Aquatic Life Stages Research Articles

A potential evolutionary trap for the extended phenotype of a nematomorph parasite

Abstract Human activities introduce new environmental cues to wild organisms, leading to maladaptive behavioral and life history decisions known as the “evolutionary trap.” This trap is thought to be a major conservation concern for free-living organisms. However, it has never been studied in endosymbionts, one of the most successful and diverse life forms on Earth. Here, we examine this trap in the extended phenotype of a parasite that exploits the visual system of hosts to alter host behavior for its benefit. Arboreal mantids infected by nematomorph parasites are drawn to horizontally polarized light, thereby inducing them to enter the water. In this study, we found that the degree of linear polarization (DOP) of reflected light served as a reliable environmental cue for identifying perennial waters, where nematomorphs can survive in their aquatic life stage without drying out. Infected mantids exhibit attraction to horizontally polarized light with higher DOP in behavioral assays and jumped into pools reflecting light with higher DOP in field experiments. The asphalt road reflected horizontally polarized light closely resembling the polarization levels observed in perennial waters, likely leading to a higher prevalence of mantids on asphalt roads compared with those found in natural arboreal habitats. In a field experiment, we observed infected mantids walking on asphalt roads more often than on cement roads. These findings imply that evolutionary traps can endanger endosymbionts beyond their hosts that directly perceive environmental cues.

Meta-analysis of the effects of microplastic on fish: Insights into growth, survival, reproduction, oxidative stress, and gut microbiota diversity

Aquatic ecosystems are primary repositories for microplastics (MPs), which pose significant risks to aquatic organisms. This study addresses the gap in understanding the effects of MPs pollution by analyzing 3,757 biological endpoints from 85 laboratory studies. Overall, our results indicate that MPs exposure significantly inhibits fish growth, survival, and reproductive ability, and increases oxidative damage, specifically, MPs exposure leads to elevated levels of malondialdehyde. However, MPs do not have a significant impact on the diversity of fish gut microbiota. Subgroup and correlation analyses indicate that the extent of various toxic effects is influenced by multiple factors, including MPs’ type, exposure pathway, size, concentration, as well as the aquatic environment or life stage of the fish. In addition, the regression analysis revealed a relationship between the magnitude of toxic effects and the size, concentration, or duration of MPs exposure. This study provides useful information for understanding the potential impacts of MPs on aquatic organisms and offers new insights for the protection and management of aquatic ecosystems.

Influence of polystyrene nanoplastics on the toxicity of haloperidol to amphibians: An in vivo and in vitro approach

Chemical pollution is a major driver for the current worldwide crisis of amphibian decline. The present study aimed to assess the influence of polystyrene nanoplastics (PS-NPLs) on the toxicity of haloperidol to aquatic life stages of amphibians, by using in vivo (tadpoles of Xenopus laevis and Pelophylax perezi) and in vitro (A6 and XTC-2 cell lines of X. laevis) biological models. Tadpoles of both species were exposed, for 96 h, to haloperidol: 0.404 to 2.05 mg l−1 (X. laevis) or 0.404 to 3.07 mg L−1 (P. perezi). The most sensitive species to haloperidol (X. laevis) was exposed to haloperidol's LC50,96h combined with two PS-NPLs concentrations (0.01 mg L−1 or 10 mg L−1); the following endpoints were monitored: mortality, malformations, body lengths and weight. In vitro cytotoxicity was assessed by exposing the two cell lines, for 72 h, to: haloperidol (0.195 to 100 mg L−1) alone and combined with 0.01 mg L−1 or 10 mg L−1 of PS-NPLs. Xenopus laevis tadpoles revealed a higher lethal and sublethal sensitivity to haloperidol than those of P. perezi, with LC50,96h of 1.45 and 2.20 mg L−1. In vitro assays revealed that A6 cell line is more sensitive haloperidol than XTC-2: LC50,72h of 13.2 mg L−1 and 5.92 mg L−1, respectively. Results also suggested a higher sensitivity of in vivo models when compared to in vitro biological. Overall, PS-NPLs did not influence haloperidol's toxicity for in vivo and in vitro biological models, except for a reduction on the incidence of malformations while increasing the lethal toxicity (at the lowest concentration) in tadpoles. These opposite interaction patterns highlight the need for a deeper comprehension of NPLs and pharmaceuticals interactions. Results suggest a low risk of haloperidol for anuran tadpoles, though in the presence of PS-NPLs the risk may be increased.

