Different Concentrations Of Chlorpyrifos Research Articles

Resilience of aerobic sludge biomass under chlorpyrifos stress and its recovery potential

Non-agricultural sources of pesticides in urban areas are responsible for their presence in domestic wastewater. Therefore, pesticides are typically found in sewage treatment plants in developed and developing countries as micro-pollutant. The presence of pesticides in the wastewater can impart stress on the aerobic sludge biomass and disrupt the functioning of the plant. However, there exists a knowledge gap regarding the resilience of aerobic sludge biomass towards stress due to the presence of pesticides in the wastewater. This study investigated the impact of chlorpyrifos (CPS) – a widely used pesticide, on sludge biomass and explored its recovery capability when CPS is discontinued in the influent. Four duplicate reactors were operated with different CPS concentrations ranging from 50 to 200 mg/L. Chemical oxygen demand (COD) removal for reactors has ranged within 18–73 % at the steady state of the stressed phase, whereas COD removal for the control reactor was 91 %. CPS stress slightly inhibited filamentous biomass growth. Biomass activity and cell viability have decreased significantly, whereas biochemical contents have varied slightly under CPS stress. The activities of the enzymes dehydrogenase and urease were significantly inhibited when compared to catalase and protease. Amplified ribosomal DNA restriction analysis reflected changes in the microbial community. The discontinuation of CPS has allowed aerobic sludge biomass to recover in its organic degradation capability (COD removal of more than 88 % at steady-state conditions of recovery phase operation), biomass growth, and cell viability. In addition, enzyme activities have retrieved to their original levels, and 78–93 % similarity of microbial community structure has been displayed between CPS-exposed and control reactor biomasses. Overall, the present study has indicated the orderly changes in the quality of aerobic sludge biomass under CPS stress through physico-chemical and biological characteristics. The study also has highlighted the self-recovery of sludge biomass characteristics stressed with different concentrations of CPS.

Effect of chlorpyrifos and its metabolites on skeletal system development of chick embryo.

Chlorpyrifos is an organophosphate insecticide used primarily to control pests on a variety of food and feed crops. Humans are directly or indirectly exposed to this pesticide through food, air, and occupation. The ill effects of chlorpyrifos on various organ systems of human has been widely documented, but less is known about its influence on human bones. To analyze the effect of chlorpyrifos and its metabolites 3,5,6-trichloro-2-pyridinol (TCPy) on the skeletal system of the chick embryo. Fertilized chick eggs were exposed to different concentrations of chlorpyrifos and its metabolite 3,5,6-TCPy on 1.5 days of incubation. The proximal phalanx of 18-day-old embryos was analyzed for defects in growth and ossification through histopathology, immunohistochemistry, angiogenesis assay, and gene expression study. Dose-dependent variations in developing bone of chick embryo were observed. Histochemical and histomorphometry studies of proximal phalanx showed increased in the growth plate length (F(9, 59)=228.9509, p=.00001) with a reduction in the total length of the phalanx (F(9, 59)=109.9905, p=.00001), decreased mineralization (F(9, 59)=224.6872, p=.00001), decreased blood islands in the bone marrow (F(9, 59)=7.7083, p=.0001) of chlorpyrifos, and 3,5,6-TCPy-exposed group. Significant downregulations in the expression patterns of the transcription factors, such as SOX9, RUNX2, and ALP, were also observed. Chlorpyrifos and its metabolite 3,5,6-TCPy exposure alters the chondrogenesis in the growth plate cartilage of long bone in chick embryo. The pesticide and its metabolite also interfere in ossification.

Effect of chlorpyrifos on the expression and regulation of salivary gland protein of Haemaphysalis longicornis

ABSTRACT The aim of this study is to explore the detoxification and homoeostasis functions of salivary glands, and to look for possible target proteins of new drugs for tick prevention and treatment. Different concentrations of chlorpyrifos to deal with the semi-engorged female Haemaphysalis longicornis, through the DIA quantitative proteomics method to quantitatively determine of all the proteins in the salivary glands, and through a variety of the depth of the bioinformatics methods for protein analysis and function mining, explain the molecular mechanism of detoxification function of salivary glands. Quantitative information from 5 normal groups and 8 different treatment groups showed that a total of 3152 proteins were identified. The expression levels of these proteins changed with the chlorpyrifos different concentration and treatment times. However, after the overall analysis, it was found that after the action of chlorpyrifos, the proteins involved in detoxification, such as glutathione S-transferase and glutamine synthetase, the heat shock proteins involved in stress, and the acid sphingomyelinase involved in maintaining the body’s homoeostasis had corresponding changes. We found and explained the functions of detoxification, stress, and homoeostasis of salivary glands. The regulatory strategies of proteins that played a key role in these functions were explored.

