Environ Sci Pollut Res (2016) 23:16671–16685 DOI 10.1007/s11356-016-6832-9 RESEARCH ARTICLE Eco-friendly drugs from the marine environment: spongeweed-synthesized silver nanoparticles are highly effective on Plasmodium falciparum and its vector Anopheles stephensi, with little non-target effects on predatory copepods Kadarkarai Murugan1,2 & Chellasamy Panneerselvam3 & Jayapal Subramaniam1 & Pari Madhiyazhagan1 & Jiang-Shiou Hwang4 & Lan Wang5 & Devakumar Dinesh1 & Udaiyan Suresh1 & Mathath Roni1 & Akon Higuchi6 & Marcello Nicoletti7 & Giovanni Benelli8,9 Received: 13 April 2016 /Accepted: 4 May 2016 /Published online: 16 May 2016 # Springer-Verlag Berlin Heidelberg 2016 Abstract Mosquitoes act as vectors of devastating pathogens (EDX), and X-ray diffraction (XRD). In mosquitocidal assays, and parasites, representing a key threat for millions of humans the 50 % lethal concentration (LC50)ofC. tomentosum extract and animals worldwide. The control of mosquito-borne dis- against Anopheles stephensi ranged from 255.1 (larva I) to eases is facing a number of crucial challenges, including the 487.1 ppm (pupa). LC50 of C. tomentosum-synthesized emergence of artemisinin and chloroquine resistance in AgNP ranged from 18.1 (larva I) to 40.7 ppm (pupa). In lab- Plasmodium parasites, as well as the presence of mosquito oratory, the predation efficiency of Mesocyclops aspericornis vectors resistant to synthetic and microbial pesticides. copepods against A. stephensi larvae was 81, 65, 17, and 9 % Therefore, eco-friendly tools are urgently required. Here, a (I, II, III, and IV instar, respectively). In AgNP contaminated synergic approach relying to nanotechnologies and biological environment, predation was not affected; 83, 66, 19, and 11 % control strategies is proposed. The marine environment is an (I, II, III, and IV). The anti-plasmodial activity of outstanding reservoir of bioactive natural products, which C. tomentosum extract and spongeweed-synthesized AgNP have many applications against pests, parasites, and patho- was evaluated against CQ-resistant (CQ-r) and CQ-sensitive gens. We proposed a novel method of seaweed-mediated syn- (CQ-s) strains of Plasmodium falciparum. Fifty percent inhib- thesis of silver nanoparticles (AgNP) using the spongeweed itory concentration (IC50)ofC. tomentosum were 51.34 μg/ml Codium tomentosum, acting as a reducing and capping agent. (CQ-s) and 65.17 μg/ml (CQ-r); C. tomentosum-synthesized AgNP were characterized by UV–Vis spectroscopy, Fourier AgNP achieved IC50 of 72.45 μg/ml (CQ-s) and 76.08 μg/ml transform infrared (FTIR) spectroscopy, scanning electron mi- (CQ-r). Furthermore, low doses of the AgNP inhibited the croscopy (SEM), energy-dispersive X-ray spectroscopy growth of Bacillus subtilis, Klebsiella pneumoniae, and Responsible editor: Philippe Garrigues * Giovanni Benelli 5 School of Life Science and Technology, Shanxi University, [email protected]; [email protected] Taiyuan 030006, China 6 Department of Chemical and Materials Engineering, National 1 Division of Entomology, Department of Zoology, School of Life Central University, No. 300 Jhongli, Taoyuan 32001, Taiwan Sciences, Bharathiar University, Coimbatore, Tamil Nadu 641046, India 7 Department of Environmental Biology, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy 2 Department of Biotechnology, Thiruvalluvar University, Serkkadu, Vellore, Tamil Nadu 632115, India 8 Insect Behaviour Group, Department of Agriculture, Food and 3 Biology Department, Faculty of Science, University of Tabuk, Environment, University of Pisa, via del Borghetto 80, Tabuk, Saudi Arabia 56124 Pisa, Italy 4 Institute of Marine Biology, National Taiwan Ocean University, 9 The BioRobotics Institute, Sant’Anna School of Advanced Studies, Keelung 20224, Taiwan Viale Rinaldo Piaggio 34, 56025 Pontedera, Italy 16672 Environ Sci Pollut Res (2016) 23:16671–16685 Salmonella typhi, using the agar disk diffusion and minimum microbial pesticides (Hemingway and Ranson 2000; inhibitory concentration protocol. Overall, C. tomentosum Aziz et al. 2016; Naqquash et al. 2016). Therefore, eco- metabolites and spongeweed-synthesized AgNP may be po- friendly tools are urgently required (Rajkumar and Jebanesan tential candidates to develop novel and effective tools in the 2008; Azizullah et al. 2014; Benelli et al. 2015a, b, c; Pavela fight against Plasmodium parasites and their mosquito vec- 2015a, b; Govindarajan and Benelli 2016). Notably, botani- tors. The employ of ultra-low doses of nanomosquitocides in cals have been used by human communities in different parts synergy with cyclopoid crustaceans seems a promising green of the world as mosquitocidals, adult repellents, and oviposi- route for effective mosquito control programs. tion deterrents, against