Pest management facing warming and chemical stresses: Multi-stress effects on the biological agent Trichogramma oleae.

Global change is affecting plant-insect interactions in agroecosystems and can have dramatic consequences on yields when causing non-targeted pest outbreaks and threatening the use of pest natural enemies for biocontrol. The vineyard agroecosystem is an interesting system to study multi-stress conditions: on the one hand, agricultural intensification comes with high inputs of copper-based fungicides and, on the other hand, temperatures are rising due to climate change. We investigated interactive and bottom-up effects of both temperature increase and copper-based fungicides exposure on the important Lepidopteran vineyard pest Lobesia botrana and its natural enemy, the oophagous parasitoid Trichogramma oleae. We exposed L. botrana larvae to three increasing copper sulfate concentrations under two fluctuating thermal regimes, one current and one future. Eggs produced by L. botrana were then exposed to T. oleae. Our results showed that the survival of L. botrana, was only reduced by the highest copper sulfate concentration and improved under the warmer regime. The development time of L. botrana was strongly reduced by the warmer regime but increased with increasing copper sulfate concentrations, whereas pupal mass was reduced by both thermal regime and copper sulfate. T. oleae F1 emergence rate was reduced and their development time increased by combined effects of the warmer regime and increasing copper sulfate concentrations. Size, longevity and fecundity of T. oleae F1 decreased with high copper sulfate concentrations. These effects on the moth pest and its natural enemy are probably the result of trade-offs between the survival and the development of L. botrana facing multi-stress conditions and implicate potential consequences for future biological pest control. Our study supplies valuable data on how the interaction between pests and biological control agents is affected by multi-stress conditions.

The dark side of organic farming: Copper sulphate compromises the life history and behaviour of the walking stick insect, Bacillus rossius.

Organic farming is considered the most sustainable form of modern soil cultivation. Yet it often relies on the use of chemical compounds that are not necessarily harmless for the surrounding wildlife. In this study, we tested the effects of realistic concentrations of copper sulphate-largely used in organic farming as a fungicide-on ecologically-relevant traits of the walking stick insect Bacillus rossius, a species commonly found in the proximity of cultivated fields across Europe. By using second-generation progeny of wild-caught parthenogenetic females bred in common gardens, we measured the impact of copper sulphate (CuSO4) on both the life-history (body condition, number of eggs, and hatching success) and behavioural traits (activity and maximum vertical speed) of the individuals. We observed strong negative effects of high, realistic concentrations of copper sulphate on most traits within 12 days of exposure, while effects were less evident at lower concentrations of the pollutant. Our results reveal that realistic concentrations of copper sulphate can compromise important traits that regulate both the survival and reproduction of animals in the wild, with such effects that are, however, dose dependent. We suggest that common practices in organic farming require further consideration on their ecological and evolutionary impact on wildlife.

Copper exposure induces mitochondrial dysfunction and hepatotoxicity via the induction of oxidative stress and PERK/ATF4 -mediated endoplasmic reticulum stress.

