Non-lethal Concentrations Research Articles

Acute copper accumulation in Mediterranean mussel and sea snail

In this study, the rate of copper accumulation in mussel (Mytilus galloprovincialis) and sea snail (Rapana venosa) organisms exposed to acutely different non-lethal copper concentrations was investigated for 5 days under laboratory conditions. All soft tissue concentrations were analyzed using an Inductively Coupled Plasma Optical Emission Spectrophotometer. At the end of accumulation, the rate of increase of copper level in mussel and snail soft tissues in different aquariums was found to be 19.19–36.93 and 2.37–7.41 times higher for mussel and snail soft tissues than before exposure, respectively. Differences in metal accumulation at the end of the experiment between species were not statistically significant (p > 0.05). In addition, bio-concentration factor and rates of accumulation values were calculated for both organisms. The corresponding bio-concentration factor values for mussels after exposure to 0.12, 0.36, and 0.60 mg L−1 copper were 406.00, 168.41, and 165.16, respectively, while the corresponding bio-concentration factor values for snails after exposure to 0.12, 0.36, and 0.60 mg L−1 copper were 276.29, 272.12, and 265.73, respectively. While accumulation rate values were 10.15, 12.26, and 20.12 for mussels exposed to 0.12, 0.36, and 0.60 mg L−1 copper, respectively, accumulation rate values were 6.85, 19.76, and 31.69 for snails exposed to 0.12, 0.36, and 0.60 mg L−1 copper, respectively.

Dietary supplementation with curcumin nanomicelles, curcumin, and turmeric affects growth performance and silver nanoparticle toxicity in Cyprinus carpio.

An 8-week feeding experiment was conducted to investigate the effects of curcumin nanomicelle, curcumin, and turmeric (Curcuma longa) on growth performances, body composition, fatty acid profile, and biochemical parameters of common carp (Cyprinus carpio), and their ameliorative effects against toxicity of silver nanoparticles (AgNPs). A total of 120 healthy carps were randomly distributed into four equal treatments. Curcumin nanomicelle, curcumin, and turmeric were each added separately to the basal diet. After the feeding trials, all treatments were exposed to a non-lethal concentration of AgNPs (0.5 mg L-1) for 96 h. Fish fed dietary turmeric showed a significantly higher weight gain. The body protein content was significantly increased in all feeding groups, while the lipid content showed a significant decrease in the turmeric-treated group. Dietary turmeric improved the concentration of saturated fatty acids (SFA) and monounsaturated fatty acid (MUFA). AgNP exposure led to increases in liver catalase (CAT) activity of carps fed with turmeric and curcumin. The lowest amount of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was obtained in fish fed with nanomicelle curcumin and curcumin diets. The lowest amount of silver accumulation in the liver of carps was found in fish fed with dietary curcumin nanomicelle. This experiment suggests that supplementation of turmeric (50 g kg-1) or curcumin (1000 mg kg-1) may play an important role in enhancing growth performances and fatty acid composition of the common carp. Moreover, administration of curcumin nanomicelle in the diet may have a potential ameliorative effect against toxicity of AgNPs.

Transcriptional response of immune-related genes in Litopenaeus vannamei cultured in recirculating aquaculture systems with elevated CO2

This short-term study evaluated the effect of non-lethal high CO2 concentration on the transcriptional response of immune-related genes of Pacific white shrimp (Litopenaeus vannamei) cultured in recirculating aquaculture systems (RAS). Two experimental groups were created: high CO2 (47.67±2.04 mg L−1) and low CO2 (2.0±1.93 mg L−1). Shrimp of 8.85±1.20 g were placed randomly at a density equivalent to 100 individuals m−3 and were monitored at 6, 12, 18, and 24 h. The transcriptional response of immune-related genes was analyzed [...]

