Molecular cloning of two chitinase genes from Serratia marcescens and their expression in Pseudomonas species

Two chitinase encoding EcoRI fragments from the enteric soil bacterium Serratia marcescens were cloned. From a genomic library of 5686 transductants, 21 expressed chitinase activity as indicated by clearing of a chitin-containing medium. The chitinase encoding clones could be divided into two groups. Four had an 18kb EcoRI fragment and 17 had a 9·4 kb EcoRI fragment. In Southern hybridization experiments the 18kb fragment showed no homology to the 9·4 kb fragment and restriction enzyme maps indicated no similarity. Triparental mating with fluorescent Pseudomonas spp. yielded transconjugants that expressed chitinase activity, inhibited growth of Fusarium oxysporum f. sp. redolens germ tubes and reduced disease of radish caused by the same fungus.     

Induction of discolored wood in Scots pine (Pinus sylvestris)

Induction of discolored wood in Scots pine (Pinus sylvestris L.) trees by treatment with ethylene, carbon dioxide, nitrogen (hypoxia) or wounding from early April to late September was investigated. All treatments induced formation of discolored wood upward and downward from the drill hole. The amount of discolored wood formed above the drill hole depended on the treatment in the following order: ethylene > carbon dioxide = nitrogen > wounding; and below the drill hole in the order: ethylene > carbon dioxide = nitrogen = wounding. Based on chemical analyses (HPLC/UV, GS/MS, LC/MS and 1H-NMR), discolored wood induced by wounding or treatment with ethylene or carbon dioxide showed compositional similarities to natural heartwood, whereas discolored wood induced by nitrogen treatment showed fewer similarities to natural heartwood. The results suggest that ethylene is an important factor controlling heartwood formation, although wounding and internal concentrations of carbon dioxide may also play a role.     

Nitrification controls N2O production rates in a frozen boreal forest soil

The winter season has been identified as a significant contributor to N₂O emissions from boreal soils, but our understanding of the processes regulating these emissions is fragmentary. We investigated potential N-sources and pathways involved in N₂O formation in a frozen boreal forest soil by labeling soil samples with ¹⁵N-containing substrates, and measured rates of ¹⁵N₂O/¹⁵N₂ formation under both oxic and anoxic conditions. Our results showed that all N₂O produced in the frozen samples originate from denitrification, but the rate-limiting factor is NO₃⁻ availability, which is largely governed by nitrification. This suggests that N₂O formation in frozen boreal soils may be sustained for a prolonged period of time, but is governed by a delicate balance of the O₂ regime.     

Influence of Disease Process and Duration on Acute Phase Proteins in Serum and Peritoneal Fluid of Horses with Colic

Background

The acute phase proteins (APP) serum amyloid A (SAA), haptoglobin, and fibrinogen are valuable blood biomarkers in equine inflammatory diseases, but knowledge of factors influencing their concentrations in blood and peritoneal fluid (PF) of horses with colic is needed.

Objectives

The objective of this study was to investigate the influence of demographics (age, sex, breed), disease process (simple obstruction, strangulating obstruction, inflammatory), disease location, disease duration, hypovolemia, and admission hospital on concentrations of APP, lactate and white blood cell counts (WBC) in horses with colic admitted to 2 referral hospitals.

Animals

The study included 367 horses with colic admitted at 2 referral hospitals.

Methods

Prospective multicenter observational study of clinical data, as well as blood and PF biomarkers. Associations between biomarker concentrations and clinical variables were analyzed using multivariate linear regression analysis.

Results

Increasing pre‐admission duration of colic was associated with increased concentrations of APP in blood and PF. Blood concentrations of SAA and fibrinogen were associated with disease process (inflammatory, strangulations, simple obstructions) in more colic duration groups (5–12 and >24 hours) than any of the other biomarkers. No relevant associations between demographic factors, hospital, or hydration status and the measured biomarkers were found.