Copper is an essential trace element, and excessive exposure could result in hepatoxicity, however, the underlying molecular mechanisms remain incompletely understood. The present study is aimed to investigate the molecular mechanisms of copper sulfate (CuSO4) exposure-induced hepatoxicity both in vivo and in vitro. In vitro, HepG2 and L02 cells were exposed to various doses of CuSO4 for 24 h. Cell viability, ROS production, oxidative stress biomarkers, mitochondrial functions, ultrastructure, intracellular calcium (Ca2+) concentration, and the expression of proteins related to mitochondrial apoptosis and endoplasmic reticulum (ER) stress were assessed. In vivo, C57BL/6 mice were treated with CuSO4 at doses of 10 and 30 mg/kg BW/day and co-treated with 4-PBA at 100 mg/kg BW/day for 35 days. Subsequently, liver function, histopathological features, and protein expression were evaluated. Results found that exposure to CuSO4 at concentrations of 100-400 µM for 24 h significantly decreased the viabilities of HepG2 and L02 cells and it was in a dose-dependent manner. Additionally, CuSO4 exposure induced significant oxidative stress and mitochondrial dysfunction in HepG2 cells, which were partially ameliorated by the antioxidant N-acetylcysteine (NAC). Furthermore, CuSO4 exposure prominently triggered ER stress, as evidenced by the upregulation of GRP94, GRP78, phosphorylated forms of PERK and eIF2α, and CHOP proteins in livers of mice and HepG2 cells. NAC treatment significantly inhibited CuSO4 exposure -induced ER stress in HepG2 cells. Pharmacological inhibition of ER stress through co-treatment with 4-PBA and the PERK inhibitor GSK2606414, as well as genetic knockdown of ATF4, partially mitigated CuSO4-induced cytotoxicity in HepG2 cells by reducing mitochondrial dysfunction and inhibiting the mitochondrial apoptotic pathway. Moreover, 4-PBA treatment significantly attenuated CuSO4-induced caspase activation and hepatoxicity in mice. In conclusion, these results reveal that CuSO4-induced hepatotoxicity involves mitochondrial dysfunction and ER stress by activating oxidative stress induction and PERK/ATF4 pathway.

Impact of copper and zinc oral chronic exposure on Carniolan honey bee survival and feeding preference.

Honey bees are important plant pollinators and honey producers. Contamination of the environment with metals can lead to a decline in honey bee populations. Copper (Cu) and zinc (Zn) salts are commonly used as fungicides and foliar fertilizers. In this study, we investigated the effects of 10-day chronic oral exposure to different concentrations of Cu (CuSO4) and Zn (ZnCl2) on survival and feeding rates of Carniolan honey bees in laboratory conditions. We found that mortality in honey bee workers increased in a concentration-dependent manner and that Cu (lethal concentration [LC50] = 66 mg/l) was more toxic than Zn (LC50 = 144 mg/l). There was no difference in the feeding rate of Cu-treated bees for the different concentrations tested, but the feeding rate decreased with the increase in Zn concentration. To determine feeding preference or avoidance for Cu and Zn, we conducted 2-choice 24-h feeding experiments. We demonstrated that honey bees preferred Zn-containing solutions compared to the control diet. A two-choice experiment with Cu showed a tendency for honey bees to be deterred by Cu at high concentrations; however, it was not statistically significant. In summary, our results suggest that honey bee workers may suffer adverse effects when exposed to ecologically relevant concentrations of Cu and Zn.

Interplay between diphenyl diselenide and copper: Impact on D. melanogaster survival, behavior, and biochemical parameters.

Copper (Cu2+) is a biologically essential element that participates in numerous physiological processes. However, elevated concentrations of copper have been associated with cellular oxidative stress and neurodegenerative diseases. Organo­selenium compounds such as diphenyl diselenide (DPDS) have in vitro and in vivo antioxidant properties. Hence, we hypothesized that DPDS may modulate the toxicity of Cu2+ in Drosophila melanogaster. The acute effects (4 days of exposure) caused by a high concentration of Cu2+ (3 mM) were studied using endpoints of toxicity such as survival and behavior in D. melanogaster. The potential protective effect of low concentration of DPDS (20 µM) against Cu2+ was also investigated. Adult flies aged 1-5 days post-eclosion (both sexes) were divided into four groups: Control, DPDS (20 µM), CuSO4 (3 mM), and the combined exposure of DPDS (20 µM) and CuSO4 (3 mM). Survival, biochemical, and behavioral parameters were determined. Co-exposure of DPDS and CuSO4 increased acetylcholinesterase (AChE) activity and the generation of reactive oxygen species (ROS as determined by DFCH oxidation). Contrary to our expectation, the co-exposure reduced survival, body weight, locomotion, catalase activity, and cell viability in relation to control group. Taken together, DPDS potentiated the Cu2+ toxicity.

Cuproptosis mediates copper-induced testicular spermatogenic cell death.

