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PubMed2003

Patulin in Italian commercial apple products.

Patulin is a mycotoxin produced by microscopic fungi belonging to the Penicillium and Aspergillus genera. The natural occurrence of patulin in four apple products marketed in Italy and purchased from the supermarket, herbalist, and retail shops was studied. Thirty-three samples of the four products had no detectable patulin contamination. The 11 positive samples had a concentration ranging between 1.4 and 74.2 microg/L with a mean of 26.7 microg/L. All vinegar samples were negative for patulin; of 10 apple-based baby foods, two samples were contaminated with 17.7 and 13.1 microg/L and both were labeled as "organic food". Comparing organic and conventional agricultural practices, no significant differences were found. Finally, optimization of extraction protocol more general and useful for juices, clarified juices, baby foods, vinegars, and purees was performed. The low incidence of the patulin level in Italian apple products is a clear parameter to judge the quality of the fruit, and the process is of a high standard.

Acetic Acid↗

PubMed1975

Influence of carbohydrate and nitrogen source on patulin production by Penicillium patulum.

A strain of Penicillium patulum, isolated from cheddar cheese, produced patulin when grown on liquid media containing lactose and milk nitrogen sources. Patulin production was affected by the temperature of incubation, the type and amount of carbohydrate, and the type of nitrogen source present. Patulin levels generally were depressed by incubation at 5 C and low carbohydrate levels. Patulin was produced at low levels in the absence of sugars at 5 C when the mold was grown on milk nitrogen sources. No patulin was detected in cultures grown on 25% casein slurries or cheddar cheese, even though growth of the mold was extensive.

Animals↗

PubMed2002

The effect of modified atmospheres and packaging on patulin production in apples.

This study was undertaken to determine the effectiveness of modified atmospheres and packaging materials on the growth of Penicillium expansum and patulin production in apples. Granny Smith apples were surface sterilized with 76% ethanol and inoculated with 0.1 ml of a 1.1 x 10(7) spore/ml P. expansum spore suspension. The apples were packaged either in polyethylene (PE) or polypropylene (PP) and treated with three different gas combinations, viz., 58% CO2/42% N2, 48% CO2/52% N2, and 88% CO2/12% N2, and were then incubated for 14 days at 25 degrees C. Fungal growth was monitored every 2 to 4 days by measuring radial growth from the point of inoculation. After the 14th day, apples were pulped, and patulin was extracted, purified, and quantified by high-performance liquid chromatography. PP did not inhibit fungal growth in any of the atmospheres tested, and it only inhibited patulin production in atmospheric gas and 58% CO2/42% N2. PE was very effective and inhibited fungal growth by four- or fivefold, depending on the modified atmosphere. Patulin production in PE-packaged apples was almost completely inhibited by all three gas combinations. Gas chromatographic analysis of the PE-packaged samples before and after the incubation period showed that CO2 levels dropped and N2 levels increased for all of the atmospheres tested. Our studies showed conclusively that PE is an excellent packaging material for the storage of apples since it inhibited the growth of P. expansum, thereby allowing <3.2 microg/ml of patulin to be produced, regardless of gaseous environment.

Carbon Dioxide↗

PubMed2005

Detection of patulin in apple juices marketed in the Tohoku district, Japan.

Patulin is a mycotoxin mainly produced by Penicillium and Aspergillus. We investigated the incidence of patulin contamination in 179 samples of apple juice and 9 samples of mixed juice (containing apple juice concentrate as an ingredient) commercially available in the Tohoku district of Japan. Patulin was detected in 3 of 143 samples containing domestic fruits and in 6 of 45 samples containing imported products and products produced in Japan using imported apple juice concentrate. Patulin analyses were carried out using high-pressure liquid chromatography with a detection limit of 4 microg/liter. The patulin content of contaminated domestic samples (three samples with concentrations ranging from 6 to 10 microg/liter), imported samples (one sample with a concentration of 15 microg/liter), and domestic samples produced containing imported concentrate (five samples with concentrations ranging from 6 to 9 microg/liter) was lower than the maximum limit of 50 microg/liter currently adopted by many countries, including Japan.