Conclusions and Clinical Importance

In horses with colic, concentrations of APP are associated mainly with disease process and duration of colic and may thus be used for assessment of disease independently of demographic or geographic factors. Serum amyloid A may be a diagnostic marker for use in colic differential diagnosis, but further evaluation is needed.     

A COMPUTED TOMOGRAPHIC STUDY OF VOLUME AND X-RAY ATTENUATION OF THE LUNGS OF BEAGLES IN VARIOUS BODY POSITIONS

The influence of body position on volume and x-ray attenuation of the lungs of three Beagles was investigated with computed tomography. In left and right lateral recumbency, the dependent lung had decreased volume and increased x-ray attenuation compared with ventral recumbency. Volume and x-ray attenuation of the nondependent lung, however, were unchanged. In dorsal recumbency, there was a vertical gradient of x-ray attenuation being increased dorsally and decreased ventrally compared with ventral recumbency where regional differences in x-ray attenuation were not found. Results indicate a risk of overlooking a mass in the dependent pulmonary parenchyma of dogs in lateral and dorsal recumbency. Regional differences in x-ray attenuation of the lungs of dogs can be avoided if the radiographs are obtained with the dog in ventral recumbency.     

Model studies on acrylamide generation from glucose/asparagine in aqueous glycerol

Acrylamide formation from asparagine and glucose in different ratios in neutral glycerol/water mixtures was found to increase with decreasing water activity (0.33 < or = aw < or = 0.71 investigated) and increasing temperature (120 degrees C < or = T < or = 160 degrees C investigated). The initial rate of acrylamide formation was found to be approximately proportional to the asparagine concentration for an excess of asparagine, but less dependent on an excess of glucose. A steady-state concentration of acrylamide was established at 160 degrees C after 1 h for aw = 0.33 (30 microg x L-1 for GLU:ASN = 10:1, 11 microg x L-1 for GLU:ASN = 1:1, and 130 microg x L-1 for GLU:ASN = 1:10) and for aw = 0.47 (15 microg x L-1 for GLU:ASN = 10:1 and 80 microg x L-1 for GLU:ASN = 1:10), suggesting a protection by glucose against acrylamide degradation. The energy of activation, as estimated from the temperature dependence of the initial rate, increased with decreasing aw despite a higher rate of formation of acrylamide at low aw. For high aw, water elimination from a reaction intermediate is suggested to be rate determining. For low aw, the increase in energy of activation (and enthalpy of activation) is accordingly counteracted by a more positive entropy of activation, in agreement with decarboxylation as rate determining at low aw.     

Bioindication and modelling of atmospheric deposition in forests enable exposure and effect monitoring at high spatial density across scales

Key message

Moss surveys provide spatially dense data on environmental concentrations of heavy metals and nitrogen which, together with other biomonitoring and modelling data, can be used for indicating deposition to terrestrial ecosystems and related effects across time and areas of different spatial extension.

Context

For enhancing the spatial resolution of measuring and mapping atmospheric deposition by technical devices and by modelling, moss is used complementarily as bio-monitor.

Aims

This paper investigated whether nitrogen and heavy metal concentrations derived by biomonitoring of atmospheric deposition are statistically meaningful in terms of compliance with minimum sample size across several spatial levels (objective 1), whether this is also true in terms of geostatistical criteria such as spatial auto-correlation and, by this, estimated values for unsampled locations (objective 2) and whether moss indicates atmospheric deposition in a similar way as modelled deposition, tree foliage and natural surface soil at the European and country level, and whether they indicate site-specific variance due to canopy drip (objective 3).

Methods

Data from modelling and biomonitoring atmospheric deposition were statistically analysed by means of minimum sample size calculation, by geostatistics as well as by bivariate correlation analyses and by multivariate correlation analyses using the Classification and Regression Tree approach and the Random Forests method.

Results

It was found that the compliance of measurements with the minimum sample size varies by spatial scale and element measured. For unsampled locations, estimation could be derived. Statistically significant correlations between concentrations of heavy metals and nitrogen in moss and modelled atmospheric deposition, and concentrations in leaves, needles and soil were found. Significant influence of canopy drip on nitrogen concentration in moss was proven.