Metamorphosis alters concentrations of bioaccumulated pharmaceuticals and endocrine disruptors in caddisflies: Implications for transfer of aquatic–terrestrial subsidies

Abstract Aquatic insects serve as vital inter‐habitat linkages between aquatic and terrestrial ecosystems, as their life cycle includes both aquatic and terrestrial stages. They facilitate the flow of energy and nutrients and play an important role in the transport of waterborne contaminants to terrestrial environments. This study focused on the under‐investigated environmental fate of emerging contaminants (ECs), particularly pharmaceuticals (PhACs) and endocrine‐disrupting compounds (EDCs), which originate from wastewater effluents and accumulate in aquatic insects. Using an in situ study of a highly abundant caddisfly species (Trichoptera), Silo nigricornis (Pictet, 1834), inhabiting a wastewater‐impacted drainage ditch, we examined the bioaccumulation and bioamplification of PhACs and EDCs across the aquatic and terrestrial life stages of the caddisfly. We observed variations in the highest concentrations of PhACs and EDCs in different sample types, including water, biofilm and different life stages of S. nigricornis. Adult S. nigricornis exhibited the highest total concentrations of ECs among the different life stages, with 17 different ECs bioaccumulated in caddisfly tissues. Most of these ECs had higher concentrations in terrestrial adults compared to aquatic larvae and pupae. The total concentration of ECs increased significantly with development, revealing a 41% higher concentration in adult terrestrial caddisflies compared to pupae and larvae. Bioamplification factors provided insights into the increased body burden of the majority of compounds during at least one stage of metamorphosis in S. nigricornis. Evidence of bioamplification was observed in both metamorphosis stages, with certain compounds, such as antibiotics (azithromycin and tilmicosin), endocrine disruptors (TCPP – tris[1‐chloro‐2‐propyl]phosphate and parabens; methylparaben, propylparaben), consistently exhibiting increased concentrations from larval to pupal and from pupal to adult stages. The observed pattern of increasing body burden of contaminants during the caddisfly life cycle emphasises the effects of metamorphosis on the concentration of PhACs and EDCs in adult holometabolous aquatic insects. Our results emphasise the importance of adult caddisflies for the transport of waterborne contaminants such as PhACs and EDCs from aquatic to terrestrial environments. Therefore, our results emphasise the need to include adult trichopterans in the assessment of habitat quality, as their influence extends beyond the boundaries of the aquatic ecosystem.

River metabolic fingerprints and regimes reveal ecosystem responses to enhanced wastewater treatment.

Although many studies have examined how improvements in wastewater treatment impact river nutrient concentrations and loads, there has been much less focus on measuring river metabolism to evaluate the wider aquatic ecosystem benefits of reducing nutrient inputs to rivers. The objectives of this study were to evaluate the effects of enhanced wastewater treatment (nitrification) on river metabolism in the Grand River, Canada's largest river draining into Lake Erie. Metabolic fingerprints and regimes (calculated from high-frequency dissolved oxygen [DO] measurements) were used to visualize whole-river ecosystem functional responses to these wastewater treatment upgrades. There was a 60% reduction in ecosystem respiration during summer, in response to reductions in effluent total ammonia inputs, causing a shift from net heterotrophy to net autotrophy, and contraction of river metabolic fingerprints. This resulted in major improvements in summer DO concentrations, with reductions in the percentage of days during summer that DO minima fell below water-quality guidelines for protection of aquatic early life stages, from 88% to ≤16%. The results also point to potential cascading impacts on coupled phosphorus and nitrogen cycles, which may generate further improvements in river water quality. During the summer, high rates of river metabolism and nutrient retention may result in measured water-column nutrient concentrations potentially underestimating nutrient pressures. This study also demonstrates the value of combining river metabolism with nutrient monitoring for a more holistic understanding of the role of nutrients in river ecosystem health and function.

Differential sensitivity of aquatic life stages of Pelophylax perezi to an acidic metal-contaminated effluent.