Protective effect of total saponins of ginseng stems and leaves (GSLS) on chlorpyrifos-induced brain toxicity in mice through the PTEN/PI3K/AKT axis.

Chlorpyrifos (CPF) is a class of toxic compounds which has been widely used in agriculture that can cause multi-organ damage to the liver, kidneys, testes, and nervous system. Currently, most studies on ginseng have concentrated on the roots and rhizomes, and less research has been conducted on the above-ground parts. Our laboratory found that ginseng stem and leaf total saponin (GSLS) features strong antioxidant activity. In this experiment, we selected different concentrations of CPF to induce hippocampal neuronal cell injury model in mice, conducted a cell survival screening test, and also selected appropriate concentrations of CPF to induce brain injury model in mice. CCK-8, flow cytometry, Elisa, Hoechst 33258 staining, Annexin V-FITC/PI staining, HE staining, Morris water maze, and qRT-PCR were adopted for detecting the effects of GSLS treatment on CPF-induced cell viability, mitochondrial membrane potential, reactive oxygen species (ROS) levels, Ca2+ concentration and GSLS treatment on CPF-induced brain injury and related signaling in mice, respectively. The effects of GSLS treatment on CPF-induced brain injury and the related signaling pathways in mice were examined. The results showed that GSLS at 60 μg/ml and 125 μg/ml concentrations elevated the viability of CPF-induced HT22 cells, increased mitochondrial membrane potential, depleted ROS, decreased Ca2+ concentration, and decreased apoptosis rate. Meanwhile, GSLS treatment significantly reduced CPF-induced escape latency in mice, elevated the number of entries into the plateau and effective area, increased the effective area and target quadrant residence time, as well as improved the pathological damage of mouse hippocampal neurons. The results of mouse brain sections demonstrated that GSLS treatment significantly increased SOD and CAT activities and lowered MDA accumulation in CPF-induced mice. qRT-PCR revealed that PTEN mRNA expression was significantly decreased with PI3K and AKT expression being significantly increased in GSLS-treated CPF-induced mice. Thus, the obtained results indicate that GSLS can effectively antagonize CPF-induced brain toxicity in mice through regulating PTEN/PI3K/AKT pathway.

Chlorpyrifos biodegradation by a Locally Isolated Bacterium Pseudomonas sp. B5-2

Chlorpyrifos has been considered as one of the most frequently used organophosphorus insecticides in agriculture worldwide. There has been considerable public concern regarding its neurotoxicity and its genotoxicity on mammals, aquatic organisms, and other non-target species. In this study 23 bacterial isolated strains from different origins were tested for their ability to resist and subsequently degrade chlorpyrifos (CP). Seven of these strains showed a resistance to up to 1000mg/L CP on solid minimal mineral salt medium (MMSM) and the strain Pseudomonas sp. B5-2 was selected for further analysis after a selection on liquid MMSM. The effect of different concentrations of CP on the growth of the selected strain was tested, and 200 mg/L was the highest concentration that supports its growth. The ability of the selected strain to eliminate CP was investigated in the presence and in the absence of glucose as an external carbon source. The results showed that the strain utilized chlorpyrifos (200 mg/L) as the sole source of carbon better than in presence of an external carbon source (glucose) with an elimination rate of 67.06% and 14.61%, respectively. These results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment.

