{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T14:49:21Z","timestamp":1792507761728,"version":"4.5.0"},"reference-count":41,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,12,7]],"date-time":"2025-12-07T00:00:00Z","timestamp":1765065600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"PAPIIT-UNAM","award":["TA200324"],"award-info":[{"award-number":["TA200324"]}]},{"name":"SECIHTI","award":["CBF-2025-I-247"],"award-info":[{"award-number":["CBF-2025-I-247"]}]},{"name":"SECIHTI","award":["CBF-2025-I-1297"],"award-info":[{"award-number":["CBF-2025-I-1297"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Parasitic infections, particularly those caused by Haemonchus contortus (H. contortus), severely impact livestock production, with growing resistance to commercial anthelmintics posing a major challenge. Green-synthesized metallic nanoparticles using Artemisia cina (A. cina), a plant with known anthelmintic and antioxidant properties, represent a promising sustainable alternative for parasite control. In this study, silver nanoparticles (AgNPs) were synthesized using an aqueous extract of A. cina to evaluate their anthelmintic activity against infective larvae (L3) of H. contortus and their effect on the expression of oxidative stress-related genes. The larval mortality bioassay was conducted in 96-well microtiter plates by incubating L3 larvae with increasing AgNP concentrations for 24 h. To investigate oxidative stress responses, larvae were exposed to sublethal concentrations of AgNPs, A. cina aqueous extract, AgNO3, and H2O2. Expression levels of SOD, GPx, and CAT genes were then quantified by RT-qPCR at multiple post-exposure time intervals. The synthesis was optimized by varying parameters such as pH, temperature, and extract volume. The nanoparticles were characterized using UV-Vis spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), and Electrophoretic Light Scattering (ELS). Overall, synthesis at pH 8 yielded small, spherical, stable, and abundant AgNPs. In vitro assays on L3 larvae showed a mortality rate of 91.33% at the highest AgNP concentration (500 \u03bcg\/mL), with lethal concentration (LC50 and LC90) values of 4.128 ppm (\u03bcg\/mL) and 17.993 \u03bcg\/mL, respectively. Relative expression analyses revealed that AgNPs induced the overexpression of the SOD gene, highlighting its role in the oxidative stress response. In contrast, the expression levels of GPx and CAT genes were markedly downregulated. These results suggest that SOD could serve as a potential biomarker of oxidative stress induced by AgNPs in combination with A. cina metabolites, influencing the infective stages of H. contortus.<\/jats:p>","DOI":"10.3390\/pathogens14121251","type":"journal-article","created":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T08:21:43Z","timestamp":1765182103000},"page":"1251","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Antiparasitic Activity of Silver Nanoparticles Synthesized from Artemisia cina Aqueous Extract Against Haemonchus contortus"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-5743-9475","authenticated-orcid":false,"given":"Lucero Itzel","family":"Hern\u00e1ndez Guerrero","sequence":"first","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9231-1556","authenticated-orcid":false,"given":"Rosa Isabel","family":"Higuera-Piedrahita","sequence":"additional","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9262-3111","authenticated-orcid":false,"given":"H\u00e9ctor Alejandro","family":"de la Cruz-Cruz","sequence":"additional","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4657-0159","authenticated-orcid":false,"given":"Jorge Alfredo","family":"Cu\u00e9llar-Ordaz","sequence":"additional","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8688-4411","authenticated-orcid":false,"given":"Mar\u00eda Eugenia","family":"L\u00f3pez-Arellano","sequence":"additional","affiliation":[{"name":"Centro Nacional de Investigaci\u00f3n Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agr\u00edcolas y Pecuarias, Carr. Fed. Cuernavaca-Cuautla 8534, Jiutepec 62550, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9634-9615","authenticated-orcid":false,"given":"Jocelyn","family":"Maza-Lopez","sequence":"additional","affiliation":[{"name":"Centro Nacional de Investigaci\u00f3n Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agr\u00edcolas y Pecuarias, Carr. Fed. Cuernavaca-Cuautla 8534, Jiutepec 