Sodium channel activation underlies transfluthrin repellency in Aedes aegypti

BackgroundVolatile pyrethroid insecticides, such as transfluthrin, have received increasing attention for their potent repellent activities in recent years for controlling human disease vectors. It has been long understood that pyrethroids kill insects by promoting activation and inhibiting inactivation of voltage-gated sodium channels. However, the mechanism of pyrethroid repellency remains poorly understood and controversial.Methodology/Principal findingsHere, we show that transfluthrin repels Aedes aegypti in a hand-in-cage assay at nonlethal concentrations as low as 1 ppm. Contrary to a previous report, transfluthrin does not elicit any electroantennogram (EAG) responses, indicating that it does not activate olfactory receptor neurons (ORNs). The 1S-cis isomer of transfluthrin, which does not activate sodium channels, does not elicit repellency. Mutations in the sodium channel gene that reduce the potency of transfluthrin on sodium channels decrease transfluthrin repellency but do not affect repellency by DEET. Furthermore, transfluthrin enhances DEET repellency.Conclusions/SignificanceThese results provide a surprising example that sodium channel activation alone is sufficient to potently repel mosquitoes. Our findings of sodium channel activation as the principal mechanism of transfluthrin repellency and potentiation of DEET repellency have broad implications in future development of a new generation of dual-target repellent formulations to more effectively repel a variety of human disease vectors.

In vitro anthelmintic efficacy of Citrullus colocynthis (L.) Schrad on Haemonchus contortus

Ethno-veterinary medicinal studies associated with traditional uses of the flora of the Cholistan desert have shown that fruits of Citrullus colocynthis are used for the treatment of helminth infections. The present research was designed to evaluate the anthelmintic efficacy of C. colocynthis against H. contortus. The in vitro anthelmintic effects of aqueous-methanol and ethyl acetate fruit extracts of C. colocynthis against H. contortus were determined through egg hatch and adult motility assays. The effect of four serial dilutions of 25 mg/mL of each extract compared to levamisol (0.55 mg/mL) and oxfendazole (three serial dilutions of 25 µg/mL) were studied. Both ethyl acetate and aqueous-methanol extracts paralyzed all adult worms 4h and 8h post-exposure at a dose of 25 mg/mL each. In the egg hatch assay, about 83.67% and 80.67% of H. contortus eggs failed to hatch with the same dose (i.e. 25 mg/mL) of ethyl acetate and CAME extracts, respectively. The results of the present study strongly support fruit extracts of C. colocynthis as a promising alternative to synthetic drugs against H. contortus. These findings will lead to further in vivo studies to investigate the bio-availability of the active ingredients of the plant and the minimum non-lethal concentration required for treatment of haemonchosis in livestock. The anthelmintic effects of C. colocynthis might be attributed to the presence of phenolic acids.

Pre-exposure to Cu2+ and CuO NPs leads to infection of caged blue mussels, Mytilus edulis L., by pathogenic microalga: Pilot study in the Lower St. Lawrence Estuary (Québec, Canada)

As evidenced from literature, exposure to non-lethal concentrations of dissolved copper (Cu2+) and copper nanoparticles (CuO NPs) promotes blue mussels susceptibility to various bacterial infections. We study whether pre-exposure (3.5 h) with CuSO4 (100 μg Cu L−1) and CuO NPs (1000 μg Cu L−1) will result in infection of M. edulis L. with pathogenic microalga Coccomyxa sp. under field conditions. In May – September 2019, cages were installed in the site Metis-sur-Mer, St. Lawrence Estuary (QC, Canada) where the native mussel population is known to be infected with the pathogen. Untreated and pre-exposed mussels were grown for up to 130 days. Only the mussels pre-exposed to copper were infected by Coccomyxa. This finding allows proposing that occurrences of Coccomyxa-infected mussels worldwide might have an association with water pollution with xenobiotics. Pre-exposure of caged mussels to copper, as a protocol monitoring for other infectious agents, can be recommended to test.

Changes in toxin production of environmental Pseudomonas aeruginosa isolates exposed to sub-inhibitory concentrations of three common antibiotics.

