A software-based sensor for combined sewer overflows

A new methodology for online estimation of excess flow from combined sewer overflow (CSO) structures based on simulation models is presented. If sufficient flow and water level data from the sewer system is available, no rainfall data are needed to run the model. An inverse rainfall-runoff model was developed to simulate net rainfall based on flow and water level data. Excess flow at all CSO structures in a catchment can then be simulated with a rainfall-runoff model. The method is applied to a case study and results show that the inverse rainfall-runoff model can be used instead of missing rain gauges. Online operation is ensured by software providing an interface to the SCADA-system of the operator and controlling the model. A water quality model could be included to simulate also pollutant concentrations in the excess flow.     

Prevalence of Clostridium botulinum in food raw materials used in REPFEDs manufactured in France

Food raw materials used in refrigerated processed foods of extended durability (REPFEDs) manufactured in France were surveyed for Clostridium botulinum types A, B and E using PCR-Enzyme-linked Immunosorbent assay (PCR-ELISA) and mouse bioassay for detection respectively of cells and toxins in enrichment broth. Portions of 25 to 50 g of food were analysed. A total of 8 out of the 102 samples of fish and shellfish, 12 out of the 143 samples of meat and poultry, 1 out of the 62 samples of aroma, sauce and gravy, 4 out of the 25 samples of thickening agents, 3 out of the 26 samples of dehydrated dairy ingredients, and none of the 65 samples of spices, herbs and dehydrated mushroom were positive for C. botulinum in PCR-ELISA, i.e., 6.6% of all the samples tested. The 28 positive samples comprised 10 type A, 10 type B, 4 with both types A and B, and 4 undetermined by PCR typing. No sample positive for type E was detected. Of the 28 samples positive in PCR-ELISA, 15 were also positive in the mouse bioassay. The MPN count was between 1 and 3 C. botulinum/kg of food, which is similar to or in the lower range of values reported in the literature.     

Multilayer polyimide film substrates for interconnections in microsystems

 Currently there is a great interest in flexible substrates for their use in the packaging of semiconductors. They allow high packaging density and permit mechanical motion of the components, if necessary. This paper describes a new substrate material which possesses a high flexibility and which can be used for the realization of interconnection cables and MCM-D. These substrates are not only highly flexible but also mechanically stable, chemically inert and can be produced with the same processes as those for rigid substrates. Received: 14 December 1998 / Accepted: 28 December 1998     

Pb1−xSnxSe-on-Si LWIR sensor arrays and thermal imaging with JFET/CMOS read-out

Long wavelength infrared (LWIR) sensor arrays were fabricated in Pb_1−xSn_xSe layers grown epitaxially on Si-substrates by MBE. A CaF2 intermediate buffer layer ≈30dgA thick was employed for compatibility reasons. The photovoltaic sensors are based on the blocking Pb-contact technique on p-type material. They were fabricated using simple wet-etching process steps only. Cut-off wave-lengths were about 10.5 μm, quantum efficiencies >60%, and resistance-aera products above 3 Ω-cm² at 90K. A demonstrational LWIR thermal imaging camera was assembled with a 256 element line array with 50 μm pitch. Low-noise signal processing was achieved with sensors with differential resistances in the 10 kOhm range by using JFET/CMOS technology. For each channel, an integrator, correlated multiple sampling and sample/hold amplifier was used before multiplexing to a common output.     

Micro-array for the identification of Shiga toxin-producing Escherichia coli (STEC) seropathotypes associated with Hemorrhagic Colitis and Hemolytic Uremic Syndrome in humans

