Sónia Soares

Quantitative detection of poultry meat adulteration with pork by a duplex PCR assay.

A species-specific duplex polymerase chain reaction (PCR) assay was developed for the simultaneous detection of pork and poultry meat species using the mitochondrial cytb and 12S rRNA as target genes for pork and poultry, respectively. By the amplification of binary reference meat mixtures, a linear normalised calibration curve was obtained using the fluorescence intensities of PCR products for pork (149 bp) and poultry (183 bp) species. The proposed method allowed the quantification of pork meat addition to poultry meat in the range of 1-75%, with a sensitivity of 0.1%. The in-house validation using samples with known amounts of pork meat (1.0%, 2.5%, 7.5%, 20.0% and 40%) evidenced a high reproducibility of the methodology (coefficient of variation from 4.1% to 7.6%). The successful application of the duplex PCR was also demonstrated by the high correlation (R(2)=0.99) obtained from regression analysis between the predicted and the actual values of pork meat addition in blind meat mixtures. The suggested methodology presents a low cost, fast, easy and reliable alternative to estimate the level of poultry meat adulteration by the addition of pork meat.

A SYBR Green real-time PCR assay to detect and quantify pork meat in processed poultry meat products

Species identification in meat products has grown in interest in recent years since these foodstuffs are susceptible targets for fraudulent labelling. In this work, a real-time PCR approach based on SYBR Green dye was proposed for the quantitative detection of pork meat in processed meat products. For the development of the method, binary meat mixtures containing known amounts of pork meat in poultry meat were used to obtain a normalised calibration model from 0.1 to 25% with high linear correlation and PCR efficiency. The method revealed high specificity by melting curve analysis, being successfully validated through its application to blind meat mixtures, which confirmed its adequacy for pork meat determination. The fully applicability of the method was further demonstrated in commercial meat products, allowing verification of labelling compliance and identification of meat species in processed foods.

Genotype analysis of Giardia isolated from asymptomatic children in northern Portugal.

ANDRE A. ALMEIDA, MARIA L. DELGADO, SONIA C. SOARES, ANTONIO O. CASTRO, MARIA J. MOREIRA, CARLA M. MENDONCA, NUNO B. CANADA and JOSE M. CORREIA DA COSTA Centro de Imunologia e Biologia Parasitaria—INSA, 4000-509 Porto, Portugal, and Centro de Estudo de Ciencia Animal—ICETA, Universidade do Porto, Porto, Portugal, and Abel Salazar Institute for Biomedical Sciences—ICBAS, Universidade do Porto, Portugal

Quantitative detection of soybean in meat products by a TaqMan real-time PCR assay.

In the present work, we propose a normalised real-time quantitative PCR assay to determine the addition of soybean to meat products. The method proved to be a powerful tool for the quantification of soybean protein (dry basis) in the range of 0.01% to 6%, being successfully in-house validated. Its application was effective in the analysis of several meat products, indicating 2% of non-compliance with the food allergen labelling legislation, and some inconsistencies when comparing the declared with estimated amounts of soybean. This work highlights the importance of efficient tools to assess labelling statements of meat products, avoiding fraudulent practices.

Genetic characterization of Cryptosporidium isolates from humans in northern Portugal.

ANDRE A. ALMEIDA, MARIA L. DELGADO, SONIA C. SOARES, ANTONIO O. CASTRO, MARIA J. MOREIRA, CARLA M. MENDONCA, NUNO B. CANADA, JOSE M. CORREIA DA COSTA and HELENA G. COELHO Centro de Imunologia e Biologia Parasitaria, INSA, Rua de S. Luis n. 16, 4000-509 Porto, Portugal and Centro de Estudo de Ciencia Animal, ICETA, Universidade do Porto, and Abel Salazar Institute for Biomedical Sciences, ICBAS, Universidade do Porto, Portugal, and Department of Infectious Diseases, Hospital of Joaquim Urbano, Rua Câmara Pestana, Porto

