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. 2014 Oct 10;9(10):e109817.
doi: 10.1371/journal.pone.0109817. eCollection 2014.

A new ochratoxin A biodegradation strategy using Cupriavidus basilensis Őr16 strain

Szilamér Ferenczi  1 , Mátyás Cserháti  2 , Csilla Krifaton  2 , Sándor Szoboszlay  2 , József Kukolya  3 , Zsuzsanna Szőke  4 , Balázs Kőszegi  4 , Mihály Albert  5 , Teréz Barna  6 , Miklós Mézes  7 , Krisztina J Kovács  1 , Balázs Kriszt  2
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Free PMC article

A new ochratoxin A biodegradation strategy using Cupriavidus basilensis Őr16 strain

Szilamér Ferenczi et al. PLoS One. .
Free PMC article

Abstract

Ochratoxin-A (OTA) is a mycotoxin with possibly carcinogenic and nephrotoxic effects in humans and animals. OTA is often found as a contaminant in agricultural commodities. The aim of the present work was to evaluate OTA-degrading and detoxifying potential of Cupriavidus basilensis ŐR16 strain. In vivo administration of OTA in CD1 male mice (1 or 10 mg/kg body weight for 72 hours or 0.5 mg/kg body weight for 21 days) resulted in significant elevation of OTA levels in the blood, histopathological alterations- and transcriptional changes in OTA-dependent genes (annexinA2, clusterin, sulphotransferase and gadd45 and gadd153) in the renal cortex. These OTA-induced changes were not seen in animals that have been treated with culture supernatants in which OTA was incubated with Cupriavidus basilensis ŐR16 strain for 5 days. HPLC and ELISA methods identified ochratoxin α as the major metabolite of OTA in Cupriavidus basilensis ŐR16 cultures, which is not toxic in vivo. This study has demonstrated that Cupriavidus basilensis ŐR16 efficiently degrade OTA without producing toxic adventitious metabolites.

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Conflict of interest statement

Competing Interests: The authors (Mihály A, Szőke Zs and Kőszegi B) are employed by a commercial company (CEVA Phylaxia Ltd and Soft Flow Hungary R&D Ltd.). This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