Acid mine drainage (AMD) involves complex mixtures of metals and hydrogen ions that can be highly toxic to the biota. Assessing the effects of AMD to aquatic stages of amphibians is key, as this group constitutes the vertebrate class with the highest proportion of species considered as threatened. Thus, the present work aimed at assessing the sensitivity of two aquatic life stages of the green frog Pelophylax perezi to an AMD originated from a cupric-pyrite mine. Embryos (Gosner stages 10-11) and tadpoles (Gosner stages 20-21) of P. perezi were exposed, for 96 h, to six AMD dilutions (1.39% to 7.5%). Endpoints involving responses at different levels of biological organization were monitored: mortality, malformations, hatching rates, body length and growth rate, enzymatic activity related with detoxification metabolism (glutathione S-transferase), and histopathologies (anatomical structures of the digestive, respiratory, and excretory systems). Embryos presented high mortality and malformation rates at AMD levels equal or above 5.36%, as well as premature hatching at 1.95% of AMD or higher. A significant reduction in body length and growth rate occurred in embryos and tadpoles exposed to 1.95% or higher levels of AMD, respectively. At the histological level, several abnormalities were observed for AMD-exposed tadpoles in a variety of tissues. One of the most noticeable histological changes occurred in the intestine that exhibited papillary epithelial hyperplasia along with a yellowish content and was more pronounced in tadpoles exposed to higher AMD levels. FEmbryos were more sensitive to lethal levels of AMD than tadpoles, suggesting embryos as a useful model life stage when performing amphibian risk assessment of mine drainage. Furthermore, AMD was highly toxic for P. perezi aquatic life stages since levels as low as 1.95% induced lethal effects. These results emphasize the importance of implementing efficient remediation methodologies for AMD, given its high toxicity.

Effects of agricultural pesticides on the susceptibility and fitness of malaria vectors in rural south-eastern Tanzania

BackgroundAgricultural pesticides may exert strong selection pressures on malaria vectors during the aquatic life stages and may contribute to resistance in adult mosquitoes. This could reduce the performance of key vector control interventions such as indoor-residual spraying and insecticide-treated nets. The aim of this study was to investigate effects of agrochemicals on susceptibility and fitness of the malaria vectors across farming areas in Tanzania.MethodsAn exploratory mixed-methods study was conducted to assess pesticide use in four villages (V1–V4) in south-eastern Tanzania. Anopheles gambiae (s.l.) larvae were collected from agricultural fields in the same villages and their emergent adults examined for insecticide susceptibility, egg-laying and wing lengths (as proxy for body size). These tests were repeated using two groups of laboratory-reared An. arabiensis, one of which was pre-exposed for 48 h to sub-lethal aquatic doses of agricultural pesticides found in the villages.ResultsFarmers lacked awareness about the linkages between the public health and agriculture sectors but were interested in being more informed. Agrochemical usage was reported as extensive in V1, V2 and V3 but minimal in V4. Similarly, mosquitoes from V1 to V3 but not V4 were resistant to pyrethroids and either pirimiphos-methyl or bendiocarb, or both. Adding the synergist piperonyl butoxide restored potency of the pyrethroids. Pre-exposure of laboratory-reared mosquitoes to pesticides during aquatic stages did not affect insecticide susceptibility in emergent adults of the same filial generation. There was also no effect on fecundity, except after pre-exposure to organophosphates, which were associated with fewer eggs and smaller mosquitoes. Wild mosquitoes were smaller than laboratory-reared ones, but fecundity was similar.ConclusionsSafeguarding the potential of insecticide-based interventions requires improved understanding of how agricultural pesticides influence important life cycle processes and transmission potential of mosquito vectors. In this study, susceptibility of mosquitoes to public health insecticides was lower in villages reporting frequent use of pesticides compared to villages with little or no pesticide use. Variations in the fitness parameters, fecundity and wing length marginally reflected the differences in exposure to agrochemicals and should be investigated further. Pesticide use may exert additional life cycle constraints on mosquito vectors, but this likely occurs after multi-generational exposures.Graphical

Bioaccumulation and bioamplification of pharmaceuticals and endocrine disruptors in aquatic insects

