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Silver nanoparticles induce histopathological alterations in juvenile Penaeus vannamei


The objective of this study was to evaluate the histopathological alterations in juvenile Penaeus vannamei caused by silver nanoparticles (AgNPs) for two types of experiments: at sublethal concentrations of 3.6 to 7.1 μg/μL of metallic silver (Ag) for a short period up to 72 h and for 2.6 to 7.9 μg of Ag/μL for the long period up to 264 h. The severity degree of the changes was evaluated and the histopathological index (Hi) was determined in both experiments using the necrosis (cellular dead) as an indicator. The pathological changes in the striated muscle, gills, antennal gland, circulatory system, heart, lymphoid organ, and connective tissue are described. The histopathological effects were similar for the two experiments without a direct relationship with the concentrations. In the short-term experiment, the values of Hi were higher (2.34 ± 0.41 at 48 hpi and 1.91 ± 0.39 at 72 hpi) compared with the long-term experiment (values between 0.57 ± 0.36 to 1.74 ± 0.57 at 264 hpi). The observed pathologies are similar to those caused by other metals, with the exception of the agglomerations of black particles in the gills, lymphoid organ, and muscle, which has not been previously reported. This work shows that silver nanoparticles cause damage to shrimp in sublethal concentrations.

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Data availability

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.


  1. Acedo-Valdez MR, Grijalva-Chon JM, Larios-Rodríguez E, Maldonado-Arce AD, Mendoza-Cano F, Sánchez-Paz JA, Castro-Longoria R (2017) Antibacterial effect of biosynthesized silver nanoparticles in Pacific white shrimp Litopenaeus vannamei (Boone) infected with necrotizing hepatopancreatitis bacterium (NHP-B) Lat. Am J Aquat Res 45(2):421–430.

    Article  Google Scholar 

  2. Akter M, Sikder MDT, Rahman MDM, Ullah AKMA, Hossain KFB, Banik S, Hosokawa T, Saito T, Kurasaki M (2018) Review. A systematic review of silver nanoparticles-induced cytotoxicity: physicochemical properties and perspectives. J Advan Res 9:1–16.

    CAS  Article  Google Scholar 

  3. Ashley P (2007) Fish welfare: current issues in aquaculture. Appl. Animal Behave Sci 104:199.235.

  4. Bahabadi MN, Delavar FH, Mirbakhsh M, Niknam K, Johari SA (2017) Assessment of antibacterial activity of two different sizes of colloidal silver nanoparticle (AgNPs) against Vibrio harveyi isolated from shrimp Litopenaeus vannamei. Aquac Int 25:463–472.

    CAS  Article  Google Scholar 

  5. Bambang Y, Thuet P, Charmantier-Daures M, Trilles JP, Charmantier G (1995) Effect of copper on survival and osmoregulation of various developmental stages of the shrimp Penaeus japonicus. Aquat Toxicol 33:125–139.

    CAS  Article  Google Scholar 

  6. Banerjee S, Kumar V, Pathak AN (2014) Drug effects on aquaculture and it’s remediation. JDDT 4(1):117–122.

    Article  Google Scholar 

  7. Banumathi B, Vaseeharan B, Suganya P, Citarasu T, Govindarajan M, Alharbi NS, Kadaikunnan S, Khaled JM, Benelli G (2017) Toxicity of Camellia sinensis-fabricated silver nanoparticles on invertebrate and vertebrate organisms: morphological abnormalities and DNA damages. J Clust Sci 28:2027–2040.

    CAS  Article  Google Scholar 

  8. Battistella S, Bonivento P, Amirante GA (1996) Hemocytes and immunological reactions in crustaceans. Ital J Zool 63(4):337–343.

    Article  Google Scholar 

  9. Bell TA, Lighter DV (1988) A handbook of normal penaeid shrimp histology. Allen Press, Lawrence, p 114

    Google Scholar 

  10. Bianchini A, Playle EC, Wood CM, Walsh OJ (2007) Short-term silver accumulation in tissues of three marine invertebrates: shrimp Penaeus duorarum, sea hare Aplysia californica, and sea urchin Diadema antillarum. Aquat Toxicol 84:182–189.

    CAS  Article  Google Scholar 

  11. Bogdanchikova N, Vázquez-Muñoz R, Huerta-Saquero A, Pena-Jasso A, Aguilar-Uzcanga G, Picos-Díaz PL, Pestryakov A, Burmistrov V, Martynyuk O, Luna-Vázquez-Gómez R, Almanza H (2016) Silver nanoparticles composition for treatment of distemper in dogs. Int J Nanotechnol 13(1–3):227–237.

    CAS  Article  Google Scholar 

  12. Borrego B, Lorenzo G, Mota-Morales J, Almanza-Reyes H, Mateos F, López-Gil E, de la Losa N, Burmistrov VA, Pestryakov AN, Brun A, Bogdanchikova N (2016) Potential application of silver nanoparticles to control the infectivity of Rift Valley fever virus in vitro and in vivo. Nanomedicine. 12(5):1185–1192.

    CAS  Article  Google Scholar 

  13. Braydich-Stolle L, Hussain S, Schlager JJ, Hofmann MC (2005) In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. Toxicol Sci 88(2):412–419.

    CAS  Article  Google Scholar 

  14. Brooking J, Davis SS, Illum L (2001) Transport of nanoparticles across the rat nasal mucosa. J Drug Target 9:267–279.

    CAS  Article  Google Scholar 

  15. Canesi L, Corsi I (2016) Effects of nanomaterials on marine invertebrates. Sci Total Environ 565:933–940.

    CAS  Article  Google Scholar 

  16. Conover WJ (2012) The rank transformation-an easy and intuitive way to connect many nonparametric methods to their parametric counterparts for seamless teaching introductory statistics courses. Wires Comput Stat 4:432–438.

    Article  Google Scholar 

  17. Conover WJ, Iman RL (1981) Rank transformations as a bridge between parametric and nonparametric statistics. Am Stat 35:124–129.

    Article  Google Scholar 

  18. Gambardella C, Costa E, Piazza V, Fabbrocini A, Magi E, Faimali M, Garaventa F (2015) Effect of silver nanoparticles on marine organisms belonging to different trophic levels. Mar Environ Res 111:41–49.

    CAS  Article  Google Scholar 

  19. Gerhardt A, de Bisthoven LJ, Mo Z, Wang C, Yang M, Wang Z (2002) Short-term responses of Oryzias latipes (Pisces: Adrianichthyidae) and Macrobrachium nipponense (Crustacea: Palaemonidae) to municipal and pharmaceutical waste water in Beijing, China: survival, behaviour, biochemical biomarker. Chemosphere 47:35–47.

    CAS  Article  Google Scholar 

  20. Gopi N, Vijayakumara S, Thayab R, Govindarajan M, Alharbie NS, Kadaikunnane S, Khalede MJ, Al-Anbre MN, Vaseeharana B (2019) Chronic exposure of Oreochromis niloticus to sub-lethal copper concentrations: effects on growth, antioxidant, non-enzymatic antioxidant, oxidative stress and non-specific immune responses. J Trace Elements Med Biol 55:170–179.

    CAS  Article  Google Scholar 

  21. Henry RP, Lucu CE, Onken H, Weihrauch D (2012) Multiple functions of the crustacean gill: osmotic/ionic regulation, acid-base balance, ammonia excretion, and bioaccumulation of toxic metals. Front Physiol 3(431):1–33.

    CAS  Article  Google Scholar 

  22. Huang S, Wang L, Liu L, Hou Y, Li L (2015) Nanotechnology in agriculture, livestock, and aquaculture in China. A review. Agron Sustain Dev 35:369–400.

    Article  Google Scholar 

  23. Ishwarya R, Vaseeharan B, Shanthi S, Ramesh S, Manogari P, Dhanalakshmi K, Vijayakumar S, Benelli G (2016) Green synthesized silver nanoparticles: toxicity against Poecilia reticulata fishes and Ceriodaphnia cornuta crustaceans. J Clust Sci.

  24. Juarez-Moreno K, Mejía-Ruiz CH, Díaz F, Reyna-Verdugo H, Red AD, Vazquez-Felix EF, Sánchez-Castrejón E, Mota-Morales JD, Pestryakovf A, Bogdanchikova N (2017) Effect of silver nanoparticles on metabolic rate, hematological response and survival of juvenile white shrimp Litopenaeus vannamei (Boone). Chemosphere 169:716–724.

    CAS  Article  Google Scholar 

  25. Kachenton S, Whangpurikul V, Kangwanrangsan N, Tansatit T, Jiraungkoorskul W (2018) Silver nanoparticles toxicity in brine shrimp and its histopathological analysis. Int J Nanosci 1-5:1850007.

    CAS  Article  Google Scholar 

  26. Kandasamy K, Alikunhi NM, Manickaswami G, Nabikhan A, Ayyavu G (2013) Synthesis of silver nanoparticles by coastal plant Prosopis chilensis (L.) and their efficacy in controlling vibriosis in shrimp Penaeus monodon. Appl Nanosci 3:65–73.

    CAS  Article  Google Scholar 

  27. Kondo M, Itami T, Takahashi Y, Fujii R, Tomonaga S (1994) Structure and function of the lymphoid organ in the kuruma prawn. Dev Comp Immunol 18(Suppl l):S109

    Google Scholar 

  28. Lekamge S, Miranda AF, Abraham A, Li V, Shukla R, Bansal V, Nugegoda D (2018) The toxicity of silver nanoparticles (AgNPs) to three freshwater invertebrates with different life strategies; Hydra vulgaris, daphnia carinata, and Paratya autraliensis. Front Environ Sci 6:162.

    Article  Google Scholar 

  29. Li N, Zhao Y, Yang J (2007) Impact of waterborne copper on the structure of gills and hepatopancreas and its impact on the content of metallothionein in juvenile giant freshwater prawn Macrobrachium rosenbergii (Crustacea: Decapoda). Arch Environ Contam Toxicol 52:73–79.

    CAS  Article  Google Scholar 

  30. Lightner DV (1996) A handbook of shrimp pathology and diagnostic procedures for diseases of cultured penaeid shrimp. J. World Aquacult. Soc., Baton Rouge

  31. Martin GG, Poole D, Poole C, Hose JE, Arias M, Reynolds L, McKrell N, Whang A (1993) Clearance of bacteria injected into the hemolymph of the penaeid shrimp, Sicyonia ingentis. J Invertebr Pathol 62(3):308–315.

    Article  Google Scholar 

  32. Martin GG, Hose JE, Minka G, Rosenberg S (1996) Clearance of bacteria injected into the hemolymph of the ridgeback prawn, Sicyonia ingentis (Crustacea: Decapoda): role of hematopoietic tissue. J Morphol 227:227–233.<227:AID-JMOR8>3.0.CO;2-5

    Article  Google Scholar 

  33. Martin GG, Quintero M, Quigley M, Khosrovian H (2000) Elimination of sequestered material from the gills of decapod crustaceans. J Crustac Biol 20:209–217.

    Article  Google Scholar 

  34. Nimmo DWR, Lightner DV, Bahner LH (1977) Effects of cadmium on shrimps Penaeus duodarum, Palaemonetes pugio and Palaemonetes vulgaris. In: Vernberg FJ, Calabresse A, Thurberg FP, Vernberg WB (eds) Physiological responses of marine biota to pollutants. Academic Press, New York, pp 131–184

    Chapter  Google Scholar 

  35. Ochoa-Meza AR, Álvarez-Sánchez AR, Romo-Quiñonez CR, Barraza A, Magallón-Barajas FJ, Chávez-Sánchez A, García-Ramos JC, Toledano-Magaña Y, Bogdanchikova N, Pestryakov A, Mejía-Ruiz CH (2019) Silver nanoparticles enhance the survival of white spot syndrome virus infected Penaeus vannamei shrimps by activation of its immunological system. Fish Shellfish Immonol 84:1083–1089.

