Evaluation of a method for assaying sulfonamide antimicrobial residues in cheese: hot-water extraction and liquid chromatography-tandem mass spectrometry

Several sulfonamide antimicrobials (SAAs) are largely used in veterinary medicine. A rapid, specific, and sensitive procedure for determining 12 SAAs in cheese is presented. The method is based on the matrix solid-phase dispersion technique followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from Mozzarella, Asiago, Parmigiano, Emmenthal, and Camembert cheese samples by 6 mL of water modified with 10% methanol and heated at 120 degrees C. The addition of methanol to hot water served to improve remarkably extraction yields of the most lipophilic SAAs, that is, sulfadimethoxine and sulfaquinoxaline. After acidification and filtration, 100 microL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, selecting two precursor-to-product ion transitions for each target compound. Methanol-modified hot water appeared to be an efficient extractant, because absolute recovery ranged between 67 and 88%. Using sulfamoxole as surrogate analyte, recovery of the 12 analytes spiked in the five types of cheese considered at the 50 ng/g level ranged between 75 and 105% with RSD not higher than 11%. Statistical analysis of the mean recovery data showed that the extraction efficiency was not affected by the type of cheese analyzed. This result indicates this method could be applied to other cheese types not considered here. The accuracy of the method was determined at three spike levels, that is, 20, 50, and 100 ng/g, and varied between 73 and 102% with relative standard deviations ranging between 4 and 12%. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to be <1 ng/g.     

Influence of harvesting method and washing on the presence of pesticide residues in olives and olive oil

The influence of the preliminary steps in olive oil production (harvesting and washing) on pesticide residues in olives and olive oil has been investigated. Analyses were performed by GC-MS/MS and revealed that endosulfan sulfate and two herbicides (diuron and terbuthylazine) were the most frequently found residues in olives and olive oil. The harvesting method has a decisive influence on herbicide concentrations found in olives. Thus, 16 and 48% of the olive samples harvested on the ground after falling from the tree presented concentrations higher than the maximum residue limit (MRL) for diuron and terbuthylazine, respectively. In olives harvested directly from the tree, diuron was not found at concentrations higher than MRL and terbuthylazine was found in only 10% of the samples. The washing step performed routinely in olive mills was effective in removing the superficial contamination by herbicides present in olives harvested on the ground. Nevertheless, even after washing, the olive oil obtained from ground olives showed herbicide residue concentrations higher than those obtained from tree olives.     

A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg

A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of 相似文献   

Soil physical quality of a Brazilian Oxisol under two tillage systems using the least limiting water range approach

Plant growth is directly affected by soil water, soil aeration, and soil resistance to root penetration. The least limiting water range (LLWR) is defined as the range in soil water content within which limitations to plant growth associated with water potential, aeration and soil resistance to root penetration are minimal. The LLWR has not been evaluated in tropical soils. Thus, the objective of the present study was to evaluate the LLWR in a Brazilian clay Oxisol (Typic Hapludox) cropped with maize (Zea mays L. cv. Cargil 701) under no-tillage and conventional tillage. Ninety-six undisturbed soil samples were obtained from maize rows and between rows and used to determine the water retention curve, the soil resistance curve and bulk density. The results demonstrated that LLWR was higher in conventional tillage than in no-tillage and was negatively correlated with bulk density for values above 1.02 g cm⁻³. The range of LLWR variation was 0–0.1184 cm³ cm⁻³ in both systems, with mean values of 0.0785 cm³ cm⁻³ for no-tillage and 0.0964 cm³ cm⁻³ for conventional tillage. Soil resistance to root penetration determined the lower limit of LLWR in 89% of the samples in no-tillage and in 46% of the samples in conventional tillage. Additional evaluations of LLWR are needed under different texture and management conditions in tropical soils.     

Geomorphological significance of lichen colonization in a present snow hollow: Hoya del Cuchillar de las Navajas, Sierra de Gredos (Spain)

This paper discusses the results of a lichenometrical and geomorphological study of one of the few remaining active snow hollows in the central region of the Iberian Peninsula. The study area, located on a glacial shoulder, is called Hoya del Cuchillar de las Navajas. A protalus rampart occurs at the base of the hollow. Our studies, conducted between 1992 and 1998, were designed to determine the geomorphological characteristics of Hoya, the mobility of the deposits, and the characteristics of the snow cover. These data formed the basis for a study of the lichen colonization on the blocks and on the wall surrounding the snow hollow. All of the lichen species found were analyzed according to their abundance, distribution and the extent of their surface cover. Measurements of the diameter of the thalli of the species Rhizocarpon geographicum were also obtained. Thalli of this species were found to require a mean snow-free growing season of at least 95 days (13.5 weeks) per year. Maximum mean thallus diameters indicate that the protalus rampart was formed during the Little Ice Age and became inactive 130 years ago.     

