Can Storage Stability and Simulated Gastrointestinal Behavior Change the Cytotoxic Effects of Concentrated Guava Leaves Extract against Human Lung Cancer Cells?

The influence of storage stability and simulated gastrointestinal behavior of different extracts of guava leaves extracts (NC: not concentrated, and C10 and C20: concentrated by nanofiltration) was evaluated based on their total phenolic compound (TPC) contents and antioxidant activity as well as on their cytotoxic effects on A549 and Vero cells. The results showed that C10 and C20 presented high stability for 125 days probably due to their high TPC contents and antioxidant activity. The simulated gastrointestinal behavior modified their TPC contents; however, after all digestion steps, the TPC values were higher than 70%, which means that they were still available to exert their bioactivities. Additionally, the cytotoxic effects of these extracts were evaluated before and after the simulated gastrointestinal behavior or under different storage conditions. C10 presented the best selectivity indices (SI) values (IC50 Vero cells/IC50 A549 cells) at both conditions suggesting that it can be considered a potential extract to be developed as a functional food due to its resistance to the gastrointestinal digestion and storage conditions tested.

An opportunity for acerola pulp (Malpighia emarginata DC) valorization evaluating its performance during the block cryoconcentration by physicochemical, bioactive compounds, HPLC-ESI-MS/MS, and multi-elemental profile analysis.

The present study evaluated the effect of cryoconcentration of pulp blocks of acerola (Malpighia emarginata DC). The study evaluated cryoconcentration in three stages. The cryoconcentrated samples, the ice fractions, and the initial pulp were evaluated for physicochemical composition, bioactive composition, and multielement profile. The cryoconcentrated sample obtained in the third stage of cryoconcentration showed the best results for the concentration factor, process efficiency, total soluble solids content, red color intensity, and increasing of the macro and micronutrients: Cu, Ca, S, Sr, K, Mn, Na, P, Mg, Fe. All stages presented good performance in the total soluble solids content, increase in the titratable acidity of the concentrates, and progressive increase in the intensity of the red color. Generally, higher levels of total phenolic and antioxidant activity were found for the 2nd and 3rd concentrates. The phenolic activity showed an increase of 166.90% in the 3rd stage concentrate compared to fresh pulp, and the antioxidant activity was 112.10% by the ABTS method and 131.60% by the DPPH method, both in the 3rd stage concentrate. The major individual polyphenols were Ferulic acid, Protocatechuic acid, and Taxifolin, with significant increases in the concentration of the compounds in the 2nd and 3rd stage concentrates. In addition, the contents of potentially toxic metals were below detection limits. During the cryoconcentration process, there was a decrease in the values ​​of vitamin C content, moisture content, density, and elements Cu, Sr, and Zn.

Bioactive compounds recovery by freeze concentration process from winemaking by-product.

Grape pomace is the main solid residue of wine industry, containing high amounts of phenolic compounds. Considering its high potential, an extraction procedure was optimized for maximal recovery of anthocyanins from grape pomace (Vitis vinifera L.) using citric acid as a generally recognized as safe (GRAS) acidulant in water. Volume of solvent (3.2-36.8 mL), time (14.4-165.6 min) and pH of solvent (1.12-4.48) were the studied variables. Furthermore, the best condition to obtain extract rich in anthocyanins was submitted to the gravitational block freeze concentration process. The performance of the process was evaluated and cryoconcentrated and ice fractions were analyzed for physicochemical properties, bioactive compounds content, and antioxidant activity. Interaction, linear, and quadratic effects for volume and pH of solvent were significant by analysis of variance (ANOVA). The experimental design allowed the prediction for maximal recovery of anthocyanins (10 mL of solvent at pH 1.8). The bioactive composition of the optimized grape pomace extract was influenced by the cryoconcentration process. After three cycles using gravitational block freeze concentration, the total phenolics and monomeric anthocyanins were approximately 4 and 5 times higher than the initial condition of the extract, respectively. Consequently, an increase in antioxidant activity was observed. The increase in the concentration of bioactive compounds reached a process efficiency of 93% (stage 1) for phenolic compounds and 91% (stage 2) for anthocyanins. Therefore, the final water-based optimized method is safe and has a low cost and the concentrated extract certainly showed higher concentrations of total phenolics and anthocyanins, compared to the initial extract. The proposed clean extraction method and cryoconcentration technique can be considered important strategies for recovering and valuing grape pomace components, improving the approach to the circular economy concept in the wine industry.

