Effects Of Larvae Research Articles

Sublethal Effects on the Biological Development of Spodoptera frugiperda (Lepidoptera: Noctuidae) When Consuming Neem

Objective: The Fall armyworm S. frugiperda was bred in laboratory and put under dose of watery extracts andoil of seed of A. indica with the aim of determining and to quantify the alterations in the biological developmentof larvae and pupae of caused by the consumption of A. indica and the doses in which the alterations happen.Design/Methodology/Approach: Two experiments were development. In one eight doses were evaluated,with watery extract of seed of A. indica in 4.0, 1.0, 0.7, 0.4, 0.1, 0.07, 0.04 and 0.01%. In the second the doses ofoil of A. indica were evaluated 0.16, 0.09, 0.02, 0.016, 0.009, 0.002, 0.0016 and 0.0009%. In both experimentsthe variables were quantified: duration of the stage of larvae and pupae, survival of the larval stage and pupae,survival of the larva stage and pupae and weight of pupae.Results: The larvae prolonged their development in three days more than the witness when consuming wateryextract of A. indica; They reduced its survival like la rvae and pupa in 18 and 27.5% respectively. In Addition,was reduced the weight of pupas. When consuming oil of A. indica, the larvae delayed their development fromthree to 15 and 19 days more than the witness.Study limitations/Implications: The study was carried out under laboratory conditions and in a single cycle;it is desirable to repeat the study for more cycles and with different populations of fall armyworm.Finding/Conclusions: The adverse effects in larvae were pronounced of three forms: 1) the duration of thelarval stage altered when staying the larvae by more days in that stage without changing pupae. 2) the survivalof the larval stage was reduced and 3) it decreased weight of pupa.

Trematocine-derived antimicrobial peptides from the Antarctic fish Trematomus bernacchaii: potent antibacterial agents against ESKAPE pathogens.

This study investigated the interaction with membrane mimetic systems (LUVs), bacterial membranes, the CD spectra, and the bactericidal activity of two designed trematocine mutants, named Trem-HK and Trem-HSK. Mutants were constructed from the scaffold of Trematocine (Trem), a natural 22-amino acid AMP from the Antarctic fish Trematomus bernacchii, aiming to increase their positive charge. The selectivity of the designed AMPs towards bacterial membranes was improved compared to Trematocine, verified by their interaction with different LUVs and their membranolytic activity. Additionally, their α-helical conformation was not influenced by the amino acid substitutions. Our findings revealed a significant enhancement in antibacterial efficacy against ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacteriaceae family) pathogens for both Trem-HK and Trem-HSK. Firstly, we showed that the selectivity of the two new designed AMPs towards bacterial membranes was greatly improved compared to Trematocine, verifying their interaction with different LUVs and their membranolytic activity. We determined that their α-helical conformation was not influenced by the amino acid substitutions. We characterized the tested bacterial collection for resistance traits to different classes of antibiotics. The minimum inhibitory and bactericidal concentration (MIC and MBC) values of the ESKAPE collection were reduced by up to 80% compared to Trematocine. The bactericidal concentrations of Trematocine mutants showed important membranolytic action, evident by scanning electron microscopy, on all tested species. We further evaluated the cytotoxicity and hemolytic activity of the mutants. At 2.5 μM concentration, both mutants demonstrated low cytotoxicity and hemolysis, indicating selectivity towards bacterial cells. However, these effects increased at higher concentrations. Assessment of in vivo toxicity using the Galleria mellonella model revealed no adverse effects in larvae treated with both mutants, even at concentrations up to 20 times higher than the lowest MIC observed for Acinetobacter baumannii, suggesting a high potential safety profile for the mutants. This study highlights the significant improvement in antibacterial efficacy achieved by increasing the positive charge of Trem-HK and Trem-HSK. This improvement was reached at the cost of reduced biocompatibility. Further research is necessary to optimize the balance between efficacy and safety for these promising AMPs.

High concentrations of phthalates affect the early development of the sea urchin Paracentrotus lividus

The toxicity of three phthalates (PAEs) - butylbenzyl phthalate (BBP), diethyl phthalate (DEP), and di-(2-ethylhexyl) phthalate (DEHP) - was tested on the Mediterranean sea urchin Paracentrotus lividus. Fertilized eggs were exposed to environmental and high PAE concentrations for 72 h. The potential toxic effects on larval development and any morphological anomalies were then assessed to estimate PAEs impact. Environmental concentrations never affected development, while high concentrations induced toxic effects in larvae exposed to BBP (EC50: 2.9 ×103 µg/L) and DEHP (EC50: 3.72 ×103 µg/L). High concentrations caused skeletal anomalies, with a slight to moderate impact for DEP/DEHP and BBP, respectively. PAE toxicity was: BBP>DEHP>DEP. In conclusion, the three PAEs at environmental concentrations do not pose a risk to sea urchins. However, PAE concentrations should be further monitored in order not to constitute a concern to marine species, especially at their early developmental stages.

Size-dependent and sex-specific negative effects of micro- and nano-sized polystyrene particles in the terrestrial invertebrate model Drosophila melanogaster

Microplastic pollution is believed to be one of the most widespread and long-lasting changes on a global scale. Our understanding that microplastics significantly impact terrestrial systems and are a global change stressor continues to grow. In the present study, we investigated the negative effect of long-term (28 days of exposure in food) polystyrene particles of micro (1.0–1.9 µm, 0.4–0.6 µm) and nano (0.04–0.06 µm) scale, in low doses, on the fruit fly – representing a common, globally distributed terrestrial invertebrate, and a model species in many fields. Our observations involved such parameters as ingestion and transfer of particles, survival, reproduction, changes in ultrastructure and tissue and cell responses in midgut epithelium (the place of direct contact with plastic), ovary, and testis in adults, and transgenerational effects in larvae. These observations may indicate possible toxic effects of the tested substances, even in low doses, that can be expected in other taxa, in terrestrial ecosystems. We observed a negative impact of polystyrene particles on the fruit fly survival, midgut, ovary, and testis, involving ultrastructural alterations, such as autophagy and/or ultimately necrosis in the midgut, triggering oxidative stress and activating processes of antioxidative protection. Despite the changes, midgut function and reproduction were not altered – spermatogenesis and oogenesis proceeded normally. The effect was size-dependent – the smaller the polystyrene particles were, the more substantial was the impact they caused. Ultrastructural changes and studied parameters, i.e., generation of ROS (overproduction of which generates oxidative stress), total glutathione concentration (involved in defense against ROS, acting in distinct pathways), and total antioxidant concentration (the oxidative defense system) showed the highest levels after exposure to the smallest nanoparticles, and vice versa. The effect was also sex-dependent, with male flies being more sensitive. Negative effects in males were more substantial and more prominent, even after contact with larger particles, compared to females. The smaller particles (0.4–0.6 µm, 0.04–0.06 µm) were transferred to the ovary and accumulated in the oocytes. In this case, a transgenerational negative effect was detected in larvae. It was characterized by size-dependent alterations, with smaller particles triggering higher levels of ROS and cellular oxidative response. Only the largest particles (1.0–1.9 µm) did not pass into the gonad and did not alter the larvae. These observations together demonstrated that polystyrene particles of micro- and nanoscale, even in a low dose, can induce numerous negative effects on terrestrial invertebrates.

Combined Effects of Cold Treatment and Phosphine in Drosophila suzukii (Diptera: Drosophilidae)

This study analyzed the effect of combined phosphine (PH3) fumigation and low-temperature treatment on the control of Drosophila suzukii. In the low-temperature, single-treatment experiment, there was no significant difference in insecticidal activity between 1 °C and 5 °C except in D. suzukii adults. Adults showed 98.0% mortality after exposure to 1 °C for 7 d. Regarding fumigation treatment with PH3 alone, adults were the most susceptible, and pupae had the highest tolerance. Combined low-temperature treatment and fumigation showed a synergistic effect, except in the egg stage, and showed slight synergistic effects in larvae and pupae regardless of the combined treatment order. In pupae, the combination group exposed to 1 °C for 24 h after exposure to PH3 for 4 h had the lowest LCT99 value, at 10.49 mg·h/L. The sorption rate of PH3 on grapes decreased to 85.49% after 4 h of exposure, with a 15% loading ratio (w/v), and there was no significant difference in various criteria of phytotoxicity in exposed grapes compared to control, even after 14 d of combined treatment. Therefore, this study indicates that combined PH3 fumigation and low-temperature treatment could be useful in D. suzukii control.

Developmental and Reproductive Impacts of Four Bisphenols in Daphnia magna

Bisphenol A (BPA) is a typical endocrine-disrupting chemical (EDC) used worldwide. Considering its adverse effects, BPA has been banned or strictly restricted in some nations, and many analogs have been introduced to the market. In this study, we selected three representative substitutes, BPS, BPF, and BPAF, along with BPA, to assess the developmental and reproductive effects on Daphnia magna. The F0 generation was exposed to bisphenols (BPs) at an environmentally relevant concentration (100 μg/L) for 21 d; then the embryo spawn at day 21 was collected. Behavior traits, the activity of antioxidant enzymes, and gene transcription were evaluated at three developmental stages (days 7, 14, and 21). Notably, body length, heart rate, and thoracic limb beating were significantly decreased, and D. magna behaved more sluggishly in the exposed group. Moreover, exposure to BPs significantly increased the antioxidant enzymatic activities, which indicated that BPs activated the antioxidant defense system. Additionally, gene expression indicated intergenerational effects in larvae, particularly in the BPAF group. In conclusion, BPA analogs such as BPF and BPAF showed similar or stronger reproductive and developmental toxicity than BPA in D. magna. These findings collectively deepen our understanding of the toxicity of BPA analogs and provide empirical evidence for screening safe alternatives to BPA.

Aquaculture rearing systems induce no legacy effects in Atlantic cod larvae or their rearing water bacterial communities

The microbial rearing quality influences the survival of marine larvae. Microbially matured water treatment systems (MMS) provide a more favourable rearing water microbiome than flow-through systems (FTS). It has previously been hypothesised, but not investigated, that initial rearing in MMS leaves a protective legacy effect in Atlantic cod larvae (Gadus morhua). We tested this hypothesis through a crossover 2 × 2 factorial experiment varying the rearing water treatment system (MMS vs FTS) and the microbial carrying capacity (+ /− added organic matter). At 9 days post-hatching, we switched the rearing water treatment system. By comparing switched and unswitched rearing tanks, we evaluated if legacy effects had been established in the larvae or their surrounding rearing water bacterial community. We analysed the bacterial communities with flow cytometry and 16S rRNA gene sequencing. We found no evidence that the initial rearing condition left a legacy effect in the communities by evaluating the bacterial community diversity and structure. Instead, the present rearing condition was the most important driver for differences in the rearing water microbiota. Furthermore, we found that MMS with high microbial carrying capacity appeared to seed a stable bacterial community to the rearing tanks. This finding highlights the importance of keeping a similar carrying capacity between the inlet and rearing water. Moreover, we reject the hypothesis that the initial rearing condition leaves a protective legacy effect in larvae, as the larval survival and robustness were linked to the present rearing condition. In conclusion, our results highlight the importance of maintaining a beneficial microbial rearing environment from hatching and throughout the larval rearing period.

Monarch Butterfly (Danaus plexippus) Life-Stage Risks from Foliar and Seed-Treatment Insecticides.

