Abstract
Mosquitoes (Diptera: Culicidae) serve as important vectors for a wide number of parasites and pathogens of huge medical and veterinary importance. Aedes aegypti is a primary dengue vector in tropical and subtropical urban areas. There is an urgent need to develop eco-friendly mosquitocides. In this study, silver nanoparticles (AgNP) were biosynthesized using neem cake, a by-product of the neem oil extraction from the seed kernels of Azadirachta indica. AgNP were characterized using a variety of biophysical methods, including UV–vis spectrophotometry, FTIR, SEM, EDX, and XRD analyses. Furthermore, the neem cake extract and the biosynthesized AgNP were tested for acute toxicity against larvae and pupae of the dengue vector Ae. aegypti. LC50 values achieved by the neem cake extract ranged from 106.53 (larva I) to 235.36 ppm (pupa), while AgNP LC50 ranged from 3.969 (larva I) to 8.308 ppm (pupa). In standard laboratory conditions, the predation efficiency of a Carassius auratus per day was 7.9 (larva II) and 5.5 individuals (larva III). Post-treatment with sub-lethal doses of AgNP, the predation efficiency was boosted to 9.2 (larva II) and 8.1 individuals (larva III). The genotoxic effect of AgNP was studied on C. auratus using the comet assay and micronucleus frequency test. DNA damage was evaluated on peripheral erythrocytes sampled at different time intervals from the treatment; experiments showed no significant damages at doses below 12 ppm. Overall, this research pointed out that neem cake-fabricated AgNP are easy to produce, stable over time, and can be employed at low dosages to reduce populations of dengue vectors, with moderate detrimental effects on non-target mosquito natural enemies.
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Abdel-Ghaffar F, Al-Quraishy S, Sobhy H, Semmler M (2008) Neem seed extract shampoo, Wash Away Louse®, an effective plant agent against Sarcoptes scabiei mite infesting dogs in Egypt. Parasitol Res 104:145–148
Abdel-Ghaffar F, Semmler M, Al-Rasheid KAS, Mehlhorn H (2009) In vitro efficacy of ByeMite® and Mite-Stop® on developmental stages of the red chicken mite Dermanyssus gallinae. Parasitol Res 105:469–471
Abdel-Ghaffar F, Al-Quraishy S, Al-Rasheid KAS, Mehlhorn H (2012) Efficacy of a single treatment of head lice with a neem seed extract: an in vivo and in vitro study on nits and motile stages. Parasitol Res 110:277–280
Al-Quraishy S, Abdel-Ghaffar F, Al-Rasheid KAS, Mehlhorn J, Mehlhorn H (2011) Effects of a neem seed extract (MiteStop®) on mallophages (featherlings) of chicken: in-vivo and in-vitro studies. Parasitol Res 110:617–622
Al-Quraishy S, Abdel-Ghaffar F, Al-Rasheid KAS, Mehlhorn J, Mehlhorn H (2012) Observations on effects of a neem seed extract (MiteStop®) on biting lice (mallophages) and bloodsucking insects parasitizing horses. Parasitol Res 110:335–339
Al-Sabti K (1995) An in vitro binucleated blocked hepatic cell technique for genotoxicity testing in fish. Mutat Res 335:109–120
Amer A, Mehlhorn H (2006a) Repellency effect of forty-one essential oils against Aedes, Anopheles and Culex mosquitoes. Parasitol Res 99:478–490
Amer A, Mehlhorn H (2006b) The sensilla of Aedes and Anopheles mosquitoes and their importance in repellency. Parasitol Res 99:491–499
Amer A, Mehlhorn H (2006c) Larvicidal effects of various essential oils against Aedes, Anopheles, and Culex larvae (Diptera, Culicidae). Parasitol Res 99:466–472
Amer A, Mehlhorn H (2006d) Persistency of larvicidal effects of plant oil extracts under different storage conditions. Parasitol Res 99:473–477