Beverages↗

PubMed1975

[Toxinogenic moulds in silage. II. -- In vitro kinetics of patulin and byssochlamic acid biosynthesis by Byssochlamys nivea Westling in liquid medium (author's transl)].

A strain of Byssochlamys nivea (E 70), isolated from an silage and in pure culture, produces two mycotoxins (patulin and byssochlamic acid). A comparison of the toxinogenesis of this fungic species on eight culture media (table I) shows that maximal patulin concentration (816 p.p.m.) is obtained on Czapek's liquid medium (Difco) enriched with yeast extract (2 p. 1000) and glucose (8 p. 1 000) ; maximal byssochlamic acid concentration (390 p.p.m.) is obtained on wet corn grains (80 p. 100 of water). The lowest amounts are found on soya meal (patulin : 30 p.p.m.; byssochamic acid : 15.2 p.p.m.). Study of patulin and byssochlamic acid biosynthesis kinetics on liquid medium at 26 degrees C reveals that maximum patulin is obtained on day 9 of culture. It precedes maximum growth of the mould, and appears to coincide with phialospores formation. Maximum production of byssochlamic acid is obtained after 60 days of culture, that is, during mould autolysis phase. Kinetics of these mycotoxin productions should be studied in silage storage conditions.

Animal Feed↗

PubMed1992

Effect of preservatives on patulin production by Penicillium expansum.

Penicillium expansum has been grown on Capek-Dox medium using glucose and fructose as carbon source. Preservatives used in fruit processing and introduced in the medium were sorbic acid, formic acid, benzoic acid, SO2 and saccharose. Sulphur dioxide had a most inhibitory effect on mycelium growth and patulin production, formic acid concentration of 0.025% increased the amount of patulin by about 30% as compared to the culture with no preservatives. However its higher concentrations inhibited synthesis of this mycotoxin. Sorbic acid concentration of 0.1% stimulated the fungus strains examined in patulin synthesis but its highest amounts were detected using 0.0125% benzoic acid increased patulin secretion from 8 to 50% as compared to the control, depending on the strain examined. Saccharose concentration up to 50% clearly decreased patulin content in the medium until its total disappearance.

Benzoates↗

PubMed1979

Protein synthesis inhibition in rat liver by the mycotoxin patulin.

Patulin, a carcinogenic mycotoxin, inhibits in vivo and in vitro protein synthesis in rat liver. The in vivo inhibition culminates 5 h after toxin administration and reaches a maximum of 65% with regard to control values; a breakdown of polysomes is associated with the translational blockage. However in in vitro systems, the cellular fractions obtained from patulin-treated rats appear equally as active in protein synthesizing ability and as sensitive to the toxin action as those prepared from control animals. The in vitro inhibition by patulin is dose related. The postmitochondrial system is less sensitive than that functioning with isolated polysomes and pH 5 enzyme. This difference might be due to the presence of soluble factor(s) that counteract patulin action. We propose that the inhibition of protein synthesis by patulin may result from an interaction of the drug with active SH groups at the membrane level (amino acid transport, ion equilibrium) and/or at the cytoplasmic level (enzymes and factors involved in the translational process).

Amino Acids↗

PubMed1977

Occurrence of patulin in fruit and vegetables.

In different varieties of apples and pears suffering from brown rot, patulin was found in about 50 p. 100 of samples investigated (about 120). Patulin levels as high as 1 g/kg rotten material were found 2-3 days after the fruit was removed from the cold store where it had been stored for 5 months. A significant diffusion into the healthy plant tissue was not observed in apples, but in peaches, tomatoes and pears. Vegetable products with natural patulin content: apples, pears, peaches, apricots, bananas, pine apples, grapes. After inoculation with Penicillium expansum, P. urticae or Byssochlamys nivea, patulin was also found in greengages, strawberries, honeydew melons, red and green paprika, tomatoes, cucumbers and carrots. The artificial infection failed in celeriac, kohlrabi, cauliflower, red cabbage, radish, horseradish, onions, squash (zuccini), potatoes and egg plants. The author further investigated the patulin synthesis as a function of the temperature as well as its oxygen requirement in various mold species and strains.