Conservation of North America's eastern monarch butterfly (Danaus plexippus) population would require establishment of milkweed (Asclepias spp.) and nectar plants in the agricultural landscapes of the north central United States. A variety of seed-treatment and foliar insecticides are used to manage early- and late-season pests in these landscapes. Thus, there is a need to assess risks of these insecticides to monarch butterfly life stages to inform habitat conservation practices. Chronic and acute dietary toxicity studies were undertaken with larvae and adults, and acute topical bioassays were conducted with eggs, pupae, and adults using 6 representative insecticides: beta-cyfluthrin (pyrethroid), chlorantraniliprole (anthranilic diamide), chlorpyrifos (organophosphate), imidacloprid, clothianidin, and thiamethoxam (neonicotinoids). Chronic dietary median lethal concentration values for monarch larvae ranged from 1.6 × 10-3 (chlorantraniliprole) to 5.3 (chlorpyrifos) μg/g milkweed leaf, with the neonicotinoids producing high rates of arrested pupal ecdysis. Chlorantraniliprole and beta-cyfluthrin were generally the most toxic insecticides to all life stages, and thiamethoxam and chlorpyrifos were generally the least toxic. The toxicity results were compared to insecticide exposure estimates derived from a spray drift model and/or milkweed residue data reported in the literature. Aerial applications of foliar insecticides are expected to cause high downwind mortality in larvae and eggs, with lower mortality predicted for adults and pupae. Neonicotinoid seed treatments are expected to cause little to no downslope mortality and/or sublethal effects in larvae and adults. Given the vagile behavior of nonmigratory monarchs, considering these results within a landscape-scale context suggests that adult recruitment will not be negatively impacted if new habitat is established in close proximity of maize and soybean fields in the agricultural landscapes of the north central United States. Environ Toxicol Chem 2021;40:1761-1777. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Comparison of the in vitro protein digestibility of Protaetia brevitarsis larvae and beef loin before and after defatting

Protein digestibility of Protaetia brevitarsis larvae before and after defatting by hexane was compared with that of beef loin in an in vitro digestion model. Larvae had higher crude protein content and 10% trichloroacetic acid (10% TCA)-soluble α-amino groups than beef. Decreases in the levels of total free sulfhydryl groups and 10% TCA-soluble α-amino groups were detected in larvae and beef after defatting (P < 0.05). Surface hydrophobicity increased after defatting in both larvae and beef, (P < 0.05) and tryptophan fluorescence intensity decreased in defatted larvae but increased in defatted beef. Levels of proteins digested into sizes under 3 and 10 kDa in larvae were higher than those in beef (P < 0.05), and defatting did not induce an effect in larvae. Therefore, in the aspect of high protein content and digestibility, larvae of P. brevitarsis can be a potential substitute of animal proteins.

In vivo synergism of ampicillin, gentamicin, ceftaroline and ceftriaxone against Enterococcus faecalis assessed in the Galleria mellonella infection model

The unfavourable safety profile of aminoglycosides and the synergistic effects observed in vitro have prompted the development of novel dual β-lactam therapies, e.g. ampicillin/ceftriaxone or ampicillin/ceftaroline, for the treatment of Enterococcus faecalis endocarditis. For comparison with in vitro chequerboard assay results, a partial chequerboard setup of ampicillin/gentamicin, ampicillin/ceftriaxone and ampicillin/ceftaroline against E. faecalis was established in the Galleria mellonella larval infection model. Discrimination of synergistic and additive interactions was based on the evaluation of larval survival, bacterial quantity in the haemolymph and a pathology score index (internal to the workgroup). Single and multiple dosing schemes based on the half-life of ampicillin were applied. Pharmacokinetic data of the antibiotics in the larvae were determined via agar plate diffusion assays. Ampicillin and ceftriaxone exhibited strain-specific synergistic interactions in the larvae under both dosing regimens, while the other two combinations showed additive effects. Ampicillin/ceftaroline was inferior to ampicillin/ ceftriaxone. Not all synergistic effects observed in vitro could be replicated in the larvae. Our results suggest superior efficacy of ampicillin/ceftriaxone for the treatment of high-inoculum enterococcal infections, for at least some strains, but question the benefit of the current standard of adding the nephrotoxic gentamicin compared with the safer ceftriaxone. This is the first study to develop a scheme for differentiation between additive and synergistic effects in larvae and apply a multiple-antibiotic dosing scheme based on the pharmacokinetics of ampicillin. The model allows the analysis of synergistic effects of antimicrobials in an in vivo setting, but the clinical correlation warrants further study.

