Origin authentication of distillers' dried grains and solubles (DDGS)--application and comparison of different analytical strategies.

In the context of products from certain regions or countries being banned because of an identified or non-identified hazard, proof of geographical origin is essential with regard to feed and food safety issues. Usually, the product labeling of an affected feed lot shows origin, and the paper documentation shows traceability. Incorrect product labeling is common in embargo situations, however, and alternative analytical strategies for controlling feed authenticity are therefore needed. In this study, distillers' dried grains and solubles (DDGS) were chosen as the product on which to base a comparison of analytical strategies aimed at identifying the most appropriate one. Various analytical techniques were investigated for their ability to authenticate DDGS, including spectroscopic and spectrometric techniques combined with multivariate data analysis, as well as proven techniques for authenticating food, such as DNA analysis and stable isotope ratio analysis. An external validation procedure (called the system challenge) was used to analyze sample sets blind and to compare analytical techniques. All the techniques were adapted so as to be applicable to the DDGS matrix. They produced positive results in determining the botanical origin of DDGS (corn vs. wheat), and several of them were able to determine the geographical origin of the DDGS in the sample set. The maintenance and extension of the databanks generated in this study through the analysis of new authentic samples from a single location are essential in order to monitor developments and processing that could affect authentication.

Novel approach for the simultaneous detection of DNA from different fish species based on a nuclear target: quantification potential.

The development of DNA-based methods for the identification and quantification of fish in food and feed samples is frequently focused on a specific fish species and/or on the detection of mitochondrial DNA of fish origin. However, a quantitative method for the most common fish species used by the food and feed industry is needed for official control purposes, and such a method should rely on the use of a single-copy nuclear DNA target owing to its more stable copy number in different tissues. In this article, we report on the development of a real-time PCR method based on the use of a nuclear gene as a target for the simultaneous detection of fish DNA from different species and on the evaluation of its quantification potential. The method was tested in 22 different fish species, including those most commonly used by the food and feed industry, and in negative control samples, which included 15 animal species and nine feed ingredients. The results show that the method reported here complies with the requirements concerning specificity and with the criteria required for real-time PCR methods with high sensitivity.

Novel approach for interlaboratory transfer of real-time PCR methods: detecting bovine meat and bone meal in feed.

The availability of robust methods for the species-specific detection of meat and bone meal (MBM) in compound feedingstuffs is an important prerequisite to enforce current and upcoming European legislation on the use of processed animal proteins in animal nutrition. Among possible methods, those based on DNA turned out to be a reliable tool for this aim, since DNA is a quite thermostable molecule able to resist severe heat treatments applied in the manufacturing of animal meals. The application of such methods by control laboratories implies that the method has been validated including an assessment of its robustness. Successful transferability between laboratories is considered an important robustness criterion of the method. However, corresponding guidelines regarding the design of such a study relevant to this field are missing. Here, we demonstrate the feasibility of an alternative concept that was applied to check for the transferability of a qualitative assay for the detection of banned MBM in feedingstuffs at trace level based on real-time PCR. The concept was based on an experimental nested design applying analysis of variance (ANOVA) that was conducted independently in two laboratories and which allows for establishing major factors influencing the result of analysis. Statistical assessment of the results confirmed the importance of the DNA extraction/purification step utilised, whereas the PCR step turned out to be a minor factor regarding the overall variability of the results. Furthermore, blind samples comprised of compound feed adulterated with MBM at 0.1% and blank compound feed were correctly classified as "positive" or "negative" samples, thus confirming fitness of purpose for the method. This approach can be of interest for other research groups working in the development of real-time PCR methods and in their use by control laboratories.

Chemometric approach for discriminating tobacco trademarks by near infrared spectroscopy.

Cigarettes are consumer products with a broad market-driven orientation. In order to satisfy the different needs of smokers, cigarette trademarks with different aroma, taste and appearance are available on the market. In this study near infrared (NIR) spectroscopy in combination with chemometrics was applied in order to distinguish cigarette trademarks by analysing the tobacco. Calibration models were developed to differentiate three tobacco trademarks and their respective producers. The developed models also allowed the discrimination of the tobacco according to their geographical origin and may serve as a tool for the detection of counterfeit tobacco.

Transferability study of a near-infrared microscopic method for the detection of banned meat and bone meal in feedingstuffs.

Near-infrared microscopy (NIRM) has been proved to be a powerful tool for the detection of banned meat and bone meal (MBM) in feed. The identification of MBM traces and its ability to differentiate animal from vegetable feed ingredients is based on the evaluation of near-infrared spectra obtained from individual particles present in the sample. This evaluation is supported by appropriate decision rules for the absorbances at specific wavelengths. Here we show that the method and the corresponding decision rules can be successfully transferred from the laboratory which constructed the decision rules to two independent laboratories that were not involved in the calibration process of the method. The analytical results from blind feed samples containing MBM (positive samples) and feed samples without MBM (negative samples) revealed a very good agreement between the three laboratories, thus demonstrating the transferability of the method.

Detection of ruminant meat and bone meals in animal feed by real-time polymerase chain reaction: result of an interlaboratory study.

The commercialization of animal feeds infected by prions proved to be the main cause of transmission of bovine spongiform encephalopathy (BSE). Therefore, feed bans were enforced, initially for ruminant feeds, and later for all feeds for farmed animals. The development and validation of analytical methods for the species-specific detection of animal proteins in animal feed has been indicated in the TSE (Transmissible Spongiform Encephalopathies) Roadmap (European Commission. The TSE (Transmissible Spongiform Encephalopathy) roadmap. URL: http://europa.eu.int/comm/food/food/biosafety/bse/roadmap_en.pdf, 2005) as the main condition for lifting the extended feed ban. Methods based on polymerase chain reaction (PCR) seem to be a promising solution for this aim. The main objective of this study was to determine the applicability of four different real-time PCR methods, developed by three National expert laboratories from the European Union (EU), for the detection and identification of cattle or ruminant species in typical compound feeds, fortified with meat and bone meals (MBM) from different animal species at different concentration levels. The MBM samples utilized in this study have been treated using the sterilization condition mandatory within the European Union (steam pressure sterilization at 133 degrees C, 3 bar, and 20 min), which is an additional challenge to the PCR methods evaluated in this study. The results indicate that the three labs applying their PCR methods were able to detect 0.1% of cattle MBM, either alone or in mixtures with different materials such as fishmeal, which demonstrates the improvement made by this technique, especially when compared with results from former interlaboratory studies.

Spectroscopy applied to feed additives of the European Union Reference Laboratory: a valuable tool for traceability.

Feed additives need to be authorised to be placed on the market according to Regulation (EU) No. 1831/2003. Next to laying down the procedural requirements, the regulation creates the European Union Reference Laboratory for Feed Additives (EURL-FA) and requires that applicants send samples to the EURL-FA. Once authorised, the characteristics of the marketed feed additives should correspond to those deposited in the sample bank of the EURL-FA. For this purpose, the submitted samples were subjected to near-infrared (NIR) and Raman spectroscopy for spectral characterisation. These techniques have the valuable potential of characterising the feed additives in a non-destructive manner without any complicated sample preparation. This paper describes the capability of spectroscopy for a rapid characterisation of products to establish whether specific authorisation criteria are met. This study is based on the analysis of feed additive samples from different categories and functional groups, namely products containing (1) selenium, (2) zinc and manganese, (3) vitamins and (4) essential oils such as oregano and thyme oil. The use of chemometrics turned out to be crucial, especially in cases where the differentiation of spectra by visual inspection was very difficult.

Differentiation of coccidiostats-containing feed additives by mid and near infrared microscopy.