Conclusion

Moss surveys should complement modelled atmospheric deposition data as well as other biomonitoring approaches and offer a great potential for various terrestrial monitoring programmes dealing with exposure and effects.

     

Mechanism of deactivation of triplet-excited riboflavin by ascorbate, carotenoids, and tocopherols in homogeneous and heterogeneous aqueous food model systems

Tocopherols (alpha, beta, gamma, and delta) and Trolox were found to deactivate triplet-excited riboflavin in homogeneous aqueous solution (7:3 v/v tert-butanol/water) with second-order reaction rates close to diffusion control [k2 between 4.8 x 10(8) (delta-tocopherol) and 6.2 x 10(8) L mol(-1) s(-1) (Trolox) at 24.0 +/- 0.2 degrees C] as determined by laser flash photolysis transient absorption spectroscopy. In aqueous buffer (pH 6.4) the rate constant for Trolox was 2.6 x 10(9) L mol(-1) s1 and comparable to the rate constant found for ascorbate (2.0 x 10(9) L mol(-1) s(-1)). The deactivation rate constant was found to be inferior in heterogeneous systems as shown for alpha-tocopherol and Trolox in aqueous Tween-20 emulsion (approximately by a factor of 4 compared to 7:3 v/v tert-butanol/water). Neither beta-carotene (7:3 v/v tert-butanol/water and Tween-20 emulsion), lycopene (7:3 v/v tert-butanol/water), nor crocin (aqueous buffer at pH 6.4, 7:3 v/v tert-butanol/water, and Tween-20 emulsion) showed any quenching on the triplet excited state of riboflavin. Therefore, all carotenoids seem to reduce the formation of triplet-excited riboflavin through an inner-filter effect. Activation parameters were based on the temperature dependence of the triplet-excited deactivation between 15 and 35 degrees C, and the isokinetic behavior, which was found to include purine derivatives previously studied, confirms a common deactivation mechanism with a bimolecular diffusion-controlled encounter with electron (or hydrogen atom) transfer as rate-determining step. DeltaH for deactivation by ascorbic acid, Trolox, and homologue tocopherols (ranging from 18 kJ mol(-1) for Trolox in Tween-20 emulsion to 184 kJ mol(-1) for ascorbic acid in aqueous buffer at pH 6.4) showed a linear dependence on DeltaS (ranging from -19 J mol(-1) K(-1) for Trolox in aqueous buffer at pH 6.4 to +550 J mol(-1) K(-1) for ascorbic acid in aqueous buffer pH 6.4). Among photooxidation products from the chemical quenching, lumicrome, alpha-tocopherol quinones and epoxyquinones, and alpha-tocopherol dimers were identified by ESI-QqTOF-MS.     

Monitoring chemical changes of dry-cured Parma ham during processing by surface autofluorescence spectroscopy

Parma hams at various processing stages were investigated by surface autofluorescence spectroscopy. Fluorescence "landscapes" of raw meat and salted (3 months), matured (11 and 12 months), and aged (15 and 18 months) Parma hams were obtained, and a three-dimensional data array (sample x emission x excitation) was used to develop a PARAFAC model including five components, which all exhibited characteristics of pure fluorophores regarding both excitation and emission spectra. The relative amount of each component related strongly to the processing stage, and sample age showed good correlation to fluorescence data (R = 0.98), with a relative error of prediction of approximately 1 month. Fluorescence measurements from samples of either semimembranosus or biceps femoris were used to predict chemical or sensory reference data, yielding good correlation for biceps femoris data, thereby enabling moisture content, sensory and instrumental color, and proteolysis value to be fairly well predicted. Overall, surface autofluorescence of Parma hams proved to hold relevant information, relating to major chemical/physical changes during processing. It is concluded that fluorescence spectroscopy has potential as an innovative method of quality control in dry-cured ham.