Arjunan NK, Murugan K, Rejeeth C, Madhiyazhagan P, Barnard DR (2012) Green synthesis of silver nanoparticles for the control of mosquito vectors of malaria, filariasis and dengue. Vector Borne Zoontic Dis 12:262–268
Ateeq B, Abul Farah M, Ahmad W (2005) Detection of DNA damage by alkaline single cell gel electrophoresis in 2,4-dichlorophenoxyacetic-acid and butachlor-exposed erythrocytes of Clarias batrachus. Ecotoxicol Environ Saf 62:348–354
Bankar A, Joshi B, Ravi Kumara A, Zinjarde S (2010) Banana peel extract mediated novel route for the synthesis of silver nanoparticles. Colloids Surf A Physicochem Eng Asp 368:58–63
Baun A, Hartmann NB, Grieger K, Kusk KO (2008) Ecotoxicity of engineered nanoparticles to aquatic invertebrates: a brief review and recommendations for future toxicity testing. Ecotoxicology 17:387–395
Benelli G (2015a) Research in mosquito control: current challenges for a brighter future. Parasitol Res 114:2801–2805
Benelli G (2015b) Plant-borne ovicides in the fight against mosquito vectors of medical and veterinary importance: a systematic review. Parasitol Res 114:3201–3212
Benelli G (2016) Plant-mediated biosynthesis of nanoparticles as an emerging tool against mosquitoes of medical and veterinary importance: a review. Parasitol Res. doi:10.1007/s00436-015-4800-9
Benelli G, Conti B, Garreffa R, Nicoletti M (2014) Shedding light on bioactivity of botanical by-products: neem cake compounds deter oviposition of the arbovirus vector Aedes albopictus (Diptera: Culicidae) in the field. Parasitol Res 113:933–940
Benelli G, Murugan K, Panneerselvam C, Madhiyazhagan P, Conti B, Nicoletti M (2015a) Old ingredients for a new recipe? Neem cake, a low-cost botanical by-product in the fight against mosquito-borne diseases. Parasitol Res 114:391–397
Benelli G, Bedini S, Cosci F, Toniolo C, Conti B, Nicoletti M (2015b) Larvicidal and ovideterrent properties of neem oil and fractions against the filariasis vector Aedes albopictus (Diptera: Culicidae): a bioactivity survey across production sites. Parasitol Res 114:227–236
Carrasco KR, Tilbury KL, Myers MS (1990) Assessment of the piscine micronucleus test as an in situ biological indicator of chemical contaminant effects. Can J Fish Aquat Sci 47:2123–2136
Cavas T, Konen S (2007) Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carassius auratus) exposed to a glyphosate formulation using the micronucleus test and the comet assay. Mutagen 22:263–268
Chandra G, Bhattacharjee I, Chatterjee SN, Ghosh A (2008) Mosquito control by larvivorous fish. Indian J Med Res 127:13–27
Countryman PI, Heddle JA (1976) The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res 41:321–332
De Miranda CG, Barreto RE, Speit G, ValenzuelaReyes VA, Volpato GL, Favero Salvadori DM (2003) Anesthesia of fish with benzocaine does not interfere with comet assay results. Mutat Res 534(1–2):165–172
Deguchi Y, Toyoizumi T, Masuda S, Yasuhara A, Mohri S, Yamada M, Inoue Y, Kinae N (2007) Evaluation of mutagenic activities of leachates in landfill sites by micronucleus test and comet assay using goldfish. Mutat Res 627:178–185
Dinesh D, Murugan K, Madhiyazhagan P, Panneerselvam C, Nicoletti M, Jiang W, Benelli G, Chandramohan B, Suresh U (2015) Mosquitocidal and antibacterial activity of green-synthesized silver nanoparticles from Aloe vera extracts: towards an effective tool against the malaria vector Anopheles stephensi? Parasitol Res 114:1519–1529