Hydrogeochemical Characteristics of Processes in the Temara Aquifer in Northwestern Morocco

The physico-chemical characteristics of the groundwater of the Temara aquifer were studied by means of piezometric mapping and determination of the ionic composition of the groundwater. In general, the agricultural activity is intense in the area, with water being pumped from numerous wells. Two aquifer formations can be distinguished which, over a wide area, are separated by layers of low permeability. The increased salinity at some points of the coastal zone is probably linked to the combined action of the washing out of Miocene marls, dissolution of carbonate rocks, agricultural pollution and seawater intrusion.     

Functional and molecular responses of soil microbial communities under differing soil management practices

The effects of soil management on some microbiological properties and soil bacterial community structure were evaluated. Two field sites with the same soil type, located on the same geographic area adjacent to one other, have received different soil management practices and cultivation. One site has been subjected for 20 years to intensive horticulture under conventional tillage and irrigation with low quality salt-rich water; the second field site has been uncultivated for a long period and was turned to organic farming practices over the last 5 years and is currently cultivated with fruit orchard. Total bacterial counts, microbial ATP, microbial community metabolic (BIOLOG^®) profiles, and DNA fingerprinting by PCR-DGGE were determined. Two-way ANOVA revealed that total bacterial counts were not significantly (P>0.3) affected by the two different management practices; ATP content was consistently and significantly (P<0.001) lower in salt-water irrigated soil than in organic soil at the three sampling times. The cluster analysis of community level physiological profiles indicated that microbial communities were much more uniform in organic soil than in irrigated one, suggesting that salt-water irrigation could have affected the size of the microbial population, its metabolic activities, as well as its composition. Molecular patterns fitted the BIOLOG^® profile diversity. In particular, at any sampling time, PCR-DGGE patterns of bacterial DNA, extracted by an indirect method, significantly discriminated irrigated from organic soil samples. The PCR-DGGE patterns of total soil DNA, extracted by a direct method, showed a moderate to significant variation among irrigated and organic soil samples. Biochemical, microbiological and molecular data contributed to evidence a significantly different response of indigenous microflora to soil management by using saline water or organic farming.     

Establishment of Retama sphaerocarpa L. seedlings on a degraded semiarid soil as influenced by mycorrhizal inoculation and sewage‐sludge amendment

A field experiment was carried out to evaluate the effectiveness of mycorrhizal inoculation with three arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge), and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and the addition of composted sewage sludge (SS) with respect to the establishment of Retama sphaerocarpa L. seedlings, in a semiarid Mediterranean area. Associated changes in soil chemical (nutrient content and labile carbon fractions), biochemical (enzyme activities), and physical (aggregate stability) parameters were observed. Six months after planting, both the addition of composted SS and the mycorrhizal‐inoculation treatments had increased total N content, available‐P content, and aggregate stability of the soil. Values of water‐soluble C and water‐soluble carbohydrates were increased only in the mycorrhizal‐inoculation treatments. Rhizosphere soil from the mycorrhizal‐inoculation treatments had significantly higher enzyme activities (dehydrogenase, protease‐BAA, acid phosphatase, and β‐glucosidase) than the control soil. In the short‐term, mycorrhizal inoculation with AM fungi was the most effective treatment for enhancement of shoot biomass, particularly with G. mosseae (about 146% higher with respect to control plants). The addition of the composted SS alone was sufficient to restore soil structural stability but was not effective with respect to improving the performance of R. sphaerocarpa plants.     

Genetic diversity of Musa diploid and triploid accessions from the Brazilian banana breeding program estimated by microsatellite markers

Banana (Musa spp.) is one of the most consumed fruits worldwide. Production is based mainly on triploid cultivars, and most genetic improvement programs aim to generate tetraploid hybrids obtained from the crossing of established triploid cultivars with a diploid parent genotype, improved or wild, exhibiting the trait of interest, normally resistance to biotic factors. Microsatellites were used to investigate the genetic variability and relationships between 58 Musa genotypes, including 49 diploids and nine triploid cultivars maintained at the Musa germplasm collection of the Brazilian dessert banana breeding program. Thirty-three primer pairs developed for banana were tested, and nine amplified reproducible and discrete fragments, producing a total of 115 alleles. The average number of alleles amplified per primer was 12.8, ranging from 10 to 15. The diploid genotypes presented the largest genetic variability, demonstrated by the large number of alleles detected, and the low similarity between the clones. The phenetic analysis clustered the triploid cultivars in a separated group, with the exception of the Nanica and Gros Michel cultivars, which showed high similarity with the diploid cultivar Mambee Thu. It was not possible to separate the wild diploid genotypes from the cultivated ones, indicating a common origin of these genotypes. A high proportion of duplicated alleles and/or loci was observed for diploid and triploid genotypes. The information gathered about the similarity between diploid and triploid accessions will help to define potential crosses to maximize the recovery of the typical fruit qualities required in Brazil (AAB, Pome and Silk dessert banana).