Cuproptosis, a novel mechanism of programmed cell death, has not been fully explored in the context of spermatogenic cells. This study investigated the potential involvement of cuproptosis in spermatogenic cell death using a mouse model of copper overload. Sixty male Institute of Cancer Research (ICR) mice were randomly divided into four groups that received daily oral gavage with sodium chloride (control) or copper sulfate (CuSO 4 ) at 50 mg kg -1 , 100 mg kg -1 , or 200 mg kg -1 , for 42 consecutive days. Mice subjected to copper overload exhibited a disruption in copper homeostasis. Additionally, significant upregulated expression of key cuproptosis factors was accompanied by a significant rise in the rates of testicular tissue cell apoptosis. Immunohistochemical analysis revealed the presence of ferredoxin 1 (Fdx1) in Sertoli cells, Leydig cells, and spermatogenic cells at various stages of testicular development, and the Fdx1-positive staining area was significantly increased in copper-overloaded mice. Mitochondrial dysfunction and decreased adenosine triphosphate levels were also observed, further implicating mitochondrial damage under cuproptosis. Further analyses revealed pathological lesions and blood-testis barrier destruction in the testicular tissue, accompanied by decreased sperm concentration and motility, in copper-overloaded mice. In summary, our results indicate that copper-overloaded mice exhibit copper homeostasis disorder in the testicular tissue and that cuproptosis participates in spermatogenic cell death. These findings provide novel insights into the pathogenic mechanisms underlying spermatogenic cell death and provide initial experimental evidence for the occurrence of cuproptosis in the testis.

Reduced toxic effects of nano­copper sulfate in comparison of bulk CuSO4 on biochemical parameters in the Rohu (Labeo rohita).

Considering the wide application of nanoparticles in various fields of life and growing concern regarding their toxic effects, the present study was designed with the aim to evaluate the potential risks of using copper sulfate nanoparticles (CuSO4-NPs) in comparison to bulk form. Nanoparticles of CuSO4, having mean size of 73 nm were prepared by ball milling method, and fingerlings of Labeo rohita were exposed to two levels, 20 and 100 µg L-1 of CuSO4 in both bulk and nano forms for 28 days and their comparative effects on the metallothioneins (MTs), heat shock proteins 70 (HSP 70), lipid profile, cholesterol (CHOL) and triglyceraldehyde (TG) levels, activities of some metabolic enzymes Alanine transaminase (ALT), Aspartate transaminase (AST) Akaline phosphatase (ALP), and genes expressions of HSP-70, TNF-α and IL1-ß were investigated. CuSO4 showed the concentration and particle type dependent effects. The over expression of HSPs and MTs, significant decreases in CHOL, TG, low density lipid (LDL) levels and ALP activity, while significant increases in high density lipid (HDL)level as well as ALT and AST activities and HSP-70, TNF-α and IL1-ß expressions were observed in response to higher concentration of both bulk and nano form of copper sulfate. At lower concentration (20 µg L-1), however, only bulk form showed toxicity. Thus, low concentrations of CuSO4-NPs pose negligible threat to freshwater fish.

Comparison of the accumulation and effects of copper pyrithione and copper sulphate on rainbow trout larvae.

Copper pyrithione (CuPT) is used as a co-biocide in new antifouling paints but its toxicity remains little known. To compare the toxicity of copper-based compounds, rainbow trout (Oncorhynchus mykiss) larvae were exposed for 8-day to CuPT and CuSO4 at equivalent copper concentrations. CuPT exposure led to the greatest accumulation of Cu in larvae. Exposure to 10 µg.L-1 CuPT induced 99% larval mortality but only 4% for CuSO4-exposed larvae. The larval development and growth were affected by CuPT (from 0.5 µg.L-1 Cu) but not by CuSO4. Lipid peroxidation was not induced by either contaminant. The expression of genes involved in oxidative stress defence, detoxification and copper transport was induced in larvae exposed to CuSO4 and CuPT but at higher concentrations for CuPT. This study highlights the marked toxicity of CuPT for early life stages of fish and raises the question of the possible environmental risks of this antifouling compound.

