmium and zinc in a soil-plant-arthropod system.

This study investigates the transfer of Cd and Zn from a soil amended with sewage sludge at rates up to 100 t ha(-1) through a multi-trophic system consisting of barley, the aphid Sitobion avenae and the larvae of the lacewing Chrysoperla carnae. Results show marked differences in the transfer of the two metals. Cadmium was freely accumulated in barley roots, but accumulation in the shoot was restricted to a concentration of around 0.22 mg kg(-1) (dry weight). This limited the transfer of Cd to higher trophic levels and resulted in no significant accumulation of Cd in S. avenae or in C. carnae. Zinc transfer in the system was largely unrestricted, resulting in significant accumulation in roots and shoots, in S. avenae and in C. carnae. Cadmium biomagnification occurred in lacewing pupae, with concentrations up to 3.6 times greater than in aphids. S. avenae biomagnified Zn by a factor of ca. 2.5 at low sludge amendment rates, but biomagnification decreased to a factor of 1.4 at the highest amendment rate. Biomagnification of Zn did not occur in C. carnae, but concentrations were up to 3.5 time higher than in soil. Results are discussed in light of the mechanisms regulating transfer of the two metals in the system.