Short communication: Antimicrobial activity of indoleacetic, gibberellic and coumaric acids against Paenibacillus larvae and its toxicity against Apis mellifera

Aim of study: To explore three isolated phytomolecules: indoleacetic acid (IAA), gibberellic acid (GA), and the secondary metabolite p-coumaric acid (CUM): (1) evaluating their toxicity against Apis mellifera larvae and adults under controlled conditions in the laboratory; (2) searching for antimicrobial activity against Paenibacillus larvae.Area of study: Honey bee larvae and adults were collected from the experimental apiary of the “Centro de Investigación en Abejas Sociales (CIAS)” (-37.9348798, -57.682817), Institute of the National University of Mar del Plata (UNMdP), Argentina.Material and methods: Paenibacillus larvae strains were isolated from beehives from different provinces of Argentina (Buenos Aires, Córdoba and Entre Ríos) showing clinical symptoms of the American foulbrood. All strains (S1, S2, S3, S4) were genotypically identified using PL5 and PL4 primers and characterized as genotype ERIC1. Then standard essays were performed to determined toxicity of phytomolecules in honey bees and antimicrobial activity through the broth microdilution method.Main results: The diet with GA, IAA and CUM did not present toxic effects in larvae or adult bees, and only CUM showed antimicrobial activity against P. larvae. In this study, we obtained in vitro values of MNIC (minimum non-inhibitory concentration) of 500 μg mL-1 and a MIC (minimum inhibitory concentration) of 650 μg mL-1 for CUM.Research highlights: The obtained results remark its potential as a natural alternative for the control of P. larvae, avoiding the problems generated by the use of synthetic antibiotics such as the resistance phenomena and the contamination of hive’s products.

Chronic Exposure to High Concentrations of Road Salt Decreases the Immune Response of Dragonfly Larvae

Salinization of freshwater ecosystems, due to the application of road salts, is recognized as a potential threat to aquatic communities. Much of the research on the impact of salinity has focused on performance metrics in vertebrates, including respiration and osmoregulation. Here we focus on immune function in of the dragonfly Anax junius, a top predator in fishless aquatic habitats. Impacts on this top predator have the potential to cascade through the community, and immune function is known to be both plastic and sensitive to stress. We injected larvae with monofilaments (simulating a parasite) and placed them in one of three environmentally relevant concentrations of deicing road salt: control (dechlorinated tap water with no added salt), low (1000 mgL-1), or high salt (3000 mgL-1), for either acute (24hr) chronic or (96hr) exposures. We hypothesized that elevated salinity would suppress the immune response and that longer exposure magnifies this effect. As predicted, chronic exposure to high salt concentrations resulted in a significantly reduced larval immune response, however, there was no detectable treatment effects in larvae exposed to low concentrations of road salt or to acute high concentrations. Our results demonstrate that prolonged exposure to high levels of road salt can compromise the immune response of dragonfly larvae. Our findings suggest that insects in aquatic environments that experience sustained environmental salt pollution will be more susceptible to parasites and pathogens, which in turn may affect the impact of this major predator on aquatic community dynamics.

Size-mediated priority and temperature effects on intra-cohort competition and cannibalism in a damselfly.