A comprehensive approach about comparison between drying technologies and powdered dairy products.

Knowledge of powdered dairy products was addressed, emphasizing powdered milk, cheese, and yogurt. These are very versatile products, and because they are within the field of innovations in developing dairy products, their production and application have been encouraged. This work aimed to reveal the possibilities of obtaining an approach to powder dairy products, the environmental and economic aspects involved, the physical properties analysis, and finally a conclusions including the findings and prospects. When comparing possible processes of powder milk obtaining the preferred comprises the following steps: fat standardization; pasteurization; evaporation using a multi-effect evaporator with the inclusion of one of two options to increase the evaporator's energy efficiency: thermal or mechanical vapor recompression; dehydration (spray drying); and adding an emulsifier (lecithin) or using the fluidized bed dryer or return of fines, resulting in a powder product with instantaneous dissolution. The following sequence is recommended for cheese powder: ingredients addition, melting, dehydration (spray drying), and finalizing with cooling. Yogurt powder has its production process initiated by agitation, ultrafiltration, and spray drying. Concerning the physical properties, they are crucial for managing dairy powder products during the final drying process and for their use as food ingredients.

An approach on detection, quantification, technological properties, and trends market of A2 cow milk.

The genetic variant A2 ß-casein integrates the casein protein group in milk and has been often associated with positive health outcomes. Therefore, this review explores the present understanding of A2 ß-casein, including detection methods and the market trends for dairy from A2 milk. Also, the interaction of A2 ß-casein with αs1-casein and κ-casein genotypes was examined in terms of technological impacts on A2 milk. A limited number of preliminary studies has aimed to investigate the sensorial and technological impacts of ß-casein variants in milk matrices, for instance, in yogurt and other derivatives. Nevertheless, considering studies carried out so far, it is concluded that the manufacture of dairy products from A2 milk is perfectly feasible, as the products presented slight differences when compared to those derived from traditional milk. In one of the works, sensitive drops in rennet coagulation time and curd firmness values were observed in cheese traits. However, it is relevant to point out that variant A of κ-casein plays a negative role in the coagulation features of milk. Therefore, alterations in the pattern of cheese-making properties are not uniquely related to ß-casein variants. Attempts to produce A2 ß-casein in laboratory (non-natural source), through biosynthesis, for example, have not been found so far. This knowledge gap offers a promising area for future studies concerning proteins and bioactive peptide production.

Dairy powder enriched with a soy extract (Glycine max): Physicochemical and polyphenolic characteristics, physical and rehydration properties and multielement composition.

This work presents the enrichment of powdered dairy compounds with soy extract, and the determination of its physical properties, rehydration characteristics, multielement profile, and polyphenolic potential. Five dairy compound formulations were developed, where milk powder was replaced by 10, 20, 30, 40, and 49 % w/w of soy extract.. Multivariate analyzes using combined PCA analyzes were used to group the samples and, thus, reveal the main characteristics associated with their physicochemical properties, bioactive composition, and multi-element profile. The protein content in the samples was not significantly affected by the addition of soy extract. There was a gradual increase in the total fat as the concentration of soy extract increased. Furthermore, with the increase in the percentage of soy extract in the dairy compounds, there was an increase in the levels of total polyphenols, total flavonols, and antioxidant capacity, and of some minerals, such as Fe, Mn, P, Cu, and Mg. The DC49 sample showed the highest values for total polyphenols (178.65 mg of gallic acid (GAE)/100 g) and total flavonols (1.51 mg of catechin/100 g). The addition of soy extract promoted the enrichment of important minerals in the samples, with an increase of up to 55 times in the Fe content and up to 40 times in the Mn content. