Caracterización citométrica y morfológica de los hemocitos de cuatro especies de crustáceos decápodos de agua dulce
DOI:
https://doi.org/10.24215/16684869e045Palabras clave:
Crustáceos, citometría de flujo, hemolinfa, análisis de hemocitosResumen
La caracterización y clasificación de los hemocitos de invertebrados es una poderosa herramienta para determinar el efecto fisiológico del estrés ambiental. Los hemocitos de cuatro especies de crustáceos decápodos Dilocarcinus pagei, Zilchiopsis collastinensis, Aegla uruguayana y Macrobrachium borellii se analizaron mediante citometría de flujo seguida de una caracterización morfológica por microscopía óptica. Basándose en las características morfológicas celulares, se identificaron tres categorías de hemocitos: granulocitos con granularidad abundante que representaban el 1,87% (D. pagei), el 5,23% (Z. collastinensis), el 0,98% (A. uruguayana) y el 2,41% (M. borellii), semigranulocitos con menor granularidad con un 2,22% (D. pagei), 4,81% (Z. collastinensis), 2,38% (A. uruguayana) y 68,19% (M. borellii) y, por último, hialinocitos casi sin granularidad con un 89,87% (D. pagei), 65,75% (Z. collastinensis), 92,33% (A. uruguayana) y 22,04% (M. borellii). El análisis del recuento total de hemocitos (células/ml) arrojó los siguientes resultados: 4,3 ± 0,58 (x106) en D. pagei, 2,3 ± 0,33 (x106) en Z. collastinensis; 3,1 ± 0,57 (x106) en A. uruguayana y 2,6 ± 0,25 (x106) en M. borellii. Este estudio demostró que la citometría de flujo es eficaz para diferenciar y contar los hemocitos circulantes, lo que la convierte en una herramienta valiosa para evaluar el estado fisiológico. Además, se observó variabilidad interespecífica en estos parámetros celulares. Los resultados contribuyen a la comprensión del sistema inmunitario innato en estas cuatro especies de crustáceos y permiten crear una línea de base para identificar condiciones de bienestar o estrés en estos animales.
Referencias
Aguirre-Guzmán, G., Sánchez-Martínez, J.G., Campa-Córdova, A.I., Luna-Gonzaléz, A. & Ascencio, F. (2009). Penaeid shrimp immune system. The Thai Journal of Veterinary Medicine, 39, 205-215.
Battison, A., Cawthorn, R. & Horney, B. (2003). Classification of Homarus americanus hemocytes and the use of differential hemocyte counts in lobsters infected with Aerococcus viridans var. homari (Gaffkemia). Journal of Invertebrate Pathology, 84, 177-197.
Bauchau, A. G. (1980). Crustaceans. In N. A. Ratcliffe and A. F. Rowley (Eds.), Invertebrate Blood Cells (pp. 385-420). Academic Press.
Buchmann, K. (2014). Evolution of innate immunity: clues from invertebrates via fish to mammals. Frontiers in Immunology, 5, 459.
Cárdenas, W., Jenkins, J. A. & Dankert, J. R. (2000). A flow cytometric approach to the study of crustacean cellular immunity. Journal of Invertebrate Pathology, 76(2), 112-119.
Carus, C. G. (1824). Von den äußern Lebensbedingungen der weiß- und kaltblutigen Thiere. Leipzig.
Carvalho, D. A., Collins, P. A., Lima-Gomes, R., Magalhães, C., Torres, M. V. & Williner, V. (2018). A comparative study of the gastric ossicles of Trichodactylidae crabs (Brachyura: Decapoda) with comments on the role of diet and phylogeny in shaping morphological traits. PeerJ, 6, e5028. https://doi.org/10.7717/peerj.5028
Cerenius, L. & Söderhäll, K. (2012). Crustacean immune responses and their implications for disease control in infectious disease in aquaculture, prevention and control. In B. Austin (Ed.), Woodhead publishing series in food science, technology and nutrition: Number 231 (pp. 69-87). Woodhead Publishing Limited.
Cheng, W. & Chen, J. C. (2001). Effects of intrinsic and extrinsic factors on the haemocyte profile of the prawn, Macrobrachium rosenbergii. Fish & Shellfish Immunology, 11(1), 53-63. https://doi.org/10.1006/fsim.2000.0293
Collins, P. A., Williner, V. & Giri, F. (2007). Littoral communities. Macrocrustaceans. In Iriondo M. H., J. C. Paggi & M. J. Parma (Eds.), The Middle Paraná River: Limnology of a subtropical wetland (pp. 277-301). Springer.
