Microbial dynamics in Mediterranean Moder humus

There is a growing interest in the links between humus forms and soil biota, and little is known about these links in Mediterranean ecosystems. Culture-independent techniques, such as DNA extraction followed by DGGE and enzyme activities, allowed us to compare microbial communities in two horizons of a forest soil in different seasonal conditions. Direct in situ lysis was applied for extraction of DNA from soil; intracellular DNA was separated from extracellular and used to represent the composition of microflora. The aims were to describe how biochemical and microbiological parameters correlate with topsoil properties in typical Mediterranean Moder humus. Changes in bacterial and fungal community composition were evident from DGGE profiles. Degrees of similarity and clustering correlation coefficients showed that the seasonal conditions may affect the composition and activity of bacterial and fungal communities in the OH horizon, while in the E horizon the two communities were hardly modified. In the same season, OH and E horizons showed a different composition of bacterial and fungal communities and different enzyme activities, suggesting similar behaviour of eubacteria and fungi relatively to all the variables analysed. Evidently, different organic carbon content in soil horizons influenced microflora composition and microbial activities involved in the P and N cycles.     

Quantitative determination of diuron in ground and surface water by time-resolved fluoroimmunoassay: seasonal variations of diuron, carbofuran, and paraquat in an agricultural area

The aim of this research is to develop an ultrasensitive time-resolved fluorescence immunoassay (TR-FIA) for herbicide diuron in water samples. This method appears to be a promising approach, instead of conventional analytical techniques, in the screening procedure of organic pollutants because it is simple, rapid, and specific, and it does not require sample preconcentration or cleanup. Lanthanide chelate used as label allows to achieve sensitivity even 10 times higher than most of the other techniques. It has been applied to monitoring diuron contamination in specimens collected along a year in an agricultural area. The water specimens were collected monthly from lake, well, and irrigation ditch in the agricultural area south of Milan. Assay was performed using diuron-specific polyclonal antibody raised in sheep; as fluorescent marker, we used rabbit antisheep IgG conjugated with a chelating molecule complexed with Eu3+. The compound 4-(3-(3,4-dichloro-phenyl)-1-methyl-ureido)-butyric acid (CPD) was synthesized and conjugated with bovine serum albumin (BSA) to prepare a solid phase. Sensitivity achieved was 20 ng L-1 below the European Community limits. Paraquat (PQ) and carbofuran (CF) presence in the same samples has been also evaluated in a similar way, using immunoassays with time-resolved revelation systems. Diuron concentration shows a peak coinciding with a peak of carbofuran during summer periods. The peak of diuron was 65 pg/mL in June and 180 pg/mL in September in ditch and lake water samples, respectively; carbofuran concentration was higher than diuron in all samples: a carbofuran peak was revealed in September and October resulting in 87 ng/mL. Herbicide paraquat was not detectable in any assayed sample.     

Pulmonary rhodococcosis in a cat

Feline Rhodococcus equi infection is rare, despite the bacteria is widespread in the environment. R equi infection is typically observed in equine species but the infection has also been reported in dogs, cats and other domestic animals. There are a few reports regarding pulmonary R equi infection in cats and the disease appears to be limited to the skin and the subcutaneous tissue. This report describes the pathological, microbiological and the virulence features associated with an acute necrosuppurative pneumonia in a cat. To the best of our knowledge, this is the first report of feline pulmonary R equi infection in Italy.     

ECHOGRAPHIC EXAMINATION OF THE STIFLE JOINT AFFECTED BY CRANIAL CRUCIATE LIGAMENT RUPTURE IN THE DOG

Ultrasound is a useful technique for the study of normal and pathologic stifle joints, in particular for soft tissue examination. The aim of this study was to evaluate sonography for examination of stifle joints affected by cranial cruciate ligament rupture. Forty-two medium to giant breed dogs were studied. Tibial compression radiography was performed. A 7.5 MHz transducer with an incorporated 2 cm thick standoff was employed. Sagittal and midsagittal images were collected. The stifle was positioned in maximum flexion during sonography. Sonographic findings were compared with pathologic findings at surgery. Ultrasound was useful in evaluating the presence of fibrous tissue within the joint due to repair processes. It was observed in 70% of stifles with radiographic evidence of chronic osteoarthritis. In 19.6% of the joints it was possible to identify the ruptured cranial cruciate ligament. Ultrasound was not an accurate test for cruciate rupture evaluation, but was specific for the soft tissue pathologic changes which were observed consequent to joint instability.     

Seasonal and interannual variability of photosynthetic capacity in relation to leaf nitrogen in a deciduous forest plantation in northern Italy

Gas exchange was measured in a forest plantation dominated by Fraxinus angustifolia Vahl. and Quercus robur L. in northern Italy, over three growing seasons that differed in water availability (2001, 2002 and 2003). The objectives were to: (1) determine variability in the photosynthetic parameters V(cmax) (maximum carboxylation capacity) and J(max) (maximum rate of electron transport) in relation to species, leaf ontogeny and drought; and (2) assess the potential of the photosynthesis-nitrogen relationship for estimating leaf photosynthetic capacity. Marked seasonal and interannual variability in photosynthetic capacity was observed, primarily caused by changes in leaf ontogeny and water stress. Relatively small differences were apparent between species. In the absence of water stress (year 2002), the seasonal patterns of V(cmax) and J(max) were characterized by a rapid increase during spring, a relatively steady state during summer and a rapid decline during autumn. In years with a moderate (year 2001) or a severe (year 2003) water stress, photosynthetic capacity decreased during the summer in proportion to drought intensity, without a parallel decline in leaf nitrogen content. The V(cmax)-nitrogen relationship was significantly affected by both leaf ontogeny and drought. As a consequence, the use of a single annual regression to predict V(cmax) from leaf nitrogen yielded good estimates only during the summer and in the absence of water stress. Irrespective of the mechanisms by which photosynthetic capacity is affected by water stress, its large seasonal and interannual variability is of great relevance for modeling the forest carbon cycle.     