Novel diagnostic tools for Asian (Apis cerana) and European (Apis mellifera) honey authentication

Honey can be produced by different species of honeybees, with two being of economic importance due to their use in apiculture, namely Apis mellifera (known as European honeybee) and Apis cerana (known as Asian honeybee). Due to the decline of the wild populations of the Asian honeybee, this honey generally attains much higher market value, being prone to adulteration. This work aims at proposing new tools, based on the use of molecular markers, for the entomological authentication of honey. To this end, new species-specific primers were designed targeting the tRNAleu-cox2 intergenic region and allowing the detection of A. cerana DNA by qualitative polymerase chain reaction (PCR). Additionally, a novel real-time PCR method with high resolution melting analysis was developed to target the 16S rRNA gene of both bee species, allowing their discrimination in different clusters. The proposed methodologies were further applied with success in the authentication of Asian and European honey samples by the identification of honeybee DNA, demonstrating the usefulness of these simple and cost-effective new approaches.

Portuguese Honeys from Different Geographical and Botanical Origins: A 4-Year Stability Study Regarding Quality Parameters and Antioxidant Activity.

Portuguese honeys (n = 15) from different botanical and geographical origins were analysed regarding their quality parameters (diastase activity, hydroxymethylfurfural content, moisture and pH), colour (L*, a*, b*) and antioxidant profile (total phenolics content, total flavonoids content, DPPH• scavenging activity, and ferric reducing power). The samples were analysed fresh and after 4-years of storage (at 25 °C and protected from light). The hydroxymethylfurfural content and diastase activity of the fresh samples were in accordance with the recommended values described in the legislation. In general, the antioxidant activity of the samples correlated more with the bioactive compounds content than with colour. The storage affected differently each individual sample, especially regarding the antioxidant profile. Nevertheless, although in general the lightness of the samples decreased (and the redness increased), after 4 years, 11 samples still presented acceptable diastase activity and hydroxymethylfurfural values.

Liquorice (Glycyrrhiza glabra): A phytochemical and pharmacological review: Liquorice (Glycyrrhiza glabra): A Review

In the last years, consumers are paying much more attention to natural medicines and principles, mainly due to the general sense that natural compounds are safe. On the other hand, there is a growing demand by industry for plants used in traditional medicine that could be incorporated in foods, nutraceuticals, cosmetics, or even pharmaceuticals. Glycyrrhiza glabra Linn. belongs to the Fabaceae family and has been recognized since ancient times for its ethnopharmacological values. This plant contains different phytocompounds, such as glycyrrhizin, 18β‐glycyrrhetinic acid, glabrin A and B, and isoflavones, that have demonstrated various pharmacological activities. Pharmacological experiments have demonstrated that different extracts and pure compounds from this species exhibit a broad range of biological properties, including antibacterial, anti‐inflammatory, antiviral, antioxidant, and antidiabetic activities. A few toxicological studies have reported some concerns. This review addresses all those issues and focuses on the pharmacological activities reported for G. glabra. Therefore, an updated, critical, and extensive overview on the current knowledge of G. glabra composition and biological activities is provided here in order to explore its therapeutic potential and future challenges to be utilized for the formulation of new products that will contribute to human well‐being.

Cryptosporidium spp. and Giardia duodenalis: A picture in Portugal

Cryptosporidium is an entero-pathogen which causes gastrointestinal disturbs. Primarily this organism infects the microvillous border of the intestinal epithelium, and to lesser extent extra intestinal epithelia, causing acute gastrointestinal disturbs (Fayer, 2004). The duration of infection and the ultimate outcome of intestinal cryptosporidiosis greatly depend on the immune status of the patient. In fact, immunologically healthy patients usually recover spontaneously in a week. The clinical signs can range from asymptomatic to acute, severe and persistent diarrhea and their potential for Cryptosporidium transmission can persist for weeks after symptoms cease (Deng et al., 2004; Fayer, 2004; Hunter and Thompson, 2005).