Environmental fate of emerging contaminants such as pharmaceuticals and endocrine disrupting compounds at the aquatic terrestrial boundary are largely unexplored. Aquatic insects connect aquatic and terrestrial food webs as their life cycle includes aquatic and terrestrial life stages, thus they represent an important inter-habitat linkage not only for energy and nutrient flow, but also for contaminant transfer to terrestrial environments. We measured the concentrations of pharmaceuticals and endocrine disrupting compounds in the larval and adult tissues (last larval stages and teneral adults) of five Odonata species sampled in a wastewater-impacted river, in order to examine their bioaccumulation and bioamplification at different taxonomic levels. Twenty different compounds were bioaccumulated in insect tissues, with majority having higher concentrations (up to 90% higher) in aquatic larvae compared to terrestrial adults (reaching 88 ng/g for 1H-benzotriazole). However, increased concentration in adults was observed for seven compounds in at least one suborder (41% of the accumulated), confirming contaminants bioamplification across the metamorphosis. Both, bioaccumulation and bioamplification differed at various taxa levels; the order (Odonata), suborder (Anisoptera and Zygoptera) and species level. Highest variability was observed between Anisoptera and Zygoptera, due to the underlying differences in their ecology. Generally, Zygoptera had higher concentrations of contaminants in both larvae and adults. Additionally, we aimed at predicting effects of contaminant properties on bioaccumulation and bioamplification patterns using the commonly used physicochemical and pharmacokinetic descriptors on both order and suborder levels, however, neither of the two processes could be consistently predicted with simple linear models. Our study highlights the importance of taxonomy in studies aiming at advancing the understanding of contaminant exchange between aquatic and terrestrial food webs, as higher taxonomic categories include ecologically diverse groups, whose contribution to “the dark side of subsidies” could substantially differ.

The wastewater micropollutant carbamazepine in insectivorous birds—an exposure estimate

Insects with aquatic life stages can transfer sediment and water pollutants to terrestrial ecosystems, which has been described for metals, polyaromatic hydrocarbons, and polychlorinated chemicals. However, knowledge of the transfer of aquatic micropollutants released by wastewater treatment plants is scarce despite some preliminary studies on their occurrence in riparian spiders. In our study, we address a major analytical gap focusing on the transfer of the micropollutant carbamazepine from the larvae to the adult midges of Chironomus riparius using an optimized QuEChERS extraction method and HPLC–MS/MS applicable to both life stages down to the level of about three individuals. We show that the uptake of carbamazepine by larvae is concentration-dependent and reduces the emergence rate. Importantly, the body burden remained constant in adult midges. Using this information, we estimated the daily exposure of insectivorous tree swallows as terrestrial predators to carbamazepine using the energy demand of the predator and the energy content of the prey. Assuming environmentally relevant water concentrations of about 1 μg/L, the daily dose per kilogram of body weight for tree swallows was estimated to be 0.5 μg/kg/day. At places of high water contamination of 10 μg/L, the exposure may reach 5 μg/kg/day for this micropollutant of medium polarity. Considering body burden changes upon metamorphosis, this study fills the missing link between aquatic contamination and exposure in terrestrial habitats showing that wastewater pollutants can impact birds’ life. Clearly, further analytical methods for biota analysis in both habitats are urgently required to improve risk assessment.Graphical abstract

How plants affect amphibian populations.

Descriptions of amphibian habitat, both aquatic and terrestrial, often include plants as characteristics but seldom is it understood whether and how those plants affect amphibian ecology. Understanding how plants affect amphibian populations is needed to develop strategies to combat declines of some amphibian populations. Using a systematic approach, we reviewed and synthesized available literature on the effects of plants on pond-breeding amphibians during the aquatic and terrestrial stages of their life cycle. Our review highlights that plant communities can strongly influence the distribution, abundance, and performance of amphibians in multiple direct and indirect ways. We found three broad themes of plants' influence on amphibians: plants can affect amphibians through effects on abiotic conditions including the thermal, hydric, and chemical aspects of an amphibian's environment; plants can have large effects on aquatic life stages through effects on resource quality and abundance; and plants can modify the nature and strength of interspecific interactions between amphibians and other species - notably predators. We synthesized insights gained from the literature to discuss how plant community management fits within efforts to manage amphibian populations and to guide future research efforts. While some topical areas are well researched, we found a general lack of mechanistic and trait-based work which is needed to advance our understanding of the drivers through which plants influence amphibian ecology. Our literature review reveals the substantial role that plants can have on amphibian ecology and the need for integrating plant and amphibian ecology to improve research and management outcomes for amphibians.