Pseudomonas aeruginosa is an environmental pathogen that can cause severe infections in immunocompromised patients. P. aeruginosa infections are typically treated with multiple antibiotics including tobramycin, ciprofloxacin, and meropenem. However, antibiotics do not always entirely clear the bacteria from the infection site, where they may remain virulent. This is because the effective antibiotic concentration and diffusion in vitro may differ from the in vivo environment in patients. Therefore, it is important to understand the effect of non-lethal sub-inhibitory antibiotic concentrations on bacterial phenotype. Here, we investigate if sub-inhibitory antimicrobial concentrations cause alterations in bacterial virulence factor production using pyocyanin as a model toxin. We tested this using the aforementioned antibiotics on 10 environmental P. aeruginosa strains. Using on-the-spot electrochemical screening, we were able to directly quantify changes in production of pyocyanin in a measurement time of 17 seconds. Upon selecting 3 representative strains to further test the effects of sub-minimum inhibitory concentration (MICs), we found that pyocyanin production changed significantly when the bacteria were exposed to 10-fold MIC of the 3 antibiotics tested, and this was strain specific. A series of biologically relevant measured pyocyanin concentrations were also used to assess the effects of increased virulence on a culture of epithelial cells. We found a decreased viability of the epithelial cells when incubated with biologically relevant pyocyanin concentrations. This suggests that the antibiotic-induced virulence also is a value worth being enclosed in regular testing of pathogens.

Can Organophosphates and Carbamates Cause Synergisms by Inhibiting Esterases Responsible for Biotransformation of Pyrethroids?

Hydrolysis catalyzed by general esterases (GEs) is the most efficient route for hydrolyzation of pyrethroid insecticides. Organophosphate (OP) and carbamate (CB) insecticides are known to inhibit GEs in addition to acetylcholinesterase (AChE), which is their main target. We hypothesize that synergies can be induced by OPs and CBs when mixed with pyrethroids, due to their inhibition of GE-dependent detoxification of pyrethroids. To test this hypothesis, we conducted mixture toxicity experiments with Daphnia magna using α-cypermethrin (α-cyp) in combination with the noninsecticidal OP tetraisopropyl pyrophosphoramide (iso-OMPA) and five AChE inhibitors diazinon, chlorpyrifos, chlorfenviphos, parathion, and aldicarb. In addition, the in vivo GE activity inhibition was measured for all compounds. Up to 10-fold synergy was found between α-cyp and iso-OMPA, and the degree of synergy correlated linearly with the inhibition of the GE activity. No synergy, however, was found in any of the insecticide mixtures nor was the GE activity inhibited within the nonlethal concentration range tested. It was concluded that the effect of the insecticides on AChE occurred at lower concentrations than their effect on GEs, making the daphnids become immobilized before any synergistic effects on mortality could be observed. The implications of the findings are discussed from a risk assessment perspective.

Physiological, Developmental, and Biomarker Responses of Zebrafish Embryos to Sub-Lethal Exposure of Bendiocarb

Bendiocarb is a broad-spectrum insecticide recommended for malaria control by the World Health Organization (WHO). Still, bendiocarb poses a toxic risk to populations of nontargeted aquatic organisms. Thus, our study was aimed to evaluate the sub-lethal effects of bendiocarb exposure on zebrafish (Danio rerio) embryos by assessing of physiological, developmental, and biochemical parameters. Bendiocarb-induced adverse effects on embryonic development, larval growth, heart rate, changes in phase II detoxifying enzyme glutathione-S-transferase (GST) activity, oxidative stress-related enzyme activities (superoxide dismutase (SOD), catalase (CAT)), and the damage-linked biomarker lipid peroxidation (LPO) in early life stage zebrafish were investigated. Our results highlight that the selected nonlethal concentrations (96 h median lethal concentration in this study was 32.52 mg/L−1) of bendiocarb inflicted adverse effects resulting in embryo deformities (96 h EC50 = 2.30 mg L−1), reduced body- and notochord length (above 0.75 and 0.39 mg L−1 bendiocarb concentrations at 96 hpf, respectively), oxidative stress, and altered heart rate (above 0.4 mg L−1 at 48 hpf) in the studied model system.

