Spinetoram is one of the most worldwidely used pesticides for its high insecticidal efficacy and low human toxicity Following the large usage of spinetoram, the ecotoxicity and environmental risks to aquatic ecosystems have call for urgent study In the present study, we investigated the combined effects of spinetoram and the harmful alga Microcystis aeruginosa in freshwater, on survival and reproduction of Daphnia pulex Acute toxicity test of spinetoram resulted in negative effects on survival, with a 48h-LC50 value of 3771 µg L–1 Under the long-time exposure to environmentally relevant concentrations (018 and 035 µg L–1) of spinetoram and a low composition of Microcystis (30%) in the diet, D pulex showed both shorter longevity and lower fecundity, the time to first brood was also increased At population level, carrying capacity was highly decreased by spinetoram and Microcystis, whereas a significant decrease of intrinsic growth rate was observed at 035 µg L–1 spinetoram with 30% Microcystis as food The present study highlighted that pesticide spinetoram had highly toxic effects on D pulex and could reduce the tolerance of D pulex to M aeruginosa, causing great effects on D pulex population in natural waterbodies
Research Article
Combined Effects of the Pesticide Spinetoram and the Cyanobacterium Microcystis on the Water Flea Daphnia Pulex
https://doi.org/10.21203/rs.3.rs-780208/v1
This work is licensed under a CC BY 4.0 License
Journal Publication
published 17 Feb, 2022
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acute toxicity
chronic exposure
Daphnia pulex
interactive effect
Microcystis aeruginosa
spinetoram
Spinosyns are a series of macrocyclic natural bio-insecticides derived from the soil actinomycete Saccharopolyspora spinosa, which have been registered in more than 100 countries to control above 200 species of crops since the launch in 1996, because of their high efficiency, low unit area usage, and short residue period (Huang et al 2009; Bacci et al 2016; Sparks et al 2020) Spinosyns act mainly on the nicotinic acetylcholine receptors (nAChRs) and γ-aminobutyric acid receptors (GABARs) in the nervous system of arthropods, through depolarizing nerve cells and cause extensive hyperactivation of central nervous system Spinetoram, the next-generation product of spinosyn insecticides launched in 2007, has large global market for its high insecticidal efficacy and low human toxicity (Galm and Sparks, 2016) Following the worldwide adoption of spinetoram, the ecotoxicity and environmental risks have call for urgent study In particular, risk assessments of spinetoram on beneficial arthropods like natural enemies are crucial because potential side effects on these non-target arthropods can greatly affect their natural populations and ecological functions (Biondi et al 2012)
Aquatic environments receive direct and indirect pesticides inputs, the enormous number of pesticides induce a series of aquatic environmental problems A great deal of research have documented the ecotoxicological risks of pesticides on the aquatic organisms and ecosystems eg (i) physiological and behavioral sublethal effects (Bownik et al 2019), (ii) long-term suppression in population levels (Dalkvist et al 2009), (iii) alternation of aquatic community composition (Vonesh and Kraus 2009) Due to the large inputs of pesticides, aquatic organisms at different trophic levels are exposed to the threat of spinetoram For example, the development (ie body length and heartbeat rate) and immune system of zebrafish embryos were negatively affected under the exposure to an effective concentration of 025 g L− 1 spinetoram (Cheng et al 2020) Spinosyn-based products, especially spinetoram, has great efficacy and selectivity to target pests (Sparks et al 2008) Nonetheless, the non-target impact of spinetoram on beneficial arthropods requires systematic risk assessments, for these including natural enemies and pollinators, play crucial roles in ecosystems (Biondi et al 2012) Recently, spinetoram as well as other spinosyn-based products are also used to control aquatic insects, such as mosquitoes at developmental stages (Mohamed et al 2021), and control ectoparasitic infestations in aquaculture raised fish (Dick et al 2013) Therefore, although aquatic arthropods are also exposed to spinetoram threat, very few studies investigated the potential effect of spinetoram on those organisms
