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Biomedical subjects

H Berrada

Publications and source records attributed to H Berrada.

7 recordsLinked to original sources

Exposure assessment of fruits contaminated with pesticide residues from Valencia, 2001- 03.

A total of 634 samples of oranges, tangerines, peaches, nectarines, khakis and watermelons were collected from an Agricultural Valencia Community Cooperative during the May 2001 to April 2003 campaigns and they were analysed for 15 pesticides among those recommended for pest treatment. A conventional multiresidue analytical procedure based on ethyl acetate extraction was used followed by gas chromatography coupled to a nitrogen phosphorus detector for routine analysis; and mass spectrometry was performed for confirmation. Recovery studies with spiked samples at 0.5 mg kg-1 for each pesticide ranged from 52% for acephate to 87% for fenthion with a standard deviation <20%. Limits of quantification ranged from 0.1 to 100 microg kg-1. A total of 43% of samples contained pesticide residues; and 5% exceeded the maximum residue levels (MRLs). Nine of the pesticides studied were found. Malathion, which was the most frequently detected, was found in 121 samples (19%) at 0.002-4.25 mg kg-1; followed by fenthion in 104 samples (16%) at 0.005-2.3 mg kg-1; and methidation in 68 samples (10%) at 0.008-1.3 mg kg-1. Khaki showed the highest contamination rates with 60% of contaminated samples that exceeded more often the MRLs; and fenthion was the pesticide more frequently detected in all the commodities studied at levels above the European MRLs. The estimated daily intakes of each pesticide calculated from the results obtained were much lower than the acceptable daily intakes.

Adult↗

Real-time quantitative PCR of Staphylococcus aureus and application in restaurant meals.

Staphylococcus aureus is considered the second most common pathogen to cause outbreaks of food poisoning, exceeded only by Campylobacter. Consumption of foods containing this microorganism is often identified as the cause of illness. In this study, a rapid, reliable, and sensitive real-time quantitative PCR was developed and compared with conventional culture methods. Real-time quantitative PCR was carried out by purifying DNA extracts of S. aureus with a Staphylococcus sample preparation kit and quantifying it in the LightCycler system with hybridization probes. The assay was linear from a range of 10 to 10(6) S. aureus cells (r2 > 0.997). The PCR reaction presented an efficiency of >85%. Accuracy of the PCR-based assay, expressed as percent bias, was around 13%, and the precision, expressed as a percentage of the coefficient of variation, was 7 to 10%. Intraday and interday variability were studied at 10(2) CFU/g and was 12 and 14%, respectively. The proposed method was applied to the analysis of 77 samples of restaurant meals in Valencia (Spain). In 11.6% of samples S. aureus was detected by real-time quantitative PCR, as well as by the conventional microbiological method. An excellent correspondence between real-time quantitative PCR and microbiological numbers (CFU/g) was observed with deviations of < 28%.

Colony Count, Microbial↗

Quantification of Listeria monocytogenes in salads by real time quantitative PCR.

A real time quantitative PCR (RTQ-PCR) was carried out purifying DNA extracts of Listeria monocytogenes using a High Pure Listeria Sample Preparation Kit and quantifying in a LightCycler system with hybridisation probes. A standard curve was constructed with serial dilutions. A range linear relationship, from 10 to 10(5)L. monocytogenes colony forming units (CFU), was observed between threshold cycle (Ct) and logarithmic concentration of the serial dilutions. The assay was linear in a range from 10 to 10(5)L. monocytogenes CFU and the coefficient of determination (r2) was >0.98. RTQ-PCR presented an efficiency of >85%. The accuracy of the PCR-based assay, expressed as % bias, ranged from 9% to 26% and the precision, expressed as % CV, ranged 9-22%. Intraday and interday variabilities were studied at 10(2) CFU/g and resulted in 12% and 14%, respectively. The proposed RTQ-PCR method and classical cultural methods were applied to analyse 77 salads from restaurants in Valencia (Spain). All culture positive samples were also RTQ-PCR positive.

Colony Count, Microbial↗

Rapid determination of ochratoxin A in cereals and cereal products by liquid chromatography.

