eISSN: 1644-4124
ISSN: 1426-3912
Central European Journal of Immunology
Current issue Archive Manuscripts accepted About the journal Editorial board Abstracting and indexing Subscription Contact Instructions for authors Ethical standards and procedures
SCImago Journal & Country Rank
vol. 34

Review paper
The immunotoxic and nephrotoxic influence of cyanotoxins to vertebrates

Anna Rymuszka
Anna Sierosławska

Centr Eur J Immunol 2009; 34 (2): 129-136
Online publish date: 2009/05/20
Article file
- The immunotoxic.pdf  [0.09 MB]
Get citation
JabRef, Mendeley
Papers, Reference Manager, RefWorks, Zotero
Toxic algae blooms are observed in many water bodies in Poland (e.g. Dobczycki Lake, Goczałkowicki Reservoir, Sulejówek Lake, Zemborzycki Reservoir, Baltic Sea) and constitute the worldwide and multifaceted research problem, having essential health implications for people and animals [1-3]. The cases of acute and chronic poisoning of aquatic organisms, including fish, farmed and domestic animals e.g. dogs, horses, cattle, birds living in the wild were reported. There are also data on intoxication of people being in contact with recreational or drinking water contaminated with cyanotoxins [4]. These substances have a various chemical structure (peptides, alkaloids) and the multidirectional mechanism of toxic action and are grouped into hepatotoxins, neurotoxins, dermatotoxin, cytotoxins, and toxins triggering other effects [5].
Long-term exposure to subclinical doses, which does not induce visible symptoms of poisoning can cause changes on the cellular level and can induce systemic dysfunctions. It is worthwhile noticing that sometimes organ changes are correlated with the decrease of the resistance to pathogens as the result of dysfunction of sensitive mechanisms of immunohomeostasis.

Nephrotoxic effects of cyanotoxins
Kidneys are the organ particularly exposed to the toxic action of xenobiotics. It is a consequence of its excretory functions and intensive blood flow through that tissue. The kidney is a complex organ where toxic changes can be observed, because many nephrotoxicants including drugs, chemicals and natural toxins and their metabolites are filtrated. The proximal tubular cells of the kidney very often can concentrate many nephrotoxins and hence are prone to the harmful effects of toxins.
There is a lot of data in the literature showing the influence of microcystins on the renal system of vertebrates (Table 1). Nephrotoxic effects of chronic administration relatively low doses (10 µg/kg i.p.) of microcystin-LR (MC–LR) and microcystin-YR (MC–YR) were studied by Milutinović et al. [20, 21]. Authors described many pathological changes in the kidneys of rats treated with MCs for 8 months. Degenerative changes in the kidneys were observed such as collapsed tufts of glomerular capillaries, the enlarged diameter of renal corpuscles, the widened Bowman’s space and proximal and distal convoluted tubules, thickened Bowman’s capsule in some renal corpuscles. Moreover, interstitial tissue was occasionally infiltrated by lymphocytes and appeared oedematous. Authors noted that the kidneys were far more affected than the liver. The cytoskeleton abnormalities and the DNA damage suggested, that the mechanisms underlying the chronic nephrotoxicity are similar at the cellular level to the mechanisms of the acute hepatoxicity of microcystins.
In the series of experiments performed by Nobre et al. [15, 18, 22, 23] it was shown that microcystin-LR can affect renal physiology. By using perfused rat kidney model authors showed an intense amount of proteinaceous material in urinary spaces following perfusion with MC-LR. Further studies confirmed that microcystin-LR promoted renal changes, such as altered vascular, glomerular and urinary parameters. MC-LR induced activation of phospholipase A2 (PLA2) and cyclooxygenase and this mechanism was similar to the mechanisms inducing hepatotoxic changes. Moreover, it was demonstrated that microcystin-LR stimulated macrophages to release the inflammatory mediators capable of promoting nephrotoxicity in the isolated perfused rat kidney. Authors examined the supernatant of macrophages stimulated in vivo by microcystin–LR and showed the presence of proinflammatory agents capable of provoking secretion of water and electrolytes (sodium, potassium and chloride). The observed collapsed filaments and other morphological changes support thesis that MCs could trigger apoptotic processes in the exposed kidney cells. That confirms the hypothesis that in vivo MCs induce cytoskeletal alterations and nuclear changes in different cells typical for appoptosis and/or necrosis [6, 13, 14, 26, 31, 35].