A shift in the relative arrival of offspring, for example a shift in hatching time, can affect competition at the intraspecific level through size-mediated priority effects, where the larger individuals gain more resources. These priority effects are likely to be affected by climate warming and the rate of intraspecific predation, that is cannibalism. In a laboratory experiment, we examined size-mediated priority effects in larvae of the univoltine damselfly, Lestes sponsa, at two different temperatures (21 and 23°C). We created three size groups of larvae by manipulating hatching time: early hatched with a large size (extra-advanced), intermediate hatched with an intermediate size (advanced) and late hatched with a small size (non-advanced). Thereafter, we reared the larvae from these groups in non-mixed and mixed groups of 12 larvae. We found strong priority and temperature effects. First, extra-advanced larvae most often had higher survival, growth and development rates than non-advanced larvae in mixed groups, compared to groups that consisted of only extra-advanced larvae. Second, temperature increased growth and development rates and cannibalism. However, the strength of priority effects did not differ between the two experimental temperatures, because there was no statistical interaction between temperature and treatments. That is, the mixed and non-mixed groups of non-advanced, advanced and extra-advanced larvae showed the same relative change in life-history traits across the two temperatures. Non-advanced and advanced larvae had similar or higher growth rate and mass in mixed groups compared to non-mixed groups, suggesting that predation from advanced larvae in the mixed group released resources for the non-advanced and advanced larvae that survived despite cannibalism risk. Thus, a thinning effect occurred due to cannibalism caused by priority effects. The results suggest that a shift in the relative arrival of offspring can cause temperature-dependent priority effects, mediated through cannibalism, growth and development, which may change the size distribution and abundance of emerging aquatic insects.

Waterborne and diet-related effects of inorganic and organic fungicides on the insect leaf shredder Chaetopteryx villosa (Trichoptera)

It is well-documented that fungicides can affect crustacean leaf shredders via two effect pathways, namely waterborne exposure and their diet (i.e., via dietary uptake of fungicides adsorbed to leaf material and an altered microorganism-mediated food quality). As a consequence of different life history strategies, the relevance of these effect pathways for aquatic shredders belonging to other taxonomic classes, for instance insects, remains unclear. Therefore, we investigated waterborne and diet-related effects in larvae of the caddisfly leaf shredder Chaetopteryx villosa (Insecta: Trichoptera) and compared our observations to previous reports on effects in adults of the crustacean leaf shredder Gammarus fossarum (Malacostraca: Amphipoda). We assessed acute waterborne effects of an organic fungicide mixture (OFM) and the inorganic fungicide copper (Cu) on the leaf consumption (n = 30) of the fourth-/fifth-instar larvae of C. villosa and their food choice (n = 49) when offered leaf material, which was either conditioned in presence or in absence of the respective fungicide(s). Moreover, the larval leaf consumption (n = 50) and physiological fitness (i.e., growth as well as lipid and protein content) were examined after subjecting C. villosa for 24 days towards the combination of both effect pathways at environmentally relevant concentrations. G. fossarum and C. villosa exhibited similar sensitivities and the same effect direction when exposed to the OFM (either waterborne or dietary pathways). Both shredders also showed the same effect direction when exposed to dietary Cu, while with regards to mortality and leaf consumption C. villosa was less sensitive to waterborne Cu than G. fossarum. Finally, as observed for G. fossarum, the combined exposure to OFM over 24 days negatively affected leaf consumption and the physiology (i.e., growth and lipid reserves) of C. villosa. While no combined Cu effects were observed for larval leaf consumption, contrasting to the observations for G. fossarum, the physiology of both shredders was negatively affected, despite partly differing effect sizes and directions. Our results suggest that C. villosa and G. fossarum are of comparable sensitivity towards waterborne and diet-related organic fungicide exposure, whereas the trichopteran is less sensitive to Cu-based waterborne fungicide exposure. However, when both pathways act jointly, organic and inorganic fungicides can affect the physiology of shredder species with completely different life history strategies. As caddisflies represent a subsidy for terrestrial consumers, these observations indicate that fungicide exposure might not only affect aquatic ecosystem functioning but also the flux of energy across ecosystem boundaries.