The freshwater cladocerans of the genus Daphnia are keystone pelagic filter feeders in the food webs of temperate ponds and lakes, and are served as an important model for ecological, ecotoxicological, biogeographical and evolutionary researches (Eads et al 2008; Seda and Petrusek, 2011; Perhar and Arhonditsis, 2015) The ability of Daphnia to suppress phytoplankton biomass, especially during the bloom of toxic cyanobacteria, indicates that the Daphnia seems to be critically important to the biomanipulation efforts in eutrophicate lakes (Chislock et al 2013) However, artificial pollutants in aquatic environments would have a significant impact on the ecological function of Daphnia (Pestana et al 2010; Liu et al 2019) The major pollutants like heavy metals and pesticides have been reported in previous studies not only affect the life-history traits but also inhibit the feeding rates of Daphnia (Hartgers et al 1999; Lessard and Frost, 2012) Whereas, artificial pollutants could suppress the grazing control of Daphnia on Microcystis, due to direct adverse effects on Daphnia or indirect interference on species interactions (Cerbin et al 2010; Yang et al 2011)
Besides artificial pollutants, intensive human activities also increase the nutrients flux in natural waterbodies, and as a result, cyanobacterial blooms occur frequently in freshwaters In freshwater aquaculture ponds, cyanobacterial blooms due to eutrophication is now becoming a major issue during pond system development (Bi et al 2019) Microcystis aeruginosa is one of the most common bloom-forming cyanobacteria in freshwater ecosystems worldwide, and it is assumed that, global warming together with eutrophication will promote Microcystis blooms (O’Neil et al 2012) In natural freshwaters, Daphnia spp play a central role in controlling Microcystis blooms (Triest et al 2016) Nonetheless, compared with green algae, Microcystis is poor in nutrient quality (Hairston et al 2001), and toxic strains cause negative effects on the fitness of zooplankton (Lei et al 2020) Additionally, study on the effects of non-toxic Microcystis on Daphnia grazing and life-history traits demonstrated that, Microcystis must contain some substances other than microcystins that have adverse effects on Daphnia (Lürling 2003)
It has been well documented that, spinosyn-based products have high toxicity risk on Daphnia (Duchet et al 2010) With the increasing global market of spinetoram, Daphnia is supposed to be under higher threat and to experience combined stress of spinetoram and Microcystis In this study, we hypothesized that the exposure to spinetoram would reduce the tolerance of Daphnia to Microcystis For this purpose, three separate experiments were conducted An acute experiment was run to assess the toxic effect of spinetoram on D pulex Whereas chronic exposure to selected concentrations of spinetoram, with or without Microcystis as food, were run to analyze the combined effects of spinetoram and Microcystis
21 Test organisms and culture conditions
A clone of D pulex, which was collected from the pond in the campus of Hohai University, Nanjing, China, and has been maintained in our laboratory since 2017, was used in this study Daphniids were cultured with a density of less than 100 individuals in 1-L beakers containing 800 mL dechlorinated tap water The suspensions of Chlorella pyrenoidosa at the stage of exponential growth were harvested by centrifugation and supplied as the food of D pulex with a density of 5×105 cells mL–1 C pyrenoidosa and a non-toxic strain of Microcystis aeruginosa (FACHB-469) were mono-cultured separately in 3 L Erlenmeyer flasks axenically using BG-11 medium and aerated with 022-μm filtrated air Erlenmeyer flasks were shaken manually 3 times a day The algae were collected by centrifugation (500 g for 10 min) and stored at 4 °C The cultivation of daphniids and algae and all the experiments were conducted in a constant temperature chamber at 25 °C, under fluorescent light at 40 μmol photons m–2s–1, with a light–dark period of 14:10 h
22 Chemical and test solution