Feeding behavior toxicity in the marine rotifer Brachionus plicatilis caused by 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47): Characteristics and mechanisms

Polybrominated diphenyl ether contamination in marine environments has received special attention due to its accumulation and magnification in the marine food web and toxicity to organisms. In the present study, a series of short-term toxicological tests were conducted with the marine rotifer Brachionus plicatilis to assess the effects on ingestion and digestive performance after treatment with 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) at nonlethal concentrations under controlled laboratory conditions and to analyze the possible mechanism. The results showed that with accumulation in rotifers, BDE-47 caused a significant decline in the filtration and feeding rates in a concentration-dependent manner. Moreover, the activities of amylase (AMS) and protease were affected, indicating that BDE-47 impaired ingestion and digestion efficiency. BDE-47 exposure did not lead to abnormal microstructures in the main digestive tract (e.g., cilia around the corona, mastax, stomach, digestive gland and esophagus), but the gastric parietal cells shrank, suggesting nutritional deficiency. BDE-47 prominently induced the occurrence of irregular mitochondria at the cilia root, and mitochondrial and isocitrate dehydrogenase activity declined, indicating mitochondrial dysfunction. Furthermore, the activity of ATPase, which catalyzes ATP hydrolysis, decreased as the BDE-47 concentration rose, implying that BDE-47 retarded rotifer ATP dynamics, inevitably interfering with cilia movement to ingest food. Additionally, a significant decline in acetylcholine esterase activity was observed, which led to a hindrance in neurotransmission involved in food intake and digestion. Altogether, our results demonstrated that nonlethal concentrations of BDE-47 could induce feeding depression in rotifers, which is mainly attributed to stymied energy metabolism and nerve conduction.

One-Two Punch Therapy for the Treatment of T-Cell Malignancies Involving p53-Dependent Cellular Senescence.

T-cell malignancies are still difficult to treat due to a paucity of plans that target critical dependencies. Drug-induced cellular senescence provides a permanent cell cycle arrest during tumorigenesis and cancer development, particularly when combined with senolytics to promote apoptosis of senescent cells, which is an innovation for cancer therapy. Here, our research found that wogonin, a well-known natural flavonoid compound, not only had a potential to inhibit cell growth and proliferation but also induced cellular senescence in T-cell malignancies with nonlethal concentration. Transcription activity of senescence-suppression human telomerase reverse transcriptase (hTERT) and oncogenic C-MYC was suppressed in wogonin-induced senescent cells, resulting in the inhibition of telomerase activity. We also substantiated the occurrence of DNA damage during the wogonin-induced aging process. Results showed that wogonin increased the activity of senescence-associated β-galactosidase (SA-β-Gal) and activated the DNA damage response pathway mediated by p53. In addition, we found the upregulated expression of BCL-2 in senescent T-cell malignancies because of the antiapoptotic properties of senescent cells. Following up this result, we identified a BCL-2 inhibitor Navitoclax (ABT-263), which was highly effective in decreasing cell viability and inducing apoptotic cell death in wogonin-induced senescent cells. Thus, the “one-two punch” approach increased the sensibility of T-cell malignancies with low expression of BCL-2 to Navitoclax. In conclusion, our research revealed that wogonin possesses potential antitumor effects based on senescence induction, offering a better insight into the development of novel therapeutic methods for T-cell malignancies.

Hearing Toxicity Induced by Tripterygium Glycosides in Zebrafish

Tripterygium glycosides (TG) is isolated from an extensively used traditional Chinese medicine herb tripterygium roots and has been extensively used in the treatment of rheumatoid arthritis, nephrotic syndrome, hyperthyroidism and other diseases due to its anti-inflammatory and immunosuppressive effects. Hearing toxicity has been recently associated with TG use in human patients. In this study, authors assessed hearing toxicity and possible molecular toxic mechanisms of TG in a whole animal model. The maximum non-lethal concentration (MNLC) of TG on the zebrafish was 21 mg/L. TG induced zebrafish hair cell loss in a dose-dependent manner (p<0.001), and the saccular otolith size reduction when treated at MNLC (p<0.01). TG treatment resulted in sound-stimulated zebrafish movement reduction (p<0.001); and the rollover zebrafish percentages were elevated as TG treatment concentrations moved up. Following TG treatment, mRNA levels of the zebrafish hearing organ development genes eya1 and val were remarkably downregulated, and the expression of apoptosis-associated genes bax and caspase3 was significantly enhanced (p<0.05). These findings confirm the hearing toxicity of TG and suggest its toxic mechanisms probably are through suppressing hearing cell development and promoting hearing cell apoptosis. Authors recommend zebrafish assay as a quick and reliable screening test of hearing toxicity for drugs and health products.

