Inhibition of uropathogenic Escherichia coli by cranberry juice: a new antiadherence assay

A combination of microplate technology and turbidity assessment for testing the adherence of P-fimbriated Escherichia coli to human uroepithelial cell line T24, validated with the addition of the known inhibitor 4-O-alpha-D-galactopyranosyl-alpha-D-galactopyranose (galabiose), resulted in a high-throughput, biologically relevant assessment of cranberry (Vaccinium macrocarpon). P-fimbriated ATCC E. coli strains 25922, 29194, and 49161 were inhibited by galabiose. ATCC 29194, a representative urine isolate containing the papGII allele (Class II fimbrial adhesin) and demonstrating the most significant inhibition in the presence of galabiose, was chosen for further testing. In this assay, a low-polarity fraction of cranberry juice cocktail demonstrated dose-dependent inhibition of E. coli adherence. Reported here, for the first time in V. macrocarpon, are 1-O-methylgalactose, prunin, and phlorizin, identified in an active fraction of cranberry juice concentrate. This in vitro assay will be useful for the standardization of cranberry dietary supplements and is currently being used for bioassay-guided fractionation of cranberry juice concentrate.     

Invertebrates and nutrients in a Mediterranean vineyard mulched with subterranean clover (Trifolium subterraneum L.)

Summary In the 25-year-old vineyard of a farm near the Maremma National Park (Central Italy), replicate plots were established with a mulch of Trifolium subterraneum L. or cultivation and two levels of fertilizer. The mai objective of this research was to analyse responses by soil invertebrates and nutrients after introducing a herbaceous component into the system; the yield was also considered. The greater availability of organic substrate and the reduced cultivation as a result of green mulching increased the soil mesofauna biomass, especially detritivores. With time, a significant difference became evident between the populations of invertebrates present in the mulched plots and those in the cultivated plots. Over 2 years, most nutrients (Ca, K, P, and N) were significantly increased in the mulched plots compared to the cultivated plots. Grape yields were consistently higher in the bare plots. No significant differences were found in fertilizer effects.     

Alkyl C and hydrophobicity in B and C horizons of an acid forest soil

Aliphatic C most probably derived from ester‐bound moieties was found to be present in sandy subsoil horizons. The hydrophobic nature of such compounds may increase their stabilization potential. Therefore, the aim of this study was to investigate the potential of aliphatic compounds in mineral soil horizons along a Dystric Cambisol profile under beech forest to increase hydrophobicity. The conceptual approach included the analyses of soil samples before and after solvent extraction and base hydrolysis for elemental and isotopic composition. Additionally, the advancing contact angle was measured to quantify hydrophobicity. Curie‐point pyrolysis GC/MS was carried out to characterize the nature of alkyl C present in subsoil samples. A close correlation between the ¹⁴C activity and the stable‐C‐isotope ratio (δ¹³C) indicates isotopic fractionation upon C stabilization in subsoils. Free lipids contributed less than 10% to the organic C found in subsoil horizons. Base hydrolysis revealed very high amounts of hydroxyalkanoic acids in the B horizons of the acid forest soil. Hydrophobicity of SOM was not found to be correlated to esterified‐ or free‐lipid content. The contact angle was in a similar range for all bulk soil horizons, suggesting greater hydrophobicity of organic matter in subsoil horizons considering their very low concentrations of organic C compared to the A horizon. The quantity and nature of pyrolysis products change with increasing depth in the soil profile. Aliphatic products cannot be detected in B and C horizons by Curie‐point pyrolysis GC/MS.     

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.     

Leaf litter-colonizing microbiota: supplementary food source or indicator of food quality for Porcellio scaber (Isopoda: Oniscidea)?

