Freshly prepared rat hepatocytes used in screening the toxicity of blue‐green algal blooms

Berg, Kjetil; Aune, Tore

doi:10.1080/15287398709530971

Cited by 22 publications

(4 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For ethical reasons, the mouse test is unsuitable for large scale and routine testing of field samples (Aune and Berg, 1986;Heinze, 1996). In recent years, in vitro toxicity tests involving the use of cultured cells have been developed to provide a substitute for the mouse bioassay (Storey et al, 1983;Aune and Berg, 1986;Berg and Aune, 1987;Heinze, 1996). The major advantage of using freshly isolated hepatocytes is their ability to maintain the activities of phase I and II drug-metabolizing enzymes, thus allowing various investigations to be performed including determination of metabolic profiles, inhibition and induction effects (Fautrel et al, 1991;Guillouzo, 1998).…”

mentioning

confidence: 99%

A comparison of in vivo and in vitro assays to assess the toxicity of algal blooms

et al. 2008

View full text Add to dashboard Cite

The toxicity of purified microcystin-LR (MC-LR) and algal material collected during the winter and summer seasons (2005/2006) from the Hartebeespoort dam, South Africa, was investigated using the enzyme-linked immunosorbent assay (ELISA), mouse bioassay, catfish primary hepatocytes (in vitro assay) and protein phosphatase inhibition (PPi) assays. Microcystis aeruginosa, known producer of microcystins, was the dominant cyanobacteria present in the water samples. Exceptionally high cell numbers per millilitre were observed, especially with the summer samples (~1.442 × 10 8 cells/ml), indicating a severe algal bloom in the dam. The toxin concentration as detected by ELISA and PPi assay in the winter and summer extracts was at least 1000 times more than the provisional guideline value (1 mg/l) set by the World Health Organization (WHO) for MC-LR in drinking water. Hepatotoxic effects and death of mice were observed after dosing with the summer extracts, while no hepatotoxic effects were observed with winter extracts. The EC 50 values obtained after exposure of the catfish primary hepatocytes for 72 h to MC-LR, winter and summer extracts was about 0.091, 0.053 and 0.014 mg/l, respectively. Similar toxicity results were obtained when the mouse bioassay and primary hepatocytes were used. Keywords: Catfish primary hepatocytes; Mouse bioassay; ELISA; Protein phosphatase inhibition; AssayIn South Africa and other parts of the world, livestock, waterfowl, wildlife and game animals have died after drinking water containing heavy blooms of blue-green algae (Steyn, 1945;Soll and Williams, 1985;Bell and Codd, 1994;Harding et al., 1995;Van Halderen et al., 1995;Kellerman et al., 2005). Records of poisoning incidents that can be attributed to cyanobacteria in South Africa date back to the 1920s, when mass mortalities of thousands of cattle, sheep, horses and rabbits around pans in the south-eastern Transvaal were reported (Steyn, 1945;Soll and Williams, 1985;Harding and Paxton, 2001). According to a report published by the Department of Water Affairs and Forestry (DWAF, 2002), many South African surface water resources exhibit high nutrient enrichments and eutrophication-related problems. About 80 dams were monitored between October 2002 and September 2003 in South Africa (Van Ginkel, 2003. Eleven per cent of the monitored dams were hypertrophic (showing serious water quality problems), 23% were eutrophic (showing increasing signs of water quality problems) and 25% were mesotrophic (showing emerging signs of water quality problems). Microcystin levels detected in the hypertrophic dams ranged from1 to 28 930 mg/l (Wicks and Thiel, 1990; DWAF, 2002;Van Ginkel, 2003). Given the current status of algal blooms in the impoundments of South Africa, sensitive and specific monitoring assays for cyanotoxins are required. Toxicity testing is important to ensure good water quality for human and animal consumption, and for recreational activities. For many years the mouse bioassay has been used to determine toxicity of blooms (Carmi...

show abstract

mentioning

confidence: 99%

A comparison of in vivo and in vitro assays to assess the toxicity of algal blooms

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Toxicity of different Microcystis-strains has been determined in mice by intraperitoneal injection of crude extracts of algal dry mass. Death occurred in a dosedependent manner within a few hours (2). In vitro testing of crude extracts correlates well with the toxicity in the mouse bioassay.…”

Section: Introductionmentioning

confidence: 63%

Cytotoxicity of Cyanobacterium Microcystis aeruginosa

Henning

Cremer

Meyer

1992

Journal of Veterinary Medicine, Series B

View full text Add to dashboard Cite

Cytotoxic effects of crude extracts and fractions of the purification steps towards Microcystin-LR (MCYST-LR) were investigated in vitro. Cytoxicity was evaluated by measure of lactate dehydrogenase liberation of Chang liver cells and by hemolysis. Crude extracts of strain PCC 7806 damaged the cells within a few minutes. In contrast, MCYST-LR did not show any detectable cytotoxic effects. The cytotoxic activity could be related to a heat-labile substance with a molecular weight of about 30,000 Da.

show abstract

“…This agrees closely with the IC5,, value determined in the V79 protein-dye binding test. Scanning electron microscopy of Microcystis toxin-treated V79 fibroblasts showed that the cells became detached from the substrate, rounded up, and large blebs appeared in the plasma membrane in a manner similar to that seen with freshly isolated hepatocytes when exposed to cyanobacterial toxin (Berg and Aune, 1987; Aune and Berg, 1986;Eriksson et al, 1988). As summarised in Table 111, all samples of natural blooms of M .…”

Section: Eodd Et Almentioning

confidence: 73%

“…The use of freshly isolated rat hepatocytes for the toxicity screening of cyanobacterial blooms has been demonstrated (Aune and Berg, 1986;Berg and Aune, 1987). We believe in uitro cytotoxicity tests hold considerable potential for the assay of cyanobacterial toxins, and here we report on our studies on the effects of toxic blooms and purified toxin on stable fibroblast cell lines.…”

Section: Introductionmentioning

confidence: 92%

Production, detection, and quantification of cyanobacterial toxins

Codd¹,

Brooks²,

Priestley³

et al. 1989

Environ. Toxicol. Water Qual.

View full text Add to dashboard Cite

Cyanobacterial blooms from several British freshwaters have been toxic by mouse bioassay each year since annual sampling began in 1981. Toxic blooms of Microcystis aeruginosa, Anabaena spp., Gloeotrichia echinulata, Oscillatoria spp., and Aphanizomenon flos‐aquae occur, with peptide toxin‐producing Microcystis and Anabaena being most often encountered. We are developing a range of detection and quantification methods for cyanobacterial peptide and alkaloid toxins to supplement the standard mouse bioassay. Both types of toxins can be readily assayed by high performance liquid chromatography, and we have developed facile high performance thin layer chromatographic procedures for their detection from natural blooms and laboratory cultures. We have also produced polyclonal and monoclonal antibodies for the assay of Microcystis toxins by enzyme‐linked immunosorbent assay and have developed in vitro fibroblast cytotoxicity assays for the toxins of Microcystis and other cyanobacteria.

show abstract

Freshly prepared rat hepatocytes used in screening the toxicity of blue‐green algal blooms

Cited by 22 publications

References 6 publications

A comparison of in vivo and in vitro assays to assess the toxicity of algal blooms

A comparison of in vivo and in vitro assays to assess the toxicity of algal blooms

Cytotoxicity of Cyanobacterium Microcystis aeruginosa

Production, detection, and quantification of cyanobacterial toxins

Contact Info

Product

Resources

About