Spinetoram was purchased from Dow AgroSciences (USA) and stored in a dark and dry place To prepare the stock solution, spinetoram was ultrasonic crushed for ten minutes and then dissolved in acetone (C995% analyticalgrade purity) The stock solution was immediately prepared prior to each experiment In all experiments, the maximum percentage of acetone in the cultural medium was below 001% (v/v) Prior to the experiments, the toxic effects of the acetone solution at a concentration of 001% (v/v) on D pulex was tested and showed no toxic effects (data not shown) Determination of the actual concentration of spinetoram in media was conducted by LC-MS/MS method, using Agilent 1290 HPLC-6495B The column was an Ascentis Express 90A C18 (30 × 150 mm, 27 μm) operated at 35°C, flow rate of 450 μL/min and the injection volume was 5 μL A prior enrichment of media was conducted using Cleanert PestiCarb/NH2 (Agela Tech China) before the concentration measurement
23 Acute exposure to spinetoram
To start a clonal culture, one ovigerous female was isolated from the D pulex population and cultured in a 100 mL beaker with 80 mL cultural medium C pyrenoidosa was supplied as food of D pulex with a final density of 5 ×105 cells mL–1 every day and renewed the cultural media every three days A continuous range of spinetoram concentrations was set: 0, 05, 1, 25, 5, 10, 20, 30, 40, 50, 75, 100, 200, 500 µg L–1 The actual concentrations of spinetoram were 0, 044, 088, 219, 428, 875, 1752, 2628, 3504, 4380, 6569, 8758, 17518 and 43795 µg L–1, respectively For each concentration, 5 replicates with 10 individuals per replicate were taken Daphniids were selected from the clonal culture randomly and cultured in 50 mL beakers with 40 mL media, and no food was offered during the acute exposure to spinetoram Mortality was recorded at 12 h, 24 h, and 48 h The death of D pulex was determined as the time when the heart stopped beating under an inverted microscope Additionally, the phenomena of rapid movement and immobilization of D pulex were recorded Dose-response curves were adopted to fit the mortality rate and immobilization rate against time, respectively A special dose-response curve was adopted to fit the mortality rate against time at 48 h, 
In the equation, M represented the mortality rate, Mm and M0 represented the maximal and minimum mortality rates, respectively C means concentration of spinetoram and k represented the slope of this curve The value of LC50 at 48 h was calculated according to the fitted curve
24 Chronic effects of spinetoram and Microcystis at individual level
Neonates of D pulex within 24 hours of birth were selected randomly and individually placed in 50 mL beakers containing 40 mL tested media A chronic 21-d toxicity experiment was carried out using a full factorial design composed of three concentrations of spinetoram (actual concentrations tested by LC-MS/MS: 0, 018 and 035 µg L–1) and two diet compositions (Ch: 100% C pyrenoidosa; Ch+Ma: 70% C pyrenoidosa + 30% M aeruginosa) with a final algal biomass of 35 mg C L–1 To maintain constant concentration of spinetoram, chemical test solutions were replaced by fresh ones every day Parameters of individual level, including survival days and reproduction were recorded The death of D pulex was determined as the time when the heart stopped beating under an inverted microscope
25 Chronic effects of spinetoram and Microcystis at population level
To investigate the long-term effects of spinetoram and Microcystis on D pulex population, a third experiment was conducted Similarly to the previous experiment set-up of the second one, we started the third experimental with a clonal culture Neonates of D pulex within 24 hours of birth in this clonal culture were raised in good diet conditions (100% C pyrenoidosa) for 3 days Then a chronic population experiment was conducted using a full factorial design similar to the second experiment, three concentrations of spinetoram (0, 018 and 035 µg L–1) and three diet compositions (Ch: 100% C pyrenoidosa; Ch+Ma: 70% C pyrenoidosa + 30% M aeruginosa; Ma: 100% M aeruginosa) with a final algal biomass of 35 mg C L–1 This experiment was established at an initial density of 5 neonates in each tested medium at 500 mL The population abundance was recorded every day, and the population growth curves were fitted by the formula, 
In the equation, r represented the intrinsic rate of population growth, K represented the carrying capacity and a was a constant
26 Statistical analysis