Collins, P. A., Carnevalt, R., Carvalho, D. & Williner, V. (2012). Dynamics of decapod crustaceans in a trophic web of continental aquatic environments in Southern South America. In J. A. Daniels (Ed.), Advances in environmental research (pp. 159-185). Nova Science Publishers, Inc.
Comité de Ética y Seguridad del Trabajo Experimental. (2016). https://santafe.conicet.gov.ar/ceyste/
Day, R. D., Fitzgibbon, Q. P. & Gardner, C. (2019). The impact of holding stressors on the immune function and haemolymph biochemistry of Southern Rock Lobsters (Jasus edwardsii). Fish & Shellfish Immunology, 89, 660-671. https://doi.org/10.1016/j.fsi.2019.03.043
de Melo, G. A. S. (2003). Manual de identificação dos Crustacea Decapoda de água doce do Brasil. Edições Loyola.
Ding, Z., Du, J., Ou, J., Li, W., Wu, T., Xiu, Y., Meng Q., Ren Q., Gu W., Xue H., Tang J. & Wang, W. (2012). Classification of circulating hemocytes from the red swamp crayfish Procambarus clarkii and their susceptibility to the novel pathogen Spiroplasma eriocheirisin vitro. Aquaculture, 356, 371-380.
Donaghy, L., Hong, H. K., Lambert, C., Park, H. S., Shim, W. J. & Choi, K. S. (2010). First characterisation of the populations and immune-related activities of hemocytes from two edible gastropod species, the disk abalone, Haliotis discusdiscus and the spiny top shell, Turbo cornutus. Fish & Shellfish Immunology, 28(1), 87-97. https://doi.org/10.1016/j.fsi.2009.10.006
dos Santos, A., Botelho, M. T., Joviano, W. R., Gomes, V., da Silva, J. R. M. C. & de Aragão Umbuzeiro, G. (2023). Characterization of hemocytes from the marine amphipod Parhyale hawaiensis (Dana 1853): Setting the basis for immunotoxicological studies. Invertebrate Biology, 142(1), e12394. https://doi.org/10.1111/ivb.12394
Du, J., Zhu, H., Ren, Q., Liu, P., Chen, J., Xiu, Y., Yao W., Meng Q., Gu W. & Wang, W. (2012). Flow cytometry studies on the Macrobrachium rosenbergii hemocytes sub-populations and immune responses to novel pathogen spiroplasma MR-1008. Fish & Shellfish Immunology, 33(4), 795-800. https://doi.org/10.1016/j.fsi.2012.07.006
Frizzera, A. C., Samartino, C. G., Santo, A. M., Bojko, J., Cremonte, F. & Vázquez, N. (2022). Haemocyte characterization in invasive Carcinus maenas (Linnaeus, 1758) (Decapoda, Portunidae) from unpolluted vs polluted sites in Argentina. Crustaceana, 95, 1083-1099.
Gargioni, R. & Barracco, M. A. (1998). Hemocytes of the palaemonids Macrobrachium rosenbergii and M. acanthurus, and of the Penaeid Penaeuspaulensis. Journal of Morphology, 236, 209-221.
Hong, Y., Yang, X., Yan, G., Huang, Y., Zuo, F., Shen, Y., Ding Y. & Cheng, Y. (2017). Effects of glyphosate on immune responses and haemocyte DNA damage of Chinese mitten crab, Eriocheir sinensis. Fish & Shellfish Immunology, 71, 19-27. https://doi.org/10.1016/j.fsi.2017.09.062
Hose, J. E., Martin, G. G., Nguyen, V. A., Lucas, J. & Rosenstein, T. (1987). Cytochemical features of shrimp hemocytes. The Biological Bulletin, 173, 178-187.
Hose, J. E., Martin, G. G. & Gerard, A. S. (1990). A decapod hemocyte classification scheme integrating morphology, cytochemistry and function. Biological Bulletin, 178, 33-45.
Jia, Z., Wang, L., Jiang, S., Sun, M., Wang, M., Yi, Q. & Song, L. (2017). Functional characterization of hemocytes from Chinese mitten crab Eriocheir sinensis by flow cytometry. Fish & Shellfish Immunology, 69, 15-25. https://doi.org/10.1016/j.fsi.2017.08.001
Jiravanichpaisal, P., Lee, B. L. & Soderhall, K. (2006). Cell mediated immunity in arthropods: hematopoiesis, coagulation, melanization and opsonization. Immunobiology, 211, 213–236.
