Chronic granulomatous inflammation in teleost fish Piaractus mesopotamicus: histopathology model study

Inflamación crónica granulomatosa en el pez teleósteo Piaractus mesopotamicus: modelo de estudio histopatológico

Objective. This study evaluated the cell kinetic and formation of granuloma during chronic inflammation induced by Bacillus Calmette-Guérin (BCG) in the skeletal muscle of Piaractus mesopotamicus, as a histopathology model to study innate immunity. Materials and methods. Sixty fish were divided in two groups: BCG-inoculated and non-inoculated fish and the inflammatory response analyzed 3, 7, 14, 21 and 33 days post-inoculation (DPI) by histopathology after hematoxylin-eosin and Ziehl-Neelsen staining. Results. 3 DPI of BCG showed a diffuse inflammatory reaction mostly composed by mononuclear cells. The inflammation continued diffuse 7 DPI initiating the cellular organization surrounding the inoculum and have continued at 14 DPI with discrete presence of epithelioid-like type cells with acidophilic cytoplasm and floppy chromatin. Higher cellular organization (21 DPI) surrounding the granuloma with intense peripheral mononuclear inflammatory infiltrate and nevertheless, an increase in the number of fibroblasts and macrophage-like cells was observed. The inflammatory process became less diffuse 33 DPI with formation of small amount of granuloma surrounded by the same type of reaction found in bigger granuloma. Both the young and old granuloma presented typical characteristic around the inoculum composed by a layer of epithelioid-like type cells, besides macrophages, some lymphocytes and abundant fibroblasts. Conclusions. This study showed the feasibility in the use of pacus to study chronic granulomatous inflammatory response induced by BCG, characterized by changes in the kinetics of inflammatory cells in skeletal muscle classifying as immune-epithelioid type, similar to granulomatous inflammation caused by M. marinum in teleost fish.