Figures

Figure 1
Figure 1. Ochratoxin-A biodegradation by Cupriavidus basilensis ŐR16 during 5 day incubation.
Continuous decrease of the OTA concentration is detected in the supernatant and pellet, while OTα concentration is increasing. Abbreviations: HPLC (supernatant OTA) – OTA concentration in the supernatant originated from the degradation experiment measured by HPLC, HPLC (supernatant OTα) – OTα concentration in the supernatant originated from the degradation experiment measured by HPLC, ELISA (supernatant) OTA – OTA concentration in the supernatant originated from the degradation experiment measured by ELISA, ELISA (pellet) OTA – OTA concentration in the pellet originated from the degradation experiment measured by ELISA. Measurements were carried out in triplicate, SD>3%.
Figure 2
Figure 2. OTA concentration in the plasma (ng/ml) analyzed by ELISA after 72 hours of treatment
(A). OTA 1 and OTA 10 groups show elevated OTA concentrations in blood plasma. The OTA 10 deg group shows increased OTA content. Abbreviations: MMS- Group treated with methyl methanesulfonate, OTA 1 and OTA 10- Groups treated with 1 and 10 mg/kg body weight Ochratoxin A, OTA 1 deg and OTA 10 deg- Groups treated with 1 and 10 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium, LB bact- Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium (Kruskal-Wallis test was used). OTA concentrations in the plasma (ng/ml) analyzed by ELISA after 21 days of treatment (B). The OTA 0.5 group shows elevated OTA blood concentrations. Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 0.5 – Group treated with 0.5 mg/kg body weight Ochratoxin A, OTA 0.5 deg – Group treated with 0.5 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium, LB bact – Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium. (One way ANOVA followed by the Tukey's post hoc test were used) Data are presented as mean ± SD (n = 7–10, **p<0.01)
Figure 3
Figure 3. Histology of the kidneys following OTA and degraded OTA administrations.
Photomicrographs showing hematoxylin-eosine stained kidney sections. Abbreviations: MMS- –methyl methanesulfonate treated animals, OTA 1 and OTA 10 – Groups treated with 1 and 10 mg/kg body weight Ochratoxin A, OTA 1 deg and OTA 10 deg – Groups treated with 1 and 10 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, OTA 0.5 – Group treated with 0.5 mg/kg body weight Ochratoxin A, OTA 0.5 deg – Group treated with 0.5 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact – Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium. Symbols: asterisk- dilated tubules with detached necrotic epithelial cells, white arrow- detached necrotic epithelial cells, black arrow- necrotic tubular cells, white arrowhead- tubular cell regeneration. Scale bar: 100 µm.
Figure 4
Figure 4. Effect of acute (72 hours) OTA treatment on gadd45 mRNA expression
(A). MMS and OTA 10 elevated the gadd45 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 1 deg and OTA 10 deg groups) not influenced the mRNA level (One way ANOVA followed by the Tukey's post hoc test were used). Effect of acute (72 hours) OTA treatment on gadd153 mRNA expression (B). OTA 10 elevated the gadd153 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 1 deg and OTA 10 deg groups) not influenced the mRNA level. (Kruskal-Wallis test was used) Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 1 and OTA 10 – Groups treated with 1 and 10 mg/kg body weight Ochratoxin A, OTA 1 deg and OTA 10 deg – Groups treated with 1 and 10 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact- Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium. Data are presented as mean ± SD (n = 7–10, * p<0.05, ***p<0.001)
Figure 5
Figure 5. Effect of acute (72 hours) OTA treatment on annexin2 mRNA expression
(A). OTA 10 elevated the annexin2 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 1 deg and OTA 10 deg groups) did not influence the mRNA level (Kruskal-Wallis test was used). Effect of acute (72 hours) OTA treatment on clusterin mRNA expression (B). OTA 10 elevated the clusterin mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 1 deg and OTA 10 deg groups) did not influence the mRNA level. (One way ANOVA followed by the Tukey's post hoc test were used) Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 1 and OTA 10 – Groups treated with 1 and 10 mg/kg body weight Ochratoxin A, OTA 1 deg and OTA 10 deg- Groups treated with 1 and 10 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact – Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium. Data are presented as mean ± SD (n = 7–10, ** p<0.01, *** p<0.001).
Figure 6
Figure 6. Effect of acute (72 hours) OTA treatment on sult1c2 mRNA expression.
OTA 10 elevated the sult1c2 mRNA level alone. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 1 deg and OTA 10 deg groups) not influenced the mRNA level. (One way ANOVA followed by the Tukey's post hoc test were used) Abbreviations: MMS- Group treated with methyl methanesulfonate, OTA 1 and OTA 10- Groups treated with 1 and 10 mg/kg body weight Ochratoxin A, OTA 1 deg and OTA 10 deg- Groups treated with 1 and 10 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium, LB bact- Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium. Data are presented as mean ± SD (n = 7–10, **** p<0.0001)
Figure 7
Figure 7. Effect of chronic (21 days) OTA treatment on gadd45 mRNA expression
(A). The OTA 0.5 mg/kg bw chronic administration elevated the gadd45 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 0.5 deg group) not influenced the mRNA levels (One way ANOVA followed by the Tukey's post hoc test were used). Effect of chronic (21 days) OTA treatment on gadd153 mRNA expression (B). The OTA 0.5 mg/kg bw chronic administration elevated the gadd153 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 0.5 deg group) did not influence the mRNA level (Kruskal-Wallis test was used). Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 0.5 – Group treated with 0.5 mg/kg body weight Ochratoxin A, OTA 0.5 deg – Group treated with 0.5 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact – Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium. Data are presented as mean ± SD (n = 7–9, *p<0.05***p<0.001)
Figure 8
Figure 8. Effect of chronic (21 days) OTA treatment on annexin2 mRNA expression
(A). The OTA 0.5 mg/kg bw chronic administration elevated the annexin2 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 0.5 deg group) did not influence the mRNA level. (One way ANOVA followed by the Tukey's post hoc test were used) Effect of chronic (21 days) OTA treatment on clusterin mRNA expression (B). The MMS and OTA 0.5 mg/kg bw chronic administration elevated the clusterin mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 0.5 deg group) did not influence the mRNA level. (One way ANOVA followed by the Tukey's post hoc test were used) Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 0.5 – Group treated with 0.5 mg/kg body weight Ochratoxin A, OTA 0.5 deg – Group treated with 0.5 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact – Cupriavidus basilensis ŐR16 in modified Luria- Bertani medium. Data are presented as mean ± SD (n = 7–9, *** p<0.001)
Figure 9
Figure 9. Effect of chronic (21 days) OTA treatment on sult1c2 mRNA expression.
The OTA 0.5 mg/kg bw chronic administration elevated the sult1c2 mRNA level. The metabolised OTA by Cupriavidus basilensis ŐR16 (OTA 0.5 deg group) did not influence the mRNA level. (One way ANOVA followed by the Tukey's post hoc test were used) Abbreviations: MMS – Group treated with methyl methanesulfonate, OTA 0.5 – Group treated with 0.5 mg/kg body weight Ochratoxin A, OTA 0.5 deg- Group treated with 0.5 mg/kg body weight Ochratoxin A + Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium, LB bact- Cupriavidus basilensis ŐR16 in modified Luria-Bertani medium. Data are presented as mean ± SD (n = 6–9, **p<0.01)
Figure 10
Figure 10. Proposed cleavage of Ochratoxin A by Cupriavidus basilensis ŐR16.
The amide bond hydrolysis (up arrow) resulting Ochratoxin α as a major degradation product.
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References

    1. van der Merwe KJ, Steyn PS, Fourie L, Scott DB, Theron JJ (1965) Ochratoxin A, a toxic metabolite produced by Aspergillus ochraceus Wilh. Nature 205: 1112–1113. - PubMed
    1. Walker R (2002) Risk assessment of ochratoxin: current views of the European Scientific Committee on Food, the JECFA and the Codex Committee on Food Additives and Contaminants. Adv Exp Med Biol 504: 249–255. - PubMed
    1. Raters M, Matissek R (2005) Study on distribution of mycotoxins in cocoa beans. Mycotoxin Res 21: 182–186. - PubMed
    1. Zimmerli B, Dick R (1995) Determination of ochratoxin A at the ppt level in human blood, serum, milk and some foodstuffs by high-performance liquid chromatography with enhanced fluorescence detection and immunoaffinity column cleanup: methodology and Swiss data. J Chromatogr B Biomed Appl 666: 85–99. - PubMed
    1. Vega FE, Posada F, Peterson SW, Gianfagna TJ, Chaves F (2006) Penicillium species endophytic in coffee plants and ochratoxin A production. Mycologia 98: 31–42. - PubMed

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Grants and funding

This study was supported by the TÁMOP-4.2.1.B-11/2/KMR-2011-0003, KTIA-AIK_12-1-2013-0017, Research Centre of Excellence-17586-4/2013/TUDPOL, TÁMOP 4.2.1/B-09/1/KONV-2010–0007 and the work was supported by grants from Hungarian Research Fund OTKA 109622 to K.J.K.; 109744 to Sz.F., and Research Centre of Excellence - 8526-5/2014/TUDPOL. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.