Magnetic nanoparticles are highly toxic to chloroquine-resistant Plasmodium falciparum, dengue virus (DEN-2), and their mosquito vectors

SURESH KUMAR SUBBIAH (A04156) / / / Kadarkarai Murugan,Jiang Wei,Mohamad Saleh Alsalhi,Marcello Nicoletti,Manickam Paulpandi,Christina Mary Samidoss,Devakumar Dinesh,Balamurugan Chandramohan,Chellasamy Paneerselvam,Jayapal Subramaniam,Chithravel Vadiva

Fabrication of nano-mosquitocides using chitosan from crab shells: impact on non-target organisms in the aquatic environment

Mosquitoes are arthropods of huge medical and veterinary relevance, since they vector pathogens and parasites of public health importance, including malaria, dengue and Zika virus. Currently, nanotechnology is considered a potential eco-friendly approach in mosquito control research. We proposed a novel method of biofabrication of silver nanoparticles (AgNP) using chitosan (Ch) from crab shells. Ch-AgNP nanocomposite was characterized by UV–vis spectroscopy, FTIR, SEM, EDX and XRD. Ch-AgNP were tested against larvae and pupae of the malaria vector Anopheles stephensi obtaining LC50 ranging from 3.18 ppm (I) to 6.54 ppm (pupae). The antibacterial properties of Ch-AgNP were proved against Bacillus subtilis, Klebsiella pneumoniae and Salmonella typhi, while no growth inhibition was reported in assays conducted on Proteus vulgaris. Concerning non-target effects, in standard laboratory considtions the predation efficiency of Danio rerio zebrafishes was 68.8% and 61.6% against I and II instar larvae of A. stephensi, respectively. In a Ch-AgNP-contaminated environment, fish predation was boosted to 89.5% and 77.3%, respectively. Quantitative analysis of antioxidant enzymes SOD, CAT and LPO from hepatopancreas of fresh water crabs Paratelphusa hydrodromous exposed for 16 days to a Ch-AgNP-contaminated aquatic environment were conducted. Notably, deleterious effects of Ch-AgNP contaminating aquatic enviroment on the non-target crab P. hydrodromous were observed, particularly when doses higher than 8–10 ppm are tested. Overall, this research highlights the potential of Ch-AGNP for the development of newer control tools against young instar populations of malaria mosquitoes, also highlighting some risks concerned the employ of nanoparticles in aquatic environments

Probing the interaction of thionine with human serum albumin by multispectroscopic studies and its in vitro cytotoxic activity toward MCF-7 breast cancer cells

The studies on protein–dye interactions are important in biological process and it is regarded as vital step in rational drug design. The interaction of thionine (TH) with human serum albumin (HSA) was analyzed using isothermal titration calorimetry (ITC), spectroscopic, and molecular docking technique. The emission spectral titration of HSA with TH revealed the formation of HSA–TH complex via static quenching process. The results obtained from absorption, synchronous emission, circular dichroism, and three-dimensional (3D) emission spectral studies demonstrated that TH induces changes in the microenvironment and secondary structure of HSA. Results from ITC experiments suggested that the binding of TH dye was favored by negative enthalpy and a favorable entropy contribution. Site marker competitive binding experiments revealed that the binding site of TH was located in subdomain IIA (Sudlow site I) of HSA. Molecular docking study further substantiates that TH binds to the hydrophobic cavity of subdomain IIA (Sudlow site I) of HSA. Further, we have studied the cytotoxic activity of TH and TH–HSA complex on breast cancer cell lines (MCF-7) by MTT assay and LDH assay. These studies revealed that TH–HSA complex showed the higher level of cytotoxicity in cancer cells than TH dye-treated MCF-7 cells and the significant adverse effect did not found in the normal HBL-100 cells. Fluorescence microscopy analyses of nuclear fragmentation studies validate the significant reduction of viability of TH–HSA-treated human MCF-7 breast cancer cells through activation of apoptotic-mediated pathways

Toxicity on Dengue Mosquito Vectors Through Myristica fragrans-Synthesized Zinc Oxide Nanorods, and Their Cytotoxic Effects on Liver Cancer Cells (HepG2)