Effects of Chlorpyrifos on biochemical characteristics of Labeorohitafish during acute and chronic exposure

The aim of present study was to examine the biochemical characteristics of Labeorohita fish during acute and chronic exposure of pesticide, chlorpyrifos (CPF). Methods:During acute exposure, fish were exposed to different concentrations of CPF ranging from 0, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04 and 0.05 mg/L for 96 hrs in glass aquaria. The 96 hrs LC50 value of CPF for Labeorohita was found to be 0.01 mg/L. During chronic exposure fish were subjected to 1/3rd, 1/5th, 1/7th and 1/9th of LC50 for 30 days. At the end of the trial, samples were collected for biochemical (Blood cells, haemoglobin, plasma glucose and plasma protein) and sent to related laboratories. Results:The present study showed the RBCs, haemoglobin and Plasma Glucose level decreases as the concentration of CPF increases. On the other hand, platelets, WBCs and Plasma glucose level increases as the concentration of CPF increases. Decrease in RBCs count and haemoglobin indicates that CPF can cause anaemia to the fish. Conclusions: It is therefore concluded that Chlorpyrifos adversely affects the major organs of the fish Labeorohita.

Chlorpyrifos degradation, biocontrol potential and antioxidant defence activation under pesticide stress by rhizosphere bacteria isolated from rhizosphere of peach (Prunus persica) plants

ABSTRACT Current experiments were done to isolate plant growth-promoting rhizobacteria (PGPR) from peach (Prunus persica) rhizosphere having biocontrol, biodegradation and defence mechanism activation against pesticide toxicity. PGPR strains (Pseudomonas and Bacillus) showed biocontrol activity against phytopathogens. Bacillus flexus PS-26 tolerated a maximum amount of chlorpyrifos and was also able to degrade different concentrations of chlorpyrifos within 30 h. Bacillus flexus PS-26 formed more biofilm, released a high amount of antioxidants and exopolysaccharides under chlorpyrifos stress, which suggested its detoxification role. The present study proved Bacillus flexus PS-26 to be a super-bioinoculant, which could protect plants against pesticide.

Which Is More Toxic? Evaluation of the Short-Term Toxic Effects of Chlorpyrifos and Cypermethrin on Selected Biomarkers in Common Carp (Cyprinus carpio, Linnaeus 1758).

The general aim of this study was to investigate the negative short-term effects of different concentrations of chlorpyrifos (CPF) and cypermethrin (CYP), based on the EU legislation (MAC-EQS) in common carp (Cyprinus carpio Linnaeus, 1758) under laboratory conditions and to compare their toxicity. The fish were exposed to the pesticides for 96 h and then different histological and biochemical biomarkers were investigated in the gills and liver, and bioaccumulation analyses were conducted. The chemical studies showed increased pesticide concentrations in the gills as the first site for pollutants compared to the liver at the 96th hour. In addition, the histological analyses showed severe alterations in the gills and liver after exposure to both tested pesticides. In the gills, we found mainly intense proliferative and, to a lesser extent, degenerative changes and alterations in the circulatory system, such as necrosis and vasodilation. In the liver, regressive and progressive lesions, as well as circulatory disturbances and inflammation, were observed. The regressive lesions showed a higher degree of expression compared to the other changes. Furthermore, we found altered enzymatic activities—catalase, glutathione reductase, and glutathione peroxidase—in the liver, compared to the control. Overall, both tested pesticides impacted the studied biomarkers in common carp, even at concentrations lower than those permitted by law. However, the results of the comparative analysis showed a relatively higher toxicity of CYP compared to CPF in the fish. Still, questions persist as to whether the observed changes are adaptive or entirely destructive. To avoid any danger or risk, these pesticides must be applied cautiously, especially near water bodies.

Biodegradation of chlorpyrifos by a Weissella confusa strain and evaluation of some probiotic traits

Pesticides play an important role in agriculture; however, their excessive use causes several problems such as pollution of ecosystems and risks to human health. The presence of microorganisms able to degrade these pollutants can reduce their negative effect. The objective of this study was to test the capacity of Weissella confusa Lb.Con to tolerate or to degrade the chlorpyrifos pesticide. The results showed the capacity of the strain to tolerate a concentration of 200μg/ml of chlorpyrifos. The strain Lb.Con has a remarkable capacity to grow in glucose-free MRS medium which contains different concentrations of chlorpyrifos. HPLC analysis showed that this strain was able to remove about 25% of chlorpyrifos. The evaluation of some probiotic properties showed that the strain Lb.Con had a remarkable resistance to the gastrointestinal conditions and a good antibacterial activity towards the pathogenic bacteria. The probiotic potential was evaluated to verify the possible use of W. confusa Lb.Con to detoxify harmful chlorpyrifos contained in food.