Elechiguerra JL, Burt JL, Morones JR, Camacho-Bragado A, Gao X, Lara HH, Yacaman JM (2005) Interaction of silver nanoparticles with HIV-1. J Nanobiotechnol 29:3–6
Fabrega J, Luoma SN, Tyler CR, Galloway TS, Lead JR (2011) Silver nanoparticles: behaviour and effects in the aquatic environment. Environ Int 37:517–531
Fayaz AM, Balaji K, Girilal MYR, Kalaichelvan PT, Venketesan R (2010) Biogenic synthesis of silver nanoparticles and their synergistic effect with antibiotics: a study against gram-positive and gram negative bacteria. Nanomed Nanotechnol Biol Med 6:103–109
Fenech M (1993) The cytokinesis-block micronucleus technique and its application to genotoxicity studies in human populations. Environ Health Perspect Suppl 101(3):101–107
Finney DJ (1971) Probit analysis. Cambridge University, London, pp 68–78
Gratz NG, Pal R (1988) Malaria vector control: larviciding. In: Wernsdorfer WH, McGregor I (eds) Malaria: principles and practice of malariology. Churchill Livingstone, Edinburgh, pp 1213–1226
Hemingway J, Ranson H (2000) Insecticide resistance in insect vectors of human disease. Annu Rev Entomol 45:371–391
Hurst TP, Brown MD, Kay BH (2004) Laboratory evaluation of the predation efficacy of native Australian fish on Culex annulirostris (Diptera: Culicidae). J Am Mosq Control Assoc 20:286–291
Jarvis TA, Miller RJ, Lenihan HS, Bielmyer GK (2013) Toxicity of ZnO nanoparticles to the copepod Acartia tonsa, exposed through a phytoplankton diet. Environ Toxicol Chem 32:1264–1269
Khandelwal N, Abhijeet S, Devendra J, Upadhyay MK, Verma HN (2010) Green synthesis of silver nanoparticles using Argimone mexicana leaf extract and evaluation of their antimicrobial activities. Digest J Nanomater Biostruct 5:483–489
Kim HC, Kim MS, Yu HS (1994) Biological control of vector mosquitoes by the use of fish predators, Moroco oxycephalus and Misgurnus anguillicandatus in the laboratory and semi field drive paddy. Korean J Entomol 24:269–284
Locher N, Al-Rasheid KAS, Abdel-Ghaffar F, Mehlhorn H (2010) In vitro and field studies on the contact and fumigant toxicity of a neem-product (Mite-Stop®) against the developmental stages of the poultry red mite Dermanyssus gallinae. Parasitol Res 107:417–423
Madhiyazhagan P, Murugan K, Naresh Kumar A, Nataraj T, Dinesh D, Panneerselvam C, Subramaniam J, Mahesh Kumar P, Suresh U, Roni M, Nicoletti M, Alarfaj AA, Higuchi A, Nicoletti M, Munusamy MA, Benelli G (2015) Sargassum muticum-synthesized silver nanoparticles: an effective control tool against mosquito vectors and bacterial pathogens. Parasitol Res. doi:10.1007/s00436-015-4671-0
Magudapathy P, Gangopadhyay PBK, Nair KGM, Dhara S (2001) Electrical transport studies of Ag nanoclusters embedded in glass matrix. Physica 299:142–146
Marimuthu S, Rahuman AA, Rajakumar G, Santhoshkumar T, Kirthi AV, Jayaseelan C, Bagavan A, Zahir AA, Elango G, Kamaraj C (2011) Evaluation of green synthesized silver nanoparticles against parasites. Parasitol Res 10:2212–2224
Mehlhorn H (ed) (2011) Nature helps. How plants and other organisms contribute to solve health problems. Parasitology Research Monographs. Springer, Berlin, pp 1–372
Mehlhorn H, Schmahl G, Schmidt J (2005) Extract of the seeds of the plant Vitex agnus castus proven to be highly efficacious as a repellent against ticks, fleas, mosquitoes and biting flies. Parasitol Res 95:363–365
Mehlhorn H, Al-Rasheid KA, Al-Quraishy S, Abdel-Ghaffar F (2012) Research and increase of expertise in arachno-entomology are urgently needed. Parasitol Res 110:259–265
Mohanpuria P, Rana NK, Yadav SK (2008) Biosynthesis of nanoparticles: technological concepts and future applications. J Nanopart Res 10:507–517