Anticancer active trifluoromethylated fused triazinones are safe for early-life stages of zebrafish (Danio rerio) and reveal a proapoptotic action

The main purpose of this investigation was to evaluate the effect of anticancer active compounds (I–VIII) on zebrafish development in order to select the safest molecules. Larval mortality, embryo hatchability and malformations were end-points used to assess the acute toxicity among embryos and larvae from compounds-/pemetrexed-treated and control groups. LC50 and MNLC (maximal non-lethal concentration) were determined. Lipophilicity-dependent structure–toxicity relationships were established. The results clearly indicated that the majority of test molecules are safe for zebrafish individuals and simultaneously are less toxic than an anticancer agent – pemetrexed. The subsequent aim of this study was to elucidate the molecular mechanism of antiproliferative activity of the most selective compounds. Substantially increased activation of caspase-6 and -8 in cancerous cell lines confirmed the proapoptotic action of molecules examined. Considering the safety for zebrafish individuals, the title compounds as inducers of apoptosis are promising drug candidates in the preclinical phase of drug development.

Cell wall distraction and biofilm inhibition of marine Streptomyces derived angucycline in methicillin resistant Staphylococcus aureus

The emergence of life threatening antibiotic resistant pathogens and its associated mortality and morbidity necessitates many new antibiotics from diverse ecological habitats. Marine sponge associated microbes are promising to provide such antimicrobial compounds. In the present study, we report antibacterial and anti-biofilm potential of the angucycline antibiotic 8-O-metyltetrangomycin from Streptomyces sp. SBRK2 isolated from a marine sponge of Gulf of Mannar, Rameswaram, India. Our screening program to tackle methicillin-resistant Staphylococcus aureus (MRSA) drug resistance from marine sponge associated actinobacteria yielded the bioactive strain SBRK2. Based on 16S rRNA gene phylogenetic analysis the isolate was found to closely related with Streptomyces longispororuber NBRC 13488T. In vitro production by agar plate fermentation, solvent based extraction, TLC, HPLC purification and LC–MS based de-replication revealed the bioactive compound as 8-O-metyltetrangomycin. The antibacterial minimum inhibitory concentrations against MRSA was identified as 2 μg/mL. Sub-inhibitory concentration of the compound 8-O-metyltetrangomycin reduced the biofilm formation of S. aureus ATCC25923 and increased the cell surface hydrophobicity index. Scanning electron microscopic observation of the sub-inhibitory concentration exposure revealed a wrinkled membrane surface and slight cellular damage shows the cell wall distracting property of the compound. Zebrafish embryo based toxicity assays exhibited 100 μg/mL of compound as maximal non-lethal concentration which had demonstrated the positive relationship in safety index. The angucycline compound 8-O-metyltetrangomycin could be a potential candidate for the development of anti-biofilm agents against drug resistant pathogens.

Zebrafish Embryonic Exposure to BPAP and Its Relatively Weak Thyroid Hormone-Disrupting Effects.

Safe endocrine-disrupting alternatives for bisphenol A (BPA) are needed because its adverse health effects have become a public concern. Some bisphenol analogues (bisphenol F and S) have been applied, but their endocrine-disrupting potential is either not negligible or weaker than that of BPA. However, the endocrine-disrupting potential of bisphenol AP (BPAP), another BPA alternative, has not yet been fully assessed. Hence, we evaluated the thyroid hormone (TH)-disrupting potency of BPAP because THs are essential endocrine hormones. Zebrafish embryos were exposed to BPAP (0, 18.2, 43.4, or 105.9 μg/L) for 120 h, and TH levels, the transcription of 16 TH-related genes, the transcriptome, development, and behavior were evaluated. In our study, a decrease in T4 level was observed only at the maximum nonlethal concentration, but significant changes in the T3 and TSHβ levels were not detected. BPAP did not cause significant changes in transcription and gene ontology enrichment related to the TH system. Developmental and behavioral changes were not observed. Despite T4 level reduction, other markers were not significantly affected by BPAP. These might indicate that BPAP has weak or negligible potency regarding TH disruption as a BPA alternative. This study might provide novel information on the TH-disrupting potential of BPAP.