Coccidiostats belong to the group of feed additives authorised within the European Union exclusively for specific preparations. These preparations not only contain one or more coccidiostats as active substance(s) but also various ingredients such as the carrier, which are included in the European legislation authorising the product. In order to allow the full traceability of the use of feed additives and to check for compliance with legal provisions, there is a strong need for analytical methods that enable the rapid characterisation of these products. This paper describes the applicability of non-destructive techniques such as mid infrared (MIR) and near infrared (NIR) microscopy supported by multivariate analysis for the characterisation of coccidiostats-containing feed additives. The application of these methods demonstrated that different feed additives could be distinguished from each other even when containing the same active substance. The use of chemometrics turned out to be crucial especially in cases where the differentiation of spectra by visual inspection was very difficult.

Calidad de vida del niño asmático y su cuidador en centros públicos y privadosde Montevideo (2009 a 2010) / Quality of life of asthmatic children and their caregivers in public and private schools Montevideo

Introducción: medir la calidad de vida en asma es importante para su tratamiento ya que refleja el estado funcional del paciente.Objetivo: conocer la calidad de vida de los niños asmáticos y de sus cuidadores en 3 centros de salud y determinar la relación entre la calidad de vida y el control de la enfermedad.Métodos: estudio descriptivo, transversal,multicéntrico con niños asmáticos entre 7 y 15 años,durante el período de marzo 2009 a julio de 2010. Se aplicaron dos cuestionarios de calidad de vida en asma (PAQLQ para el niño y PACQLQ para el cuidador) de Juniper previamente validado en Uruguay. Se consideró alteración de la calidad de vida a un promedio menor de 5 puntos. Se clasificó a los niños en asmáticos controlados y no controlados según GINA. Se analizó la relación entre la calidad de vida y el control de la enfermedad. Se utilizaron promedio, mediana y porcentajes. Para el análisis estadístico se utilizó c2 considerando como significativo un p < 0,05.Resultados: se encuestaron 195 pacientes y cuidadores pero la clasificación según los criterios de Gina se realizó en 174 pacientes. Presentaron afectación de la calidad de vida global el 28,7% de los niños y el 60% de los cuidadores. El 70% se clasificó como asma controlada. La afectación de la calidad de vida se presentó en el 73% de los paciente con asma no controlada (p<0,05).No hubo diferencias en cuanto a la edad (niños versus adolescentes) y sexo . La afectación de la calidad de vida en los tres dominios(limitación de actividades, función emocional y síntomas) se asoció a falta de control del asma en los niños. Los cuidadores presentaron mayor afectación de la calidad de vida global, de la función emocional y limitación de actividades que los niños asmáticos.Conclusiones: en el grupo analizado, la mayoría delos niños no presentaron afectación de la calidad de vida. Existe relación entre el control de la enfermedad y la calidad de vida. La percepción de la enfermedad entre niños...

Nevo displásico / Dysplastic nevus

Os autores fazem revisäo da literatura sobre nevo displásico, focalizando os aspectos clínicos, histopatológicos e dermatoscópicos. Consideram que a associaçäo desses dados é fundamental para o diagnóstico. Pöem em evidência as controvérsias que existem sobre a conceituação de nevo displásico e os percentuais de risco de malignizaçäo

Fatores que influenciam sobrre o prognóstico do melanoma maligno / Factors that influenced the prognosis of malignant melanoma

O Grupo de Melanoma de Clínica Dermatológica da Santa Casa está completando nove anos de atividades. Buscando avaliar seus resultados, analisou os assim chamados fatores de prognóstico do melanoma, a saber: espessura de Breslow, níveis de Clark, estádio, número de linfonodos regionais com metástases, tipo histológico, localizaçäo anatômica, áreas de risco, idade, sexo, ulceraçäo da lesäo inicial, associaçäo com nevo adquirido preexistente realizaçäo de biópsia. Para comparaçäo de ressultados, foi utilizada a casuística publicada pelo Grupo da New York University Medical Center, que conta com 1.130 pacientes. A casuística que é objeto do presente trabalho é pequena, composta por 67 pacientes. Estes, entretanto vêm sendo acompanhados com regularidade, a cada três ou seis meses, ininterruptamente, sempre pelo mesmo o grupo muldisciplinar. Os resultados que temos obtido se aproximam daqueles do grupo do New York University