The results were presented as means±1 standard error To test the effects of spinetoram and Microcystis on survival and reproduction parameters, two-way ANOVAs were performed The population growth was fitted according to logistic regression, and the significance of fitted logistic regression curves with observations were assessed through analysis of variances Two-way ANOVAs were performed to analyze the significant effects of spinetoram and Microcystis on carrying capacity and population growth rate All analyses were performed with SigmaPlot 140 Statistical significance was established at α < 005
31 Acute toxic effect of spinetoram on D pulex
The exposure to spinetoram stimulated rapid movement of D pulex, second antennae flapped quicker than in control medium The response of D pulex movement to spinetoram presented both concentration effect and temporal effect As shown in Table 1, at 12 h, most of individuals showed quick and nondirectional swimming trajectory in the treatments with spinetoram concentration above 1752 µg L− 1 After 24 hours, observation of rapid movement decreased to 088 µg L− 1, and at 48 h, the concentration which could induce rapid movement was as low as 044 µg L− 1 However, enhancement of movement was followed by loss of motion ability, which indicated the paralytic effect of spinetoram on D pulex At 12 h, immobilization of D pulex was observed at the spinetoram concentration of 2628 µg L− 1, and the concentration decreased to 428 and 088 µg L− 1 at 24 h and 48 h, respectively (Table 1)
| Observation of rapid movement | Observation of immobilization | |
|---|---|---|
| 12 h | 1752 µg L− 1 | 2628 µg L− 1 |
| 24 h | 088 µg L− 1 | 428 µg L− 1 |
| 48 h | 044 µg L− 1 | 088 µg L− 1 |
Increasing trends were determined in mortality rate and immobilization rate with both the increasing concentration of spinetoram and time (Fig. 1) During the first 24 hours, mortality rate was below 50% under all concentrations of spinetoram (Fig. 1A) According to the fitted dose-response curve, the value of 48h-LC50 equals to 3771 µg L− 1 The value of EC50 of immobilization rate was also calculated according to dose-response curves and the EC50 values at 24 h and 48 h were 1976 and 319 µg L− 1, respectively (Fig. 1C, D)
32 Chronic effects of spinetoram and Microcystis at individual level
During the 21-day chronic experiment, the survivorship of D pulex was affected by both spinetoram and Microcystis (Fig. 2) Generally, individuals fed on Chlorella as the only food survived for longer time than these fed on a mixed diet containing 30% Microcystis at all concentrations of spinetoram In the Chlorella groups, 90% individuals cultured in medium without spinetoram survived until the end of experiment, whereas mortality occurred on 5th day and 2nd day when exposed to 018 and 035 µg L− 1 spinetoram (Fig. 2A) However, 30% Microcystis in diet caused a negative effect on survival, the survival rate of individuals cultured with no spinetoram decreased to 80% at the end of experiment Mortality was observed on 3rd day and 2nd day and 21-day mortality rate decreased to 40% and 30% at 018 and 035 µg L− 1 spinetoram, respectively (Fig. 2B)
Besides survivorship, the reproduction was also affected significantly by spinetoram and Microcystis In both diet groups, exposure to spinetoram increased the time to first brood, especially at higher spinetoram concentration (035 µg L− 1) (Fig. 3A), and the spinetoram concentration had a significantly effect on the time to first brood (Table 2) Fewer offspring was produced by D pulex at first brood (Fig. 3B) and per brood (Fig. 3C) The result of Two-way ANOVA showed a significant effect of spinetoram concentration on offspring number of the first brood, and the number of offspring per brood was affected significantly by both spinetoram concentration and food composition (Table 2) Fewer broods were reproduced by D pulex under the exposure to spinetoram and Microcystis as compared to the control group (Fig. 3D), but no statistical significance was found As for the average number of total offspring produced by one female, significant decrease was found at 035 µg L− 1 spinetoram treatments of both food groups (Fig. 3E), and both spinetoram concentration and food showed significant effects The interval of molting showed different trend in two food groups (Fig. 3F), but the concentration of spinetoram had no significant effect (Table 2)
| Traits | Factors | SS | DF | MS | F (DFn, DFd) | p |
