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In a context of inclusive school, this paper aims to demonstrate good practices to implement, to promote autistic students' academic success in regular classes during the primary to secondary school transition. The information gathered is based on an action research conducted during two years in three schools from two different Quebec administrative regions. Stakeholders (n=25), autistic students (n=11), and their parents (n=10), have helped to outline the salient aspects of the practices implemented to promote an inclusive education for all. Our conclusions show that it is imperative to provide professional development on understanding the scheme of thinking of autistic students, to regular classroom stakeholders. It is also important that decision-makers give time to set up collaboration between primary and secondary level resources, as well as those of community and rehabilitation networks. [ABSTRACT FROM AUTHOR]

A method has been developed for the detection in beef trim of priority Shiga toxin-producing E. coli (STEC) strains, defined as E. coli possessing the virulence factors stx1 and/or stx2 and intimin (eae), with O serogroups O26, O45, O103, O11 1, O121, O145, or O157. The method is based on recovery of the target bacteria by overnight enrichment in a broth optimized for recovery of O157 and non-O157 STEC, followed by screening using multiplex PCR techniques targeting (i) stx1 stx2, and eae (STE PCR) and (ii) gene sequences associated with the seven priority O serogroups (Poly O PCR), and then direct plating of broth samples positive in both STE and Poly O PCR onto Rainbow agar. Colonies on agar media were screened batchwise for STEC by the STE PCR, and presumptive isolates were characterized using a multiplex PCR and cloth-based hybridization array system targeting key virulence and O serogroup-specific markers. Using one representative strain of each priority O serogroup individually inoculated in beef trim samples, the method exhibited a limit of detection approaching 1 to 2 viable STEC cells per 65 g. None of the uninoculated trim samples produced positive results with either of the screening PCR procedures or on analysis of colonies recovered on plating media. STEC-negative samples were readily identified by screening PCR within 24 h, with a turnaround time of fewer than 4 days for confirmation of positives. The inclusivity and exclusivity characteristics of the screening PCR techniques were verified using a total of 65 different priority STEC strains: 24 nonpriority STEC, 15 non-STEC bacteria, and only those strains bearing the targeted characteristics produced screening PCR-positive results. [ABSTRACT FROM AUTHOR]

A cloth-based hybridization array system (CHAS) was developed for the identification of foodborne colony isolates of seven priority enterohemorrhagic Escherichia coli (EHEC-7) serogroups targeted by U.S. food inspection programs. Gene sequences associated with intimin; Shiga-like toxins 1 and 2; and the antigenic markers O26, O45, O103, O111, O121, O145, and O157 were amplified in a multiplex PCR incorporating a digoxigenin label, and detected by hybridization of the PCR products with an array of specific oligonucleotide probes immobilized on a polyester cloth support, with subsequent immunoenzymatic assay of the captured amplicons. The EHEC-7 CHAS exhibited 100% inclusivity and 100% exclusivity characteristics with respect to detection of the various markers among 89 different E. coli strains, with various marker gene profiles and 15 different strains of non-E. coli bacteria. [ABSTRACT FROM AUTHOR]

Abstract: At Cape Vera (Devon Island, Nunavut, Canada), a seabird colony of northern fulmars (Fulmarus glacialis) congregates and releases nutrients through the deposition of guano to the coastal terrestrial environment, thus creating nutrient-fertilized habitats important to insects, birds, and mammals. Here we determined whether mercury was similarly enriched in various terrestrial food web components in this High Arctic coastal ecosystem due to seabird inputs. Stable isotopes (δ 15N, δ 13C) were used to identify trophic linkages and possible routes of contaminant transfer in the food web. Values of δ 15N were significantly higher in lichens and certain plants collected closer to the bird colony, demonstrating a gradient of seabird influence, and were higher at Cape Vera than our reference site at Cape Herschel, on eastern Ellesmere Island, an area relatively unaffected by seabirds. In contrast, δ 13C showed little variation among terrestrial species, suggesting minimal influence by seabirds. Concentrations of total mercury (THg) in primary producers and phyto/zooplankton were not significantly correlated with distance from the seabird colony or δ 15N values, and were similar to other taxa from the High Arctic. Our results provide novel data on THg in several Arctic taxa where concentrations have not been reported previously. Moreover, the analyses indicate that δ 15N is significantly enriched in the adjacent environment by guano fertilization, but our study was unable to show an enrichment of THg and δ 13C in the terrestrial food web near the seabird colony. [Copyright &y& Elsevier]