Population connectivity of aquatic insects in a dam‐regulated, desert river

AbstractHumans have exaggerated natural habitat fragmentation, negatively impacting species dispersal and reducing population connectivity. Habitat fragmentation can be especially detrimental in freshwater populations, whose dispersal is already constrained by the river network structure. Aquatic insects, for instance, are generally limited to two primary modes of dispersal: downstream drift in the aquatic juvenile life stages and flight during the terrestrial winged adult stage. Yet the impacts of large hydropower dams can make rivers uninhabitable for incoming (drifting) juvenile insects, with remaining refugia found only in tributaries. The ability of adult aquatic insects to traverse such river stretches in search of suitable tributary habitat likely depends on factors such as species‐specific dispersal ability and distance between tributaries. To explore the intersection of natural and human‐induced habitat fragmentation on aquatic insect dispersal ability, we quantified population genetics of three taxa with varying dispersal abilities, a caddisfly (Hydropsychidae, Hydropsyche oslari), a mayfly (Baetidae: Fallceon quilleri), and a water strider (Veliidae: Rhagovelia distincta), throughout tributaries of the Colorado River in the Grand Canyon, Arizona, USA. Using 2bRAD reduced genome sequencing and landscape genetics analyses, we revealed a strong pattern of isolation by distance among mayfly populations. This contrasts with caddisfly and water strider populations, which were largely panmictic. Analysis of thousands of informative single nucleotide polymorphisms showed that realized dispersal ability may not be accurately predicted by species traits for these widespread species. Principal components analysis revealed a strong division between caddisfly populations upstream and downstream of Havasu Creek (279 km through the 390 km study reach), suggesting that the geography of the Grand Canyon imposes a dispersal barrier for this species. Our use of genetic tools in the Grand Canyon to understand population structure has enabled us to elucidate dispersal barriers for aquatic insects. Ultimately, these data may be useful in informing effective conservation management plans for understudied organisms of conservation interest.

Could species-focused suppression of Aedes aegypti, the yellow fever mosquito, and Aedes albopictus, the tiger mosquito, affect interacting predators? An evidence synthesis from the literature.

The risks of Aedes aegypti and Aedes albopictus nuisance and vector‐borne diseases are rising and the adverse effects of broad‐spectrum insecticide application have promoted species‐specific techniques, such as sterile insect technique (SIT) and other genetic strategies, as contenders in their control operations. When specific vector suppression is proposed, potential effects on predators and wider ecosystem are some of the first stakeholder questions. These are not the only Aedes vectors of human diseases, but are those for which SIT and genetic strategies are of most interest. They vary ecologically and in habitat origin, but both have behaviorally human‐adapted forms with expanding ranges. The aquatic life stages are where predation is strongest due to greater resource predictability and limited escape opportunity. These vectors' anthropic forms usually use ephemeral water bodies and man‐made containers as larval habitats; predators that occur in these are mobile, opportunistic and generalist. No literature indicates that any predator depends on larvae of either species. As adults, foraging theory predicts these mosquitoes are of low profitability to predators. Energy expended hunting and consuming will mostly outweigh their energetic benefit. Moreover, as adult biomass is mobile and largely disaggregated, any predator is likely to be a generalist and opportunist. This work, which summarizes much of the literature currently available on the predators of Ae. aegypti and Ae. albopictus, indicates it is highly unlikely that any predator species depends on them. Species‐specific vector control to reduce nuisance and disease is thus likely to be of negligible or limited impact on nontarget predators. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Long-lasting effects of experimental flow intermittency on alpine stream macroinvertebrates (Val Roseg, Switzerland)

