[Fusaria and their mycotoxins in hay and grass silage from grassland with different intensities of management]. / Nachweis von Fusarien und deren Mykotoxinen in Futterkonservaten aus Grünlandbeständen mit differenzierter Bewirtschaftungsintensität.

Conservation forage (17 hay and 18 grass silage samples) from 15 farms with different intensities of grassland management in the Federal State of Brandenburg were examined for contamination with fusaria and their mycotoxins. The numbers of culturable filamentous fungi in hay were determined by plate counting andFusarium isolates were classified taxonomically. The mycotoxins Zearalenone (ZEA) and Deoxynivalenol (DON) were extracted from hay as well as silage by different procedures and detected chromatographically (HPLC). The numbers of filamentous fungi in the hay samples were 10(2) and 10(6) CFU/g FM independently of intensive or extensive management. Only fourFusarium species were identified.Fusarium culmorum, a potential toxin producing species, was most frequently detected (52% of all isolates). ZEA was found in two hay and four silage samples (6-66 µg/kg), DON in three hay and seven silage samples (63-1290 µg/kg). There were no differences between forage samples of extensive and intensive cultivated grassland of the year 2003 regarding numbers of fusaria and the content of their mycotoxins.

Description of Microbacterium foliorum sp. nov. and Microbacterium phyllosphaerae sp. nov., isolated from the phyllosphere of grasses and the surface litter after mulching the sward, and reclassification of Aureobacterium resistens (Funke et al. 1998) as Microbacterium resistens comb. nov..

The taxonomic position of a group of coryneform bacteria isolated from the phyllosphere of grasses and the surface litter after sward mulching was investigated. On the basis of restriction analyses of 16S rDNA, the isolates were divided into two genotypes. According to the 16S rDNA sequence analysis, representatives of both genotypes were related at a level of 99.2% similarity and clustered within the genus Microbacterium. Chemotaxonomic features (major menaquinones MK-12, MK-11 and MK-10; predominating iso- and anteiso-branched cellular fatty acids; G+C content 64-67 mol%; peptidoglycan-type B2beta with glycolyl residues) corresponded to this genus as well. DNA-DNA hybridization studies showed a reassociation value of less than 70% between representative strains of both subgroups, suggesting that two different species are represented. Although the extensive morphological and physiological analyses did not reveal any differentiating feature for the genotypes, differences in the presence of the cell-wall sugar mannose enabled the subgroups to be distinguished from one another. DNA-DNA hybridization with type strains of closely related Microbacterium spp. indicated that the isolates represent two individual species, which can also be differentiated from previously described species of Microbacterium on the basis of biochemical features. As a result of phenotypic and phylogenetic analyses, the species Microbacterium foliorum sp. nov., type strain P 333/02T (= DSM 12966T = LMG 19580T), and Microbacterium phyllosphaerae sp. nov., type strain P 369/06T (= DSM 13468T = LMG 19581T), are proposed. Furthermore, the reclassification of Aureobacterium resistens (Funke et al. 1998) as Microbacterium resistens (Funke et al. 1998) comb. nov. is proposed.

A taxonomic study of bacteria isolated from grasses: a proposed new species Pseudomonas graminis sp. nov.

The taxonomic position of a yellow-pigmented group of bacteria, isolated from the phyllosphere of grasses was investigated. Results obtained from restriction analysis of amplified 16S rDNA with seven endonucleases (CfoI, HaeIII, AluI, HinfI, MspI, Sau3A and ScrFI) showed identical restriction patterns for each enzyme of all isolates studied, which suggests that all strains belong to the same species. The grass isolates displayed the characteristics of the genus Pseudomonas. They were Gram-negative, aerobic and rod-shaped with polar flagella. Isolates were catalase-positive and oxidase-negative, and unable to oxidize or ferment glucose with the production of acid. The isolates did not reduce nitrate to nitrite but were able to utilize a wide range of compounds individually as a sole carbon source, with preference being given to the utilization of monosaccharides. The disaccharides tested were not utilized as substrates. The DNA base compositions of the tested strains ranged from 60 to 61 mol% G+C. The major isoprenoid quinone of each was ubiquinone Q-9 and hydroxy fatty acids were represented by 3-hydroxydodecanoic acid and 2-hydroxydodecanoic acid. Comparison of 16S rDNA sequences showed that the bacteria were members of the genus Pseudomonas, with similarity values between 91.5 and 97.7%. DNA-DNA hybridization studies with closely related neighbours revealed a low level of homology (< 27%), indicating that the isolates represent an individual species. On the basis of phenotypic and phylogenetic analyses a new species, Pseudomonas graminis sp. nov. (type strain DSM 11363T), is proposed.