Mukherjee P, Roy M, Mandal BP, Dey GK, Mukherjee PK, Ghatak J, Tyagia K, Kale SP (2008) Green synthesis of highly stabilized nanocrystalline silver particles by a non-pathogenic and agriculturally important fungus T. asperellum. Nanotech 19:075103
Mulla MS (1961) Mosquito control investigations with emphasis on integration of chemical and biological control in mosquito abatement. 29th Proc Pap Ann Con Cal Mosq Control Assoc 1–4
Murrell S, Wu SC, Butler M (2011) Review of dengue virus and the development of a vaccine. Biotechnol Adv 29:239–247
Murugan K, Benelli G, Ayyappan S, Dinesh D, Panneerselvam C, Nicoletti M, Hwang JS, Mahesh Kumar P, Subramaniam J, Suresh U (2015a) Toxicity of seaweed-synthesized silver nanoparticles against the filariasis vector Culex quinquefasciatus and its impact on predation efficiency of the cyclopoid crustacean Mesocyclops longisetus. Parasitol Res. doi:10.1007/s00436-015-4417-z
Murugan K, Benelli G, Panneerselvam C, Subramaniam J, Jeyalalitha T, Dinesh D, Nicoletti M, Hwang JS, Suresh U, Madhiyazhagan P (2015b) Cymbopogon citratus-synthesized gold nanoparticles boost the predation efficiency of copepod Mesocyclops aspericornis against malaria and dengue mosquitoes. Exp Parasitol 153:129–138
Murugan K, Priyanka V, Dinesh D, Madhiyazhagan P, Panneerselvam C, Subramaniam J, Suresh U, Chandramohan B, Roni M, Nicoletti M, Alarfaj AA, Higuchi A, Munusamy MA, Khater HF, Messing RH, Benelli G (2015c) Enhanced predation by Asian bullfrog tadpoles, Hoplobatrachus tigerinus, against the dengue vector Aedes aegypti in an aquatic environment treated with mosquitocidal nanoparticles. Parasitol Res. doi:10.1007/s00436-015-4582-0
Murugan K, Aamina Labeeba M, Panneerselvam C, Dinesh D, Suresh U, Subramaniam J, Madhiyazhagan P, Hwang JS, Wang L, Nicoletti M, Benelli G (2015d) Aristolochia indica green-synthesized silver nanoparticles: a sustainable control tool against the malaria vector Anopheles stephensi? Res Vet Sci 102:127–135
Murugan K, Dinesh D, Paulpandi M, Meqbel Althbyani AD, Subramaniam J, Madhiyazhagan P, Wang L, Suresh U, Mahesh Kumar P, Mohan J, Rajaganesh R, Wei H, Kalimuthu K, Parajulee MN, Mehlhorn H, Benelli G (2015e) Nanoparticles in the fight against mosquito-borne diseases: bioactivity of Bruguiera cylindrica-synthesized nanoparticles against dengue virus DEN-2 (in vitro) and its mosquito vector Aedes aegypti (Diptera: Culicidae). Parasitol Res. doi:10.1007/s00436-015-4676-8
Murugan K, Sanoopa CP, Madhiyazhagan P, Dinesh D, Subramaniam J, Panneerselvam C, Roni M, Suresh U, Nicolettib, Alarfajc AA, Munusamyc MA, Higuchi A, Kumar S, Haribalan P, Ahn YJ, Benelli G (2015f) Rapid biosynthesis of silver nanoparticles using Crotalaria verrucosa leaves against the dengue vector Aedes aegypti: what happens around? An analysis of dragonfly predatory behaviour after exposure at ultra-low doses. Nat Prod Res. doi:10.1080/1478641920151074230
Nagajyothi PC, Sreekanth TVM, Lee JL, Lee KD (2014) Mycosynthesis: antibacterial, antioxidant and antiproliferative activities of silver nanoparticles synthesized from Inonotus obliquus (Chaga mushroom) extract. J Photochem Photobiol B Biol 130:299–304
Nicoletti M, Serafini M, Aliboni A, D’Andrea A, Mariani S (2010) Toxic effects of neem cake extracts on Aedes albopictus larvae. Parasitol Res 107:89–84
Oberdorster E, Zhu S, Michelle Blickley T, McClellan-Green P, Haasch ML (2006) Ecotoxicology of carbon-based engineered nanoparticles: effects of fullerene (C60) on aquatic organisms. Carbon 44:1112–1120