Food Microbiology↗

PubMed1973

Patulin production in apples decayed by Penicillium expansum.

Sixty isolates of Penicillium expansum were tested for patulin production in decaying apples. All the isolates were found to produce the mycotoxin patulin as determined by thin-layer chromatography. Since patulin is known to be stable in many apple products, the results indicate that apple products made partially from apples decayed by P. expansum will contain patulin which may present a health hazard. The results also suggest that patulin may be important in the decay of apples by P. expansum.

Chromatography, Thin Layer↗

PubMed1989

Patulin-induced ion flux in cultured renal cells and reversal by dithiothreitol and glutathione: a scanning electron microscopy (SEM) X-ray microanalysis study.

Patulin (PAT), a compound produced by certain species of Aspergillus, Penicillium, and Byssochlamys, is frequently found associated with agricultural commodities. PAT has many effects on membrane function, including the inhibition of the isolated Na+-K+ ATPase. In this study, a scanning electron microscope equipped with an energy dispersive spectroscopy X-ray microanalysis system was used to examine individual cultured renal epithelial cells (LLC-PK1) in order to determine the effects of PAT on the relative intracellular ion concentrations. The estimated EC50 (60 min) for both sodium influx and potassium efflux was between 10 and 50 microns for ouabain. For PAT, the EC50 (60 min) was 250 microns for sodium influx and 100 microns for potassium efflux. However, 1 mM patulin at 240 min caused complete reversal of the sodium and potassium content of cells, and 1 mM ouabain at 240 min did not. The effect of patulin on sodium and potassium flux was both concentration and time dependent and was reversed by dithiothreitol and glutathione. PAT (250 microM) but not ouabain (250 microM) induced massive blebbing of LLC-PK1 cells. Thus, the interaction of PAT with cellular membranes involves both alterations in the regulation of intracellular ion content and the cytoskeleton. We hypothesize that patulin alters intracellular ion content via Na+-K+ ATPase and non-Na+-K+ ATPase mechanisms.

Animals↗

PubMed2002

The mycotoxins citrinin, gliotoxin, and patulin affect interferon-gamma rather than interleukin-4 production in human blood cells.

Exposure to molds diminishes the numbers of T-helper type 1 (Th1) cells in the peripheral blood of children and is a risk factor for the development of allergic diseases (results of LARS: Leipzig Allergy Risk Children Study, Mueller et al. 2002). We hypothesized that mycotoxins are responsible for this effect and therefore investigated the influence of citrinin, gliotoxin, and patulin on human peripheral blood mononuclear cells (PBMC). CD3/CD28-stimulated PBMC of healthy donors were incubated for 24 h with the mycotoxins in serial dilutions and triplicates. Vitality and proliferation were tested using the MTT assay and T-cell function by the expression of cytokines (ELISA, intracellular cytokine staining, and real-time polymerase chain reaction (RT-PCR) for interferon-gamma (IFN-gamma) and interleukin-4 (IL-4). The cytokine secretion was inhibited at concentrations 2-130 times lower compared to vitality (ELISA versus MTT assay). The strongest inhibition of cytokine expression was found for IFN-gamma: 8.3 microg/mL citrinin, 34.2 ng/mL gliotoxin, and 64.8 ng/mL patulin caused a 50% inhibition of the IFN-gamma release (50% inhibitory dose, ID(50)). For IL-4 release the corresponding ID(50) values were 21.6 microg/mL citrinin, 82.8 ng/mL gliotoxin, and 243.2 ng/mL patulin. Furthermore, 3 ng/mL patulin caused a significant increase of IL-4 but a significant suppression of IFN-gamma. On the mRNA level, after 24 h an unaltered or enhanced IL-4 was observed compared to a reduced IFN-gamma expression. Using a method of intracellular cytokine staining, we were able to show that the described effects are caused by a reduction of the number of IFN-gamma-producing T lymphocytes rather than by a reduced functional capacity of the single cell. We suggest that mycotoxins primarily cause stronger inhibition of IFN-gamma-producing Th1 cells, which may lead to T-cell polarization toward the Th2 phenotype and may raise the risk for the development of allergies.