Effect of chlorpyrifos and carbofuran on morphology, behavior and acetylcholinesterase activity of earthworm (Eisenia fetida)

Organophosphates (OP) and carbamates when applied in agricultural fields are the potential environmental polluters and toxicants for soil flora and fauna. Upon exposure to such chemical pesticides, morphology, behaviour and acetylcholinesterase (AChE) activity of earthworms (Eisenia fetida) is altered. The present study dealt with the comparative analysis of toxic effects induced by chlorpyrifos (OP) and carbofuran (carbamate) exposure on morphology, behavior and AChE activity using standard filter paper contact toxicity method. The LC50 of chlorpyrifos and carbofuran for 24 hr was 0.25% and 5.13%, respectively, marking chlorpyrifos as more toxic pesticide than carbofuran. The treated worms exhibited abnormal morphological symptoms such as excessive mucus secretion, depigmentation, deformity, loss of metameric segments and damaged clitellum. Variation in behaviour such as reduced activity, sluggish movements and flattened posture marked the stress induced due to pesticide toxicity even at lower doses. Restlessness along with jerky movements was observed on exposure to higher concentrations of chlorpyrifos, whereas lower concentrations resulted in avoidance behavior toward pesticide coated glass vials. Thereafter, worms were exposed to different concentrations of chlorpyrifos (0.13%, 0.25% and 0.38%) and carbofuran (2.57%, 5.13% and 7.70%) for 24h and 48h to assess sub-acute and acute toxicity effects of these pesticides on AChE. The enzyme activity after 48h was 94.14%, 60.21% and 40.44% for the worms exposed to 0.125%, 0.25% and 0.375% chlorpyrifos, respectively. The enzyme activity after 48h was 98.17%, 93.92% and 79.25% for the worms exposed to 2.57%, 5.13% and 7.70% carbofuran, respectively. Time and dose dependent significant (p<0.05) decrease in the levels of AChE in pre-clitellar region of earthworms was observed as compared to control when worms were exposed to chlorpyrifos and carbofuran. Alteration in behavioural response of the earthworms may be attributed to the decline in AChE activity of pre-clitellar region. However, chlorpyrifos was more potent inhibitor of AChE activity in Eisenia fetida as compared to that of carbofuran. Such alterations indicate the potential health risk of these pesticides’ exposure at high concentrations to E. fetida.

Sensitive SERS Characterization and Analysis of Chlorpyrifos and Aldicarb Residues in Animal Feed using Gold Nanoparticles

The spectroscopic method based on surface-enhanced Raman spectroscopy (SERS) technique combined with chemometric methods was developed for simple, cost-effective, and efficient analysis of chlorpyrifos (CPF) and aldicarb (ALD) pesticide residues in animal feed. Animal feeds free from the pesticides were spiked at different concentrations of CPF (0-20 mg/kg) and aldicarb (0-100 µg/kg). Gold nanoparticles were mixed with sample extract for SERS measurement. A significant spectral difference induced by the presence and different level of CPF and ALD concentration in animal feed was observed between the pesticide spiking groups. Different chemometric models applied on training datasets showed excellent classification rates (100 percent) while the models on external validation dataset exhibited lower correct classification rates (50.0-76.7 percent) with no false-negative error. The selected chemometric models for CPF and ALD quantification also showed a high predictive ability and performance. The developed models displayed no statistical significant difference between model predicted and reference values in the external validation dataset (p < 0.01). The study results indicate that the SERS spectroscopic method could be an effective and efficient analytical tool for pesticide analysis in highly complex animal feed matrices for screening at a point of sampling to improve food and feed safety.
 https://doi.org/10.21423/jrs-v08lee

Leukocyte Profiles in Odontophrynus carvalhoi (Amphibia: Odontophrynidae) Tadpoles Exposed to Organophosphate Chlorpyrifos Pesticides