a wide number of mosquito species (e.g., Benelli 2015a, b). People entering into regions where Keywords Anopheles stephensi . Codium tomentosum . dengue, malaria, or yellow fever risks exist may protect them- Malaria . Mosquito-borne diseases . Plasmodium falciparum . selves by use of plant-derived repellents (Mehlhorn et al. Mesocyclops aspericornis . Anti-bacterial activity 2012;AmerandMehlhorn2006a, b). On the other hand, people living in endemic regions have to protect themselves by several strategies at the same time, since infection rates of Introduction mosquitoes may be extremely high (Amer and Mehlhorn 2006c, d; Semmler et al. 2009;Benelli2015a). Arthropods are dangerous vectors of deadly pathogens and Biological control of mosquito vectors using predatory parasites (Mehlhorn et al. 2012; Benelli 2015a). Among them, copepods, tadpoles, and fishes also received attention mosquitoes (Diptera: Culicidae) represent a key threat for mil- (Marten 2000; Murugan et al. 2015a, b, c; Subramaniam lions of humans and animals worldwide. Their medical and et al. 2016a, b). Predation is an important factor for the veterinary importance is mainly due to the fact that their act as maintenance of trophic equilibrium of ecological commu- vectors for a number of pathogens and parasites of public nities, including aquatic ones. Good examples are odonate health relevance, including malaria, avian malaria, yellow fe- young instars, water bugs, tadpoles, fishes, crabs, and co- ver, dengue, Japanese encephalitis, Zika virus, Rift Valley pepods acting as natural enemies of mosquito young in- fever, Western equine encephalomyelitis, bancroftian and stars (Bowatte et al. 2013; Murugan et al. 2015d, e, f, g brugian filariae, canine heartworm disease (Dirofilaria 2016). Cyclopoid copepods are crucial invertebrate pred- immitis), and setariosis (Setaria spp.) (Benelli and Mehlhorn ators in zooplankton communities, and are frequently 2016; Benelli et al. 2016). present in lakes, drains, ponds, and reservoirs (Chang Malaria is caused by Plasmodium parasites, mainly and Hanazato 2003; Kumar and Hwang 2006). The Plasmodium falciparum, Plasmodium vivax, Plasmodium cyclopoid Mesocyclops aspericornis is a generalist preda- ovale, and Plasmodium malariae. In addition, periodic reports tor that preys on a wide spectrum of food types, ranging highlighted the presence of simian malaria parasites found in from rotifers, cladocerans, mosquito larvae, and fish humans, most of them implicating Plasmodium knowlesi. young instars (Kumar et al. 2012). More generally, several Plasmodium parasites are vectored to vertebrates through the species of copepods, including M. aspericornis, Mesocyclops bites of infected Anopheles mosquitoes, which bite mainly guangxiensis, Mesocyclops longisetus,andMesocyclops between dusk and dawn. According to the latest estimates, thermocyclopoides, have been reported as potential biological there were about 198 million cases of malaria in 2013 and control agents against mosquitoes (Rawlins et al. 1997; an estimated 584,000 deaths. In Africa, most deaths occur Manrique-Saide et al. 1998; Schaper 1999; Murugan et al. among children living where a child dies every minute from 2015b). malaria, although mortality rates among children have been Nanotechnology has the potential to revolutionize a wide reduced by an estimated 58 % since 2000 (Mehlhorn 2008; array of applications in the fields of catalysis, sensors, opto- WHO 2016). In addition, besides the fall of malaria infection electronics, magnetic devices, drug delivery, anti-microbials, rates worldwide, with special reference to sub-Saharan Africa, pest management, and parasitology (Scrinis and Lyons 2007; 2015 was an annus mirabilis for malaria control, due to the Haverkamp 2010, Govindarajan et al. 2016; Kumar et al. Nobel Prize to the Chinese scientist Youyou Tu for the dis- 2016). The utilization of plants for nanoparticle synthesis covery of artemisinin and the development of the first vaccine can be advantageous over other biological processes, because against P. falciparum malaria [i.e., RTS,S/AS01 (RTS,S)] it eliminates the elaborate process of maintaining cell cultures (Benelli and Mehlhorn 2016). and can also be suitably scaled up for large-scale nanoparticle However, malaria prevention and control is facing a synthesis (Shankar et al. 2004; Chandramohan et al. number of crucial challenges, including the emergence 2016;Jaganathanetal.2016). Recently, several plants have of artemisinin and chloroquine resistance in Plasmodium been screened for efficient and rapid extracellular synthesis of parasites (Benelli and Mehlhorn 2016), as well as the metal nanoparticles with different mosquitocidal and anti- presence of mosquito strains resistant to synthetic