Analysis and assessment of heavy metal pollution in suspended solids and sediments of the river Danube.

The Joint Danube Survey (JDS)--a comprehensive monitoring survey to assess the environmental pollution status of the river Danube--was carried out in 2001. Samples were taken at 74 positions along the river from Neu-Ulm (River-km 2589) down to the Danube Delta at the Black Sea (River-km 0) and in 24 main tributaries and anabranches. Besides other biological and chemical parameters, concentrations of Al, As, Cd, Cr, Cu, Fe, Pb, Mn, Hg, Ni, and Zn were determined in sediments and suspended solids. Lowest heavy metal concentrations were measured around River-km 1800. After an increase down to River-km 1000 (the Irongate Reservoir), a constant level or a slight decrease could be found down to and in the Danube Delta. Very high element concentrations were determined at only a few stations of the river Danube and in some tributaries. An evaluation of the pollution status of the river was carried out by enrichment factors (EFs) calculated using adapted background concentrations of heavy metals. Except single sampling sites and some tributaries, the pollution of the river Danube by As, Cr, Cu, Pb, Hg, Ni, and Zn can be regarded as rather low. However, elevated concentrations of Cd were found in both investigated matrices, particularly in the lower stretch of the river Danube beginning at the Irongate.

[The detection of Alternaria mycotoxins in laboratory culture]. / Nachweis von Alternaria-Mykotoxinen in Laborkulturen.

Analytical methods are described for detection of the Alternaria mycotoxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT) and tenuazonic acid (TeA) in natural and semisynthetic laboratory cultures. After extraction and purification of the crude extract by column chromatography on silica gel the qualitative and quantitative analyses were carried out by thin layer (TLC)- and high performance liquid chromatography (HPLC). HPLC separations were achieved using a Hypersil ODS column with methanol/water containing a complexing agent as eluent. Detection at 340 nm (AOH, AME, ALT) and 280 nm (TeA), respectively, has proved to be favourably. AME and TeA were produced in high purity and high yields as standard substances by two Alternaria strains. The identity of the toxins could be confirmed by EI-, CI- and FAB-mass spectrometry.

[Occurrence of Fusarium strains and their mycotoxins in corn silage. 7. Formation of deoxynivalenol (DON) in a silage corn plot artificially inoculated with Fusarium culmorum and the effect of silaging on the stability of the DON formed]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 7. Mitteilung. Bildung von Deoxynivalenol (DON) in einem künstlich mit Fusarium culmorum infizierten Silomaisbestand sowie der Einfluss der Silierung auf die Stabilität des gebildeten DON.

The formation of deoxynivalenol in a maize plot inoculated with Fusarium culmorum was studied over a growing season. Already three weeks after inoculation 4.9 mg/kg of DON were measured in the infected ears. The toxin concentration increased continuously up to harvest after eight weeks to a value of 261 mg/kg. Ensilage experiments in laboratory scale silos have shown that the DON content of naturally contaminated corn-cob-mix was not reduced during the ensilage process. It was concluded that infection of maize plants by toxin-producing Fusarium species followed by DON production in the field seems to be the most probable way of contamination of maize silage with this mycotoxin.

[Occurrence of Fusarium strains and their mycotoxins on corn silage. 6. Formation of zearalenone and trichothecenes (type A) by indigenous Fusarium isolates]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 6. Mitteilung: Bildung von Zearalenon und Trichothecenen (Typ A) durch einheimische Fusarium-Isolate.

A total of 399 indigenous Fusarium strains mainly isolated from silage maize were tested for the production of zearalenone and type A trichothecenes by thin-layer chromatography and biological assays. About 45% of the isolates examined were capable of producing different levels of zearalenone and trichothecene toxins on a cracked corn substrate. The majority of these strains (75%) produced zearalenone only and no trichothecenes type A. The results of the biological tests indicated a higher rate of toxin-positive extracts than chemical analysis. Isolates of nine out of seventeen Fusarium species examined produced one or several mycotoxins looked for. The most important toxin producers were F. culmorum and F. crookwellense (zearalenone) and F. sporotrichioides (trichothecenes type A), respectively. F. avenaceum, the species most frequently isolated from silage maize, produced neither zearalenone nor trichothecenes but avenacein Y a antibiotic compound. First results of a study of the production of type B trichothecenes have shown that indigenous F. culmorum isolates were capable of producing high levels of deoxynivalenol.

[Occurrence of Fusarium species and their mycotoxins in corn silage. 5. Fusarium infestation in corn silage]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 5. Mitteilung. Untersuchungen zum Fusarium-Befall von Silomais.

The internal mycoflora of silage maize plants was examined in several years. Average Fusarium infection of the maize plants was 10.4% and leaf sheaths and stalks were preferably colonized by the fungi. During the growing season an increase in total colonization of the maize plant by field fungi as well as by species of the genus Fusarium was observed 17 different Fusarium species were isolated from silage maize plants. Most frequently occurring species (more than 85%) in descending order were F. avenaceum, F. culmorum, F. equiseti, F. sporotrichioides, F. crookwellense, F. oxysporum, F. sambucinum var. coeruleum.