A simple cloth-based hybridization array system (CHAS) was developed for the identification of toxin genes associated with major foodborne pathogenic bacteria, including toxigenic Escherichia coli, Vibrio cholerae and Salmonella spp. Bacterial isolates were subjected to a multiplex polymerase chain reaction incorporating digoxigenin (DIG)–deoxyuridine triphosphate and primers targeting a variety of toxin genes (verotoxin, Salmonella enterotoxin, cholera toxin and heat-labile/stable enterotoxin), followed by hybridization of the amplicons with an array of probes immobilized on polyester cloth and subsequent immunoenzymatic assay of the bound DIG label. This system provided sensitive and specific detection of the different target toxin gene markers in a variety of bacterial isolates, exhibiting the expected patterns of reactivity with a panel of bacteria having defined toxigenicity profiles. The CHAS is a cost-effective tool facilitating the determination of the potential toxigenic profile of bacteria in the food microbiology laboratory, thus contributing valuable information to the risk-assessment process in the microbiological analysis of foods. [ABSTRACT FROM AUTHOR]

The growth characteristics of 96 shiga toxin-producing Escherichia coli (STEC) strains representing 36 different O-types (including priority O types O26, O45, O103, O111, O121, O145 and O157) on commercial and in-house agar media were studied. The ability of the strains to grow on agar media with varying selective supplement formulations was evaluated using MacConkey Agar (MAC); Rainbow® Agar O157 (RBA); Rainbow® Agar O157 with manufacturer-recommended selective supplements (RBA-NT); Rainbow® Agar O157 with USDA-recommended selective supplements (RBA-USDA); CHROMagar STEC™ (CH STEC); Tryptone Bile agar containing cefixime and tellurite (TBA-CT); Tryptone Bile agar containing cefixime, tellurite, eosin and methylene blue (TBA-EM); and VTEC agar. All of the strains were able to grow on MAC, RBA and VTEC agar, whereas a number of strains (including some non-O157 priority O types) were unable to grow on the highly selective media CH STEC, RBA-NT, RBA-USDA, TBA-EM and TBA-CT. Only RBA-NT and CH STEC exhibited significant inhibition of background flora from ground beef enrichment. Significant inhibition of background flora from beef trim enrichment was observed with RBA-NT, RBA-USDA, CH STEC, TBA-EM and VTEC agar. With exception of E. coli O157, several different colony morphologies were observed on the differential plating media among strains of the same O type, indicating that this colony morphology is not a reliable means of identifying target STEC. These results suggest that an approach to maximize the recovery of target STEC from beef enrichment cultures is dual plating on lesser (RBA, MAC, VTEC agar) and more highly (RBA-NT, CH STEC) selective agars. [ABSTRACT FROM AUTHOR]

Nalidixic acid-resistant (NalR) mutants of Salmonella enterica serovar Berta and Escherichia coli O157:H7 were derived from wild-type laboratory cultures to serve as distinguishable control strains for routine use in food microbiology testing programs. The prevalence of the NalR phenotype among different bacteria was verified using panels of related and unrelated strains with the ability to grow vigorously on plating media containing nalidixic acid, being restricted to the NalR mutants. The NalR phenotype was stable in both mutant strains over several generations in the absence of selective pressure and enabled their differentiation from wild-type bacteria on the basis of their ability to grow on plating media containing nalidixic acid. A similar approach for the development of a distinguishable Listeria monocytogenes control strain was not possible due to the inherent resistance of this organism to nalidixic acid. Instead, an L. monocytogenes isolate with rare genotypic and serologic features was identified as a possible candidate to serve as a unique and distinguishable positive control strain. [ABSTRACT FROM AUTHOR]