The Combined Use of Copper Sulfate and Trichlorfon Exerts Stronger Toxicity on the Liver of Zebrafish.

In aquaculture, copper sulphate and trichlorfon are commonly used as disinfectants and insecticide, sometimes in combination. However, improper use can result in biotoxicity and increased ecological risks. The liver plays a crucial role in detoxification, lipid metabolism, nutrient storage, and immune function in fish. Selecting the liver as the main target organ for research helps to gain an in-depth understanding of various aspects of fish physiology, health, and adaptability. In the present study, zebrafish were exposed to Cu (0.5 mg/L) and Tri (0.5 mg/L) alone and in combination for 21 days. The results demonstrate that both Cu and Tri caused hepatocyte structure damage in zebrafish after 21 days of exposure, with the combination showing an even greater toxicity. Additionally, the antioxidant and immune enzyme activities in zebrafish liver were significantly induced on both day 7 and day 21. A transcriptome analysis revealed that Cu and Tri, alone and in combination, impacted various physiological activities differently, including metabolism, growth, and immunity. Overall, Cu and Tri, either individually or in combination, can induce tissue damage by generating oxidative stress in the body, and the longer the exposure duration, the stronger the toxic effects. Moreover, the combined exposure to Cu and Tri exhibits enhanced toxicity. This study provides a theoretical foundation for the combined use of heavy metal disinfectants and other drugs.

Effects of copper sulfate algaecide on the cell growth, physiological characteristics, the metabolic activity of Microcystis aeruginosa and raw water application.

Harmful cyanobacteria blooms (HCBs) occurred frequently and become a serious scientific challenge. Copper sulfate (CuSO4) is a broad-spectrum chemical algaecide to control algae blooms. Herein, the Microcystis aeruginosa was exposed to different CuSO4 (0.0, 0.2 and 0.5 mg/L) to assess the variations in algal physiological process and metabolic profiles. The results indicated that exposure to CuSO4 of 0.5 mg/L at 72 h could significantly inhibit the cell growth and photosynthetic capacity of M. aeruginosa, including chl-a content and chlorophyll fluorescence parameters. Plasma membrane damage causing cell lysis of M. aeruginosa increased the K+ release. The increase of SOD and CAT suggested that CuSO4 treatment caused oxidative stress in algal cells. Different doses of CuSO4 modified the carbon metabolic potential, algal cells had their unique metabolic mode thereby. Moreover, the research further verified that CuSO4 would also inhibit algal growth and change algal community structure in site-collected water application. Overall, laboratory results of M. aeruginosa to CuSO4 and site-collected water application of algal responses to CuSO4 might be conducive to uncovering the controlling mechanism of algae and the potential effect of carbon cycling in an ecological environment.

Differences in toxicity and accumulation of metal from copper oxide nanomaterials compared to copper sulphate in zebrafish embryos: Delayed hatching, the chorion barrier and physiological effects.

The mechanisms of toxicity of engineered nanomaterials (ENMs) to the early life stages of freshwater fish, and the relative hazard compared to dissolved metals, is only partially understood. In the present study, zebrafish embryos were exposed to lethal concentrations of copper sulphate (CuSO4) or copper oxide (CuO) ENMs (primary size ∼15 nm), and then the sub-lethal effects investigated at the LC10 concentrations over 96 h. The 96 h-LC50 (mean ± 95% CI) for CuSO4 was 303 ± 14 µg Cu L-1 compared to 53 ± 9.9 mg L-1 of the whole material for CuO ENMs; with the ENMs being orders of magnitude less toxic than the metal salt. The EC50 for hatching success was 76 ± 11 µg Cu L-1 and 0.34 ± 0.78 mg L-1 for CuSO4 and CuO ENMs respectively. Failure to hatch was associated with bubbles and foam-looking perivitelline fluid (CuSO4), or particulate material smothering the chorion (CuO ENMs). In the sub-lethal exposures, about 42% of the total Cu as CuSO4 was internalised, as measured by Cu accumulation in the de-chorionated embryos, but for the ENMs exposures, nearly all (94%) of the total Cu was associated with chorion; indicating the chorion as an effective barrier to protect the embryo from the ENMs in the short term. Both forms of Cu exposure caused sodium (Na+) and calcium (Ca2+), but not magnesium (Mg2+), depletion from the embryos; and CuSO4 caused some inhibition of the sodium pump (Na+/K+-ATPase) activity. Both forms of Cu exposure caused some loss of total glutathione (tGSH) in the embryos, but without induction of superoxide dismutase (SOD) activity. In conclusion, CuSO4 was much more toxic than CuO ENMs to early life stage zebrafish, but there are subtle differences in the exposure and toxic mechanisms for each substance.