|---|---|---|---|---|---|---|
| Individual level | ||||||
| Time to first brood | Interaction | 168 | 2 | 08402 | F (2, 28) = 02450 | 07843 |
| Spinetoram concentration | 7206 | 2 | 3603 | F (2, 28) = 1051 | 00004 ** | |
| Food composition | 384 | 1 | 384 | F (1, 28) = 1120 | 0299 | |
| Offspring number of first brood | Interaction | 577 | 2 | 2885 | F (2, 28) = 06815 | 05141 |
| Spinetoram concentration | 3425 | 2 | 1713 | F (2, 28) = 4046 | 00286 * | |
| Food composition | 1344 | 1 | 1344 | F (1, 28) = 3176 | 00856 | |
| Offspring per brood | Interaction | 2408 | 2 | 1204 | F (2, 28) = 09108 | 0965 |
| Spinetoram concentration | 4104 | 2 | 2052 | F (2, 28) = 1552 | 00577 | |
| Food composition | 1173 | 1 | 1173 | F (1, 28) = 8869 | 00831 | |
| Number of brood | Interaction | 009821 | 2 | 00491 | F (2, 28) = 003564 | 04138 |
| Spinetoram concentration | 8717 | 2 | 4358 | F (2, 28) = 3164 | < 00001 ** | |
| Food composition | 4449 | 1 | 4449 | F (1, 28) = 3229 | 00059 ** | |
| Total offspring per female | Interaction | 8388 | 2 | 4194 | F (2, 28) = 02649 | 07692 |
| Spinetoram concentration | 1965 | 2 | 9824 | F (2, 28) = 6205 | 00059 ** | |
| Food composition | 9963 | 1 | 9963 | F (1, 28) = 6293 | 00182 * | |
| Interval of molting (d) | Interaction | 02666 | 2 | 01333 | F (2, 52) = 09713 | 03853 |
| Spinetoram concentration | 01246 | 2 | 006229 | F (2, 52) = 04539 | 06376 | |
| Food composition | 07782 | 1 | 07782 | F (1, 52) = 5671 | 00209 * | |
| Population level | ||||||
| Carrying capacity | Interaction | 392993 | 4 | 98248 | F (4, 18) = 1653 | < 00001** |
| Spinetoram concentration | 666427 | 2 | 333213 | F (2, 18) = 5606 | < 00001** | |
| Food composition | 1467725 | 2 | 733862 | F (2, 18) = 1235 | < 00001** | |
| Population growth rate | Interaction | 0004114 | 4 | 0001028 | F (4, 18) = 7864 | 00008** |
| Spinetoram concentration | 003612 | 2 | 001806 | F (2, 18) = 1381 | < 00001** | |
| Food composition | 03056 | 2 | 01528 | F (2, 18) = 1168 | < 00001** |
33 Chronic effects of spinetoram and Microcystis at population level
During the first days of the experiment, D pulex population were composed by immature individuals and, afterwards, following the first reproduction, population kept growing exponentially, except in the 100% Microcystis treatment (Fig. 4) Generally, at three spinetoram concentrations (0, 018, 035 µg L− 1), the populations of D pulex fed on mixed food (70% Chlorella + 30% Microcystis) grew slower than those fed on 100% Chlorella However, feeding on 100% Microcystis could not support the population establishment of D pulex (Fig. 4) The population growth was fitted following the logistic growth curve in both food groups at three spinetoram concentrations
To compare the growth dynamics of different treatments, carrying capacity and intrinsic growth rate were analyzed Carrying capacity was affected significantly by both food and spinetoram concentration (Table 2) It significantly decreased with the increasing spinetoram concentrations, and showed significant decreases in mixed food groups at all three spinetoram concentrations (Fig. 5A) The carry capacity was higher than 800 inds L− 1 when the population of D pulex was raised in medium with no spinetoram and fed on 100% Chlorella However, the carry capacity decreased to about 110 inds L− 1 under the exposure of 035 µg L− 1 spinetoram and fed on 30% Microcystis As for the population of D pulex feeding on 100% Microcystis, the carry capacity was much lower than other groups, and there was no significant difference of carry capacity among the three spinetoram concentrations (Fig. 5A) Besides, a significant interaction between spinetoram concentration and food composition was also found (Table 2) The population growth rate was significantly affected by the concentration of spinetoram and food composition (Table 2), especially in the mixed food group (Fig. 5B) In the groups fed on 100% Chlorella, the growth rate in two spinetoram concentrations were significantly lower than that in no spinetoram treatment Under mixed food and total Microcystis, growth rate decreased with increasing spinetoram concentration (Fig. 5B) In the groups fed on 100% Microcystis, the growth rate was below zero when exposed to spinetoram (Fig. 5B), indicating a negative growth of D pulex population