Rao et al. [19] suggests, that the observed cytotoxic effects leading to apoptosis were induced by generation of reactive oxygen species and caspase activation of cyanobacterial neurotoxins-anatoxin a in non-neuronal cells [19]. The results of this study showed that anatoxin-containing cell-free extracts from Anabena flos aquae and purified anatoxin-a induced concentration dependent cytotoxicity and apoptosis in African green monkey kidney cells (Vero). The authors observed morphological changes typical for apoptosis as plasma membrane blebbing, cell shrinkage, condensed chromatin, nuclear fragmentation and formation of DNA-containing apoptotic bodies. Several comparative studies have shown that microcystins develop the same cytotoxic response [8-10].
It was revealed by the immunostaining that the injected conjugates can accumulate in the kidneys [36]. It might be thus speculated that in the conditions of chronic exposure to MC-LR accumulation of its metabolites in the kidneys and changes in their physiology may occure. Kotak et al. [11] indicated that kidney tubular epithelial cells in fish were affected after acute exposure by interperitoneal injection of MC-LR 400 µg/kg and 1000 µg/kg. Similarly, Radbergh et al. [7] have shown degenerative changes in the tubular epithelial cells, glomeruli and interstitial tissue in kidneys of carp intraperitoneally exposed to MC-LR with the LD50 ranging from 80 to between 300 and 550 µg/kg. The studies also indicated that fish can tolerate higher doses of the toxin and have a longer survival period compared to mice. Probably the uptake of microcystin to the kidney may be dependent on body temperature.
Fisher and Dietrich [16] found microcystin-induced alterations in kidney tissues of carp when Microcystis aeruginosa (PCC 7806) amounting to an equivalent of 400 µg MC-LR/kg bw were directly administered to the fish stomach. In the kidney degenerative changes were observed in the renal proximal tubules, a segment known for its high capacity of active protein and peptide reabsorption. Moreover, the studies on the mechanism of cell toxicity showed that cyanotoxins induce oxidative stress in tissues of vertebrates and the potential alterations of the antioxidant status [25, 27, 30, 34].

Immunotoxic effects of cyanotoxins
Many substances present in the aquatic environment at relatively low concentrations demonstrate toxic action on the immune cells and organs of fish and higher vertebrates. Immune system, together with other systems e.g. nervous and endocrine, takes part in regulating homeostasis, so cyanotoxins, which have multidirectional nature of the action can also induce directly or indirectly dysfunctions of the immune system. The immunotoxic effects of cyanotoxins are summarised in Table 2. Immunosuppression was also confirmed in our study, which determined the influence of microcystin-LR and anatoxin-a to the basic functions of the fish immune cells [56, 57, 60].
The obtained findings showed the inhibition of the viability of lymphocytes and phagocytes isolated from rainbow trout by the toxin in the time and concentration dependent manner. Microcystin-LR suppressed the examined functions of immune cells (metabolic activity of phagocytes, proliferative response of lymphocytes) more distinctively compared to anatoxin-a (in press). Moreover, we noted that phagocytes are more sensitive to microcystin-LR than lymphocytes. It is interesting, that extracts containing the cyanotoxins are more immunotoxic, than the pure form toxins (unpublished).
Moreover, other authors describe the decrease of the total number of white blood cells, including T lymphocytes (particularly cytotoxic T cell), B cells, mielocytes and the lowered value of the phagocytic index after the exposure to microcystins [37, 47, 59].