62550, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6403-5216","authenticated-orcid":false,"given":"Abraham","family":"M\u00e9ndez-Albores","sequence":"additional","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5978-2375","authenticated-orcid":false,"given":"Alma","family":"V\u00e1zquez-Dur\u00e1n","sequence":"additional","affiliation":[{"name":"Facultad de Estudios Superiores Cuautitl\u00e1n, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuautitl\u00e1n Izcalli 54714, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Flay, K.J., Hill, F.I., and Muguiro, D.H. (2022). A Review: Haemonchus contortus Infection in Pasture-Based Sheep Production Systems, with a Focus on the Pathogenesis of Anaemia and Changes in Haematological Parameters. Animals, 12.","DOI":"10.3390\/ani12101238"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1038\/nature06722","article-title":"A New Class of Anthelmintics Effective against Drug-Resistant Nematodes","volume":"452","author":"Kaminsky","year":"2008","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5403","DOI":"10.2147\/IDR.S332378","article-title":"Anthelmintic Resistance and Its Mechanism: A Review","volume":"14","author":"Fissiha","year":"2021","journal-title":"Infect. Drug Resist."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.pt.2014.04.009","article-title":"Chasing Helminths and Their Economic Impact on Farmed Ruminants","volume":"30","author":"Charlier","year":"2014","journal-title":"Trends Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1080\/00480169.2009.36902","article-title":"Managing Anthelmintic Resistance: Modelling Strategic Use of a New Anthelmintic Class to Slow the Development of Resistance to Existing Classes","volume":"57","author":"Leathwick","year":"2009","journal-title":"N. Z. Vet. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.vetpar.2009.04.015","article-title":"The Role of Targeted Selective Treatments in the Development of Refugia-Based Approaches to the Control of Gastrointestinal Nematodes of Small Ruminants","volume":"164","author":"Kenyon","year":"2009","journal-title":"Vet. Parasitol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1136\/vr.102512","article-title":"Practices to Optimise Gastrointestinal Nematode Control on Sheep, Goat and Cattle Farms in Europe Using Targeted (Selective) Treatments","volume":"175","author":"Charlier","year":"2014","journal-title":"Vet. Rec."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.smallrumres.2011.10.016","article-title":"Nutritional Manipulation of Sheep and Goats for the Control of Gastrointestinal Nematodes under Hot Humid and Subhumid Tropical Conditions","volume":"103","author":"Hoste","year":"2012","journal-title":"Small Rumin. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1093\/jas\/skab096.066","article-title":"Multi-Species Grazing for Control of Gastrointestinal Nematodes\u2014A Review","volume":"99","author":"Burke","year":"2021","journal-title":"J. Anim. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.vetpar.2016.05.015","article-title":"Breeding for Resistance to Gastrointestinal Nematodes\u2014The Potential in Low-Input\/Output Small Ruminant Production Systems","volume":"225","author":"Zvinorova","year":"2016","journal-title":"Vet. Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1186\/1743-8977-3-11","article-title":"The Potential Risks of Nanomaterials: A Review Carried out for ECETOC","volume":"3","author":"Borm","year":"2006","journal-title":"Part. Fibre Toxicol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.glohj.2023.02.008","article-title":"Applications of Nanotechnology in Medical Field: A Brief Review","volume":"7","author":"Haleem","year":"2023","journal-title":"Glob. Health J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e20210195","DOI":"10.1590\/0103-8478cr20210195","article-title":"Nanotechnology in Veterinary Medicine: A Review","volume":"52","author":"Ianiski","year":"2022","journal-title":"Ci\u00eancia Rural."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5","DOI":"10.2174\/2211738508666200524233724","article-title":"Nanotechnological Improvement of Veterinary Anthelmintics","volume":"9","author":"Sanabria","year":"2021","journal-title":"Pharm. Nanotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Hill, E.K., and Li, J. (2017). Current and Future Prospects for Nanotechnology in Animal Production. J. Anim. Sci. Biotechnol., 8.","DOI":"10.1186\/s40104-017-0157-5"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.yexmp.2008.12.004","article-title":"Nanoparticle-Based Targeted Drug Delivery","volume":"86","author":"Singh","year":"2009","journal-title":"Exp. Mol. Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Bajwa, H.U.R., Khan, M.K., Abbas, Z., Riaz, R., ur Rehman, T., Abbas, R.Z., Aleem, M.T., Abbas, A., Almutairi, M.M., and Alshammari, F.A. (2022). Nanoparticles: Synthesis and Their Role as Potential Drug Candidates for the Treatment of Parasitic Diseases. Life, 12.","DOI":"10.3390\/life12050750"},{"key":"ref_18","first-page":"2843","article-title":"Advances in Nanotechnology and the Benefits of Using Cellulose Nanofibers in Animal Nutrition","volume":"14","author":"Sotomaior","year":"2021","journal-title":"Vet. World"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rashid, M.M.O., Ferdous, J., Banik, S., Islam, M.R., Uddin, A.H.M.M., and Robel, F.N. (2016). Anthelmintic Activity of Silver-Extract Nanoparticles Synthesized from the Combination of Silver Nanoparticles and M. Charantia Fruit Extract. BMC Complement. Altern. Med., 16.","DOI":"10.1186\/s12906-016-1219-5"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Hern\u00e1ndez-G\u00f3mora, A., Lara-Carrillo, E., Robles-Navarro, J., Scougall-Vilchis, R., Hern\u00e1ndez-L\u00f3pez, S., Medina-Sol\u00eds, C., and Morales-Luckie, R. (2017). Biosynthesis of Silver Nanoparticles on Orthodontic Elastomeric Modules: Evaluation of Mechanical and Antibacterial Properties. Molecules, 22.","DOI":"10.3390\/molecules22091407"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Goel, V., Das Singla, L., and Choudhury, D. (2020). Cuminaldehyde Induces Oxidative Stress-Mediated Physical Damage and Death of Haemonchus contortus. Biomed. Pharmacother., 130.","DOI":"10.1016\/j.biopha.2020.110411"},{"key":"ref_22","unstructured":"Higuera-Piedrahita, R.I. (2016). Efecto de Artemisia Cina 30CH y Artemisia cina En Extracto Etan\u00f3lico Contra La Infecci\u00f3n Natural e Inducida Por Haemonchus contortus, Universidad Nacional Aut\u00f3noma de M\u00e9xico."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Woerdenbag, H., van Uden, W., and Pras, N. (1997). Artemisia cina. Adverse Effects of Herbal Drugs, Springer.","DOI":"10.1007\/978-3-642-60367-9_2"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Sakipova, Z., Giorno, T.B.S., Bekezhanova, T., Siu Hai Wong, N., Shukirbekova, A., Fernandes, P.D., and Boylan, F. (2020). Pharmacological Evaluation of Artemisia cina Crude CO2 Subcritical Extract after the Removal of Santonin by Means of High Speed Countercurrent Chromatography. Molecules, 25.","DOI":"10.3390\/molecules25122728"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1007\/s11686-021-00364-w","article-title":"In Vitro Nematocidal Effect and Anthelmintic Activity of Artemisia cina Against Haemonchus contortus in Gerbils and Relative Expression of Hc29 Gene in Transitional Larvae (L3\u2013L4)","volume":"66","author":"Zamilpa","year":"2021","journal-title":"Acta Parasitol."},{"key":"ref_26","first-page":"46","article-title":"Estr\u00e9s Oxidativo y Da\u00f1o Vascular En Procesos de Hipoxia. Malondialdeh\u00eddo (MDA) Como Biomarcador de Da\u00f1o Oxidativo","volume":"2","year":"2014","journal-title":"Rev. Electr\u00f3nica Biomed."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Consales, C., Merla, C., Marino, C., and Benassi, B. (2012). Electromagnetic Fields, Oxidative Stress, and Neurodegeneration. Int. J. Cell Biol., 2012.","DOI":"10.1155\/2012\/683897"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.dsr.2013.07.014","article-title":"Oxidative Stress in Deep Scattering Layers: Heat Shock Response and Antioxidant Enzymes Activities of Myctophid Fishes Thriving in Oxygen Minimum Zones","volume":"82","author":"Lopes","year":"2013","journal-title":"Deep Sea Res. Part I Oceanogr. Res. Pap."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Flores Jim\u00e9nez, N.G., Zamorano, M.M., Res\u00e9ndiz-Gonz\u00e1lez, G., Mercado-M\u00e1rquez, C., Morales-\u00c1lvarez, J.F., Cu\u00e9llar-Ordaz, J.A., de-la-Cruz Cruz, H.A., Adams-V\u00e1zquez, O., L\u00f3pez-Arellano, M.E., and D\u00edaz-Torres, R. (2024). Biochemical Parameters, Oxidative Stress Biomarkers, and Anatomopathological Changes in Wistar Rats Treated with 3\u2032-Demethoxy-6-O-Demethylisoguaiacin and Norisoguaiacin. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-61903-9"},{"key":"ref_30","first-page":"347","article-title":"Ricerche Sulla Tecnica Di Coltura Delle Larve Infestive Degli Strongili Gastro-Intestinali Del Bovino","volume":"9","author":"Corticelli","year":"1963","journal-title":"Acta Med. Vet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1186\/s13071-022-05531-0","article-title":"Comparative Analysis of the Anthelmintic Efficacy of European Heather Extracts on Teladorsagia circumcincta and Trichostrongylus colubriformis Egg Hatching and Larval Motility","volume":"15","author":"Shepherd","year":"2022","journal-title":"Parasit. Vectors"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Maza-Lopez, J., Camas-Pereyra, R., L\u00f3pez-Arellano, M., and Contreras-Ochoa, C. (2023). Selection of Molecules with Immunological Potential from Excretory and Secretory Products from the Nematode Haemonchus placei by Cell Proliferation and Gene Expression Assays. Bio Protoc., 13.","DOI":"10.21769\/BioProtoc.4702"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"110255","DOI":"10.1016\/j.vetpar.2024.110255","article-title":"Molecular Study of the Transcription Factor SKN-1 and Its Putative Relationship with Genes That Encode GST and Antioxidant Enzymes in Haemonchus contortus","volume":"331","year":"2024","journal-title":"Vet. Parasitol."},{"key":"ref_34","first-page":"4128152","article-title":"Oxidative Stress Indexes for Diagnosis of Health or Disease in Humans","volume":"2019","year":"2019","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"S\u00e1nchez-Mendoza, A.E., Res\u00e9ndiz-Gonz\u00e1lez, G., Rico-Mej\u00eda, E., de la Cruz-Cruz, H.A., Ram\u00edrez-Rico, G., Cu\u00e9llar-Ordaz, J.A., Montiel-Sosa, J.F., L\u00f3pez-Arellano, M.E., Reyes-Guerrero, D.E., and Dom\u00ednguez-Delgado, C.L. (2025). Oxidative Stress on Haemonchus contortus Larvae Exposed to Alternative Treatment with Artemisia cina N-Hexane Extract and Cinaguaiacin Metabolites. Vet. Sci., 12.","DOI":"10.3390\/vetsci12050467"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Reyes-Guerrero, D.E., Jim\u00e9nez-Jacinto, V., Alonso-Morales, R.A., Alonso-D\u00edaz, M.\u00c1., Maza-Lopez, J., Camas-Pereyra, R., Olmedo-Ju\u00e1rez, A., Higuera-Piedrahita, R.I., and L\u00f3pez-Arellano, M.E. (2023). Assembly and Analysis of Haemonchus contortus Transcriptome as a Tool for the Knowledge of Ivermectin Resistance Mechanisms. Pathogens, 12.","DOI":"10.3390\/pathogens12030499"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Sun, W., and Shahrajabian, M.H. (2023). Therapeutic Potential of Phenolic Compounds in Medicinal Plants\u2014Natural Health Products for Human Health. Molecules, 28.","DOI":"10.3390\/molecules28041845"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1186\/s13046-018-0897-x","article-title":"Vitexin Compound 1, a Novel Extraction from a Chinese Herb, Suppresses Melanoma Cell Growth through DNA Damage by Increasing ROS Levels","volume":"37","author":"Liu","year":"2018","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Higuera-Piedrahita, R.I., Dolores-Hern\u00e1ndez, M., de la Cruz-Cruz, H.A., L\u00f3pez-Arellano, R., Gives, P.M., Olmedo-Ju\u00e1rez, A., Cu\u00e9llar-Ordaz, J.A., Gonz\u00e1lez-Cortazar, M., Ble-Gonz\u00e1lez, E.A., and L\u00f3pez-Arellano, M.E. (2023). 3\u2032-Demethoxy-6-O-Demethylisoguaiacin and Norisoguaiacin Nematocidal Lignans from Artemisia cina against Haemonchus contortus Infective Larvae. Plants, 12.","DOI":"10.3390\/plants12040820"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"105920","DOI":"10.1016\/j.actatropica.2021.105920","article-title":"Anthelmintic Activity of a Nanoformulation Based on Thiophenes Identified in Tagetes patula L. (Asteraceae) against the Small Ruminant Nematode Haemonchus contortus","volume":"219","author":"Politi","year":"2021","journal-title":"Acta Trop."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Agustina, T.E., Handayani, W., and Imawan, C. (2021, January 24\u201325). The UV-VIS Spectrum Analysis from Silver Nanoparticles Synthesized Using Diospyros maritima Blume Leaves Extract. Proceedings of the 3rd KOBI Congress, International and National Conferences, Online.","DOI":"10.2991\/absr.k.210621.070"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1251\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,8]],"date-time":"2025-12-08T08:39:59Z","timestamp":1765183199000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1251"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,7]]},"references-count":41,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["pathogens14121251"],"URL":"https:\/\/doi.org\/10.3390\/pathogens14121251","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,7]]}}}