The use of photometric test kits for investigations of soil enzyme activities

The objective was to test the applicability of different photometric test kits for the determination of potential and actual nitrification and the activities of urease, invertase, and cellobiase in soil. Three water affected grassland sites, influenced by the maritime climate were investigated. The results obtained by the test kits and by standard methods were similar. Soil organic matter content was found as most important factor determining the enzyme activities on these three grassland sites with similar pH‐values. At the site with the largest soil organic matter content the highest activities for urease, invertase, and cellobiase were found. However, this was not true for the nitrification rates.     

Distribution of Al and Heavy Metals in Bulk Soil and Aggregates at Three Sites Contaminated by the Emissions of a Central Slovak Al Smelter

Large spoils of metal-rich filtercake from the Port Kembla BHP steelworks were characterised by certain physical, chemical and biological parameters. The vegetation was assessed for potential metal hyperaccumulators and the presence of arbuscular mycorrhizae in the rhizospheres. Fresh filtercake (<2 yr old) was charcoal in colour and had a fine (clay loam) texture. Its composition was distinctly different to the older filtercake (3–4 yr) which was similar to the reference soil, being lighter in colour, loamy textured, more aggregated and with higher organic matter. Variability between the two filtercake types also occurred for electrical conductivity and pH. Total trace metal concentrations (Cd, Cu, Pb and Zn) were high and although only minor proportions were in bioavailable forms, the greater percentages in the older filtercake suggested temporal changes in metal speciation. Fresh filtercake was devoid of any plant life and fungal communities. The older material, however, was able to sustain a reasonable environment for plant, bacterial and fungal growth. The vegetation was dominated by Ricinus communis, along with species belonging to the families Asteraceae, Solanaceae, Malvaceae and Apiaceae. Rhizosphere of all these plants harboured mycorrhizae belonging to the Gigaspora and Glomus strains and while trace metal accumulation was observed no plant was a hyperaccumulator. The reference site was principally inhabited by certain Poaceae, Asteraceae and Verbenaceae species. All were mycorrhizal (principally Glomus strains), but no plant was represented on the contaminated site. Here also trace metal accumulation was taking place. The absence of hyperaccumulation in the filtercake was considered a consequence of low levels of bioavailable trace metals. However, as such metal forms may increase over time, particularly for copper and lead, cultivation of known hyperaccumulators could become worthwhile after prolonged exposure (>4 yr). Among the naturally colonising plants, Ricinus communis and Sonchus oleraceus are regarded the most suitable options for zinc and cadmium removal, respectively. Arbuscular mycorrhizal infection (particularly with Glomus and Gigaspora strains) should be promoted for optimum soil reclamation.     

Nature’s Lab for Derivatization: New and Revised Structures of a Variety of Streptophenazines Produced by a Sponge-Derived Streptomyces Strain

Eight streptophenazines (A–H) have been identified so far as products of Streptomyces strain HB202, which was isolated from the sponge Halichondria panicea from the Baltic Sea. The variation of bioactivities based on small structural changes initiated further studies on new derivatives. Three new streptophenazines (I–K) were identified after fermentation in the present study. In addition, revised molecular structures of streptophenazines C, D, F and H are proposed. Streptophenazines G and K exhibited moderate antibacterial activity against the facultative pathogenic bacterium Staphylococcus epidermidis and against Bacillus subtilis. All tested compounds (streptophenazines G, I–K) also showed moderate activities against PDE 4B.     

Isolation,Structure Elucidation and Total Synthesis of Lajollamide A from the Marine Fungus Asteromyces cruciatus

The marine-derived filamentous fungus Asteromyces cruciatus 763, obtained off the coast of La Jolla, San Diego, USA, yielded the new pentapeptide lajollamide A (1), along with the known compounds regiolone (2), hyalodendrin (3), gliovictin (4), ¹N-norgliovicitin (5), and bis-N-norgliovictin (6). The planar structure of lajollamide A (1) was determined by Nuclear Magnetic Resonance (NMR) spectroscopy in combination with mass spectrometry. The absolute configuration of lajollamide A (1) was unambiguously solved by total synthesis which provided three additional diastereomers of 1 and also revealed that an unexpected acid-mediated partial racemization (2:1) of the L-leucine and L-N-Me-leucine residues occurred during the chemical degradation process. The biological activities of the isolated metabolites, in particular their antimicrobial properties, were investigated in a series of assay systems.     

A Collaborative Evaluation of LC-MS/MS Based Methods for BMAA Analysis: Soluble Bound BMAA Found to Be an Important Fraction

Exposure to β-N-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer’s disease and Parkinson’s disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D₃BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%–32%), implying that D₃BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery (<10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis.     