The mechanism of toxic action of microcystins is the blockade of the activity of protein phosphatases serine (PP1) and threonine (PP2) what leads to hyperphosphorylation of plasmic and cellular cytoskeleton proteins. The disorders of the intracellular homeostasis can result in the uncontrolled proliferation and as a consequence induce carcinogenesis [19, 41, 44, 50, 52, 54, 55, 59, 61].
The activation of the phagocytic cells is connected with a sequence of changes in their cytoskeleton. Our studies showed that phagocytes may be the target cells for toxic effects of the microcystin-LR in the fish immune system [57, 60]. The toxin at the concentrations environmentally relevant caused the increased production of reactive forms of oxygen in phagocytes and modulation of the phagocytosis. The disorders of the metabolic activity of these cells can result from the direct effects of the toxin on their cytoskeleton which influence the activation and process of phagocytosis.
Our studies indicated that microcystin-LR is more suppressive to B lymphocytes, than to T cells [56]. Differences in the response of the two lymphocyte populations to the cyanotoxin probably result from the action of cytokines – essential regulatory proteins of the immune system, as well as nervous and endocrine systems. This hypothesis is confirmed by studies carried out on mammalian immune cells. These studies showed that microcystin-LR influenced the production of cytokines: interleukin-2 (IL-2) and interleukin-6 (IL-6) responsible for lymphocyte functioning [45]. Moreover, the influence of this toxin on the expression of cytokines such as: IL-1b, IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-g, as well as on the nitric oxide synthase activity in phagocytes was observed [39, 42, 48, 49, 51, 52, 58].
In summary, our research and the observations of other authors suggested that cyanotoxins may induce nephrotoxic and immunotoxic effects and changes in physiology of vertebrates.

1. Owsianny PM, Stefaniak K, Kokociński M (2003): Occurrence of potentially toxic Peridiniopsis polonicum (Dinophyta) in lakes of the Wielkopolska region (Poland). Acta Bot Warmiae et Masuriae 3: 87-99.
2. Mazur H, Pliński M (2003): Cyanobacterial toxins in fresh and brackish waters of Pomorskie Province (Northern Poland). Oceanol Hydrobiol Stud 32: 15-26.
3. Pawlik-Skowrońska B, Skowroński T, Pirszel J, Adamczyk A (2004): Realtionship between cyanobacterial bloom composition and anatoxin-a and microcystin occurrence in the eutrophic dam reservoir (SE Poland). Pol J Ecol 52: 479-490.
4. Ibelings BW, Chorus I (2007): Accumulation of cyanobacterial toxins in freshwater “seafood” and its consequences for public health: A review. Environ Pollut 150: 177-192.
5. Wiegand C, Pflugmacher S (2005): Ecotoxicological effects of selected cyanobacterial secondary metabolites: a short review. Toxicol Appl Pharmacol 203: 201-218.
6. Hooser SB, Beasley VR, Lovell RA et al. (1989): Toxicity of microcystin LR, a cyclic heptapeptide hepatotoxin from Microcystis aeruginosa, to rats and mice. Vet Pathol 26: 246-252.
7. Radbergh CMI, Bylund G, Eriksson JE (1991): Histological effects of microcystin-LR, a cyclic peptide toxin from the cyanobacterium (blue-green alga) Microcystis aeruginosa, on common carp (Cyprinus carpio L.). Aquat Toxicol 20: 131-146.
8. Wickstrom ML, Khan SA, Haschek WM et al. (1995): Alterations in microtubules, intermediate filaments, and microfilaments induced by MCLR in cultured cells. Toxicol Pathol 23: 326-337.
9. Khan SA, Ghosh S, Wickstrom M et al. (1995): Comparative pathology of microcistin-LR in cultured hepatocytes, fibroblasts, and renal epithelial cells. Nat Toxins 3: 119-128.