Mass spectrometry: a tool for on-line monitoring of animal cell cultures.

The magnetic sector mass spectrometer is able to detect oxygen uptake and carbon dioxide production rates from animal cell cultivations performed in 101 bioreactors. Such data have not been available with the use of classic exhaust gas analysis applying paramagnetic analyzers and infra-red sensors due to the insensitivity of the apparatus available. In the course of the present work we were able to demonstrate, that the oxygen uptake rate correlates to the number of viable cells. Additionally oxygen uptake rates supplied on-line information about the actual physiology of the cells: When the rates changed during the cultivation process, this immediately indicated the occurrence of limitations of components in the medium. The information could be useful in timing key events, such as performing splits or harvesting the bioreactor.

[Primary hyperparathyroidism: studies of 4,000 urinary calculi patients treated with extracorporeal shockwave lithotripsy]. / Primärer Hyperparathyreoidismus: Untersuchungen bei 4000 mit der ESWL behandelten Harnsteinpatienten.

The incidence of primary hyperparathyroidism was determined in 4000 renal stone formers treated by ESWL at our institution from 1983 to 1990. Based on repeated measurements of serum calcium and serum parathyroid hormone an incidence of 2.8% was found. In 60% of patients with primary hyperparathyroidism stone disease occurred for the first time. The majority of patients (68%) were more than 50 years of age. 56% of patients were men, 44% were women.

Industrial production of monoclonal antibodies and therapeutic proteins by dialysis fermentation.

A novel and powerful fermentation method is reported for the large-scale growth of mammalian cells and their secreted products. The system described illustrates many of the advantages of conventional batch fermentation processes but in addition has been shown to yield cell densities in excess of 1 x 10(7) cells/ml with concomitant increase in product concentration.

The production of L-serine with a methylotrophic microorganism using the L-serine pathway and coupling with an L-tryptophan-producing process.

The cultural conditions for the production of enhanced formation of L-serine (up to 7 g/L) are described with the methylotrophic bacterium Pseudomonas 3 ab (DSM 672). The batch process is divided into three parts: (1) the biomass production phase, (2) substrate limitation period, and (3) L-serine accumulation phase. The initial specific production rate of q(p) = 0.1 g L-serine/g dry wt/h is based on the inhibition of the L-serine pathway. This is accomplished by high precursor concentrations (glycine) and a pH shift to pH 8.5. The enzymatic background is discussed. Furthermore, a coupling of the L-serine process with a L-tryptophan-producing process is demonstrated.

Continuous production of L-tryptophan from indole and L-serine by immobilized Escherichia coli cells.

Escherichia coli B 10, which has high activity of tryptophan synthetase, was grown in a 50-L batch culture in order to determine in which growth phase the cells have the highest specific tryptophan productivity. Accordingly, whole cells of the stationary phase were used for immobilization in polyacrylamide beads. After immobilization, these immobilized cells had 56% activity of tryptophan synthetase compared with that of free cells. First, the properties of immobilized cells were investigated. Next, discontinuous productions of L-tryptophan were carried out by using immobilized cells. In discontinuous production of L-tryptophan by the batch, the activity remaining of immobilized cells was 76-79% after 30 times batchwise use. In continuous production of L-tryptophan with a continuous stirred tank reactor (CSTR), the activity remaining of the immobilized cells was 80% after continuous use for 50 days. The maximum productivity of L-tryptophan in this CSTR system was 0.12 g tryptophan L(-1) h(-1).

Estimation of glomerular filtration rate in conscious dogs following a bolus of creatinine. Comparison with simultaneously determined inulin clearance.

The renal clearance of creatinine was measured following orogastric administration in conscious dogs. Values of creatinine clearance were compared with simultaneously determined values of inulin clearance, when urine volume, glomerular filtration rate and volume status were acutely altered by a variety of experimental manoeuvres. At urine volumes greater than 20 microliters X min-1 X kg-1, creatinine clearance was not significantly different from inulin clearance. At low urine volumes there was some evidence of creatinine reabsorption. It is concluded that the bolus creatinine technique provides reliable estimates of glomerular filtration rate and is particularly applicable to long-term studies in conscious dogs.