Photosynthesis-nitrogen relationships: interpretation of different patterns between Pseudotsuga menziesii and Populus x euroamericana in a mini-stand experiment

We compared photosynthesis-nitrogen relationships of one broad-leaved (poplar; Populus x euroamericana (Dole) Guinier) and one conifer (Douglas-fir; Pseudotsuga menziesii (Mirb.) Franco) species. Plants were grown in large pots to allow free root development and were kept well watered. We determined effects of low, intermediate and high nitrogen supply rates on area-based leaf nitrogen (Na) and chlorophyll concentrations, leaf mass per area (LMA), light-saturated photosynthesis (Amax), maximum carboxylation (Vcmax) and electron transport rate (Jmax), photosynthetic nitrogen-use efficiency (PNUE), and proportions of leaf N in active Rubisco (PR), bioenergetic pools (PB) and the light-harvesting complex (PLH). Nitrogen supply significantly affected leaf Na. Leaf mass per area did not differ between species and was unaffected by the N treatments. In both species, there was a positive correlation between leaf Na and chlorophyll concentration, and between leaf Na and the photosynthetic parameters Amax, Jmax and Vcmax. At comparable leaf Na, however, poplar showed twofold higher PNUE and a threefold steeper slope of the Amax- nitrogen relationship than Douglas-fir. Leaf Na was negatively correlated with PNUE in Douglas-fir but not in poplar. Leaf Na was also negatively correlated with PR, PB and PLH in Douglas-fir, whereas in poplar, a negative correlation was found only for PLH. Parameter PR was significantly higher in poplar than in Douglas-fir. The ratio of CO2 concentration in the intercellular space to that in ambient air was higher in poplar than in Douglas-fir. Overall, our data suggest that differences in the photosynthesis-nitrogen relationship and PNUE between Douglas-fir and poplar primarily reflect a different investment of N to active Rubisco, and possibly a different constraint to CO2 diffusion.     

The effect of pharmaceutical waste-fungal biomass,treated to degrade DNA,on the composition of eubacterial and ammonia oxidizing populations of soil

The aim of this work was to study variations in the composition of eubacteria and ammonia-oxidizing populations of soil, both determined by denaturing gradient gel electrophoresis (DGGE), after the addition of a pharmaceutical fungal biomass, treated to degrade its DNA. This waste can be used as an amendment. The fungal biomass waste was added at three rates: 0.05, 0.1, and 1% per dry weight of soil. Control soil, without any amendment, was also investigated. Total DNA was extracted, purified, and amplified by using either universal (eubacteria) or specific (amoA) primers. Amplicons were separated by DGGE. Sequencing was also carried out to better assess the diversity of ammonia oxidizing bacteria. Changes in the composition of eubacterial community were detected after 3 days only in the soil treated with the highest dose, while the ammonia oxidizing population responded more promptly (after 1 day) with evident modifications at level of Nitrosolobus like sequences.     

Carbon dioxide efflux and concentrations in two soils under temperate forests

The carbon dioxide efflux to the atmosphere and the concentrations at various depths in two soils were measured, for more than a year, under pure stands of silver fir ( Abies alba Mill.) and European beech ( Fagus sylvatica L.) in central Italy. Microbial biomass and activity at the monitored depths were determined in the laboratory and the CO₂ evolved from incubated samples was submitted to radiocarbon analysis to assess the mean residence time of the organic matter degraded by microorganisms. The CO₂ efflux showed similar trends in the two soils, with highest values in October and lowest in January. The efflux depended more on air and soil temperatures than soil moisture, and was related to these variables better under fir than under beech. In both soils, the CO₂ concentration increased with depth: in the top horizon it was low and similar to that of the atmosphere, while in the deeper horizons it often amounted to considerable values (up to more than 1% by volume in the BC horizon under fir). The subsoil of the fir stand generally showed much higher CO₂ concentrations than that of the beech. The basal respiration as determined in the laboratory was at a maximum in the topsoil and decreased sharply downwards. Therefore, the high CO₂ concentrations measured in the field at the bottom of the profiles--where roots were few, and microbial biomass and available C pool were at a minimum--appeared to be due more to slow diffusivity of the soil matrix rather than to heavy release of the gas by the biota. The organic matter respired by microorganisms in incubated soil samples showed positive values of j¹⁴C that revealed a recent synthesis. The estimated mean residence time increased with depth, suggesting a generally higher degree of stabilisation of the organic pool in the subsoil.     

On the Different Sensitivity of White Clover Clones to Ozone: Physiological and Biochemical Parameters in a Multivariate Approach

This study analyses the physiological and biochemical basis of chronic ozone exposure (60 ppb for 56 days, 5 h day^?1) on NC-S (sensitive) and NC-R (resistant) white clover clones. Analyses were performed after 0, 14, 28 and 56 days of fumigation which corresponded to AOT40 s of 0, 1400, 2800 and 5600 ppb.h, respectively. NC-S exhibited foliar injury and had a decreased content of photosynthetic pigments, while peroxidized lipids and solute leakage increased, indicating that the plants were subjected to membrane damage. The multivariate approach identified five groups. The NC-R group, with the exception of samples at 0 days of exposure and treated for the longest time period (and thus at the highest dose), and NC-S controls after 28 and 56 days were associated with photosynthetic pigments variables. Ascorbate peroxidase was twinned with NC-R treated at the highest dose. Guaiacol peroxidase and solute leakage was mildly linked with NC-S following ozone treatment for 56 days (AOT40 = 5600 ppb.h).