Response of Sabella spallanzanii to multiple stressors. The combined effect of infection and copper sulphate.

The aim of this work is to study the immune responses of the polychaete Sabella spallanzanii after exposure to copper sulphate, an immunomodulating agent in marine organisms, and the multiple stresses caused by Escherichia coli infection, to validate the species as a model organism in marine-coastal biomonitoring programmes. Polychaetes were housed in laboratory and divided into five experimental groups: 1. Control (no microinjected), 2. filtered seawater + TBS injection (control of point 3), 3. filtered seawater + E. coli injection (control of point 4), 4. CuSO4 + TBS injection (control of point 5), and 5. CuSO4 + E. coli injection. The immune variables, esterase and alkaline phosphatase activity, cytotoxicity and detoxifying/antioxidant enzymes such as glutathione peroxidase were evaluated in total body extracts of the animals. Moreover, toll-like receptor, allograft inflammatory factor-1, lysozyme and haemagglutinating activity were investigated to highlight possible interactions. Indeed, the results of this work demonstrate the immunomodulating effect of copper sulphate on S. spallanzanii total body extracts related to oxidative stress and inflammatory markers.

Ecotoxicology and geostatistical techniques employed in subtropical reservoirs sediments after decades of copper sulfate application.

Spatial distribution linked to geostatistical techniques contributes to sum up information into an easier-to-comprehend knowledge. This study compares copper spatial distribution in surface sediments and subsequent categorization according to its toxicological potential in two reservoirs, Rio Grande (RG) and Itupararanga (ITU) (São Paulo-Brazil), where copper sulfate is applied and not applied, respectively. Sediments from 47 sites in RG and 52 sites in ITU were collected, and then, copper concentrations were interpolated using geostatistical techniques (kriging). The resulting sediment distributions were classified in categories based on sediment quality guides: threshold effect level and probable effect level; regional reference values (RRVs) and enrichment factor (EF). Copper presented a heterogenic distribution and higher concentrations in RG (2283.00 ± 1308.75 mg/kg) especially on the upstream downstream, associated with algicide application as well as the sediment grain size, contrary to ITU (21.81 ± 8.28 mg/kg) where a no-clear pattern of distribution was observed. Sediments in RG are predominantly categorized as "Very Bad", whereas sediments in ITU are mainly categorized as "Good", showing values higher than RRV. The classification is supported by the EF categorization, which in RG is primarily categorized as "Very High" contrasting to ITU classified as "Absent/Very Low". Copper total stock in superficial sediment estimated for RG is 4515.35 Ton of Cu and for ITU is 27.45 Ton of Cu.

Copper sulfate induced toxicological impact on in-vivo zebrafish larval model protected due to acacetin via anti-inflammatory and glutathione redox mechanism.