The determination of Shiga toxin (ST) subtypes can be an important element in the risk characterization of foodbome ST producing Escherichia coli (STEC) isolates for making risk management decisions. ST subtyping methods include PCR techniques based on electrophoretic or pyrosequencing analysis of amplicons and in silico techniques based on whole genome sequence analysis using algorithms that can be readily incorporated into bioinformatics analysis pipelines for characterization of isolates by their genetic composition. The choice of technique will depend on the performance characteristics of the method and an individual laboratory's access to specialized equipment or personnel. We developed two whole genome sequence-based ST subtyping tools: (t) an in silico PCR algorithm requiring genome assembly to replicate a reference PCR-based method developed by the Statens Serum Institut (SSI) and (ii) an assembly-independent routine in which raw sequencing results are mapped to a database ot known ST subtype sequence variants (V-Typer). These tools were evaluated alongside the SSI reference PCR method and a recently described PCR-based pyrosequencing technique. The V-Typer method results corresponded closely with the reference method in the analysis of 67 STEC cultures obtained from a World Health Organization National Reference Laboratory. In contrast, the in silico PCR method failed to detect ST subtypes in several cases, a result which we attribute to assemblyinduced errors typically encountered with repetitive gene sequences. The V-Typer can be readily integrated into bioinformatics protocols used in the identification and characterization of foodbome STEC isolates. [ABSTRACT FROM AUTHOR]

Genomic antimicrobial resistance (AMR) prediction tools have the potential to support foodborne illness outbreak investigations through their application in the analysis of bacterial genomes from causative strains. The AMR marker profile of a strain of interest, initially identified in outbreak-associated clinical samples, may serve as the basis for customization of selective enrichment media, facilitating its recovery from samples in a food safety investigation. Different possibilities for AMR analyses include the use of comprehensive AMR gene databases such as the Comprehensive Antibiotic Resistance Database, which can be mined with in-house bioinformatics alignment tools (e.g., Antimicrobial Resistance Marker Identifier), or publicly available tools based on clinically relevant acquired AMR gene databases (e.g., ResFinder). In combination with a previously reported pipeline (SigSeekr) designed to identify specific DNA sequences associated with a particular strain for its rapid identification by PCR, it should be possible to deploy custom recovery and identification tools for the efficient detection of priority pathogens such as Shiga toxigenic Escherichia coli (STEC) outbreak strains within the time frame of an active investigation. Using a laboratory STEC strain as a model, trimethoprim resistance identified by both Antimicrobial Resistance Marker Identifier and ResFinder was used as the basis for its selective recovery against a background of commensal E. coli bacteria in ground beef samples. Enrichment in modified tryptic soy broth containing trimethoprim greatly enhanced the recovery of low numbers of model strain cells inoculated in ground beef samples, as verified by the enumeration of colonies on plating media using a strain-specific PCR method to determine the recovery efficiency for the target strain. We discuss the relative merits of different AMR marker prediction tools for this purpose and describe how such tools can be utilized to good effect in a typical outbreak investigation scenario. [ABSTRACT FROM AUTHOR]

We describe the translation of a cloth-based hybridization array system (CHAS), a colorimetric DNA detection method that is used by food inspection laboratories for colony screening of pathogenic agents, onto a microfluidic chip format. We also introduce an articulated centrifugal platform with a novel fluid manipulation concept based on changes in the orientation of the chip with respect to the centrifugal force field to time the passage of multiple components required for the process. The platform features two movable and motorized carriers that can be reoriented on demand between 0 and 360° during stage rotation. Articulation of the chip can be used to trigger on-the-fly fluid dispensing through independently addressable siphon structures or to relocate solutions against the centrifugal force field, making them newly accessible for downstream transfer. With the microfluidic CHAS, we achieved significant reduction in the size of the cloth substrate as well as the volume of reagents and wash solutions. Both the chip design and the operational protocol were optimized to perform the entire process in a reliable, fully automated fashion. A demonstration with PCR-amplified genomic DNA confirms on-chip detection and identification of Escherichia coli O157:H7 from colony isolates in a colorimetric multiplex assay using rfbO157, fliCH7, vt1, and vt2 genes. [ABSTRACT FROM AUTHOR]