Anti-Bacterial Agents↗

PubMed1984

The interaction between additives, yeast and patulin production in grass silage.

Both laboratory-prepared and sterile farm silage was found to support growth of Paecilomyces sp. and patulin production. The formation of patulin was affected by the levels of yeast present in the silage, and it was found that there was an inverse relationship between yeast population levels and patulin concentration. The commercial silage additive, "Sylade' had a greater lethal effect on yeast and fungi than "Add F', the latter allowing the formation of patulin by Paecilomyces sp. in the silage.

Animal Feed↗

PubMed1992

Action of gamma-irradiated patulin on Saccharomyces cerevisiae.

The action of gamma-irradiated patulin on Saccharomyces cerevisiae LOCK 2144 in liquid culture was studied. Patulin irradiated in an aqueous solution with doses ranging over 0.34-1.36 kGy inhibited the yeast growth in a proportion to the concentration of undestroyed toxin. Patulin disappearance in the growth medium occurred between 12-72 h of incubation at 30 degrees C. The patulin content did not essentially change in the period of log phase of yeast growth which is accompanied by rapid glucose uptake.

Chromatography, High Pressure Liquid↗

PubMed1979

Stability of patulin at ph 6.0-8.0 and 25 degrees c.

Patulin was tested for stability at pH values of 6.0, 6.5, 7.0, 7.5, and 8.0, using Sörensen's phosphate buffer at 25 degrees C. Patulin was determined by high-performance liquid chromatography. When the percentage of patulin remaining was plotted versus reaction time, apparent first-order reaction plots were obtained. Reaction rate constants for disappearance of patulin ranged from 1.1 x 10(-2) h at pH 8.0-5.3 x 10(-4) h at pH 6.0. Values for half-life were calculated and ranged from 64 h at pH 8.0 to 1310 h at pH 6.0.

Hydrogen-Ion Concentration↗

PubMed2007

The sequence of the isoepoxydon dehydrogenase gene of the patulin biosynthetic pathway in Penicillium species.

Interest in species of the genus Penicillium is related to their ability to produce the mycotoxin patulin and to cause spoilage of fruit products worldwide. The sequence of the isoepoxydon dehydrogenase (idh) gene, a gene in the patulin biosynthetic pathway, was determined for 28 strains representing 12 different Penicillium species known to produce the mycotoxin patulin. Isolates of Penicillium carneum, Penicillium clavigerum, Penicillium concentricum, Penicillium coprobium, Penicillium dipodomyicola, Penicillium expansum, Penicillium gladioli, Penicillium glandicola, Penicillium griseofulvum, Penicillium paneum, Penicillium sclerotigenum and Penicillium vulpinum were compared. Primer pairs for DNA amplification and sequencing were designed from the P. griseofulvum idh gene (GenBank AF006680). The two introns present were removed from the nucleotide sequences, which were translated to produce the IDH sequences of the 12 species for comparison. Phylogenetic relationships among the species were determined from rDNA (ITS1, 5.8 S, ITS2 and partial sequence of 28S rDNA) and from the idh nucleotide sequences minus the two introns. Maximum parsimony analysis showed trees based on rDNA and idh sequences to be congruent. It is anticipated that the genetic information obtained in the present study will aid in the design of probes, specific for patulin biosynthetic pathway genes, to identify the presence of these mycotoxigenic fungi.