Antibiotic tetracycline enhanced the toxic potential of photo catalytically active P25 titanium dioxide nanoparticles towards freshwater algae Scenedesmus obliquus

Titanium dioxide nanoparticles (TiO2 NPs) often co-exist with the other co-contaminants like antibiotics. The antibiotics can potentially modify the toxic effects of the co-contaminants like the NPs in the environment. Hence, the present study aims to understand the toxic potential of a binary mixture of tetracycline (TC) and TiO2 NPs to a model freshwater alga – Scenedesmus obliquus. Since, TiO2 NPs are known to be photo-catalytically active, non-irradiated (NI-TiO2 NPs), UVA pre-irradiated (UVA-TiO2 NPs), and UVB pre-irradiated (UVB-TiO2 NPs) TiO2 NPs was mixed separately with TC and their toxicity evaluated. It was observed that the cell viability for the three experimental groups decreased significantly (p < 0.001) with respect to the individual NPs-treated algae. Abbott’s model suggested that the interaction between TC and Ni-TiO2 NPs was additive for all the concentrations of NI-TiO2 NPs tested. However, in the case of both the UV pre-irradiated NPs, the interaction was additive for the lower concentration (1.56 μM) and synergistic for both the higher concentrations (3.13, and 6.26 μM). At the concentrations tested the cell membrane damage and intracellular uptake of NPs increased significantly (p < 0.05) for the mixture in comparison with the individual NPs treated algae. This study suggested that even a non-lethal concentration of TC (EC10 = 0.135 μM) increased the toxic potential of the TiO2 NPs significantly and when this antibiotic was used in combination with the UV pre-irradiated NPs, toxicity even increased to a higher level.

The Latest Time Point of Retreatment (LTPR) as a Novel Method to Determine Antibacterial Effects for Binary Use of Cold Atmospheric Plasma and Conventional Agents.

Multi-resistant microorganisms are a long-standing problem for public healthcare, as inactivating those resistant pathogens with conventional antibiotics or antiseptics often no longer achieves the expected clinical success. The aim of this in vitro study was to investigate the antibacterial efficacy of binary combinations of conventional antibacterial agents with cold atmospheric plasma (CAP), when both are applied in non-lethal concentrations. In this study, Enterococcus faecalis biofilms were treated with CAP in binary combinations with benzalkonium chloride (BAC), chlorhexidine (CHX), or ciprofloxacin (CIP), respectively, which were applied in different sequences. In order to evaluate effects of binary use of two different antibacterial approaches, the so-called latest time point of retreatment (LTPR) was defined. For this purpose, regrowth curves of the bacteria were measured following the respective treatment combinations. LTPR is defined as the time component of the inflection point of a normalized regrowth curve and allows the rating and interpretation of single or binary treatments with different agents or approaches. Furthermore, LTPR designates the latest time point where a retreatment appears to be appropriate for preventing regrowth of the bacteria in case the first treatment was not lethal. Here in our study, the binary combination of 10 min CAP with BAC, CHX, or CIP leads to higher LTPRs as compared to single treatments for both sequences of application. Overall, the combination of two antimicrobial approaches is an effective alternative for inactivating bacteria in biofilms instead of a single treatment. Thus, LTPR provides a novel promising way to determine antibacterial effects for single or binary use of given antimicrobial approaches.