Fluazinam impairs oxidative phosphorylation and induces hyper/hypo-activity in a dose specific manner in zebrafish larvae

Fluazinam is a pyridinamine fungicide that induces oxidative stress and mitochondrial damage in cells, and it has been reported to be neurotoxic. To characterize the biological effects of fluazinam, we assessed mitochondrial bioenergetics, dopamine system expression, and behavior of early life staged zebrafish (0.01 μM–0.5 μM). Fluazinam at environmentally-relevant levels did not induce sub-lethal effects in larvae, but at the LC50 (0.5 μM), fluazinam decreased basal and ATP-linked respiration significantly in embryos. As mitochondria are directly related to redox homeostasis and apoptosis, the expression of genes related to oxidative stress and apoptosis were measured. Superoxide dismutase 2 (sod2), heat stock protein 70 (hsp70), bcl2-associated X protein (bax), and caspase 9 (casp9) mRNA levels were up-regulated by 0.5 μM fluazinam. Taken together, there was evidence for mitochondrial dysfunction and oxidative damage at the highest concentration of fluazinam (0.5 μM) tested. As there are reports for fluazinam-induced neurotoxicity in dopamine synthesizing cells, transcriptional targets in the dopamine system were assessed in the zebrafish. Tyrosine hydroxylase 1 (th1) and dopamine receptor 2a (drd2a) mRNA levels were decreased by 0.5 μM fluazinam, suggesting that this fungicide may affect the dopaminergic system. To further assess the potential for fluazinam-mediated neuromodulation, the dark photokinesis response was assessed in larvae following exposure. Larvae exposed to 0.1 μM fluazinam showed hyperactivity, while larvae exposed to 0.2 and 0.3 μM showed hypo-activity. This study demonstrates that fluazinam disrupts mitochondrial bioenergetics in zebrafish, inducing an oxidative stress response, and aberrant behaviors in larvae that are dose dependent.

Nanotoxicological and teratogenic effects: A linkage between dendrimer surface charge and zebrafish developmental stages

This article reports novel results about nanotoxicological and teratogenic effects of the PAMAM dendrimers DG4 and DG4.5 in zebrafish (Danio rerio). Zebrafish embryos and larvae were used as a rapid, high-throughput, cost-effective whole-animal model. The objective was to provide a more comprehensive and predictive developmental toxicity screening of DG4 and DG4.5 and test the influence of their surface charge. Nanotoxicological and teratogenic effects were assessed at developmental, morphological, cardiac, neurological and hepatic level. The effect of surface charge was determined in both larvae and embryos. DG4 with positive surface charge was more toxic than DG4.5 with negative surface charge. DG4 and DG4.5 induced teratogenic effects in larvae, whereas DG4 also induced lethal effects in both zebrafish embryos and larvae. However, larvae were less sensitive than embryos to the lethal effects of DG4. The platform of assays proposed and data obtained may contribute to the characterization of hazards and differential effects of these nanoparticles.

Analysis of the acute response of Galleria mellonella larvae to potassium nitrate

Potassium nitrate (E252) is widely used as a food preservative and has applications in the treatment of high blood pressure however high doses are carcinogenic. Larvae of Galleria mellonella were administered potassium nitrate to establish whether the acute effects in larvae correlated with those evident in mammals. Intra-haemocoel injection of potassium nitrate resulted in a significant increase in the density of circulating haemocytes and a small change in the relative proportions of haemocytes but haemocytes showed a reduced fungicidal ability. Potassium nitrate administration resulted in increased superoxide dismutase activity and in the abundance of a range of proteins associated with mitochondrial function (e.g. mitochondrial aldehyde dehydrogenase, putative mitochondrial Mn superoxide dismutase), metabolism (e.g. triosephosphate isomerase, glyceraldehyde 3 phosphate dehydrogenase) and nitrate metabolism (e.g. aliphatic nitrilase, glutathione S-transferase). A strong correlation exists between the toxicity of a range of food preservatives when tested in G. mellonella larvae and rats. In this work a correlation between the effect of potassium nitrate in larvae and mammals is shown and opens the way to the utilization of insects for studying the in vivo acute and chronic toxicity of xenobiotics.