Johansson, M. W., Keyser, P., Sritunyalucksana, K. & Söderhäll, K. (2000). Crustacean haemocytes and haematopoiesis. Aquaculture, 191, 45-52.
Jussila, J., Jago, J., Tsvetnenko, E., Dunstan, B. & Evans, L. H. (1997). Total and differential haemocyte counts in western rock lobsters (Panulirus cygnus George) under post-harvest stress. Marine and Freshwater research, 48, 863-868.
Kaluza (version 2.2) Software. (2023). Beckman Coulter Inc.. https://www.beckman.com/flow-cytometry software/kaluza
Kruangkum, T., Söderhäll, K. & Söderhäll, I. (2025). The hematopoietic tissue of the freshwater crayfish, Pacifastacus leniusculus: organization and expression analysis. Cell and Tissue Research, 399(3), 303-322. http://dx.doi.org/10.1007/s00441-024-03943-1
Koiwai, K., Alenton, R. R. R., Shiomi, R., Nozaki, R., Kondo, H. & Hirono, I. (2017). Two hemocyte sub-populations of kuruma shrimp Marsupenaeus japonicus. Molecular Immunology, 85, 1-8.
Lee, S. Y. & Söderhäll, K. (2002). Early events in crustacean innate immunity. Fish & Shellfish Immunology, 12(5), 421-437. https://doi.org/10.1006/fsim.2002.0420
Li, F., Chang, X., Xu, L. & Yang, F. (2018). Different roles of crayfish hemocytes in the uptake of foreign particles. Fish & Shellfish Immunology, 77, 112-119. https://doi.org/10.1016/j.fsi.2018.03.029
Lin, X., Söderhäll, K. & Söderhäll, I., (2011). Invertebrate hematopoiesis: An astakine dependent novel hematopoietic factor. Journal of Immunology, 186(4), 2073-2079. http://dx.doi.org/10.4049/jimmunol.1001229
Liu, M. J., Liu, S. & Liu, H. P. (2021). Recent insights into hematopoiesis in crustaceans. Fish and Shellfish Immunology Reports, 2, 100040. https://doi.org/10.1016/j.fsirep.2021.100040
Lorenzon, S., Francese, M., Smith, V. J. & Ferrero, E. A. (2001). Heavy metals affect the circulating haemocyte number in the shrimp Palaemon elegans. Fish & shellfish Immunology, 11(6), 459-472. https://doi.org/10.1006/fsim.2000.0321
Lv, S., Xu, J., Zhao, J., Yin, N., Lu, B., Li, S., Chen, Y. & Xu, H. (2014). Classification and phagocytosis of circulating haemocytes in Chinese mitten crab (Eriocheir sinensis) and the effect of extrinsic stimulation on circulating haemocytes in vivo. Fish & Shellfish Immunology, 39(2), 415-422. http://dx.doi.org/10.1016/j.fsi.2014.05.036
Mauro, M., Arizza, V., Arculeo, M., Attanzio, A., Pinto, P., Chirco, P. Badalamenti, G., Tesoriere, L. & Vazzana, M. (2022). Haemolymphatic parameters in two aquaculture crustacean species Cherax destructor (Clark, 1836) and Cherax quadricarinatus (Von Martens, 1868). Animals, 12, 543.
Mix, M. C. & Sparks, A. K. (1980). Hemocyte classification and differential counts in the Dungeness crab, Cancer magister. Journal of Invertebrate Pathology, 35, 134-143.
Morrone, J. J. & Lopretto, E. C. (1994). Distributional patterns of freshwater Decapoda (Crustacea: Malacostraca) in southern South America: A panbiogeographic approach. Journal of Biogeography, 21(1), 97-109.
Nobili, G. (1896). Viaggio del Dr. Alfredo Borlli nel Chaco Boliviano e nella Repubblica Argentina. Bolletino dei Musei di Zoologia ed Anatomia comparata della R. Università di Torino, 11, 265-267.
Oliver, J. D., Loy, J. D., Parikh, G. & Bartholomay, L. (2011). Comparative analysis of hemocyte phagocytosis between six species of arthropods as measured by flow cytometry. Journal of Invertebrate Pathology, 108, 126-130.
Parrinello, D., Sanfratello, M. A., Celi, M. & Vazzana, M. (2015). Hemocyte types and some plasmatic properties of two edible crabs Cancer borealis and Cancer pagurus. Invertebrate Survival Journal, 12, 195-202.