Dengue is an arbovirus mainly vectored by Aedes mosquitoes. Its prevention and control depends to effective vector control measures. Cancer causes millions of death every year. Most of the anticancer drugs have high toxicity and low specificity of action, leading to systemic toxicity and severe side effects. Thus, the development of effective tools is a priority. We fabricated zinc oxide nanoparticles using the Myristica fragrans extract as a reducing and stabilizing agent. Nanoparticles were studied using UV–vis spectrophotometry, Fourier transform infrared spectroscopy, X-ray diffraction, zeta potential, dynamic light scattering, energy dispersive X-ray analysis, field emission scanning electron microscopy and transmission electron microscopy. ZnO nanorods were highly effective against A. aegypti young instars, with LC50 ranging from 3.44 (larva I) to 14.63 ppm (pupa). Nanorods showed adult LC50 of 15.004 ppm. ZnO nanorods exhibited dose-dependent cytotoxicity against human hepato-cancer cells (HepG2). After 48 and 24 h of incubation, the IC50 were 20 and 22 μg/ml, respectively. Nanorods triggered the induction of apoptosis. Overall, this study highlights that the possibility to employ M. fragrans-synthesized ZnO nanorods in mosquito control, as well as in the development of novel chemotherapeutic agents with reduced systemic toxicity

Nanofabrication of Graphene Quantum Dots with High Toxicity Against Malaria Mosquitoes, Plasmodium falciparum and MCF-7 Cancer Cells: Impact on Predation of Non-target Tadpoles, Odonate Nymphs and Mosquito Fishes

Recently, it has been highlighted an overlooked connection between the biting activity of Anopheles mosquitoes and the spread of cancer. The excellent physico-chemical properties of graphene quantum dots (GQDs) make them a suitable candidate for biomedical applications. We focused on the toxicity of GQDs against Plasmodium falciparum and its vector Anopheles stephensi, and their impact on predation of non-target mosquito predators. Biophysical methods, including UV–vis, photoluminescence, FTIR and Raman spectroscopy, XRD analysis and TEM, confirmed the effective GQD nanosynthesis. LC50 against A. stephensi ranged from 0.157 (larva I) to 6.323 ppm (pupa). The antiplasmodial activity of GQDs was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of P. falciparum. IC50 were 82.43 (CQ-s) and 85.17 μg/ml (CQ-r). In vivo experiments conducted on Plasmodium berghei infecting albino mice showed moderate activity of GQDs if compared to chloroquine. Concerning non-target effects, the predation efficiency of Gambusia affinis, Anax immaculifrons and Hoplobatrachus tigerinus post-treatment with GQDs was enhanced. Lastly, GQDs were toxic against MCF-7 breast cancer cell lines with an IC50 = 24.81 μg/ml, triggering apoptosis in treated cells. Overall, we highlighted the multipurpose potential of GQDs for the development of newer drugs in the fight against Anopheles vectors, Plasmodium parasites and breast cancer cells

Hydrothermal synthesis of titanium dioxide nanoparticles: mosquitocidal potential and anticancer activity on human breast cancer cells (MCF-7)

