Determination of the More Adequate Modified Agglutination Test Cut‐off for Serodiagnosis of Toxoplasma gondii Infection in Sheep

Toxoplasmosis is one of the most common food borne disease world‐wide. Among food animals, sheep seems to having higher prevalence of Toxoplasma gondii infection. However, there is no consensus about the best cut‐off for serodiagnosis in sheep. To estimate the more adequate cut‐off value of Modified Agglutination Test (MAT) for serodiagnosis in sheep, a commercial ELISA kit was used as a golden standard. Evaluation of the optimal sensitivity and specificity was calculated using Youden’s J‐statistics. Values obtained were used to estimate the prevalence of sheep toxoplasmosis. One thousand four hundred and sixty seven blood samples were collected randomly from 160 farms from northern Portugal, representing approximately 10% of the ovine population from the region. All sera were tested for anti‐T. gondii antibodies using the MAT. One hundred nine sheep (7.4%) presented a MAT titer ≥1 : 80; 45 (3.0%) a MAT titer of 1 : 40; 97 (6.6%) a MAT titer of 1 : 20 and 1216 (83.0%) a MAT titer ≤1 : 20. The best Youden’s J‐statistic was obtained at 1 : 20 titer (0.752), with 86.15% of sensitivity and 89.09% of specificity with negative and positive predictive values of 90.32% and 84.48% respectively, suggesting that the 1 : 20 was the most appropriate cut‐off for serodiagnosis of toxoplasmosis in sheep. Assuming this cut‐off, the prevalence of toxoplasmosis in the studied population was 17.1% and 92 (57.5%) of the 160 studied flocks having one or more positive sheep. Those results indicate that toxoplasmosis in Portugal should be considered in the differential diagnosis of abortions in sheep and neurological signs in lambs. Furthermore, while Portugal produces ovine meat for internal consumption and for exportation, isolation of T. gondii from ovine meat and further characterization of the isolates will be needed to understand the risk that ovine toxoplasmosis may represent for human health.     

Response surface optimization of antioxidants extraction from chestnut (Castanea sativa) bur

The chestnut bur, a forest waste product from chestnut processing in the food industry, was studied as a potential source of natural antioxidants. Extractions were performed using aqueous solutions of methanol or ethanol. Experiments were planned according to an incomplete 3³ factorial design to study the influence of temperature (25-75 °C), time (30-120 min) and solvent concentration (50-90%) on extraction yield and on extract properties: total phenols content, antioxidant activity (using the FRAP, DPPH and ABTS methods) and average molecular weights. All dependent variables were influenced by temperature and solvent concentration whereas the influence of time was almost negligible. Using the response surface methodology the optimal extraction conditions were selected: the highest temperature assayed (75 °C), the lowest solvent concentration (50%) and an extraction time of 75 min for the methanolic extractions and of 30 min for the ethanolic ones. Under those conditions the values predicted for extraction yield and total phenols content were 18.95% and 36.32 g GAE/100 g extract for the methanolic extract and 17.95% and 26.11 g GAE/100 g extract for the ethanolic ones. Methanolic extracts showed superior total phenols content and antioxidant properties and slightly higher extraction yields than ethanolic extracts; however, ethanol is recommended for food applications due to its GRAS (Generally Recognized as Safe) qualification. Gallic acid esters of glucose, ellagic acid and small proportions of quercetin-3-β-d-glucoside, phenolic compounds with demonstrated antioxidant properties, were identified in chestnut bur extracts by RP-HPLC-ESI-TOF.