Penicillium verrucosum, P. nordicum and Aspergillus carbonarius are three important ochratoxin A producing species. P. verrucosum is in addition able to produce citrinin. It has been shown earlier that P. nordicum is adapted to NaCl rich environments like salt rich dry cured foods or even salines. In this organism, the biosynthesis of ochratoxin A plays an adaptive role in this habitat. P. verrucosum generally can be found on cereals, but occasionally also on salt rich dry cured foods. In contrast A. carbonarius usually cannot be found in NaCl rich environments, but it occurs in another environment with high concentration of solutes, e.g., in sugar rich substrates like grapes and grape juices. Usually osmotic challenging conditions activate the HOG MAP kinase signal cascade, which in turn activates various osmo-regulated genes. In the current analysis, it could be demonstrated that in case of P. nordicum and P. verrucosum the NaCl induced production of ochratoxin A is correlated to the phosphorylation status of the HOG MAP kinase. Just the opposite was true for A. carbonarius. In this case, also higher amounts of NaCl in the medium lead to an increased phosphorylation status of HOG, but no increase in ochratoxin biosynthesis was observed. In contrast to the Penicillia, higher NaCl concentrations lead to a rapid cessation of growth by A. carbonarius. High glucose concentrations have much less impact on growth and the phosphorylation of HOG.
1 September 2013 in International Journal of Food Microbiology
It has previously been shown that the biosynthesis of the mycotoxins ochratoxin A and B and of citrinin by Penicillium is regulated by light. However, not only the biosynthesis of these mycotoxins, but also the molecules themselves are strongly affected by light of certain wavelengths. The white light and blue light of 470 and 455 nm are especially able to degrade ochratoxin A, ochratoxin B and citrinin after exposure for a certain time. After the same treatment of the secondary metabolites with red (627 nm), yellow (590 nm) or green (530 nm) light or in the dark, almost no degradation occurred during that time indicating the blue light as the responsible part of the spectrum. The two derivatives of ochratoxin (A and B) are degraded to certain definitive degradation products which were characterized by HPLC-FLD-FTMS. The degradation products of ochratoxin A and B did no longer contain phenylalanine however were still chlorinated in the case of ochratoxin A. Citrinin is completely degraded by blue light. A fluorescent band was no longer visible after detection by TLC suggesting a higher sensitivity and apparently greater absorbance of energy by citrinin. The fact that especially blue light degrades the three secondary metabolites is apparently attributed to the absorption spectra of the metabolites which all have an optimum in the short wave length range. The absorption range of citrinin is, in particular, broader and includes the wave length of blue light. In wheat, which was contaminated with an ochratoxin A producing culture of Penicillium verrucosum and treated with blue light after a pre-incubation by the fungus, the concentration of the preformed ochratoxin A reduced by roughly 50% compared to the control and differed by > 90% compared to the sample incubated further in the dark. This indicates that the light degrading effect is also exerted in vivo, e.g., on food surfaces. The biological consequences of the light instability of the toxins are discussed.
Aspergillus flavus and A. parasiticus are the main causes of aflatoxin contamination in various foods, particularly grains, as they can thrive in environments with lower water activity and higher temperatures. The growth of Aspergillus and the formation of the mycotoxins aflatoxin and cyclopiazonic acid are strongly influenced by environmental stimuli and can be reduced by modulating parameters such as water activity, pH, temperature and light during the storage. This study has two objectives—on the one hand, to assess how global warming and an increase in exposure to sunlight affect growth and mycotoxin formation, and on the other hand, how the findings from these experiments can be used to reduce fungal growth and mycotoxin formation in stored foods. Using growth substrates with two different water activities (aw 0.95, aw 0.98), together with a light incubation device consisting of different chambers equipped with diodes emitting visible light of five different wavelengths (455 nm, 470 nm, 530 nm, 590 nm, 627 nm) plus white light, we analyzed the growth and mycotoxin formation of selected Aspergillus flavus and A. parasiticus isolates. It was shown that light with a wavelength of 455/470 nm alone, but especially in combination with a lower water activity of aw 0.95, leads to a significant reduction in growth and mycotoxin formation, which was accompanied by reduced transcriptional activity of the responsible mycotoxin biosynthetic genes. Therefore, these results can be used to significantly reduce the growth and the mycotoxin formation of the analyzed fungi during storage and to estimate the trend of fungal infestation by Aspergillus flavus and A. parasiticus in water activity- and light exposure-equivalent climate change scenarios. Mycotoxin-producing aspergilli can be effective and sustainably inhibited using a combination of short-wave light and lowered water activity in the substrate. A higher annual mean temperature accompanying climate change may lead to an increased spread of aflatoxin-producing fungi in areas that were previously too cold for them. On the other hand, there will be regions in the world where contamination with aflatoxin-producing fungi will be reduced due to increased drought and sun exposure.
Nanoparticles are ubiquitous in the environment. They originate from anthropogenic or natural sources or they are intentionally produced for different purposes. There exist manifold applications of nanoparticles in modern life leading unavoidably to a confrontation and interaction between nanomaterial and living organisms. Based on their wide distribution tending to increase steadily, the influence of particles based on silica and silver, exhibiting nominal sizes between 0.65 nm and 200 nm, on the physiology of the mycotoxigenic filamentous fungus Penicillium verrucosum was analyzed. The applied concentration and time-point, the size and the chemical composition of the particles was shown to have a strong influence on growth and mycotoxin biosynthesis. On microscopic scale it could be shown that silver nanoparticles attach to the mycelial surface. Moreover, silver nanoparticles with 0.65 nm and 5 nm in size were shown to internalize within the cell, form agglomerates in the cytoplasm and associate to cell organelles.