Changing weather patterns and receding glaciers are predicted to increase flow intermittency in alpine streams. If aquatic macroinvertebrate communities largely comprise taxa adapted to perennial flows, an increase in flow intermittency substantially reduces biodiversity and affects functional processes. We conducted a before-after-control-impact field experiment to examine how macroinvertebrate communities in an alpine headwater stream responded to and recovered from a repeated experimental increase in flow intermittency. Flow in one channel was manipulated to simulate increased summer intermittency (June–September) over two consecutive years, whilst an adjacent channel served as a control. We monitored the density of benthic macroinvertebrates, periphyton and organic matter at approximately monthly intervals over three years during the snow-free period. Before manipulation, both channels had similar ecological properties. The flow manipulation reduced the overall macroinvertebrate density, and especially the proportional rheophile density, across both years. Recovery of the macroinvertebrate community following experimental flow intermittency took more than a year, and longer than our study period. This could be due to long aquatic life stages, dispersal limitation and biotic interactions. We conclude that climate-induced changes in alpine stream flow regimes can lead to a fundamental shift in macroinvertebrate assemblages through local extinctions, mostly of rheophilic species.

Improving estimations of life history parameters of small animals in mesocosm experiments: A case study on mosquitoes

AbstractMesocosm experiments enable researchers to study animal dynamics, but determining accurate estimates of survival and development rates of different life stages can be difficult, especially as the subjects may be hard to sample and mortality rates can be high. We propose a new methodology for estimating such parameters.We used an experimental set‐up with 48 aquatic mesocosms, each with 20 first instar mosquito larvae and under 1 of 12 treatments with varying temperatures and nutrient concentrations. We took daily subsamples of the aquatic life stages as well as counting the emerging adults. We developed a method to estimate the survival and development probabilities at each life stage, based on optimising a matrix population model. We used two different approaches, one assuming the difference between predictions and observations was normally distributed, and the other using a combination of a normal and a multinomial distribution. For each approach, the resulting optimisation problem had around 100 parameters, making conventional gradient descent ineffective with our limited number of data points. We solved this by computing the formal derivatives of our matrix model.Both approaches proved effective in predicting mosquito populations over time, also when compared against a separate validation dataset, and the two approaches produced similar results. They also both predicted similar trends in the survival and development probabilities for each life stage, although there were some differences in the actual values. The approach which only used the normal distribution was considerably more computationally efficient than the mixed distribution approach.This is an effective approach for determining the survival and development rates of small animals in mesocosm experiments. We have not found any other reliable methodology for estimating these parameters, especially not from incomplete data or when there are many different experimental treatments. This methodology enables researchers to gain a much more detailed understanding of the life cycles of small animals, potentially leading to advances in a wide range of areas, for example in mosquito‐borne disease risk or in considering the effects of biodiversity loss or climate change on different species.

Exploring the community structure of Afrotropical macroinvertebrate traits and ecological preferences along an agricultural pollution gradient in the Kat River, Eastern Cape, South Africa

• We identified signature traits and ecological preferences of agricultural pollution. • Traits and ecological preferences responded differentially to agricultural pollution. • Shredders, crawlers and a preference for macrophytes were sensitive to agriculture. • Predators, swimmers, haemoglobin and medium body size were tolerant to agriculture. Agricultural activities impact riverine ecosystem structure, function, and processes. In the Afrotropical regions, research on agricultural effects on macroinvertebrate trait distribution is sparse. In this study, we investigated the spatial and temporal changes in the community structure of macroinvertebrate traits along an agricultural disturbance gradient in an Afrotropical River system. Physicochemical variables were sampled alongside macroinvertebrates at eight sites in the dry (winter and spring) and wet (summer and autumn) periods of 2018–2019. We grouped the sites into four categories using the percentage of agricultural land-use cover within each drainage area. Our results showed that agricultural pollution exhibited varying effects on traits and ecological preferences, with traits such as a predatory lifestyle, medium body-size (>10–20 mm), active swimming, possession of spiracles and haemoglobin, and adult aquatic life stage increasing with the pollution gradient. These traits were positively associated with nutrients (PO 4 + -P, NO 2 + -N, NH 4 + -N and NO 3 + -N), salinity, turbidity and temperature and were deemed tolerant of agricultural pollution. Shredding, crawling, and a preference for macrophytes as food showed strong positive associations with the least disturbed sites and were negatively associated with increasing nutrients, salinity, turbidity and water temperature. As such, these three traits were considered sensitive to agricultural pollution. The identified indicator traits can be used to predict the survival and distribution patterns of organisms under the impact of agriculture-induced stress.