Вплив згодовування плідникам райдужної форелі ганаміновіту на виживання заплідненої ікри та передличинок

The aim of research was a study the effect of influence of feeding by feed supplement of Ganaminovit the breeding stock of rainbow trout on the state of fertilized eggs and pre-larvae. The task of research was to study influence of Ganaminovit on the survival of fertilized eggs and on the productive effect of larvae in the conditions of the closed water-supply. The breeding stock of rainbow trout were fed by the extruded mixed fodders with Ganaminovit in 2015 during two months to and after spawning and in 2016 before spawning. The influence of feeding rainbow trout breeding stock, along with extruded feed vitamin supplements – Ganaminovit on the survival rate of fertilized eggs and pre-larvae for the mass of 0.3 g. It is established that in conditions of closed water system in the pre-spawning and after-spawning periods of the breeding stock of rainbow trout requires additional vitamin supplements, other than those included in the modern extruded food. The introduction supplements for two months, before incubation, promotes more rapid recovery of the body in after-spawning period and enhances immunity and high survival rates of eggs and of pre-larvae for the mass of 0.3 g. Mortality of eggs in the control group of breeding stock at the stage of the eye was 61.7%, and the increase of weight of the larvae during transition to active feeding in 2015–0,041 g. In 2016, these figures were higher and amounted to respectively 79.4% of and 0.052 g.

&lt;b&gt;Impact of Brazilian fish species at early developmental stages on plankton communities and water chemical parameters

Effect of larvae of Astyanax altiparanae (AA), Piaractus mesopotamicus (PM), Leporinus obtusidens (LO) and Prochilodus lineatus (PL) on plankton communities and water quality were evaluated. Twenty 500 L tanks were used into a completely randomized design with five treatments and four replications, containing 350 individuals of each species per tank. One treatment control had no larvae (CT). Every six days, were collected periphyton and water to determine the concentrations of chlorophyll and total phosphorus. Plankton samples (20 mm mesh net) were taken every three days. Fish affected plankton communities, which showed lower densities of total zooplankton in AA tanks than in LO tanks. Increased density of rotifers and reduced density of copepods were found in AA, PM and PL tanks in relation to the CT. Higher density of testate amoebae was observed in AA, PM and LO tanks. As to biovolume, higher values for total plankton were registered in the CT and LO tanks compared with AA, PM and PL tanks; higher values for rotifers were observed in PL tanks than in the CT. Copepod biovolume was higher in CT and LO tanks. Biovolume of cladoceran and testate amoebae were not affected. Plankton´s chlorophyll concentration was higher in tanks with fish. Higher concentrations of dissolved oxygen were found in AA and PM tanks. It can be concluded that juveniles of these fish species differently affect the plankton community and chemical parameters of the water.

Stressing biological samples with pulsed magnetic fields: physical aspects and experimental results

Magnetic field effects are diffused among living organisms. They are mainly studied with static or extremely low frequency fields, while scarce information is available for pulsed fields. This work is devoted to the study of the interaction between Drosophila melanogaster, both adults and larvae, and pulsed magnetic fields. We exposed the organisms to a peak field of 0.4 T, lasting for about 2 μ s, within an ad hoc designed copper coil. Adult individuals didn't present any deregulation of repetitive sequences in the germ line of Drosophila. Instead, we noticed a marked magnetic field effect in larvae. Polytene chromosomes coming from treated individuals showed the presence of heat shock puffs; the same organisms revealed also an upregulation of the genes encoding for the Hsp70 protein. These observations suggest that the larvae underwent an oxidative stress caused by the modulation of free radicals' yield induced by the magnetic field through a radical pair mechanism.