1. Whipps CM, Lieggi C, Wagner R. Mycobacteriosis in Zebrafish Colonies. ILAR 2012; 53(2):95-105.
2. Waltzek TB, Cortés-Hinojosa G, Wellehan JF Jr, Gray GC. Marine mammal zoonoses: a review of disease manifestations. Zoonoses Public Health 2012; 59:521-535.
3. Ulrichs T, Lefmann M, Reich M, Morawietz K, Roth A, Brinkmann V,et al. Modified immunohistological staining allows detection of Ziehl–Neelsen-negative Mycobacterium tuberculosis organisms and their precise localization in human tissue. J Pathol 2005; 205:633–640.
4. Kazda J, Pavlik I, Falkinham JO. III, Hruska K. The Ecology of Mycobacteria: Impact on Animal’s and Human’s Health. 1ed., Netherlands: Springer; 2009.
5. Jacobs JM, Stine CB, Baya AM, Kent ML. A review of mycobacteriosis in marine fish. J Fish Dis 2009; 32:119–130.
6. Kent ML, Whipps CM, Matthews JL, Florio D, Watral V, Bishop-Stewart JK, et al. Mycobacteriosis in zebrafish (Danio rerio) research facilities. Comp Biochem Physiol C Toxicol Pharmacol 2004; 138:383-390.
7. Manrique WG, Claudiano GS, Figueiredo MAP, Petrillo TR, Marcusso PF, Gimeno EJ, et al. Lectinhistochemical staining of granuloma induced by bacillus Calmette-Guérin in Piaractus mesopotamicus. Rev MVZ Córdoba 2013; 18(3):3753-3758.
8. Manrique WG, da Silva Claudiano G, de Castro MP, Petrillo TR, Figueiredo MAP, de Andrade Belo MA, et al. Expression of cellular components in granulomatous inflammatory response in Piaractus mesopotamicus Model. PLoSONE 2015; 10(3):e0121625.
9. Belo MAA, Souza DGF, Faria VP, Prado EJR, Moraes FR, Onaka EM. Haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorynchus curemai. J Fish Biol 2013; 82:1403-1410.
10. Belo MAA, Moraes FR, Yoshida L, Prado EJR, Moraes JRE, Soares VE, et al. Deleterious effects of low level of vitamin E and high stocking density on the hematology response of pacus, during chronic inflammatory reaction. Aquaculture 2014; 422-423: 124-128.
11. Belo MAA, Schalch SHC, Moraes FR, Soares VE, Otobon IA, Moraes JER. Effect of dietary supplementation with vitamin E and stocking density on macrophage recruitment and giant cell formation in the teleost fish, Piaractus mesopotamicus. J Comp Pathol 2005; 133:146-154.
12. Belo MAA, Moraes JRE, Soares VE, Martins ML, Brum CD, Moraes FR. Vitamin C and endogenous cortisol in foreign-body inflammatory response in pacus. Pesqui Agropecu Bras 2012; 47:1015-1021.
13. Sakabe R, Moraes FR, Belo MAA, Moraes JER, Pilarski F. Kinects of chronic inflammation in Nile tilapia supplemented with essential fatty acids n-3 and n-6. Pesqui Agropecu Bras 2013; 48:313-319.
14. Onwueme KC, Vos CJ, Zurita J, Ferreras JA, Quadri LEN. The dimycocerosate ester polyketide virulence factors of mycobacteria. Prog Lipid Res 2005; 44:259–302.
15. Cambier CJ, Takaki KK, Larson RP, Hernandez RE, Tobin DM, Urdahl KB, et al. Mycobacteria manipulate macrophage recruitment through coordinated use of membrane lipids. Nature 2014; 505:218–222.
16. Haenen OLM, Evans JJ, Berthe F. Bacterial infections from aquatic species: potential for and prevention of contact zoonoses. Rev Sci Tech Off Int Epiz 2013; 32(2):497-507.
17. Andersen P, Kaufmann SH. Novel vaccination strategies against tuberculosis. Cold Spring Harb Perspect Med 2014; 2;4(6)pii: a018523.
18. Sado RY, Matushima ER. Histopathological, immunohistochemical, and ultra-structural evaluation of chronic inflammatory response of robalo (Centropomus spp.) to BCG. Braz J Vet Res Anim Sci 2007; 44:58-64.
19. Oksanen KE, Myllymäki H, Ahava MJ, Mäkinen L, Parikka M, Rämet M. DNA vaccination boosts Bacillus Calmette-Guérin protection against mycobacterial infection in zebrafish. Dev Comp Immunol 2016; 9:54(1):89-96.
20. Ishikawa CM, Matushima ER, Oliveira de Souza CW, Timenetsky J, Ranzani-Paiva MT. Micobacteriose em peixes. Bol Inst Pesca 2001; 27:231-242.
21. Castro MP, Moraes FR, Fujimoto RY, Cruz C, Belo MAA, Moraes JRE. Acute toxicity by water containing hexavalent or trivalente chromium in native Brazilian fish, Piaractus mesopotamicus: Anatomopathological alterations and mortality. Bull Environ Contam Toxicol 2014; 92:213-219.
22. Castro MP, Claudiano GS, Petrillo TR, Shimada MT, Belo MAA, Marzocchi-Machado CM, et al. Acute aerocystitis in Nile tilapia bred in net cages and supplemented with chromium carbochelate and Saccharomyces cerevisiae. Fish Shellfish Immunol 2014; 31:284-290.
23. Ishikawa CM. Quantificação bacteriana e avaliação das lesões em peixes da espécie Oreochromis niloticus (tilápia do Nilo) inoculados experimentalmente com Mycobacterium marinum ATCC 927. São Paulo. Dissertação [Mestrado em Epidemiologia Experimental Aplicada às Zoonoses] – Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo; 1998.
24. Ramos P. Visceral granuloma in sea bream (Sparus aurata L.) with presumptive etiology of mycobacteriosis. Rev Port Ci Vet 2000; 95(536):185-191.
25. Manrique WG, Claudiano GS, Petrillo TR, Castro MP, Figueiredo MAP, Belo MAA, et al. Response of splenic melanomacrophage centers of Oreochromis niloticus to inflammatory stimuli by BCG and foreign body. J Appl Ichthyol 2014; 30:1001-1006.
26. Romano LA, Sampaio LA, Tesser MB. Micobacteriose por Mycobacterium marinum em “linguado” Paralichthys orbignyanus e em “barber goby” Elacatinus figaro: diagnóstico histopatológico e imuno-histoquímico. Pesq Vet Bras 2012; 32(3):254-258.
27. Gauthier DT, Rhode MW, Vogelbein WK, Kator H, Ottinger CA. Experimental mycobacteriosis in striped bass Morone saxatilis. Dis Aqua Organ 2003; 54:105-117.
28. Mariano M. The experimental granuloma. A hypothesis to explain the persistence of the lesion. Rev Inst Med Trop São Paulo 1995; 37:161-176.