10. Khan SA, Wickstrom ML, Haschek WM et al. (1996): Microcystin-LR and kinetics of cytoskeletal reorganization in hepatocytes, kidney cells, and fibroblasts. Nat Toxins 4: 206-214.
11. Kotak BG, Semalulu S, Fritz DL et al. (1996): Hepatic and renal pathology of intraperitoneally administered microcystin-LR in rainbow trout (Oncorhynchus mykiss). Toxicon 34: 517-525.
12. Bhattacharya R, Sugendran K, Dangi RS, Rao PV (1997): Toxicity evaluation of freshwater cyanobacterium Microcystis aeruginosa PCC 7806. II. Nephrotoxicity in rats. Biomed Environ Sci 10: 93-101.
13. Rao PVL, Bhattacharya R, Parida MM et al. (1998): Freshwater cyanobacterium Microcystis aeruginosa (UTEX 2385) induced DNA damage in vivo and in vitro. Environ Toxicol Pharmacol 5: 1-6.
14. Vajcova V, Navaratil S, Palikova M (1998): The effect of intraperitoneally applied pure microcystin-LR on haematological and morphological indices of silver carp (Hypopthalmichthys molitrix val.). Acta Vet Brno 67: 281-287.
15. Nobre ACL, Jorge MCM, Menezes DB et al. (1999): Effects of microcystin-LR in isolated perfused rat kidney. Braz J Med Biol Res 32: 985-988.
16. Fischer WJ, Dietrich DR (2000): Pathological and biochemical characterization of microcystin – induced hepatopancreas and kidney damage in carp (Cyprinus carpio). Toxicol Appl Pharmacol 164: 73-81.
17. Beasley VR, Lovell RA, Holmes KR et al. (2000): Microcystin-LR decreases hepatic and renal perfusion, and causes circulatory shock, severe hypoglycemia, and terminal hyperkalemia in intravascularly dosed swine. J Toxicol Environ Health A 61: 281-303.
18. Nobre AC, Coelho GR, Coutinho MC et al. (2001): The role of phospholipase A2 and cyclooxygenase in renal toxicity induced by microcystin-LR. Toxicon 39: 721-724.
19. Rao PVL, Bhattacharya R, Gupta N et al. (2002): Involvement of caspase and reactive oxygen specie in cyanobacterial toxin anatoxin-a-induced cytotoxicity and apoptosis in rat thymocytes and Vero cells. Arch Toxicol 76: 227-235.
20. Milutinovič A, Sedmark B, Horvat-Znidaršič I, Šuput D (2002): Renal injuries induced by chronic intoxication with microcystins. Cell Mol Biol Lett 7: 139-141.
21. Mulutinović A, Zivin M, Zorc-Pleskovič R et al. (2003): Nephrotoxic effects of chronic administration of microcystins-LR and -YR. Toxicon 42: 281-288.
22. Norbe AC, Martins AM, Havt A et al. (2003): Renal effects of supernatants from rat peritoneal macrophages activated by microcystin-LR: role protein mediators. Toxicon 41: 377-381.
23. Norbe ACL, Nunes-Monteiro SM, Monteiro MCSA et al. (2004): Microcystin-LR promote intestinal secretion of water and electrolytes in rats. Toxicon 44: 555-559.
24. Molina R, Moreno I, Pichardo S et al. (2005): Acid and alkaline phosphatase activities and pathological changes induced in tilapia fish (Oreochromis sp.) exposed subchronically to microcystins from toxic cyanobacterial blooms under laboratory conditions. Toxicon 46: 725-735.
25. Jos A, Pichardo S, Prieto AI et al. (2005): Toxic cyanobacterial cells containing microcystins induce oxidative stress in exposed tilapia fish (Oreochromis sp.) under laboratory conditions. Aquat Toxicol 72: 261-271.
26. Moreno I, Pichardo S, Jos A et al. (2005): Antioxidation enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon 45: 395-402.