Panneerselvam C, Murugan K, Kovendan K, Mahesh Kumar P, Subramaniam J (2013) Mosquito larvicidal and pupicidal activity of Euphorbia hirta Linn. (Family: Euphorbiaceae) and Bacillus sphaericus against Anopheles stephensi (L) (Diptera: Culicidae). (Diptera: Culicidae). Asian Pac J Trop Med 6:102–109
Park J, Kim S, Yoo J, Lee JS, Park JW, Jung J (2014) Effect of salinity on acute copper and zinc toxicity to Tigriopus japonicus: the difference between metal ions and nanoparticles. Mar Pollut Bull 85:526–531
Patil CD, Patil SV, Borase HP, Salunke BK, Salunkhe RB (2012a) Larvicidal activity of silver nanoparticles synthesized using Plumeria rubra plant latex against Aedes aegypti and Anopheles stephensi. Parasitol Res 110:1815–1822
Patil CD, Borase HP, Patil SV, Salunkhe RB, Salunke BK (2012b) Larvicidal activity of silver nanoparticles synthesized using Pergularia daemia plant latex against Aedes aegypti and Anopheles stephensi and nontarget fish Poecillia reticulata. Parasitol Res 111:555–562
Pavela R (2015) Essential oils for the development of eco-friendly mosquito larvicides: a review. Ind Crops Prod 76:174–187
Prakash P, Gnanaprakasam P, Emmanuel R, Arokiyarajb S, Saravanan M (2013) Green synthesis of silver nanoparticles from leaf extract of Mimusops elengi, Linn for enhanced antibacterial activity against multi drug resistant clinical isolates. Coll Surf B Biointerfaces 108:255–259
Rai M, Yadav A, Gade A (2009) Silver nanoparticles as a new generation of antimicrobials. Biotechnol Adv 27:76–83
Rajan R, Chandran K, Harper SL, Yun SI, Kalaichelvan PT (2015) Plant extract synthesized nanoparticles: an ongoing source of novel biocompatible materials. Ind Crop Prod 70:356–373
Rao JV, Kavitha P (2010) In vitro effects of chlorpyrifos on the acetylcholinesterase activity of euryhaline fish, Oreochromis mossambicus Z. Naturforsch C 65:303–306
Rawani A, Ghosh A, Chandra G (2013) Mosquito larvicidal and antimicrobial activity of synthesized nano-crystalline silver particles using leaves and green berry extract of Solanum nigrum L (Solanaceae: Solanales). Acta Trop 128:613–622
Roni M, Murugan K, Panneerselvam C, Subramaniam J, Nicoletti M, Madhiyazhagan P, Dinesh D, Suresh U, Khater HF, Wei H, Canale A, Alarfaj AA, Munusamy MA, Higuchi A, Benelli G (2015) Characterization and biotoxicity of Hypnea musciformis synthesized silver nanoparticles as potential eco-friendly control tool against Aedes aegypti and Plutella xylostella. Ecotoxicol Environ Saf. doi:10.1016/jecoenv201507005
Sathyavathi R, Balamurali Krishna M, Venugopal Rao S, Saritha R, Narayana Rao D (2010) Biosynthesis of silver nanoparticles using Coriandrum sativum leaf extract and their application in nonlinear optics. Adv Sci Lett 3:1–6
Schmahl G, Al-Rasheid KAS, Abdel-Ghaffar F, Klimpel S, Mehlhorn H (2010) The efficacy of neem seed extracts (Tresan®, MiteStop®) on a broad spectrum of pests and parasites. Parasitol Res 107:261–269
Semmler M, Abdel-Ghaffar F, Al-Rasheid KAS, Mehlhorn H (2009) Nature helps: from research to products against blood sucking arthropods. Parasitol Res 105:1483–1487
Shankar SS, Rai A, Ahmad A, Sastry M (2004) Biosynthesis of silver and gold nanoparticles from extracts of different parts of the Geranium plant. Appl Nanotechnols 1:69–77
Soni N, Prakash S (2014) Microbial synthesis of spherical nanosilver and nanogold for mosquito control. Ann Microbiol 64:1099–1111