Comparative Microbiota Composition Across Developmental Stages of Natural and Laboratory-Reared Chironomus circumdatus Populations From India.

Chironomids are aquatic insects that undergo a complete metamorphosis of four life stages. Here we studied, for the first time, the microbiota composition of Chironomus circumdatus, a tropical midge species, both from the Mula and Mutha Rivers in Pune, India and as a laboratory-reared culture. We generated a comparative microbial profile of the eggs, larvae and pupae, the three aquatic life stages of C. circumdatus. Non-metric multidimensional scaling analysis (NMDS) demonstrated that the developmental stage had a more prominent effect on the microbiota composition compared to the sampling location. Notably, the microbiota composition of the egg masses from the different sampling points clustered together and differed from laboratory culture larvae. Proteobacteria was the dominant phylum in all the environmental and laboratory-reared egg masses and pupal samples, and in the laboratory-reared larvae, while Fusobacteria was the dominant phylum in the larvae collected from the field environment. The most abundant genera were Cetobacterium, Aeromonas, Dysgonomonas, Vibrio, and Flavobacterium. The ten amplicon sequence variants (ASVs) that most significantly contributed to differences in microbiota composition between the three sampled locations were: Burkholderiaceae (ASVs 04 and 37), C39 (Rhodocyclaceae, ASV 14), Vibrio (ASV 07), Arcobacter (ASV 21), Sphaerotilus (ASV 22), Bacteroidia (ASVs 12 and 28), Flavobacterium (ASV 29), and Gottschalkia (ASV 10). No significant differences were found in the microbial richness (Chao1) or diversity (Shannon H’) of the three sampled locations. In contrast, significant differences were found between the microbial richness of the three life stages. Studying the microbiota of this Chironomus species may contribute to a better understanding of the association of C. circumdatus and its microbial inhabitants.

Health risks of sulfentrazone exposure during zebrafish embryo-larvae development at environmental concentration

Knowledge about the negative effects and mechanism of sulfentrazone (SUL) on aquatic early life stages is still limited. Here we investigated the lethal and sub-lethal effects of SUL during zebrafish embryo-larvae development. Results demonstrated that the 96 h and 120 h-LC50 of SUL to embryonic zebrafish was 2.02 mg/L, and the 30 d-LC50 was 0.899 mg/L after embryos exposed to SUL for 30 d. High concentrations of SUL delayed yolk sac absorption, disordered the hatching and heart rate during zebrafish embryonic stage, while 0.0100–0.100 mg/L SUL had no phenotypic changes on embryonic development, but decreased the body weight of larvae after 30 d exposure. RNA-seq identified 321, 394 and 727 differentially expressed genes in larvae after embryos exposed to 0.0100 mg/L, 0.0400 mg/L and 0.400 mg/L SUL for 30 d, found that the transcriptional profiles involved in heart development and endocrine disruption were simultaneously influenced by different concentrations of SUL, such as adrenergic signaling in cardiomyocytes, cardiac muscle contraction, cell adhesion molecules and steroid biosynthesis. Biochemical analysis showed that SUL increased the levels of E2, T3 and TSH, induced the activities of mitochondrial complex IV, cytochrome c oxidase, Ca2+-ATPase, total Na+K+-ATPase and Ca2+Mg2+-ATPase, and decreased ATP formation after embryos exposed to SUL for 5 d and 30 d. Further comprehensive analysis demonstrated that SUL caused more significantly alteration on the transcript, level or activity of the key elements involved in heart development and endocrine disruption after 30 d exposure, indicated long-term SUL exposure might cause more negative effects on zebrafish at doses below the presumed no-observed-adverse-effect level during early life development. The results inferred the environmental concentration of SUL might cause potential cardiac and endocrine health risk in zebrafish later life stages, also facilitated a better understanding of the sub-lethal effects and molecular mechanism of SUL on aquatic organism.