We analyzed the toxic effects of the organophosphate insecticide chlorpyrifos (Klorpan® 480 EC formulation) on the blood leukocyte profiles of Odontophrynus carvalhoi tadpoles, a neotropical species endemic to Brazil, exposed to four different concentrations of that insecticide (10, 100, 200, and 400 μg L−1) for 192 h. The tadpoles were anesthetized and their blood was collected by cardiac puncture, with subsequent cell smears and staining with May–Grunwald–Giemsa and Wright. The numbers and types of leukocytes were determined after different periods of exposure (24 h, 48 h, 96 h, and 192 h) by analyzing 100 leukocytes from each tadpole (under 1000× magnification). The tadpoles exposed to different concentrations of chlorpyrifos exhibited reductions in their numbers of lymphocytes, monocytes, and basophils, but increases in the numbers of neutrophils and eosinophils. As such, exposure to chlorpyrifos altered cell type proportions, which could alter the capacity of those tadpoles to respond to environmental stresses, make them more susceptible to infection by diverse pathogens, and thus compromise their chances of survival. That scenario would be even more serious for species having restricted distributions, as population declines would be more severe and impacting. Therefore, the use of chlorpyrifos constitutes a serious risk for anuran amphibians that occur in or near agricultural fields by contributing to immunosuppression and threatening their survival.

Chlorpyrifos induces the apoptosis and necroptosis of L8824 cells through the ROS/PTEN/PI3K/AKT axis

Excessive chlorpyrifos (CPF) in the environment causes toxicity to nontarget organisms by triggering oxidative stress. Phosphatase and tensin homolog deleted on chromosome ten (PTEN) plays an important role in controlling apoptosis and necrosis by negatively regulating the phosphatidylinositol 3-kinase/threonine kinase (PI3K/AKT) pathway. However, the effects of different concentrations of CPF on grass fish liver cell injury and the role of the ROS/PTEN/PI3K/AKT axis remain poorly understood. In this study, L8824 cells treated with different concentrations of CPF (0, 40, 60, or 80 μM) were used as the research object. The results showed that the median inhibitory concentration (IC50) was 112.226 μM. As the CPF concentrations increased, the ROS and MDA levels increased, and the T-AOC levels and SOD/GPx/GST activities decreased. As PTEN expression increased, PI3K/AKT, BCL-2, and Caspase-8 expression dramatically decreased. Conversely, RIPK1/RIPK3/MLKL and Bax/Cyt-c/Caspase-3 expression increased. Additionally, necroptosis increased in a dose-dependent manner, while apoptosis first increased and then decreased. In conclusion, our study showed that CPF could trigger oxidative stress and induce apoptosis and necroptosis in fish liver cells by regulating the ROS/PTEN/PI3K/AKT axis, and the type of damage induced was dose-dependent. These results are meaningful for toxicological studies of CPF and efforts to protect the ecosystem.

Obesogenic effects of chlorpyrifos and its metabolites during the differentiation of 3T3-L1 preadipocytes

Chlorpyrifos (CPF) is an organophosphorus pesticide widely and extensively used in agriculture in more than one hundred countries and found ubiquitously in the environment. The present study was aimed at providing a better understanding of the obesogenic potential of CPF and its metabolites, as well as to evaluate their effects on the adipocyte differentiation process. For it, during the initial differentiation process, 3T3-L1 mouse preadipocytes were exposed to different concentrations of CPF, CPF-oxon (CPO), or 3,5,6-trichloropyridinol (TCP), which did not affect cell survival. Results showed how CPF and, to a lesser extent, its metabolite TCP, had a positive metabolic influence over the adipogenic process by fostering an increase in the number of differentiated 3T3-L1 preadipocytes, and by enhancing the capacity to store lipid droplets. These processes seem to occur through the upregulation of the transcription factors CCAAT/enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ), which are related to a significant higher expression of the fatty acid-binding protein 4 (FABP4) adipokine. Based on this finding, CPF exposure could be one of the many factors that contributes to the worldwide increase in the incidence of obesity. However, additional investigations are clearly needed.