Subarani S, Sabhanayakam S, Kamaraj C (2013) Studies on the impact of biosynthesized silver nanoparticles (AgNPs) in relation to malaria and filariasis vector control against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae). Parasitol Res 112:487–499
Subramaniam J, Murugan K, Panneerselvam C, Kovendan K, Madhiyazhagan P, Mahesh Kumar P, Dinesh D, Chandramohan B, Suresh U, Nicoletti M, Higuchi A, Hwang JS, Kumar S, Alarfaj AA, Munusamy MA, Messing RH (2015) Eco-friendly control of malaria and arbovirus vectors using the mosquitofish Gambusia affinis and ultra-low dosages of Mimusops elengi-synthesized silver nanoparticles: towards an integrative approach? Environ Sci Pollut Res. doi:10.1007/s11356-015-5253-5
Sujitha V, Murugan K, Paulpandi M, Panneerselvam C, Suresh U, Roni M, Nicoletti M, Higuchi A, Madhiyazhagan P, Subramaniam J, Dinesh D, Vadivalagan C, Chandramohan B, Alarfaj AA, Munusamy MA, Barnard DR, Benelli G (2015) Green-synthesized silver nanoparticles as a novel control tool against dengue virus (DEN-2) and its primary vector Aedes aegypti. Parasitol Res 114:3315–3325
Suman TY, Radhika Rajasree SR, Kanchanab A, Beena Elizabethc SS (2013) Biosynthesis, characterization and cytotoxic effect of plant mediated silver nanoparticles using Morinda citrifolia root extract. Colloids Surf B Biointerface 106:74–78
Suresh U, Murugan K, Benelli G, Nicoletti M, Barnard DR, Panneerselvam C, Mahesh Kumar P, Subramaniam J, Dinesh D, Chandramohan B (2015) Tackling the growing threat of dengue: Phyllanthus niruri-mediated synthesis of silver nanoparticles and their mosquitocidal properties against the dengue vector Aedes aegypti (Diptera: Culicidae). Parasitol Res 114:1551–1562
Theodorakis CW, D’Surney SJ, Shugart LR (1994) Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis. Environ Toxicol Chem 13(7):1023–1031
Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221
Tietze NS, Hester PG, Olson MA, Hallmon CF, Shaffer KR (1994) Acute toxicity of mosquito control compounds to Cyprinodon variegates and Menidia beryllina laboratory and field tests. J Fla Anti-Mosq 65:37–44
Verkerk RHJ, Wright DJ (1993) Biological activity of neem seed kernel extract and synthetic azadirachtin against larvae of Plutella xylostella. Pestic Sci 37:83–91
Vinayaga Moorthi P, Balasubramanian C, Mohan S (2015) An improved insecticidal activity of silver nanoparticle synthesized by using Sargassum muticum. Appl Biochem Biotechnol 175:135–140
Walldorf V, Mehlhorn H, Al-Quraishy S, Al-Rasheid KAS, Abdel-Ghaffar F, Mehlhorn J (2012) Treatment with a neem seed extract (MiteStop®) of beetle larvae parasitizing the plumage of poultry. Parasitol Res 110:623–627
WHO (2015) Dengue and severe dengue. Fact sheet N°117. World Health Organization, Geneva
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We thank the financial support by King Saud University, Saudi Arabia, Vice Deanship of Research Chairs: Research Chairs.
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The authors declare no conflicts of interest. G. Benelli is an Editorial Board Member of Parasitology Research. This does not alter the author’s adherence to all the Parasitology Research policies on sharing data and materials.
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Chandramohan, B., Murugan, K., Panneerselvam, C. et al. Characterization and mosquitocidal potential of neem cake-synthesized silver nanoparticles: genotoxicity and impact on predation efficiency of mosquito natural enemies. Parasitol Res 115, 1015–1025 (2016). https://doi.org/10.1007/s00436-015-4829-9
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DOI: https://doi.org/10.1007/s00436-015-4829-9