Amino Acid Sequence↗

PubMed1991

The mechanism of patulin's cytotoxicity and the antioxidant activity of indole tetramic acids.

In LLC-PK1 cells exposed to patulin (50 microM), lipid peroxidation, abrupt calcium influx, extensive blebbing, and total LDH release appeared to be serially connected events with each representing a step in the loss of structural integrity of the plasma membrane. The aforementioned patulin-induced events were prevented by concurrent incubation with butylated hydroxytoluene, deferoxamine, and cyclopiazonic acid, a fungal metabolite. Patulin also caused depletion of nonprotein sulfhydryls, increased 86Rb+ efflux, dome collapse, and eventually the loss of cell viability. These events were not prevented by antioxidants, results consistent with the hypothesis that they were also serially connected but occurring parallel to those previously mentioned. The earliest events observed in patulin-treated cells were the decrease in nonprotein sulfhydryls and increase in 86Rb+ efflux (5 min) which occurred before statistically significant alterations in protein-bound sulfhydryls. The increased potassium efflux (86Rb+ efflux) occurred via a pathway distinct from BaCl2, quinine, or tetraethylammonium sensitive potassium channels. This is the first published report of the antioxidant activity of indole tetramic acids (cyclopiazonic acid and cyclopiazonic acid imine). The protective effect of tetramic acids in LLC-PK1 cells was restricted to indole tetramic acids, and their prevention of lipid peroxidation did not involve iron chelation. The results of this study demonstrate that cyclopiazonic acid is a potent inhibitor of azide-insensitive, ATP-dependent, a23187-sensitive calcium uptake by the lysate of LLC-PK1 cells. This result is consistent with the hypothesis that the endoplasmic reticulum calcium transport ATPase is a sensitive target for cyclopiazonic acid in LLC-PK1 cells. These findings raise the interesting possibility that the antioxidant activity of indole tetramic acids may involve multiple novel mechanisms: surface charge alterations on the cytoplasmic surface of plasma membranes, alterations in calcium permeability in the plasma and endoplasmic reticulum membrane, and inhibition of the calcium-dependent ATPase of the endoplasmic reticulum.

Adenosine Triphosphate↗

PubMed1982

Patulin, a further clastogenic mycotoxin, is negative in the SCE assay in Chinese hamster V79-E cells in vitro.

Patulin is a potent inducer of chromatid-type aberrations in Chinese hamster V79-E cells, but loses its activity when 9000 g supernatant of rat-liver homogenate is added. The narrow dose range of patulin clastogenicity shows a quantitative relationship between absolute amount of mycotoxin applied and the number of indicator cells treated. Within a dose range permitting survival of V79-E, patulin does not induce an increase of the SCE rate. It is suggested that patulin clastogenicity is caused by interaction with chromosomal proteins and that DNA is not the virtual target of this mycotoxin.

Animals↗

PubMed1988

Subacute toxicity study of patulin in the rat: effects on the kidney and the gastro-intestinal tract.

Female and male Wistar rats were given drinking-water containing patulin at concentrations of 0, 24, 84 or 295 mg/litre citrate buffer (1 mM) for 4 wk. Compared with the controls, there were decreases in food and liquid intake by rats given the mid and high doses of patulin and the body weights of rats in the high-dose group were also decreased. The creatinine clearance in the high-dose group was lowered, although morphological glomerular damage was not observed. In the high-dose group, fundic ulcers in the stomach and enlargement and activation of the pancreatico-duodenal lymph nodes were noticed, while villous hyperaemia in the duodenum was observed in the mid- and high-dose groups. From the present data it can be concluded that high doses of patulin, administered via the drinking-water, caused effects on the kidney and gastro-intestinal tract. Since no changes in the relative weight or histological appearance of the adrenal glands were observed, it is suggested that the fundic ulcers in the stomach were caused by a direct effect of patulin and not indirectly (i.e. by stress).

Animals↗