27. Prieto AI, Jos A, Pichardo S et al. (2006): Differential oxidative stress responses to microcystins LR and RR in intraperitoneally exposed tilapia fish (Oreochromis sp.). Aquat Toxicol 77:
28. Jayaraj R, Deb U, Bhaskar AS et al. (2007): Hepatoprotective efficacy of certain flavonoids against microcystin induced toxicity in mice. Environ Toxicol 22: 472-479.
29. Carvalho EG, Sotero-Santos RB, Martinez CBR et al. (2007): Kidney histology of mice after seven days oral intake of cyanobacterial extract. J Braz Soc Ecotoxicol 2: 39-43.
30. Li L, Xie P, Chen J (2007): Biochemical and ultrastructural changes of the liver and kidney of the phytoplanktivorous silver carp feeding naturally on toxic Microcystis blooms in Taihu Lake, China. Toxicon 49: 1042-1053.
31. La-Salete R, Oliveira MM, Palmeira CA et al. (2008): Mitochondria a key role in microcystin-LR kidney intoxication. J Appl Toxicol 28: 55-62.
32. Gaudin J, Huet S, Jarry G, Fessard V (2008): In vivo DNA damage induced by the cyanotoxin microcystin-LR: comparison of intraperitonial and oral administration by use of the comet assay. Mutat Res 652: 65-71.
33. Andrinolo D, Sedan D, Telese L et al. (2008): Hepatic recovery after damage produced by sub-chronic intoxication with the cyanotoxin microcystin-LR. Toxicon 51: 457-467.
34. Atencio L, Moreno I, Jos A et al. (2009): Effects of dietary selenium on the oxidative stress and pathological changes in tilapia (Oreochromis niloticus) exposed to a microcystin-producing cyanobacterial water bloom. Toxicon 53: 269-282.
35. Dias E, Andrade M, Alverca E et al. (2009): Comparative study of the cytotoxic effect of microcystin-LR and purified extracts from Microcystis aeruginosa on a kidney cell line. Toxicon 53: 487-495.
36. Ito E, Takai A, Kondo F et al. (2002): Comparison of protein phosphatase inhibitory activity and apparent toxicity of microcystins and related compounds. Toxicon 40: 1017-1025.
37. Palikova M, Kovaru F, Navratil S et al. (1998): The effects of pure microcystin LR and biomass of blue-green algae on selected immunological indices of carp (Cyprinus carpio L.) and silver carp (Hypophthalmichthys molitrix Val.). Acta Vet Brno 67: 265-272.
38. Hermández M, Macia M, Padilla C, Del Campo FF (2000): Modulation of human polymorphonuclear leukocyte adherence by cyanopeptide toxins. Environ Res 84: 64-68.
39. Rocha MFG, Sidrim JJC, Soares AM et al. (2000): Supernatants from macrophages stimulated with microcystin-LR induce electrogenic intestinal response in rabbit ileum. Pharmacol Toxicol 87: 46-51.
40. Yea SS, Kim HM, Jeon YJ et al. (2000): Suppression of IL-2 and IL-4 gene expression by nodularin through the reduced NF-AT binding activity. Toxicol Lett 114: 215-224.
41. Mankiewicz J, Tarczyńska M, Fladmark KE et al. (2001): Apoptotic effect of cyanobacterial extract on rat hepatocytes and human lymphocytes. Environ Toxicol 16: 225-233.
42. Yea SS, Kim HM, Oh HM et al. (2001): Microcystin-induced down-regulation of lymphocyte functions through reduced IL-2 mRNA stability. Toxicol Lett 122: 21-31.
43. Shen PP, Zhao SW, Zheng WJ et al. (2003): Effects of cyanobacteria bloom on some parameters of immune function in mice. Toxicol Lett 143: 27-36.
44. Lankoff A, Krzowski Ł, Głąb J et al. (2004): DNA damage and repair in human peripheral blood lymphocytes following treatment with microcystin-LR. Mutat Res 559: 132-142.