Control strains of bacterial pathogens such as Escherichia coli O157:H7 are commonly processed in parallel with test samples in food microbiology laboratories as a quality control measure to assure the satisfactory performance of materials used in the analytical procedure. Before positive findings can be reported for risk management purposes, analysts must have a means of verifying that pathogenic bacteria (e.g., E. coli O157:H7) recovered from test samples are not due to inadvertent contamination with the control strain routinely handled in the laboratory environment. Here, we report on the application of an in-house bioinformatic pipeline for the identification of unique genomic signature sequences in the development of specific oligonucleotide primers enabling the identification of a common positive control strain, E. coli O157:H7 (ATCC 35150), using a simple PCR procedure. [ABSTRACT FROM AUTHOR]

Non-O157 enterohemorrhagic Escherichia coli in priority serogroups O26, O45, O103, O111, O121, and O145 are increasingly recognized as important human pathogens. In the present study, a panel of monoclonal antibodies (MAbs) to the lipopolysaccharide O antigens of E. coli in serogroups O26, O45, O103, O111, O121, and O145 was produced. The specificity was evaluated by examining the reactivity of the MAbs with 50 E. coli strains and 42 non-E. coli bacteria, and several MAbs highly specific for E. coli strains in each of the six non-O157 priority serogroups were identified. The use of these highly specific MAbs may be of considerable value for determining whether an E. coli isolate belongs to one of the six priority non-O157 serogroups, for developing specific detection assays for these organisms, and for characterizing the lipopolysaccharide O antigens of isolates in these serogroups. [ABSTRACT FROM AUTHOR]

A simple immunoenzymatic enterohemorrhagic Escherichia coli (EHEC) colony check (ECC) assay was developed for the presumptive identification of priority EHEC colonies isolated on plating media from enrichment broth cultures of foods. With this approach, lipopolysaccharide extracted from a colony is spotted on the grid of a polymyxin-coated polyester cloth strip, and bound E. coli serogroup O26, O45, P103, O111, O121, O145, and O157 antigens are subsequently detected by sequential reactions with a pool of commercially available peroxidase-conjugated goat antibodies and tetramethylbenzidine substrate solution. Each strip can accommodate up to 15 colonies, and test results are available within 30 min. Assay performance was verified using colonies from a total of 73 target EHEC isolates covering the range of designated priority serogroups (all of which were reactive), 41 nontarget E. coli isolates including several nontarget Shiga toxin-producing E. coli serogroups (all unreactive), and 33 non-E. coli strains (all unreactive except two bacterial strains possessing O-antigenic structures in common with those of the priority EHEC). The ECC assay was reactive with target colonies grown on several types of selective and nonselective plating media designed for their cultivation. These results support the use of the ECC assay for high-throughput screening of colonies isolated on plating media for detecting priority EHEC strains in foods. [ABSTRACT FROM AUTHOR]

A rapid method was devised for the selective enrichment and detection of multi-antibiotic-resistant Salmonella enterica subsp. enterica serotype Typhimurium (S. Typhimurium) DT104 in poultry. The target organism was pre-enriched in trypticase soy broth supplemented with chloramphenicol, followed by plating on modified semisolid Rappaport Vassiliadis medium and sampling a portion of the swarming growth. The presence of S. Typhimurium DT104 was determined by testing the growth using a cloth-based hybridization array system (CHAS) targeting antibiotic resistance and other marker genes associated with this organism. In this CHAS, a multiplex polymerase chain reaction incorporating digoxigenin–dUTP was used to simultaneously amplify seven target gene sequences, with subsequent rapid detection of the amplicons by hybridization with an array of probes immobilized on polyester cloth and immunoenzymatic assay of the bound label. This procedure permitted the rapid determination of S. Typhimurium DT104 in chicken carcass rinse, powdered egg and feed samples inoculated with different levels of the target organism. PRACTICAL APPLICATIONS The combined selective enrichment and CHAS procedure will enable the rapid screening of poultry and related products for the presence of multidrug resistant Salmonella Typhimurium DT104. The convenience of this approach lies in the fact that samples containing salmonellae requiring more comprehensive analysis can be rapidly identified on the basis of a simple preliminary enrichment procedure involving examination of MSRV plating media for characteristic swarming growth. The swarming growth is readily sampled and analyzed by the DT104 CHAS to determine the presence of key marker genes associated with the multidrug resistant pathogen, obviating the need for time- and labor-consuming conventional purification and antimicrobial susceptibility testing procedures. [ABSTRACT FROM AUTHOR]