Mosquito vectors (Diptera: Culicidae) are responsible for transmission of serious diseases worldwide. Mosquito control is being enhanced in many areas, but there are significant challenges, including increasing resistance to insecticides and lack of alternative, cost-effective, and eco-friendly products. To deal with these crucial issues, recent emphasis has been placed on plant materials with mosquitocidal properties. Furthermore, cancers figure among the leading causes of morbidity and mortality worldwide, with approximately 14 million new cases and 8.2 million cancer-related deaths in 2012. It is expected that annual cancer cases will rise from 14 million in 2012 to 22 million within the next two decades. Nanotechnology is a promising field of research and is expected to give major innovation impulses in a variety of industrial sectors. In this study, we synthesized titanium dioxide (TiO2) nanoparticles using the hydrothermal method. Nanoparticles were subjected to different analysis including UV–Vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), zeta potential, and energy-dispersive spectrometric (EDX). The synthesized TiO2 nanoparticles exhibited dose-dependent cytotoxicity against human breast cancer cells (MCF-7) and normal breast epithelial cells (HBL-100). After 24-h incubation, the inhibitory concentrations (IC50) were found to be 60 and 80 μg/mL on MCF-7 and normal HBL-100 cells, respectively. Induction of apoptosis was evidenced by Acridine Orange (AO)/ethidium bromide (EtBr) and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) staining. In larvicidal and pupicidal experiments conducted against the primary dengue mosquito Aedes aegypti, LC50 values of nanoparticles were 4.02 ppm (larva I), 4.962 ppm (larva II), 5.671 ppm (larva III), 6.485 ppm (larva IV), and 7.527 ppm (pupa). Overall, our results suggested that TiO2 nanoparticles may be considered as a safe tool to build newer and safer mosquitocides and chemotherapeutic agents with little systemic toxicity

Fighting arboviral diseases: low toxicity on mammalian cells, dengue growth inhibition (in vitro), and mosquitocidal activity of Centroceras clavulatum-synthesized silver nanoparticles

Dengue is a mosquito-borne viral disease that has rapidly spread in all regions of the world in recent years. Female mosquitoes, mainly Aedes aegypti, transmit dengue. Approximately 3,900 million people, in 128 countries, are at risk of dengue infection. Recently, a focus has been provided on the potential of green-synthesized nanoparticles as inhibitors of the production of dengue viral envelope (E) protein in Vero cells and downregulators of the expression of dengue viral E gene. Algae are an outstanding reservoir of novel compounds, which may help in the fight against mosquito-borne diseases. In this research, silver nanoparticles (AgNP) were rapidly synthesized using a cheap extract of the alga Centroceras clavulatum. AgNP were characterized by UV–vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffraction (XRD). In mosquitocidal assays, LC50 values of C. clavulatum extract against A. aegypti larvae and pupae were 269.361 ppm (larva I), 309.698 ppm (larva II), 348.325 ppm (larva III), 387.637 ppm (larva IV), and 446.262 ppm (pupa). C. clavulatum extract also exhibitedmoderate antioxidant activity, both in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging assays. LC50 values of C. clavulatum-synthesized AgNP were 21.460 ppm (larva I), 23.579 ppm (larva II), 25.912 ppm (larva III), 29.155 ppm (larva IV), and 33.877 ppm (pupa). Furthermore, C. clavulatum-synthesized AgNP inhibited dengue (serotype dengue virus type-2 (DEN-2)) viral replication in Vero cells. Notably, 50 μg/ml of green-synthesized AgNP showed no cytotoxicity on Vero cells while reduced DEN-2 viral growth of more than 80 %; 12.5 μg/ml inhibited viral growth ofmore than 50 %. Cellular internalization assays highlighted that untreated infected cells showed high intensity of fluorescence emission, which denotes high level of viral internalization. Conversely, AgNP-treated infected cells showed reduced levels of fluorescence, failing to show significant viral load. Overall, our study showed that alga-mediated synthesis of metal nanoparticles may be considered to develop newer, safer, and cheap tools in the fight against the dengue virus, serotype DEN-2, and its vector A. aegypti, with little cytotoxicity on mammalian cells

Genetic deviation in geographically close populations of the dengue vector Aedes aegypti (Diptera: Culicidae): influence of environmental barriers in South India