45. Lankoff A, Carmichael WW, Grasman KA, Yuan M (2004): The uptake kinetics and immunotoxic effects of microcystin-LR in human and chicken peripheral blood lymphocytes in vitro. Toxicology 204: 23-40.
46. Wright PFA, Harford A, O’Halloran K (2004): Immunomodulation of head kidney cell functions in Murray cod by microcystin-LR. Toxicol Appl Pharmacol 197: 284-294.
47. Palikova M, Navratil S, Krejci R et al. (2004): Outcomes of repeated exposure of the carp (Cyprinus carpio L.) to cyanobacteria extract. Acta Vet Brno 73: 259-265.
48. Chen T, Zhao X, Liu Y et al. (2004): Analysis of immunomodulating nitric oxide iNOS and cytokines mRNA in mouse macrophages induced by microcystin-LR. Toxicology 197: 67-77.
49. Shi Q, Cui J, Zhang J et al. (2004): Expression modulation of multiple cytokines in vivo by cyanobacteria blooms extract from Taihu lake, China. Toxicon 44: 871-879.
50. Teneva I, Mladenov R, Popov N, Dzhambazov B (2005): Cytotoxicity and apoptotic effects of microcystin-LR and anatoxin-a in mouse lymphocytes. Folia Biol Praha 51: 62-67.
51. Chen T, Shen P, Zhang J, Hua Z (2005): Effects of microcystin-LR on patterns of iNOS and cytokine mRNA expression in macrophages in vitro. Environ Toxicol 20: 85-91.
52. Goncalves EAP, Dalboni MA, Peres AT et al. (2006): Effect of microcystin on leukocyte viability and function. Toxicon 47: 774-779.
53. Kujbida P, Hatanaka E, Campa A et al. (2006): Effects of microcystins on human polymorphonuclear leukocytes. Biochem Bioph Res Commun 341: 273-277.
54. Zhang H, Zhang J, Chen Y (2006): Sensitive apoptosis induced by microcystins in the crucian carp (Carassius auratus) lymphocytes in vitro. Toxicol In Vitro 20: 560-566.
55. Zhang H, Zhang J, Chen Y, Zhu Y (2007): Influence of intracellular Ca(2+), mitochondria membrane potential, reactive oxygen species, and intracellular ATP on the mechanism of microcystin-LR induced apoptosis in (Carassius auratus) lymphocytes in vitro. Environ Toxicol 22: 559-564.
56. Rymuszka A, Sierosławska A, Bownik A, Skowroński T (2007): In vitro effects of pure microcystin-LR on the lymphocyte proliferation in rainbow trout (Oncorhynchus mykiss). Fish Shell Immunol 22: 289-292.
57. Sierosławska A, Rymuszka A, Bownik A, Skowronski T (2007): The influence of microcystin-LR on fish phagocytic cells. Hum Exp Toxicol 26: 603-607.
58. Kujbida P, Hatanaka E, Campa A et al. (2008): Analysis of chemokines and reactive oxygen species formation by rat and human neutrophils induced by microcystin-LA, -YR and -LR. Toxicon 51: 1274-1280.
59. Wei LL, Sun BJ, Nie P (2008): Ultrastructural alteration of lymphocytes in spleen and pronephros of grass carp (Ctenopharyngodon idella) experimentally exposed to microcystin-LR. Aquaculture 280: 270-275.
60. Rymuszka A, Sierosławska A, Bownik A, Skowroński T (2008): Immunotoxic potential of cyanotoxins on the immune system of fish. Cent Eur J Immunol 33: 150-152.
Copyright: © 2009 Polish Society of Experimental and Clinical Immunology This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) License (http://creativecommons.org/licenses/by-nc-sa/4.0/), allowing third parties to copy and redistribute the material in any medium or format and to remix, transform, and build upon the material, provided the original work is properly cited and states its license.
Quick links
© 2020 Termedia Sp. z o.o. All rights reserved.
Developed by Bentus.
PayU - płatności internetowe