[The occurrence of Fusarium strains and their mycotoxins in corn silage. 4. The occurrence of zearalenone in the first cut zone of horizontal silos]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 4. Mitteilung. Untersuchungen zum Vorkommen von Zearalenon in der Anschnittzone eines Horizontalsilos.

Studies of the occurrence of zearalenone in the cutting surface of a horizontal silo were carried out over 12 days. On all sampling days the silage contained less than 0.1 mg/kg zearalenone (detection limit of the thin-layer chromatographic procedure). Zearalenone could not be detected in silage put into interim store over a period of 17 days either. The results document that zearalenone-producing Fusarium species have no considerable share in the decay of maize silage.

[The occurrence of Fusarium varieties and their mycotoxins on silo corn. 1. A method for the determination of zearalenone in corn and corn silage by high performance liquid chromatography (HPLC) with fluorescence detection]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 1. Mitteilung. Methode zur Bestimmung von Zearalenon in Mais und Maissilage durch Hochleistungs-Flüssigkeitschromatographie (HPLC) mit Fluoreszenzdetektion.

A method for the determination of zearalenone in maize and maize silage was developed which distinguishes itself by the effective and fast cleaning of the extracts with the help of a silica gel minicolumn. The samples were extracted with chloroform/methanol (9 + 1) and cleaned on a silica gel minicolumn after acid-base partition. The zearalenone was quantitatively determined optionally by means of high-performance liquid chromatography (HPLC) with fluorescence detection (excitation wavelength 236 nm, emission filter 418 nm) or thin-layer chromatography (TLC), p-methoxybenzene diazonium fluoroborate and aluminium chloride were used as detection chemicals. The limits of detection are 0.01 mg/kg (HPLC) and 0.1 mg/kg resp. (TLC), the average recovery is 81%. The method was used for the determination of zearalenone in grain maize, CCM silage and silage from whole maize plants.

[The occurrence of Fusarium varieties and their mycotoxins in silo corn. 2. The formation of zearalenone in the field by artificial infection of silo corn with Fusarium culmorum (W. G. Smith) Sacc]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 2. Mitteilung. Bildung von Zearalenon auf dem Feld durch künstliche Infektion von Silomais mit Fusarium culmorum (W. G. Smith) Sacc.

The formation of zearalenone in a maize plot artificially infected with Fusarium culmorum was studied. The zearalenone concentration steeply increased only in the 8th week after inoculation and reached a maximum value of ca. 7 ppm, whereas zearalenone could not be detected in the control variants. The crude nutrient and dry matter content was not significantly influenced by the fungal infection. The infected crop showed average ear dry weights distinctly lower than that of the control variants (P less than or equal to 0.001). Apart from zearalenone, the mycotoxin deoxynivalenol was qualitatively detected in the infected maize. The toxicological relevance of the ascertained zearalenone content with regard to the health of dairy cattle and pigs was discussed.

[The occurrence of Fusarium varieties and their mycotoxins in silo corn. 3. The effect of silaging on the zearalenone content of CCM corn]. / Vorkommen von Fusarium-Arten und ihren Mykotoxinen auf Silomais. 3. Mitteilung. Der Einfluss der Silierung auf den Zearalenongehalt von CCM-Mais.

The influence of silaging on zearalenone already formed and on the growth of Fusarium culmorum in naturally contaminated CCM maize was studied. The zearalenone content remained approximately constant (13.35 +/- 2.94 mg/kg, n = 59) over the whole test period (12 weeks), whereas Fusarium culmorum could not longer be detected after 11 days. Thus the hypothesis that zearalenone survives the silaging process unchanged is experimentally strongly supported. The study confirms the view that zearalenone detected in maize silages is probably already formed in the field or during intermediate storing. Relations between the fermentation process and the toxin content of the silage could not be ascertained.

Production of sterigmatocystin by Aspergillus versicolor isolated from roughage.

A total of 69 samples of hay and straw collected during the winter period of 1984/85 were surveyed for their contamination by Aspergillus versicolor. The percentage of A. versicolor-positive samples was 14.5%. Nineteen A. versicolor strains mainly isolated from roughage were tested for the production of sterigmatocystin. All of the isolates examined were capable of producing different levels of sterigmatocystin on a cracked corn substrate. The majority of these strains were highly toxigenic; 53% of the isolates produced more than 500 mg/kg of sterigmatocystin. These findings suggest that corn is a very suitable substrate for sterigmatocystin production and that particularly in the surface layers of feed stocks and corn silos such toxigenic strains of A. versicolor can produce considerable growth and possibly sterigmatocystin, too.

Formation of Avenacein Y by Fusarium avenaceum Fries Sacc. isolates from Germany and pathogenicity of the isolates to cereal seedlings.

Twelve isolates ofFusarium avenaceum Fries Sacc. originating from diseased corn plants from Germany produced Avenacein Y in amounts ranging from 0.001 to 1.6 g/kg of wheat grain. The isolates proved most pathogenic to triticale seedlings, less pathogenic to rye seedlings and least to wheat. Pathogenicity of the isolates was not correlated with their ability to produce Avenacein Y.