Mosquitoes are vectors of devastating pathogens and parasites, causing millions of deaths every year. Dengue is a mosquito-borne viral infection found in tropical and subtropical regions around the world. Recently, dengue transmission has strongly increased in urban and semiurban areas, becoming a major international public health concern. Aedes aegypti (Diptera: Culicidae) is a primary vector of dengue. Shedding light on genetic deviation in A. aegypti populations is of crucial importance to fully understand their molecular ecology and evolution. In this research, haplotype and genetic analyses were conducted using individuals of A. aegypti from 31 localities in the north, southeast, northeast and central regions of Tamil Nadu (South India). The mitochondrial DNA region of cytochrome c oxidase 1 (CO1) gene was used as marker for the analyses. Thirty-one haplotypes sequences were submitted to GenBank and authenticated. The complete haplotype set included 64 haplotypes from various geographical regions clustered into three groups (lineages) separated by three fixed mutational steps, suggesting that the South Indian Ae. aegypti populations were pooled and are linked with West Africa, Columbian and Southeast Asian lineages. The genetic and haplotype diversity was low, indicating reduced gene flow among close populations of the vector, due to geographical barriers such as water bodies. Lastly, the negative values for neutrality tests indicated a bottle-neck effect and supported for low frequency of polymorphism among the haplotypes. Overall, our results add basic knowledge to molecular ecology of the dengue vector A. aegypti, providing the first evidence for multiple introductions of Ae. aegypti populations from Columbia and West Africa in South India

Nanofabrication of Graphene Quantum Dots with High Toxicity Against Malaria Mosquitoes, Plasmodium falciparum and MCF-7 Cancer Cells: Impact on Predation of Non-target Tadpoles, Odonate Nymphs and Mosquito Fishes

Recently, it has been highlighted an overlooked connection between the biting activity of Anopheles mosquitoes and the spread of cancer. The excellent physico-chemical properties of graphene quantum dots (GQDs) make them a suitable candidate for biomedical applications. We focused on the toxicity of GQDs against Plasmodium falciparum and its vector Anopheles stephensi, and their impact on predation of non-target mosquito predators. Biophysical methods, including UV–vis, photoluminescence, FTIR and Raman spectroscopy, XRD analysis and TEM, confirmed the effective GQD nanosynthesis. LC50 against A. stephensi ranged from 0.157 (larva I) to 6.323 ppm (pupa). The antiplasmodial activity of GQDs was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of P. falciparum. IC50 were 82.43 (CQ-s) and 85.17 μg/ml (CQ-r). In vivo experiments conducted on Plasmodium berghei infecting albino mice showed moderate activity of GQDs if compared to chloroquine. Concerning non-target effects, the predation efficiency of Gambusia affinis, Anax immaculifrons and Hoplobatrachus tigerinus post-treatment with GQDs was enhanced. Lastly, GQDs were toxic against MCF-7 breast cancer cell lines with an IC50 = 24.81 μg/ml, triggering apoptosis in treated cells. Overall, we highlighted the multipurpose potential of GQDs for the development of newer drugs in the fight against Anopheles vectors, Plasmodium parasites and breast cancer cells

DNA barcoding and molecular evolution of mosquito vectors of medical and veterinary importance

Mosquitoes (Diptera: Culicidae) are a key threat for millions of people worldwide, since they act as vectors for devastating pathogens and parasites. The standard method of utilisation of morphological characters becomes challenging due to various factors such as phenotypical variations. We explored the complementary approach of CO1 gene-based identification, analysing ten species of mosquito vectors belonging to three genera, Aedes, Culex and Anopheles from India. Analysed nucleotide sequences were found without pseudo genes and indels; they match with high similarity in nucleotide Basic Local Alignment Search Tool (BLASTn) search. The partial CO1 sequence of Anopheles niligricus was the first time record submitted to National Center for Biotechnology Information (NCBI). Mean intra- and interspecies divergence was found to be 1.30 and 3.83 %, respectively. The congeneric divergence was three times higher than the conspecifics. Deep intraspecific divergence was noted in three of the species, and the reason could be explained more accurately in the future by improving the sample size across different locations. The transitional and transversional substitutions were tested individually. Ts and Tv substitutions in all the 1st, 2nd and 3rd codons were estimated to be (0.44, 99.51), (40.35, 59.66) and (59.16, 40.84), respectively. Saturation of the sequences was resolved, since both the Ts and Tv exhibited a linear relationship suggesting that the sequences were not saturated. NJ and ML tree analysis showed that the individuals of the same species clustered together based on the CO1 sequence similarity, regardless of their collection site and geographic location. Overall, this study adds basic knowledge to molecular evolution of mosquito vectors of medical and veterinary importance and may be useful to improve biotechnological tools employed in Culicidae control programmes