A micro-array has been developed, based on the GeneDisc® array, for the genetic identification of 12 O-types and 7 H-types of Shiga toxin-producing Escherichia coli (STEC) including the most clinically relevant enterohemorrhagic E. coli (EHEC) serotypes. The genes selected for determination of the O antigens (rfbE_O157, wzx_O26, wzx_O103, wbd1_O111, ihp1_O145, wzx_O121, wzy_O113, wzy_O91, wzx_O104, wzy_O118, wzx_O45, and wbgN_O55) and H-types (fliC_H2, fliC_H7, fliC_H8, fliC_H11, fliC_H19, fliC_H21, and fliC_H28) showed a high specificity and concordance with serology. The micro-array also had a high specificity for EHEC-associated virulence factors, including Shiga toxins 1 and 2 (stx1 and stx2), intimin (eae), enterohemolysin (ehxA), serine protease (espP), catalase peroxidase (katP), the type II secretion system (etpD), subtilase cytotoxin (subA), autoagglutinating adhesin (Saa) and type III secreted effectors encoded in the genomic islands OI-122 (ent/espL2, nleB, and nleE) and OI-71 (nleF, nleH1-2, and nleA). The eae gene was detected in all typical EHEC strains, and the pattern of nle genes encoded in OI-71 and OI-122 was found to be closely associated with certain serotypes of typical EHEC and emerging EHEC strains. Virulence plasmid associated genes such as katP, espP, and etpD were more common in EHEC than in STEC strains; this supports their association with virulence. This array constitutes a valuable approach for the identification of STEC strains with a high potential for human virulence.     

Evaluation of a polymerase chain reaction-based test for detecting Salmonella spp. in food samples: Probelia Salmonella spp

A commercially available polymerase chain reaction (PCR) kit was evaluated for the detection of Salmonella spp. in food samples. The test combines PCR amplification and sandwich hybridization of the amplified DNA in microtiter plates. The sensitivity and specificity was evaluated with 52 Salmonella strains and 51 non-Salmonella strains and showed that the test was entirely reliable. The threshold sensitivity was 10(2) CFU/ml. The limit of detection of dead cells that determines the minimum detection level of dead cells in food samples was superior to 10(6) CFU/25 g, a level rarely achieved in naturally contaminated samples. After an 18-h pre-enrichment step, the test could detect viable Salmonella in artificially contaminated food samples, even for the lower contamination level (3 CFU/25 g). There was complete agreement between the PCR test and the ISO 6579 bacteriological reference method with artificially contaminated samples. Regarding the accuracy of the results obtained from 253 naturally or noncontaminated foods and from 32 artificially contaminated foods, the agreement percentage was 99.6%. The fidelity of the technique was evaluated in a collaborative study with eight European laboratories and showed a correlation of 98.4%.     

Semi-automated sample preparation for plasma proteomics

The discovery of new biomarkers will be an essential step to enhance our ability to better diagnose and treat human disease. The proteomics research community has recently increased its use of human blood (plasma/serum) as a sample source for these discoveries. However, while blood is fairly non-invasive and readily available as a specimen, it is not easily analyzed by liquid chromatography (LC)/mass spectrometry (MS), because of its complexity. Therefore, sample preparation is a crucial step prior to the analysis of blood. This sample preparation must also be standardized in order to gain the most information from these valuable samples and to ensure reproducibility. We have designed a semi-automated and highly parallel procedure for the preparation of human plasma samples. Our process takes the samples through eight successive steps before analysis by LC/MS: (1) receipt, (2) reformatting, (3) filtration, (4) depletion, (5) concentration determination and normalization, (6) digestion, (7) extraction, and (8) randomization, triplication, and lyophilization. These steps utilize a number of different liquid handlers and liquid chromatography (LC) systems. This process enhances our ability to discover new biomarkers from human plasma.     

Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test

A duplex PCR procedure was evaluated for the detection of Clostridium perfringens in food and biological samples and for the identification of enterotoxigenic strains. This method uses two sets of primers which amplify in the same reaction two different DNA fragments simultaneously: the 283-bp C. perfringens phospholipase C gene fragment and the 426-bp enterotoxin gene fragment. Internal primers within the two primer sets confirmed the specificity of the method by DNA-DNA hybridization with the PCR products. No cross-reaction was observed with other Clostridium species or with other bacteria routinely found in food. The detection level was approximately 10(5) C. perfringens cells per g of stool or food sample. When overnight enrichment culture was used, 10 C. perfringens cells per g was detected in 57 artificially contaminated food samples. The duplex PCR is a rapid, sensitive, and reliable method for the detection and identification of enterotoxigenic C. perfringens strains in food samples. A slide latex agglutination test was also evaluated as a rapid, simple technique for the detection of C. perfringens enterotoxin in stool samples.