Spinetoram, one of the most worldwidely used pesticides, not only caused toxic effects on D pulex at both individual and population levels, but also affected the tolerance to Microcystis, in our study Increasing mortality of D pulex was caused by the increasing concentration of spinetoram Whereas the negative effect on longevity, as well as reproduction of D pulex was aggravated when fed on mixed food (70% Chlorella + 30% Microcystis) under long-time exposure Results of this present study demonstrated an interactive effect of spinetoram and Microcystis on the population of D pulex
In the present study, the adverse influence on the survival of D pulex caused by spinetoram was assessed During the acute experiment of direct exposure to spinetoram, under a wide concentration range, the mortality rate was lower than 50% during the first 24 hours (Fig. 1A) Nonetheless, mortality rate showed a quick increase at 48 h, especially the individuals exposed to spinetoram at concentrations higher than 875 µg L− 1 (Fig. 1B) For evaluating both the efficacy on target insects and the potential ecological risk impact on beneficial arthropods of spinetoram, acute toxic studies have been reported previously It could be found in previous studies that the 48 h-LC50 value to diamondback moth, Plutella xylostella, was ranged from 0131 to 1001 mg L− 1 (Li et al 2015), the values of LC50 of spinetoram to the third instar larvae of Lampides boeticus at 24, 48, and 72 h was 671, 223 and 128 mg L− 1 (Sanjeevikumar and Muthukrishnan, 2017) Our experiment figured out the value of 48h-LC50 was approximately 4306 µg L− 1 According to the toxicity categories based on LC50, spinetoram could be classified as “highly toxic pesticide” to Daphnia Besides, the study of cytotoxicity on human liver cells demonstrated that spinetoram could inhibit the proliferation of human liver HepG2 cells and induce the oxidative DNA damage (Zhang et al 2019) Therefore, because spinetoram occupies a great market share, the ecological risk assessment should be taken into serious consideration
For the complex and varied environmental factors faced by Daphnia in natural waterbodies, our present study assumed a mixed threat to D pulex, spinetoram and Microcystis, that is real phenomena in freshwater Results turned out that, over a longer time-scale, both spinetoram and Microcystis had adverse effect on the survival and reproduction of D pulex, and these two factors had interactive effects Fecundity of D pulex was inhibited by both spinetoram and Microcystis, that less offspring were produced under the effects of single and mixed factors Previous studies demonstrated the adverse effect of Microcystis on fecundity of Daphnia mostly reflected in the smaller size of each brood and smaller body size of offspring (Lürling 2003; Huang et al 2020; Li et al 2020) As for spinetoram, the main target sites in arthropods are nAChRs and GABARs in the nervous system, most studies focused on the acute toxic effect of spinetoram on insects (Dissanayaka et al 2020; Sammani et al 2020) However, widely application of spinetoram in agriculture along with the surface run-off, the environmental concentration of spinetoram was stable at relatively low concentrations Our study focused on the long-term exposure to spinetoram, and figured out reproduction of D pulex was also sensitive to spinetoram even at very low concentrations
The impact of spinetoram and Microcystis on the reproduction of D pulex in our study was not only in the fecundity but also in the time to brood Time to first brood was delayed with the increasing spinetoram concentration (Fig. 3A) Exposure to pollutions often caused increased time to maturity along with the decreased fecundity of Daphnia, which might be due to the delayed development and shortage in energy storage (Reynaldi et al 2006; Saebelfeld et al 2017) Usually, Daphnia has high phenotypic plasticity in life history under complex environments (Otte et al 2019; Gu et al 2020) Slight delay in first brood was also observed in D pulex fed on Microcystis (Fig. 3A), which was similar to previous studies (Wang et al 2019; Lei et al 2020; Zhou et al 2020) Resources and energy might be reallocated from somatic growth to reproduction so that Daphnia reaches maturity earlier under the influence of spinetoram and Microcystis in the present study