Evaluation of cytotoxic potential of chlorpyrifos using Lens culinaris Med as efficient bioindicator

The aim of this study was to evaluate the cytotoxic effect of different concentrations of chlorpyrifos (CPF), using L. culinaris apical cells as a biological indicator. L. culinaris seeds were exposed to different concentrations of chlorpyrifos (0, 1, 3, 5, 7, 8, 10 and 15 mg L−1) and a control solution based on distilled water. Subsequently, root growth was measured during 24, 48 and 72 h. Therefore, the mitotic index (MI) and the number of cellular abnormalities were determined at 72 h. According to the obtained results, a decrease in root size was observed in the concentrations of T5 (8 mg L−1) and T6 (10 mg L−1). On the other hand, it was evidenced that, through all the evaluated concentrations, the inhibition of mitosis in the concentrations of T5 (8 mg L−1), T6 (10 mg L−1) and T7 (15 mg L−1) was greater than 50%. Additionally, a variety of chromosomal abnormalities were reported, such as Micronuclei, sticky chromosomes in anaphase, chromosome disruption, irregular anaphase, nucleus absence, nuclear lesions, chromosomes grouped in metaphase, anaphase bridges, metaphase sticky chromosomes, present in all concentrations evaluated. Consequently, the presence of micronuclei in the concentrations of 8 mg L−1, 10 mg L−1 and 15 mg L−1 indicates that the CPF is a highly cytotoxic substance to L. culinaris. Therefore, L. culinaris is a plant species that offers a feasible experimental model to be implemented in laboratory studies with the purpose to evaluate the cytotoxic effect of pesticides.

Chlorpyrifos induced oxidative stress to promote apoptosis and autophagy through the regulation of miR-19a-AMPK axis in common carp

Chlorpyrifos (CPF) has become a mainly pollution in water environment. Micro-RNAs (miRNAs) play an important part in the development of apoptosis and autophagy. However, the potential mechanism of CPF induced kidney toxicity and the roles of miRNAs are still unclear. To explore the underlying mechanism, the kidney of common carp exposed to different concentrations of CPF for 40 days was used as a research object. We found that CPF could damage the ultrastructure and function of kidney; and also caused antioxidant system disorder. CPF inhibited the mRNA level of miR-19a which improved AMP-activated protein kinase (AMPK). Furthermore, the detection of apoptosis and autophagy relative genes showed that the expressions of TSC complex subunit 2 (TSC2), light chain 3 (LC3), Dynein, tumor protein 53 (p53), Bcl-2 associated X protein (Bax), caspase-3 and caspase-9 were enhanced and the expressions of nechanistic target of rapamycin (mTOR), Ras homolog mTORC1 binding (Rheb) and B-cell lymphoma (Bcl-2) were reduced in dose-dependent way. Taken together, we conclude that CPF causes oxidative stress and miR-19a-AMPK axis disorder, thereby promotes apoptosis and autophagy in common carp kidney. Our study will provide theoretical basis for toxicology research and environmental protection of CPF.

Effects of long-term chlorpyrifos exposure on mortality and reproductive tissues of Banded Gourami (Trichogaster fasciata)

This study assessed the long-term toxicity of chlorpyrifos on survival and reproduction of Banded Gourami by using mortality, gonado-somatic index (GSI) and histopathological observations as endpoints. Adult fish were exposed to five different concentrations of chlorpyrifos (0, 15, 50, 150, 500 µg/L) in 15 PVC tanks for 15, 30, 45, 60 and 75 days. Results showed that all male and female fish died after 15 days of 500 µg/L chlorpyrifos exposure. No consistent significant effect was observed for both male and female GSI. Furthermore, results showed dose- and time-dependent histopathological alterations for both ovary and testes. The 60-d No Observed Effect Concentration (NOEC) for most histopathological alterations of Banded Gourami ovary and testes was 50 μg/L, while 60-d NOEC for mortality of both male and female fish was < 15 μg/L. The results show that the long-term exposure to chlorpyrifos not only affect the reproductive tissues of Banded Gourami at exposure concentrations but also cause their mortality. Future studies should evaluate effects at lower concentrations.

Chlorpyrifos activates cell pyroptosis and increases susceptibility on oxidative stress-induced toxicity by miR-181/SIRT1/PGC-1α/Nrf2 signaling pathway in human neuroblastoma SH-SY5Y cells: Implication for association between chlorpyrifos and Parkinson's disease.