Lindgomycin,an Unusual Antibiotic Polyketide from a Marine Fungus of the Lindgomycetaceae

An unusual polyketide with a new carbon skeleton, lindgomycin (1), and the recently described ascosetin (2) were extracted from mycelia and culture broth of different Lindgomycetaceae strains, which were isolated from a sponge of the Kiel Fjord in the Baltic Sea (Germany) and from the Antarctic. Their structures were established by spectroscopic means. In the new polyketide, two distinct domains, a bicyclic hydrocarbon and a tetramic acid, are connected by a bridging carbonyl. The tetramic acid substructure of compound 1 was proved to possess a unique 5-benzylpyrrolidine-2,4-dione unit. The combination of 5-benzylpyrrolidine-2,4-dione of compound 1 in its tetramic acid half and 3-methylbut-3-enoic acid pendant in its decalin half allow the assignment of a new carbon skeleton. The new compound 1 and ascosetin showed antibiotic activities with IC₅₀ value of 5.1 (±0.2) µM and 3.2 (±0.4) μM, respectively, against methicillin-resistant Staphylococcus aureus.     

Detection of rhodococcus equi by microbiological culture and by polymerase chain reaction in samples of tracheobronchial secretions of foals

The goal of the present study was to investigate whether new PCR-methods would improve diagnostic of R. equi. In a first step, sensitivity and specificity of the PCR-methods in respect to the"gold standard" microbiological culture were determined. Secondly, sensitivity and specificity of both microbiological methods were evaluated in respect to the clinical diagnosis. The tracheobronchial secretions of 48 foals with pulmonary abscesses and of 37 healthy foals were evaluated by bacteriological culture as well as by four PCR-methods: aceA-, ideR-, vapA- and VP-PCR. In respect to the"gold standard" microbiological culture, the sensitivity of most PCR methods lay between 63.9 and 69.4 % except the vapA-PCR (27.8 %).The specificity of all PCR methods in this comparison was between 98 to 100 %. In this analysis, clinical diagnosis had a low sensitivity (66.7 %) and a low specificity (51.0 %). In respect to the clinical diagnosis, microbiological culture sensitivity was 50.0 % and specificity 67.7 9%. In this analysis, sensitivity rates of aceA-, ideR and VP-PCR methods lay between 33.3 and 37.5 %, sensitivity of the vapA-PCR was lower (10.4 %).The specificity of all PCR methods ranged from 78.4 to 86.5 %. In conclusion, these results show that the diagnostic potential of the microbiological methods"Culture" and "PCR" is different and that for the diagnosis of R. equi-pneumonia in foals the combination of microbiological culture with PCR should be used for examination of samples of the airways of foals.     

Relative ileal amino acid flows and microbial counts in intestinal effluents of Goettingen Minipigs and Saddleback pigs are not different

We explored the suitability of Goettingen Minipigs as models to measure ileal AA digestibility and evaluate dietary proteins for conventional pigs. Further, a potential for secondary ileal microbial colonization 5 mo after establishing end-to-end ileorectal anastomosis was investigated. Goettingen Minipigs (BW 18 kg) and Saddleback pigs (BW 27 kg) fitted with end-to-end ileorectal anastomosis were fed six diets based on barley and oilseed meals and three diets based on wheat and milk powder differing in total and ileal digestible lysine. Apparent ileal digestibilities of CP (N x 6.25) and of 20 AA were determined. No differences (P = 0.062 to 0.982) were found in AA apparent ileal digestibilities between breeds. Therefore, Minipigs are a reasonable model to estimate apparent ileal digestibility of AA for evaluation of dietary proteins. However, the apparent ileal digestibility of CP (P = 0.048) was higher in Minipigs than in Saddleback pigs (barley and oilseed meals-based diets 70% vs. 66%; wheat and milk powder-based diets 80% vs. 77%), which is probably due to a smaller contribution of non-AA-nitrogen in the ileal effluent of Goettingen Minipigs. For lysine, the apparent ileal digestibilities (means of both breeds) ranged from 78 to 85% in wheat and milk powder-based, and 70 to 78% in barley and oilseed-based diets. Experimentally derived concentrations of ileally digestible lysine confirmed the values predicted from a published table. Microbial counts were not affected by breed as shown for lactobacilli, with 9.1+/-0.2 and 9.1+/-0.2 (P = 0.977), enterococci with 4.8+/-0.3 and 5.6+/-0.4 (P = 0.162), and yeasts with 4.6+/-0.3 and 4.6+/-0.4 (P = 0.906) log cfu/g effluent for Goettingen Minipigs and Saddleback pigs, respectively. The counts did not change over 5 mo, suggesting that no secondary microbial colonization occurred in pigs with end-to-end ileorectal anastomosis.