Pretzmann, G. (1968). Die Familie Trichodactylidae (Milne-Edwards 1853) Smith 1870 (Vorläufige Mitteilung). Entomologisches Nachrichtenblatt, 15(7-8), 70-76.
Qyli, M., Aliko, V. & Faggio, C. (2020). Physiological and biochemical responses of Mediterranean green crab, Carcinus aestuarii, to different environmental stressors: Evaluation of hemocyte toxicity and its possible effects on immune response. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 231, 108739.
R Core Team. (2023). R: A language and environment for statistical computing. R Foundation for Statistical Computing. https://www.R-project.org/
Russo, J., Brehelin, M. & Carton, Y. (2001). Haemocyte changes in resistant and susceptible strains of D. melanogaster caused by virulent and avirulent strains of the parasitic wasp Leptopilina boulardi. Journal of Insect Physiology, 47, 167-172.
Schmitt, W. L. (1942). The freshwater decapod Crustacea of the Rio de la Plata region. Proceedings of the United States National Museum, 92(3167), 1-47.
Sequeira, T., Vilanova, M., Lobo-Da-Cunha, A., Baldaia, L. & Arala-Chaves, M. (1995). Flow cytometric analysis of molt-related changes in hemocyte type in male and female Penaeus japonicus. The Biological Bulletin, 189, 376-380.
Shapiro, H. M. (2003). Practical flow cytometry. John Wiley & Sons, Inc.
Smith, V. J., Swindlehurst, R. J., Johnston, P. A. & Vethaak, A. D. (1995). Disturbance of host defence capability in the common shrimp, Crangon crangon, by exposure to harbour dredge spoils. Aquatic Toxicology, 32, 43-58.
Sneddon, L. U. (2015). Pain in aquatic animals. The Journal of Experimental Biology, 218, 967-976.
Söderhäll, I. (2016). Crustacean hematopoiesis. Developmental & Comparative Immunology, 58, 129-141.
Söderhäll, I. & Söderhäll, K. (2022). Blood cell formation in crustaceans. Fish & Shellfish Immunology, 131, 1335-1342. http://dx.doi.org/10.1016/j.fsi.2022.10.008
Stimpson, W. M. (1861). Notes on certain decapod Crustacea. Proceedings of the Academy of Natural Sciences of Philadelphia, 13, 372-375.
Sung, H. H. & Sun, R. (2002). Use of monoclonal antibodies to classify hemocyte subpopulations of tiger shrimp (Penaeus monodon). Journal of Crustacean Biology, 22, 337-344.
Taylor, S., Landman, M. J. & Ling, N. (2014). Flow cytometric characterization of freshwater crayfish hemocytes for the examination of physiological status in wild and captive animals. Journal of Aquatic Animal Health, 21(3), 195-203. http://dx.doi.org/10.1577/H09-003.1
Williner, V. (2010). Foregut ossicles morphology and feeding of the freshwater anomuran crab Aegla uruguayana (Decapoda, Aeglidae). Acta Zoologica, 91, 408-415.
Yifei, Y., Zhixiong, Z., Luna, C., Qihui, C., Zuoyuan, W., Xinqi, L., Zhexiang, Z. F. & Xiujuan, Z. (2024). Marine pollutant Phenanthrene (PHE) exposure causes immunosuppression of hemocytes in crustacean species, Scylla paramamosain. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 275, 109761.
Zar, J. H. (1996). Biostatistical analysis. Prentice Hall.
Zhang, Z. F., Shao, M. & Kang, K. H. (2006). Classification of haematopoietic cells and haemocytes in Chinese prawn Fenneropenaeus chinensis. Fish & Shellfish Immunology, 21(2), 159-169. https://doi.org/10.1016/j.fsi.2005.11.003
Zhou, Y. L., Gu, W. B., Tu, D. D., Zhu, Q. H., Zhou, Z. K., Chen, Y. Y. & Shu, M. A. (2018). Hemocytes of the mud crab Scylla paramamosain: Cytometric, morphological characterization and involvement in immune responses. Fish & Shellfish Immunology, 72, 459-469. https://doi.org/10.1016/j.fsi.2017.10.055
Descargas
Publicado
Número
Sección
Licencia
Derechos de autor 2025 Paula Sofía Reyes, Carlos Leandro Negro, Diana Alberto, María Florencia Viozzi, Veronica Williner
Esta obra está bajo una licencia internacional Creative Commons Atribución-NoComercial-CompartirIgual 4.0.