Different explanations have been proposed for why terrestrial isopods prefer, and gain from, feeding on microbially inoculated food materials. In the present study, no-choice feeding experiments are used to test three contrasting, but not mutually exclusive, hypotheses. (1) The digestion and nutritional utilization of microbial cells enhance the nutritive value of leaf litter; (2) extracellular digestive enzymes of microbial origin promote digestion of leaf litter; (3) leaf litter-colonizing microbiota serve as indicators for easily digestible leaf litter of high nutritive value. Predictions derived from these hypotheses are compared with the results of a study with experimentally manipulated leaf litter, serving as food sources for the common woodlouse Porcellio scaber. Leaf litter-colonizing microbiota increased consumption of leaf litter by isopods and contributed somewhat to biomass gain of isopods, but not to the assimilation of ingested food. Thus, the present results coincide with predictions derived from hypothesis (3) suggesting that leaf litter-colonizing microbiota stimulate consumption by indicating high food quality. Their positive effects on assimilation and growth, however, are context-specific, being stronger in case of high-quality food than in case of low-quality food.     

Purification and structural characterization of 3-hydroxypropionaldehyde and its derivatives

The compound 3-hydroxypropionaldehyde (3-HPA), together with HPA hydrate and HPA dimer, in aqueous solution forms a system with interesting chemical properties. Therefore, 3-HPA has attracted attention by the chemical industry for use as a precursor in the production of plastics, acrylic acid, and 1,3-propanediol and by the food industry, in using 3-HPA-producing Lactobacillus reuteri as a probiotic. To produce 3-HPA in high yield from glycerol, L. reuteri was used as a biotransformation system. A convenient chromatographic purification method was developed, and purified 3-HPA was analyzed using electrospray ionization mass spectrometry and (13)C NMR. Quantitative (13)C NMR revealed a concentration-dependent distribution of the three compounds forming the HPA system. At concentrations above 1.4 M, the HPA dimer was predominant. However, at concentrations relevant for biological systems, HPA hydrate was the most abundant, followed by the aldehyde form. Our results indicate that the dimeric form with expected antibiotic properties should not be the active form.     

Effect of exposure to soil-bound polycyclic aromatic hydrocarbons on milk contaminations of parent compounds and their monohydroxylated metabolites

The aim of this study was to determine the transfer kinetics of soil-bound polycyclic aromatic hydrocarbons to milk in lactating cows. Soil (500 g/day) fortified with fluorene (104 microg/g dry soil), phenanthrene (82 microg/g), pyrene (78 microg/g), and benzo[a]pyrene (33 microg/g) was administered to three dairy cows via a rumen cannulas for 28 consecutive days. Parent compounds and their major metabolites in milk were measured using gas chromatography-mass spectrometry. Secretion of parent compounds in milk did not increase significantly (P > 0.05) over the control values measured before supply. Target monohydroxylated metabolites were not detected in control samples, but 2-hydroxy fluorene, 3-hydroxy phenanthrene, and 1-hydroxy pyrene were present in milk by the second day of dosing. The highest concentrations of metabolites in milk (31-39 ng/mL) were for 1-hydroxy pyrene at days 7 and 14 of dosing. The observed plateaus for 3-hydroxy phenanthrene and 2-hydroxy fluorene were lower (respectively, 0.69 and 2.79 ng/mL) but significantly increased in comparison to the control samples. Contrarily, 3-hydroxy benzo[a]pyrene was not detected in milk at any sampling time. These results suggested a notable metabolism of the parent compounds after their extraction from soil during the digestive transfer. Thus, the metabolization of fluorene and pyrene can lead to higher concentrations of metabolites than of parent compounds in milk. Despite the absence of a significant transfer of parent PAHs to milk, the appearance of metabolites raises the questions of their impact on human health.     

Surgical approach for tentorial meningiomas in cats: a review of six cases

The surgical technique for removal of tentorial meningiomas is described on six cats using a unilateral temporal supracerebellar transtentorial approach. Complete gross tumour resection was achieved in four of six cats. In one cat, only subtotal resection was achieved. One cat died shortly after surgery because of extensive cerebral haemorrhage. The surgical approach, combined with cisternal or ventricular cerebrospinal fluid puncture and an open-window technique (tumour fenestration and enucleation) provided sufficient visibility and tumour accessibility without excessive manipulation of the brain parenchyma. In all patients, a postoperative transient worsening of the clinical signs was observed. The neurological signs resolved with time with the exception of blindness in two cats. All five surviving cats were monitored for a mean follow-up time of 19 months (median 20 months; range 6-30 months). All patients died or were euthanased because of tumour regrowth within the follow-up period. Although challenging, surgical treatment is a useful therapeutic measure in the treatment of cats presenting with tentorial meningiomas.     