Effects of chlorantraniliprole insecticide on innate immune response of silver catfish (Rhamdia quelen) naturally infected with Aeromonas hydrophila

It is well documented today that pesticides, used in crop production, may modulate the immune system of healthy fish. However, there is still only limited information regarding the effects of these anthropogenic stressors in conjunction with natural stressors (pathogens), on the innate immune responses of freshwater fish. Thus, the aim of this investigation was to compare the innate immune response of two groups of fish (Rhamdia quelen), naturally infected with Aeromonas hydrophila, exposed and unexposed to a non-lethal concentration of chlorantraniliprole (CAP) insecticide (0.0 and 1.3 μg/L/24 h). Unhealthy fish exposed to CAP showed significant higher total leukocyte counts and neutrophils percentage compared to non-exposed infected fish). However, the monocytes and eosinophils percentage significantly decreased in fish exposed to CAP. Furthermore, lysozyme activity values measured in plasma, skin mucus, gill and intestine significantly reduced in fish exposed to CAP. The CAP-induced immunomodulation may interfere on the ability of the animal to heal or fight the infection, and possible contribute to the spread of bacterial infection in fish production or environment.

Phenolic Profile of Herbal Infusion and Polyphenol-Rich Extract from Leaves of the Medicinal Plant Antirhea borbonica: Toxicity Assay Determination in Zebrafish Embryos and Larvae.

Antirhea borbonica (A. borbonica) is an endemic plant from the Mascarene archipelago in the Indian Ocean commonly used in traditional medicine for its health benefits. This study aims (1) at exploring polyphenols profiles from two types of extracts—aqueous (herbal infusion) and acetonic (polyphenol rich) extracts from A. borbonica leaves—and (2) at evaluating their potential toxicity in vivo for the first time. We first demonstrated that, whatever type of extraction is used, both extracts displayed significant antioxidant properties and acid phenolic and flavonoid contents. By using selective liquid chromatography–tandem mass spectrometry, we performed polyphenol identification and quantification. Among the 19 identified polyphenols, we reported that the main ones were caffeic acid derivatives and quercetin-3-O-rutinoside. Then, we performed a Fish Embryo Acute Toxicity test to assess the toxicity of both extracts following the Organisation for Economic Cooperation and Development (OECD) guidelines. In both zebrafish embryos and larvae, the polyphenols-rich extract obtained by acetonic extraction followed by evaporation and resuspension in water exhibits a higher toxic effect with a median lethal concentration (LC50: 5.6 g/L) compared to the aqueous extract (LC50: 20.3 g/L). Our data also reveal that at non-lethal concentrations of 2.3 and 7.2 g/L for the polyphenol-rich extract and herbal infusion, respectively, morphological malformations such as spinal curvature, pericardial edema, and developmental delay may occur. In conclusion, our study strongly suggests that the evaluation of the toxicity of medicinal plants should be systematically carried out and considered when studying therapeutic effects on living organisms.

Influence of Non-lethal Doses of Natural Insecticides Spinetoram and Azadirachtin on Helicoverpa punctigera (Native Budworm, Lepidoptera: Noctuidae) Under Laboratory Conditions.

Helicoverpa punctigera (native budworm) is an important pest species in crops across Australia. From the third instar onward, this species causes severe damage to crop plants: therefore, caterpillars need to be managed at an early stage of their development. In our experiment, we raised H. punctigera on an artificial diet, which included different concentrations of the natural insecticides Spinetoram and Azadirachtin. The survival of the larvae, growth and body mass gain was recorded over 17 days. Only caterpillars raised on lowest toxin concentrations survived and molted successfully to the fifth instar, but had slower growth and body mass gain compared to the insecticide-free control group. Caterpillars fed on higher toxin concentrations never molted to the next instar or died in the first few days. To test how the toxins influence physiological conditions including metabolic rate and water loss, surviving fifth instar larvae were exposed to thermolimit respirometry: starting at 25°C following a constant increasing temperature ramping rate of 0.25°C–1, until reaching the critical thermal maxima (CTmax). Caterpillars raised on a non-lethal dose of insecticides had higher metabolic rates and lost more water compared to the control group. Insects that have seem to consume more energy per mg tissue and have a higher water loss at high temperatures. Non-lethal concentrations of insecticides on pest insects physiology may reduce their impact on crops and may enable more targetted insecticide application.