    CAS  Article  Google Scholar 

  36. Ramamoorthy S, Kannaiyan P, Moturi M, Devadas T, Muthuramalingam J, Natarajan L, Arunachalam N, Ponniah AG (2013) Antibacterial activity of zinc oxide nanoparticles against Vibrio harveyi. Indian J Fish:107–112

  37. Rather MA, Sharma R, Aklakur M, Ahmad S, Kumar N, Khan M, Ramya VL (2011) Nanotechnology: a novel tool for aquaculture and fisheries development. A prospective mini-review. Fish Aquac J. (2011): FAJ-16

  38. Romo-Quiñonez CR, Álvarez-Sánchez AR, Álvarez-Ruiz P, Chávez-Sánchez MC, Bogdanchikova N, Pestryakov A, Mejia-Ruiz CH (2020) Evaluation of a new Argovit as an antiviral agent included in feed to protect the shrimp Litopenaeus vannamei against White Spot Syndrome Virus infection. PeerJ 8:e8446.

    Article  Google Scholar 

  39. Rusaini, Owens L (2010) Insight into the lymphoid organ of penaeid prawns: a review. Fish Shellfish Immunol 29:367–377.

    CAS  Article  Google Scholar 

  40. Sabu AS, Sanil NK, Nammalwar P (2017) Effect of cadmium in the gills of green tiger prawn, Penaeus semisulcatus. Int J Fish Aquac 5(1):467–478

    Google Scholar 

  41. Selvaraj B, Subramanian K, Gopal S, Renuga PS (2014) Nanotechnology as a novel tool for aquaculture industry: a review. World J Pharma Sci 2(9):1089–1096

    Google Scholar 

  42. Sibaja-Luis AI, Ramos-Campos EV, Luiz de Oliveira J, Fernandes-Fraceto L (2019) Trends in aquaculture sciences: from now to use of nanotechnology for disease control. Rev Aquac 11:119–132.

    Article  Google Scholar 

  43. Sivaramasamy E, Zhiwei W, Li F, Xiang J (2016) Enhancement of vibriosis resistance in Litopenaeus vannamei by supplementation of biomastered silver nanoparticles by Bacillus subtilis. J Nanomed Nanotechnol 7:1–10.

    CAS  Article  Google Scholar 

  44. Soegianto A, Charmantier-Daures M, Trilles J, Charmantier G (1999) Impact of copper on the structure of gills and epipodites of the shrimp Penaeus japonicus (Crustacea: Decapoda). J Crustac Biol 19:209–223.

    Article  Google Scholar 

  45. Sonnenholzner S, Rodríguez J, Pérez F, Betancourt I, Echeverría F, Calderón J (2002) Supervivencia y respuesta inmune de camarones juveniles L. vannamei desafiador por vía oral a WSSV a diferentes temperaturas. El Mundo Acuícola 8:50–55

    Google Scholar 

  46. Tsai JR, Lin HC (2014) Functional anatomy and ion regulatory mechanisms of the antennal gland in a semi-terrestrial crab, Ocypode stimpsoni. Biol Open 3:409–417.

    Article  Google Scholar 

  47. Untersteiner H, Gretschel G, Puchner T, Napetschnig S, Kaiser H (2005) Monitoring behavioral responses to the heavy metal cadmium in the marine shrimp Hippolyte inermis leach (Crustacea: Decapoda) with video imaging. Zool Stud 44(1):71–80

    CAS  Google Scholar 

  48. Van de Braak CBT, Botterblom MHA, Taverne N, van Muiswinkel WB, Rombout JHWM, van der Knaap WPW (2002) The roles of the haemocytes and the lymphoid organ in the clearance of injected Vibrio bacteria in Penaeus monodon shrimp. Fish Shellfish Immunol 13:293–309.

    Article  Google Scholar 

  49. Vaseeharan B, Ramasamy P, Chen JC (2010) Antibacterial activity of silver nanoparticles (AgNPs) synthesized by tea leaf extracts against pathogenic Vibrio harveyi and its protective efficacy on juvenile Feneropenaeus indicus. Lett Appl Microbiol 50(4):352–356.