Copper sulfate (CuSO4) as industrial effluent is intentionally or unintentionally released into water bodies and accumulates in the fish. Because of its numerous applications, CuSO4 can be hazardous to non-target creatures, producing direct alterations in fish habitats. Acacetin is a flavonoid present in all vascular plants that are extensively dispersed in plant pigments and responsible for many natural hues. However, the impact of acacetin on mitigating the toxic effect of CuSO4 in the in-vivo conditions is not known. The toxicity of acacetin was determined by measuring the survival, deformities and heart rate after treatment with various concentrations to larvae. The protective effect of acacetin was also observed in CuSO4 exposed zebrafish larvae by reducing malformation, mortality rate and oxidative stress. Meanwhile, the acacetin-protected larvae from CuSO4 effects through the molecular mechanism by suppressing pro-inflammatory genes (COX-2, TNF-α and IL-1) and upregulating antioxidant genes (GPx, GST and GR). Overall, our findings suggest that acacetin can act as a protective barrier against CuSO4-induced inflammation in an in-vivo zebrafish larval model.

Inhalation toxicity of copper compounds: Results of 14-day range finding study for copper sulphate pentahydrate and dicopper oxide and 28-day subacute inhalation exposure of dicopper oxide in rats.

Inhalation exposure to copper may occur during a range of occupational activities and the purpose of this study was to characterise the toxicological response to repeated inhalation of two copper compounds, representative of copper substances in large-scale production/use. Crl:CD(SD) rats were repeatedly exposed to aerosols of dicopper oxide (Cu2O) or copper sulphate pentahydrate (CuSO4.5 H2O) for 14-days as part of a range finding study at normalised copper doses of 0.18, 0.71, 1.78 and 8.9 mg/m3 Cu. Within a 28-days main study (Cu2O only), animals were repeatedly exposed to 0.2, 0.4, 0.8 and 2.0 mg/m3 Cu2O following OECD TG 412. The main study also consisted of satellite groups exposed for 1-, 2- or 3- weeks as well as a 13-week post-exposure recovery period group. Repeated exposure for 14-days to both copper compounds, normalised for copper content, led to an acute influx of polymorphonuclear leukocytes (neutrophils) and macrophages whilst only CuSO4.5 H2O exposure resulted in epithelial hyperplasia. This differential response may reflect the highly dissolvable nature of CuSO4.5 H2O in lung lining fluid leading to a release of copper ions at the epithelial surface whilst Cu2O is relatively indissolvable at neutral pH. In the 28-day study with Cu2O, an increase in cellularity was also evident in both histological and BALF samples and was dose-related with minimal to mild (neutrophilic) inflammation observed > 0.4 mg/m3 in the lung tissue sections and significant increases from 0.2 mg/m3 in BALF. There were no minimal haematological findings, no clinical findings and systemic organs were unaffected by inhalation exposure to dicopper oxide. The lung cellular response was limited to alveolar histiocytosis and neutrophil influx with no evidence of epithelial hyperplasia or fibrosis and all lung biomarkers returned to control levels within the post-exposure recovery period. Interestingly, the satellite groups showed that this acute cellular response followed a biphasic rather than monotonic pattern with a peak in lung biomarkers between weeks 1-3 and reduction thereafter. This reduction in lung biomarkers occurred during continued exposure and may indicate an adaptive response to copper exposure. Overall, these results show that repeated exposure to copper compounds results in an acute cellular response with no associated pathology and which fully resolved after the cessation of exposure. Therefore, the cellular response is evidence of a controlled and adaptive response associated with the removal of Cu2O from the alveolar surface.

Impact of copper sulfate on survival, behavior, midgut morphology, and antioxidant activity of Partamona helleri (Apidae: Meliponini).

Copper sulfate (CuSO4) is widely used in agriculture as a pesticide and foliar fertilizer. However, the possible environmental risks associated with CuSO4 use, particularly related to pollinating insects, have been poorly studied. In this study, we evaluated both lethal and sublethal effects of CuSO4 on the stingless bee Partamona helleri. Foragers were orally exposed to five concentrations of CuSO4 (5000, 1666.7, 554.2, 183.4, 58.4 µg mL-1), and the concentration killing 50% (LC50) was estimated. This concentration (142.95 µg mL-1) was subsequently used in behavioral, midgut morphology, and antioxidant activity analyses. Bee mortality increased with the ingestion of increasing concentrations of CuSO4. Ingestion at the estimated LC50 resulted in altered walking behavior and damage to the midgut epithelium and peritrophic matrix of bees. Furthermore, the LC50 increased the catalase or superoxide dismutase activities and levels of the lipid peroxidation biomarker malondialdehyde. Furthermore, the in situ detection of caspase-3 and LC3, proteins related to apoptosis and autophagy, respectively, revealed that these processes are intensified in the midgut of treated bees. These data show that the ingestion of CuSO4 can have considerable sublethal effects on the walking behavior and midgut of stingless bees, and therefore could pose potential risks to pollinators including native bees. Graphical abstract.