In vitro modulatory effect of ω-3 polyunsaturated fatty acid (EPA and DHA) on phagocytosis and ROS production of goat neutrophils

An in vitro study was carried out to examine the influence of two fish-oil-derived long chain ω-3 polyunsaturated fatty acids (PUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on goat polymorphonuclear leukocytes (PMN). Twelve Saanen healthy goats were used as blood donors. Neutrophils were isolated from blood and incubated with increasing concentration of EPA and DHA (25, 50, 100, 200 μM). Control samples were incubated in the absence of ω-3 PUFAs. Phagocytosis was evaluated by fluorescein-labeled Escherichia coli incorporation, while extracellular Reactive Oxygen Species (ROS) production was determined by cytochrome c reduction assay, which was selected among the others due to its specificity for extracellular superoxide anion release. Phagocytic activity was significantly increased by EPA (P < 0.05) and DHA (P < 0.01). Treating PMN with EPA does not affect extracellular ROS production which is, on the contrary, down-regulated by DHA. This effect was increased in experimental conditions which mimic pro-inflammatory challenges (stimulation with PMA).This study demonstrates that EPA and DHA may have beneficial effect on neutrophil function by increasing their phagocytosis activity and, in the meanwhile, decreasing the tissue damages due to extracellular release of ROS.     

Production of carbon dioxide and nitrous oxide in alkaline saline soil of Texcoco at different water contents amended with urea: A laboratory study

Soil of the former lake Texcoco is alkaline saline with pH often >10 and electrolytic conductivity (EC) >70 dS m^?1 with rapidly changing water contents. Little is known how fertilizing this area with urea to vegetate the soil would affect emissions of carbon dioxide (CO₂) and dynamics of N. Texcoco soil with electrolytic conductivity (EC) 2.3 dS m^?1 and pH 8.5 (TEXCOCO A soil), EC 2.0 dS m^?1 and pH 9.0 (TEXCOCO B soil) and 200 dS m^?1 and pH 11.2 (TEXCOCO C soil) was amended with or without urea and incubated at 40% of water holding capacity (WHC), 60% WHC, 80% WHC and 100% WHC, while emissions of nitrous oxide (N₂O) and CO₂ and dynamics of ammonium (NH₄⁺), nitrite (NO₂^?) and nitrate (NO₃^?) were monitored for 7 days. An agricultural soil served as control (ACOLMAN soil). The emission of CO₂ increased in the urea amended soil 1.5 times compared to the unamended soil, it was inhibited in TEXCOCO C soil and was >1.2 larger in soil incubated at 40%, 60% and 80% WHC compared to soil incubated at 100% WHC. The emission of N₂O increased in soil added with urea compared to the unamended soil, was similar in TEXCOCO A and B soils, but was <0.2 mg N kg^?1 soil day^?1 in TEXCOCO C soil and generally larger in soil incubated at 60% and 80% WHC compared to soil incubated at 40% and 100% WHC. The water content of the soil had no significant effect on the mean concentration of NH₄⁺, but addition of urea increased it in all soils. The concentration of NO₂^? was not affected by the water content and the addition of urea except in TEXCOCO A soil where it increased to values ranging between 20 and 40 mg N kg^?1. The concentration of NO₃^? increased in the ACOLMAN, TEXCOCO A and TEXCOCO B soil amended with urea compared to the unamended soil, but not in the TEXCOCO C soil. It decreased with increased water content, but not in TEXCOCO C soil. It was found that the differences in soil characteristics, i.e. soil organic matter content, pH and EC between the soils had a profound effect on soil processes, but even small changes affected the dynamics of C and N in soil amended with urea.