Tree Swallows (Tachycineta bicolor) Nesting in Agricultural Settings: Going Deep into Pesticide Exposure

Photo 1. A female tree swallow (Tachycineta bicolor) hangs on one of the 400 nest boxes of our study system. Tree swallows feed on aerial insects, mainly diptera and insects with an aquatic life stage, from which they can be exposed to various pesticides. Photograph credit: Marie-Christine Poisson. Photo 2. This young tree swallow is likely only a few moments away from fledging, which they usually do at 20–22 days old. All individuals in our study system, including nestlings, are identified with unique band numbers. Photograph credit: Marie-Christine Poisson. These photographs illustrate the article “Assessing pesticides exposure effects on the reproductive performance of a declining aerial insectivore” by Marie-Christine Poisson, Daniel R. Garrett, Audrey Sigouin, Marc Bélisle, Dany Garant, Lounès Haroune, Jean-Philippe Bellenger, and Fanie Pelletier published in Ecological Applications. https://doi.org/10.1002/eap.2415.

Polymethylmethacrylate nanoplastics can cause developmental malformations in early life stages of Xenopus laevis

Polymethylmethacrylate (PMMA) production has increased almost 20% over the last years. With its release into the aquatic environment, its breakdown or degradation to nano dimensions (nanoplastics-NPLs) due to biological and physical/mechanical action is, theoretically, anticipated. The occurrence of PMMA-NPLs in aquatic ecosystems may thus cause adverse effects particularly to early life stages of amphibians, which may be in contact with PMMA-NPLs suspended in the water column or deposited in upper layers of the sediments. Accordingly, this work aimed at assessing the effects of PMMA-NPLs to aquatic early life stages of the model anuran species Xenopus laevis. To attain this objective, two types of toxicity assays were carried out by exposing embryos [Nieuwkoop and Faber (NF) stage 8-11] or tadpoles (NF 45) to three concentrations of PMMA-NPLs (1, 100 and 1000 μg/L): i) 96-h embryo teratogenicity assay, where survival, malformation, and total body length (BL) of embryos were assessed; and ii) 48-h feeding rate assay, where survival, feeding (FR), malformations and growth rates (body weight-BW and BL) of tadpoles were evaluated. PMMA-NPLs exposure had no significant effects on mortality, malformations of X. laevis embryos but BL was lower at 1000 μg PMMA-NPLs/L. In tadpoles, no effects on survival or FR were observed after exposure to PMMA-NPLs, but significant changes occured in BW and BL. Moreover, anatomical changes in the abdominal region (externalization of the gut) were observed in 62.5% of the tadpoles exposed to 1000 μg PMMA-NPLs/L. Despite the lack of knowledge regarding the environmental levels of NPLs, it is expected that sediments constitute a sink for these contaminants, where they can become available for organisms that, like tadpoles, feed on the organic matter at the surface of sediments. Considering the continuous release and subsequent accumulation of PMMA, the malformations obtained in the feeding assays suggest that, in the future, these nano-polymers may constitute a risk for aquatic life stages of amphibians.

Few Impacts of Introduced Cutthroat Trout (Oncorhynchus clarki) on Aquatic Stages of Boreal Toads (Anaxyrus boreas boreas)

Introduced salmonids can impact aquatic ecosystems through direct predation and indirect effects. We explored the effects of introduced Cutthroat Trout (Onchoryncus clarki) on Boreal Toad (Anaxyrus boreas boreas) survival and habitat use during two aquatic life stages, the embryo and tadpole, at Boreal Toad breeding sites with and without Cutthroat Trout. We found no difference in embryo survival and higher tadpole survival at the site with Cutthroat Trout. Cutthroat Trout are unlikely to use the shallow areas where Boreal Toad eggs are deposited; however, during the tadpole stage, Cutthroat Trout and tadpoles overlap broadly in near-shore aquatic habitats. Frequency of tadpole habitat use is lower in Cutthroat Trout–used areas, but we observed no behavioral or temporal avoidance of Cutthroat Trout by tadpoles. Our results suggest that Cutthroat Trout do not have a negative effect on Boreal Toad embryo or tadpole survival in wild settings and that Cutthroat Trout presence does not preclude tadpoles from using habitats.