Lupine flour, protein, and fiber have become common ingredients in food products. The association of lupine-related allergic incidents with peanut allergy is a cause for concern as the latter may bring about severe reactions. In this study, a hybridization probe-based real-time PCR assay for the detection of lupine DNA in foods was developed. Particular attention was paid to the specificity of the method, which was verified by analysis of DNA extracts from more than 50 potential food ingredients such as legumes, cereals, seeds, nuts, spices, fruits, and meat. The limit of detection of the method was determined as 0.1 mg/kg. The successful detection of the presence/absence of lupine DNA in 20 samples proved the suitability of the assay for the analysis of frequently encountered food matrices.
A risk of cross-contamination exists when preparing a gluten-free (GF) meal in kitchen facilities that usually handle gluten-containing (GC) foods. Cross-contamination with gluten may occur during the preparation or cooking process; however, published data are lacking on gluten cross-contamination from kitchenware. This study was conducted to determine whether cross-contamination occurs through shared domestic kitchenware and, if so, which cleaning method is most reliable for avoiding this cross-contamination. Kitchenware (wooden spoon, colander, ladle, and knife) previously used to cook and/or prepare GC foods was used for the preparation of GF foods (bread and pasta). The gluten concentration of the GF foods was then determined using an established enzyme-linked immunosorbent assay. A PCR assay was also used to detect the presence of wheat ω-gliadin DNA in the food samples. Three cleaning methods were assessed to determine the concentrations of gluten and wheat DNA in GF foods cooked with utensils cleaned directly after the preparation of GC foods. Contrary to our expectations, gluten was not detected in relevant and quantifiable amounts in our samples (<20 mg/kg). The cleaning method used did not influence gluten concentrations: all samples contained <10 mg/kg. Based on PCR analyses, the only sample with lower cycle threshold ( C) values (i.e., higher concentration of wheat DNA) was from the contaminated ladle used to serve GF pasta. This outcome led to the hypothesis that shared ladles pose a higher risk for contamination of GF foods than do shared wooden spoons, colanders, or knives. Cross-contamination with gluten in a kitchen environment may occur, but kitchen utensils used for preparing GC pasta and for cutting GC bread should not pose a relevant problem to patients with celiac disease, at least in a domestic environment.
AIMS OF THE STUDY: Wastewater-based epidemiology has contributed significantly to the comprehension of the dynamics of the current COVID-19 pandemic. Its additional value in monitoring SARS-CoV-2 circulation in the population and identifying newly arising variants independently of diagnostic testing is now undisputed. As a proof of concept, we report here correlations between SARS-CoV-2 detection in wastewater and the officially recorded COVID-19 case numbers, as well as the validity of such surveillance to detect emerging variants, exemplified by the detection of the B.1.1.529 variant Omicron in Basel, Switzerland. METHODS: From July 1 to December 31, 2021, wastewater samples were collected six times a week from the inflow of the local wastewater treatment plant that receives wastewater from the catchment area of the city of Basel, Switzerland, comprising 273,075 inhabitants. The number of SARS-CoV-2 RNA copies was determined by reverse transcriptase-quantitative PCR. Spearman’s rank correlation coefficients were calculated to determine correlations with the median seven-day incidence of genome copies per litre of wastewater and official case data. To explore delayed correlation effects between the seven-day median number of genome copies/litre wastewater and the median seven-day incidence of SARS-CoV-2 cases, time-lagged Spearman’s rank correlation coefficients were calculated for up to 14 days. RNA extracts from daily wastewater samples were used to genotype circulating SARS-CoV-2 variants by next-generation sequencing. RESULTS: The number of daily cases and the median seven-day incidence of SARS-CoV-2 infections in the catchment area showed a high correlation with SARS-CoV-2 measurements in wastewater samples. All correlations between the seven-day median number of genome copies/litre wastewater and the time-lagged median seven-day incidence of SARS-CoV-2 cases were significant (p<0.001) for the investigated lag of up to 14 days. Correlation coefficients declined constantly from the maximum of 0.9395 on day 1 to the minimum of 0.8016 on day 14. The B.1.1.529 variant Omicron was detected in wastewater samples collected on November 21, 2021, before its official acknowledgement in a clinical sample by health authorities. CONCLUSIONS: In this proof-of-concept study, wastewater-based epidemiology proved a reliable and sensitive surveillance approach, complementing routine clinical testing for mapping COVID-19 pandemic dynamics and observing newly circulating SARS-CoV-2 variants.
Influenza infections are challenging to monitor at the population level due to a high proportion of mild and asymptomatic cases and confounding of symptoms with other common circulating respiratory diseases, including COVID-19. Alternate methods capable of tracking cases outside of clinical reporting infrastructure could improve monitoring of influenza transmission dynamics. Influenza shedding into wastewater represents a promising source of information where quantification is unbiased by testing or treatment-seeking behaviors. We quantified influenza A and B virus loads from influent at Switzerland's three largest wastewater treatment plants, serving about 12% of the Swiss population. We estimated trends in infection incidence and the effective reproductive number Re in these catchments during a 2021/22 epidemic and compared our estimates to clinical influenza surveillance data. We showed that wastewater-based incidence is better aligned with catchment-level confirmed cases than national ILI, and that only the wastewater data capture a peak in incidence in December 2021. We further estimated Re to have been below 1.05 after introduction of work from home measures in December 2021 and above 0.97 after these measures were relaxed in two out of three catchments based on wastewater data. The third catchment yielded qualitatively the same results, although with wider confidence intervals. The confirmed-case data yielded comparatively less precise estimates that include 1 before and during the period of measures. On the basis of this research we developed an online dashboard for wastewater-based influenza surveillance in Switzerland where we will continue to monitor the onset and dynamics of the 2022/23 flu season.
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