Assessment of reproductive toxicity of carbofuran and copper sulfate in male Nile tilapia (Oreochromis niloticus).

This study's hypothesis is that carbofuran and copper sulfate have a synergistic harmful impact on the fertility of male Nile tilapia. Hence, this study was designed to assess the toxic reproductive outcome of carbofuran, copper sulfate, and their mixture in male Nile tilapia. Sixty male Nile tilapia fishes were separated into four groups (15 fish/group). The control group; carbofuran group, was given dechlorinated tap water containing 0.02 mg/L (1/10 dose of LC50) carbofuran; copper group was given dechlorinated tap water containing 4.0 mg/L (1/10 dose of LC50) copper sulfate; carbofuran + copper sulfate group received dechlorinated tap water containing 0.02 mg/L carbofuran plus 4.0 mg/L copper sulfate. After 6 weeks, results revealed a significant rise in testicular malondialdehyde levels and a significant decrease in testicular reduced glutathione contents among all experimental groups compared to the control group. Testicular testosterone levels were significantly declined in copper and combined groups compared to the control. The seminal evaluation using computer-assisted sperm analysis showed a significant decline in the progressive motility percentage, motile ratio percentage, sperm concentration, curvilinear velocity, straight-line velocity, average path velocity, and wobble in all intoxicated groups, particularly, the combined group. The histopathology of testes in all intoxicated groups revealed a detachment of the basal membrane of some seminiferous tubules, and others were free from spermatogonia and spermatozoa with interstitial eosinophilic granular cell infiltration. Testicular lesions were more severe in the combined group. Finally, it was concluded that carbofuran and copper sulfate exerted a negative effect on the reproductive function of male Nile tilapia, and they have a synergistic harmful impact on the fertility of male Nile tilapia.

Protective effect of surfactin on copper sulfate-induced inflammation, oxidative stress, and hepatic injury in zebrafish.

Surfactin, an antibacterial peptide, produced by various Bacillus subtilis strains, have broad-spectrum antibacterial and immune-enhancing functions. In this study, we investigated the anti-inflammatory, antioxidant, and hepatoprotective effect of surfactin on zebrafish (Danio rerio) larvae following their exposure to copper sulfate (CuSO4 ). The mature AB wild-type and a transgenic line of zebrafish larvae that expressed enhanced GFP (EGFP) named Tg (Lyz:EGFP) were exposed to 0, 20, 40, and 60 µg/mL surfactin after incubation with 3.2 µg/mL CuSO4 for 2 h from 72 h postfertilization (hpf). Different endpoints, such as migration of GFP-labeled neutrophils, analysis of inflammatory cytokines and transaminases, markers of oxidation, expression of certain genes, and histological changes of liver, were studied to evaluate the function of surfactin. The protein expression levels of NF-κBp65, TNF-α, cyclooxygenase-2 (COX-2), and iNOS were determined in murine macrophage RAW 264.7 cells by western blotting. Our results show that surfactin reduced migration of neutrophils and relieved hepatic injury. In addition, surfactin reduced the index levels of inflammatory factors, oxidative stress response, and improved hepatic function. Surfactin also significantly inhibited the expression of IL-1ß, IL-8, TNF-α, nitric oxide, NF-κBp65, COX-2, and iNOS, and increased the expression of IL-10. Thus, our results demonstrate that surfactin has anti-inflammatory, antioxidant, and hepatoprotective activities. Surfactin has potential as a novel inflammation and immune adjustment.