A new method based on extraction with octylsilica (C8) followed by liquid chromatography coupled with fluorescence detection (LC-FLD) was studied to determine ochratoxin A (OTA) from cereals and cereal products. Optimization of different parameters, such as type and amount of solid phase, type and volume of eluent and amount of sample were carried out. Recovery of OTA from rice samples spiked at 10 ng/g level was of 86% with relative standard deviation of 5%. The limits of detection and quantification of the proposed method were 0.25 and 0.75 ng/g, respectively. Furthermore, LC-FLD after of OTA methylation and liquid chromatography coupled to mass spectrometry with an electrospray interface were used for confirmation of OTA in all studied samples. The proposed method was applied to 62 samples of cereals and cereal products and the presence of OTA was found in seven samples.

Chromatography, Liquid↗

Application of solid-phase microextraction for determining phenylurea herbicides and their homologous anilines from vegetables.

Residues of metobromuron, monolinuron and linuron herbicides and their aniline homologous were analyzed in carrots, onions and potatoes by solid-phase microextraction (SPME) performed with a polyacrylate fiber. A juice was obtained from food samples that were further diluted, and an aliquot was extracted after sodium chloride (14%) addition and pH control. At pH 4 only the phenylureas were extracted. A new extraction at pH 11 allowed the extraction of phenylureas plus homologous aniline metabolites. Determination was carried out by gas chromatography with nitrogen-phosporus detection (NPD) the identity of the determined compounds was studied by gas chromatography-mass spectrometry. Limits of quantification (LOQs) obtained with NPD and MS (selected-ion monitoring) were in the microg/kg order allowing determination of maximum residue levels (MRLs) established in the Spanish regulations. MRLs ranged from 0.02 to 0.1 mg/kg depending on the kind of food and herbicide. Under the proposed conditions matrix effects were low enough to permit calibration with samples proceeding from ecological (non-pesticide treated) crops. Twelve commercial samples of each carrots, onions and potatoes were analyzed and only three samples of potatoes contained residues of linuron at levels below MRLs.

Aniline Compounds↗

[Prosthetic hip joint infection caused by Listeria monocytogenes].

The authors report an unusual case of prosthetic hip joint infection caused by Listeria monocytogenes. The patient, an 87-year-old lady who had undergone a right total hip replacement 10 years previously, presented with pain and restriction of hip motion three weeks after an episode of abdominal pain. Aspiration of the joint yielded a dark fluid, from which Listeria Monocytogenes type 4-b was isolated. Blood cultures remained negative. After prolonged antibiotic therapy, symptoms gradually resolved. A few months later, pain recurred with radiological signs of loosening of the femoral component. One-stage revision arthroplasty was performed combined with antibiotic treatment. The patient remains asymptomatic at one year follow-up. Laboratory data and x-ray control are normal. Prosthetic hip joint infection with Listeria monocytogenes is uncommon; few cases have been reported. The literature review shows that prolonged antibiotic therapy alone may be used in patients for whom removal of the prosthesis is not desirable, although revision arthroplasty or prosthesis removal remains necessary in the other cases.

Abdominal Pain↗

Indirect analysis of urea herbicides from environmental water using solid-phase microextraction.

We described here a solid-phase microextraction procedure used to extract six urea pesticides-- chlorsulfuron, fluometuron, isoproturon, linuron, metobromuron and monuron--from environmental samples. Two polydimethylsiloxanes and a polyacrylate fiber (PA) are compared. The extraction time, pH control, addition of NaCl to the water and the influence of organic matter such as humic acid on extraction efficiency were examined to achieve a sensitive method. Determination was carried out by gas chromatography with nitrogen-phosphorus detection. The proposed method requires the extraction of 2 ml of sample (pH 4, 14.3%, w/v, NaCl) for 60 min with the PA fiber. The limits of detection range from 0.04 for linuron to 0.1 microg/l for fluometuron and monuron and the relative standard deviations at the 1 microg/l level are between 15% and 9%. The apparent fiber-water distribution constants (Kfw) calculated in the proposed conditions were in the order of 10(3). Phenylurea herbicides were indirectly determined in the form of their derived anilines and chlorsulfuron in the form of an aminotriazine as confirmed by gas chromatography-mass spectrometry. Natural waters were utilized to validate the final procedure. However, a unequivocal identification in unknown environmental samples should be done by LC-MS. The presence of dissolved organic matter such as humic acid produces losses during the extraction step. Adding sodium chloride to the sample compensates for this effect.

Herbicides↗