Because of the adverse effects caused by both spinetoram and Microcystis, population dynamics of D pulex showed significant changes (Fig. 5A) Generally, carrying capacity of D pulex population was highly decreased by both spinetoram and Microcystis, and these two factors had significant interactive effect The potential explanation of the reduced carrying capacity in D pulex population might be due to the energy allocation between development and the elimination of toxic effects of spinetoram and bad food conditions caused by Microcystis Lower intrinsic growth rate of D pulex population was observed at the higher spinetoram with 30% Microcystis as food (Fig. 5B), but other treatments had no significant difference Decline in carrying capacity of D magna population has also been observed in the condition of microplastics-induced food limitation (Bosker et al 2019) Research on the effects of cadmium showed lower population growth rate and population biomass of D manga (Connon et al 2008) Researches show the profound influence of artificial pollutants on Daphnia populations at a long term, which might also disrupt the ecological functions of Daphnia such as consuming Microcystis blooms Extensive application of pesticides is now gradually influencing the structure and biodiversity of both terrestrial and aquatic ecosystems For spinetoram, the application disrupts the structure and the abundance of surface-active arthropod fauna causing considerable changes in the ground dwelling biodiversity of this ecosystem (Lefkaditis et al 2017) As for aquatic ecosystems, very few studies focused on the risk assessment of spinetoram The findings of our study indicated that spinetoram not only affected the fitness of D pulex but also reduced the tolerance of D pulex to Microcystis
With the worldwide use of spinetoram, resistance has been frequently demonstrated in various species of pests such as Plutella xylostella (Lima Neto et al 2016), environmental concentration of spinetoram in waterbodies is assumed increasing in the future Results of our study showed the interactive effects of spinetoram and Microcystis on D pulex, suggesting the consideration of complexed factors in the environmental risk assessment of pesticides Given the importance of Daphnia in freshwater food webs, the interactive effects caused by spinetoram and Microcystis can potentially impact the ecological function of D pulex, and cause great influence on aquatic ecosystems
In the present study, we exposed D pulex to the pesticide spinetoram, and tested the fitness-related traits Survival and reproduction of D pulex showed significant reduction under the acute and chronic exposure to spinetoram Simultaneously, we demonstrated the interactive effects of spinetoram and the main bloom-forming phytoplankton, M aeruginosa, on the individual fitness and population dynamics of D pulex This highlighted that spinetoram had severe toxic effects on D pulex and could reduce the tolerance of D pulex to M aeruginosa, that would cause great effects on D pulex population in natural waterbodies
Acknowledgements
The authors would like to thank the funding institutions and Reviewers
Ethics approval and consent to participate
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Consent for publication
Not applicable
Availability of data and materials
The datasets during the current study are available from the corresponding author on reasonable request
Competing interests
The authors declare no competing interests
Funding
The present study is supported by the earmarked fund for Jiangsu Agricultural Industry Technology System (JATS [2020] 303), Fundamental research Funds for the Central Universities (2017B05014), Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX20_0523), the Fundamental Research Funds for the Central Universities (B200203136)
Authors information
Affiliations
Department of Marine Biology, College of Oceanography, Hohai University, 1 Xikang Road, Nanjing 210098, China
Qiutong Shen, Xuanhe Jia, Yihe Zhan, Xuexia Zhu & Tianheng Gao
Shanghai Cremo Laboratory Co, 688 Qiushi Road, Shanghai, 201512, China
Bangping Li
Contributions
Conceptualization, data curation, and writing: QS, XJ and YZ; review and editing: XZ and TG; methodology and formal analysis: BL; funding acquisition: QS and TG All authors have read and agreed to the published version of the manuscript
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