Allicin ameliorates aluminium- and copper-induced cognitive dysfunction in Wistar rats: relevance to neuro-inflammation, neurotransmitters and Aß(1-42) analysis.

Alzheimer's disease (AD) is a multifactorial neurological disorder associated with neuropathological and neurobehavioral changes, like cognition and memory loss. Pathological hallmarks of AD comprise oxidative stress, formation of insoluble ß-amyloid (Aß) plaques, intracellular neurofibrillary tangles constituted by hyperphosphorylated tau protein (P-tau), neurotransmitters dysbalanced (DA, NE, 5-HT, GABA and Glutamate) and metal deposition. Chronic exposure to metals like aluminium and copper causes accumulation of Aß plaques, promotes oxidative stress, neuro-inflammation, and degeneration of cholinergic neurons results in AD-like symptoms. In the present study, rats were administered with aluminium chloride (200 mg/kg p.o) and copper sulfate (0.5 mg/kg p.o) alone and in combination for 28 days. Allicin (10 and 20 mg/kg i.p) was administered from day 7 to day 28. Spatial and recognition memory impairment analysis was performed using Morris water maze, Probe trial, and Novel Object Recognition test. Animals were sacrificed on day 29, brain tissue was isolated, and its homogenate was used for biochemical (lipid peroxidation, nitrite, and glutathione), neuro-inflammatory (IL-1ß, IL-6 and TNF- α), neurotransmitters (DA, NE, 5-HT, GABA and Glutamate), Aß(1-42) level, Al concentration estimation, and Na+/K+-ATPase activity. In the present study, aluminium chloride and copper sulfate administration increased oxidative stress, inflammatory cytokines release, imbalanced neurotransmitters' concentration, and promoted ß-amyloid accumulation and Na+/K+-ATPase activity. Treatment with allicin dose-dependently attenuated these pathological events via restoration of antioxidants, neurotransmitters concentration, and inhibiting cytokine release and ß-amyloid accumulation. Moreover, allicin exhibited the neuroprotective effect through antioxidant, anti-inflammatory, neurotransmitters restoration, attenuation of neuro-inflammation and ß-amyloid-induced neurotoxicity.

Soil invertebrate toxicity and bioaccumulation of nano copper oxide and copper sulphate in soils, with and without biosolids amendment.

The fate, toxicity and bioaccumulation of copper oxide nanoparticles (nCuO) was investigated in soil, with and without biosolids amendment, through chronic exposures using the earthworm, Eisenia andrei, and the collembolan, Folsomia candida. The effects of copper sulphate (CuSO4) were included so as to compare the behavior of nCuO to a readily soluble counterpart. The fate of nCuO was evaluated through characterization of dissolved and nano-particulate fractions (via single particle ICP-MS) as well as extractable Cu2+ throughout the duration of select tests. Neither Cu form was particularly toxic to F. candida, but effects on E. andrei reproduction were significant in all treatments (IC50 range: 98 - 149 mg Cu kg-1 dry soil). There were no significant differences in toxicity between the Cu forms, nor in extractable Cu2+ activities, indicative that particle dissolution within the soil and, subsequent activity of Cu2+ was likely the primary mode of toxicity in the nCuO exposures. The presence of biosolids did not significantly alter toxicity of nCuO, but did affect Cu2+ activity over time. Bioaccumulation of total Cu in E. andrei when exposed to nCuO (kinetic bioaccumulation factor (BAFk): 0.80 with biosolids and 0.81 without) was lower than exposure to CuSO4 (BAFk: 2.31 with biosolids and 1.12 without). Enhanced dark-field hyperspectral imaging showed accumulation of nCuO along the epidermis and gut of E. andrei, with trace amounts observed in muscle and chloragogenous tissue, providing evidence of nCuO translocation within the organism. The present study demonstrates that the current risk assessment approach for trace metals in the environment, based on substance solubility and bioavailability of the dissolved free ion, are applicable for nCuO exposure to soil invertebrates, but that the rate of particle dissolution in different soil environments is an important factor for consideration.