The insecticide exposure has been linked to Parkinson's disease (PD). In the present study, we used a most widely used cell line in study of PD, the SH-SY5Y cells, to investigate mechanisms of chlorpyrifos (CPF) induced cell toxicity and the possible roles of cell pyroptosis and oxidative stress in SH-SY5Y cells, as well as role of miR-181/SIRT1/PGC-1α/Nrf2 signaling pathway in this process. SH-SY5Y cells were treated with different concentrations of CPF. Cell viability was measured using CCK-8 assay. Cell pyroptosis was determined by immunofluorescence of caspase-1 and TUNEL assay. The miR-181 (has-miR-181-5p) level was determined by qRT-PCR. Expression of SIRT1, PGC-1α, Nrf2, and pyroptosis related proteins NLRP3, caspase-1, IL-1β, and IL-18 was determined by both qRT-PCR and Western blotting. Cell viability was found to be decreased with the increased CPF concentrations. The pyroptosis related proteins, ROS levels, as well as level of caspase-1 and the TUNEL positive cells were all significantly up-regulated by CPF. Meanwhile, expression of miR-181 and pyroptosis proteins was also enhanced, while the SIRT1/PGC-1α/Nrf2 signaling was inhibited by CPF. Knockdown of Nrf2 significantly up-regulated the expression of pyroptosis related proteins, ROS level, caspase-1, and the TUNEL positive cells, while over-expression of Nrf2 resulted in opposite results. The expression of PGC-1α and Nrf2 was significantly down-regulated when SIRT1 was inhibited, while over-expressed SIRT1 led to increased PGC-1α and Nrf2 levels. Besides, miR-181 promoted the CPF induced activation of pyroptosis and oxidative stress, as well as down-regulated SIRT1/PGC-1α/Nrf2 signaling, while inhibition of miR-181 led to opposite results. Chlorpyrifos could inhibit cell proliferation, activate cell pyroptosis and increase susceptibility on oxidative stress-induced toxicity by elevating miR-181 through down-regulation of the SIRT1/PGC-1α/Nrf2 pathway in human neuroblastoma SH-SY5Y cells. This study might give deeper insights for mechanisms of CPF induced toxicity and might give some novel research targets for PD treatment.

Designing of 3D Sensor Chamber for Plasmonic-based Toxic Sensor Detection

Plasmonic sensor implementing an optical phenomenon called Localized Surface Plasmon Resonance (LSPR) resulting from the interaction of free electron with electromagnetic field of light at the metal nanoparticles surface. In this study, the plasmonic sensor has been developed for toxic detection in solution form. This system consists of five components which are the light source, duplex fiber optic, sensor chamber, spectrometer and computer. The sensor chamber has been specially designed using SolidWork software and printed using 3D printer with polylactic acid (PLA) material. The sensing activity was done in the sensor chamber with a sliding drawer which is used to place the sensing material or sample. OceanView software was used to analyze the recorded spectrum from the spectrometer. For this project, the experiment of the plasmonic sensor was carried out using targeted analyte namely chlorpyrifos with deionized (DI) water was set as a reference medium. Gold nanoparticles with nanospheres shape used as sensing materials. The sensing parameters are based on changing its intensity and resonance peak position. This plasmonic sensor was compared with UV-VIS spectrometer data to make sure it standardize and function correctly. Besides, the sensing process toward different concentrations of chlorpyrifos from 7.15 mM to 28.60 mM have been done. As a conclusion, the plasmonic sensor was successfully developed for toxic detection in solution form.

Effects of the insecticide chlorpyrifos, on hatching, mortality and morphology of Duttaphrynus melanostictus embryos

In an attempt to assess the effects of chlorpyrifos [O,O-diethyl O-(3,5,6-trichloropyridin-2-yl) phosphorothioate], the second largest selling insecticide in India, studies were made with reference to some non-target organisms. The present study was undertaken to evaluate the effects in the embryos of Duttaphrynus melanostictus caused by the commercial formulations of chlorpyrifos (Tricel, chlorpyrifos, 20% EC). The LC50 value for Duttaphrynus melanostictus embryos after 48 h (h) of treatment with chlorpyrifos was found to be 57.525 ppm. The mortality of the embryo was significantly affected by different concentrations of chlorpyrifos when compared with the control groups. An increase in concentration of chlorpyrifos resulted in the simultaneous decrease of the hatching percentage and an increase in the morphological abnormalities such as compression of the embryo, reduced body size and curling of tail.