    CAS  Article  Google Scholar 

  50. Vaseeharan B, Sargunar GC, Lin CY, Chen CJ (2012) Green synthesis of silver nanoparticles through Calotropis gigantea leaf extracts and evaluation of antibacterial activity against Vibrio alginolyticus. Nanotech Dev 2(1):12–16.

    CAS  Article  Google Scholar 

  51. Vazquez-Muñoz R, Avalos-Borja M, Castro-Longoria E (2014) Ultrastructural analysis of Candida albicans when exposed to silver nanoparticles. PLoS One 9:e108876.

    CAS  Article  Google Scholar 

  52. Wu JP, Chen HC, Huang DJ (2009) Histopathological alterations in gills of white shrimp, Litopenaeus vannamei (Boone) after acute exposure to cadmium and zinc. Bull Environ Contam Toxicol 82:90–95.

    CAS  Article  Google Scholar 

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The authors thank CONACyT for the support granted to the Basic Science Project number 258607 and Russian grand N18-29-24037. We also thank Tomsk Polytechnic University Competitiveness Enhancement Program, Project VIU-RS. Also, important recognition is given to the technicians Victor Joaquín Álvarez López and José Antonio Velázquez Garay, for their support during the experiments.


All sources of funding for the research were declared. The National Council for Science and Technology (CONACyT) granted MCCHS (CONACyT grant number 258607), while the Russian project N18-29-24037 supplied the silver nanoparticles of the project.

Author information




MCCHS: leadership responsible for the research activity planning and execution, including mentorship external to the core team. Management and coordination responsibility for research activity planning. Planning with the group the experimental design and methodology. Responsible together with the histology group for the histopathology results. Preparation and translation of the work to be published together with the histopathology group

SAR: from the histopathology group. Responsible for the histology processing as well as for the analysis and description of the histopathology observed in the treated shrimps. Responsible for the analysis of the data and application of statistics. Preparation of the work to be published in conjunction with the leader and RLO

RLO: histopathology group. Responsible for the histology processing as well as for the analysis and description of the histopathology observed in the treated shrimps, together SAR and MCCHS. Analysis of the data and application of statistics. Preparation of the work to be published together with the leader and SAR

LMR: participate actively in the design of the experiments and in the execution of the tests. Contributes in obtaining the best shrimp juveniles and its better management during the experiments, provision of study materials, reagents, materials, and instrumentation

MAFN: participate actively in the design of the experiments and in the execution of the tests. Verification, as a part of the activity of the overall replication/reproducibility of results/experiments. Analysis of the data and application of statistics. Specially, critical review, commentary, and revision

CHRM: participate actively in the design of the experiments and in the execution of the tests. Critical review, commentary, or revision of the paper

AP: donation of the silver nanoparticles

NB: critical review, commentary, and revision of the paper

Cristina Chávez led the project, got the financial, formal analysis, investigation, data curation, writing original draft, and visualization, and changes was realized by all the authors.

Corresponding author

Correspondence to María-Cristina Chávez-Sánchez.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures were performed according to Mexican guidelines and policies stated in the NOM-062-ZOO-1999 (these guidelines apply mostly to mammalian species but we applied the same principles regarding animal welfare and care) and British guidelines for fish welfare reported by Ashley (2007).

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• Sublethal concentrations of silver nanoparticles caused 23% mortality to shrimp

• Silver nanoparticles at sublethal levels cause pathological damage to shrimp

• Silver nanoparticles produce necrosis to different tissues and organs

• The treated shrimp could mitigate through their immune system the effects of AgNps

Responsible editor: Philippe Garrigues

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Chávez-Sánchez, MC., Abad-Rosales, S., Lozano-Olvera, R. et al. Silver nanoparticles induce histopathological alterations in juvenile Penaeus vannamei. Environ Sci Pollut Res 28, 8224–8234 (2021).

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  • Silver nanoparticles
  • Pathological alterations
  • Sublethal concentrations
  • Penaeus vannamei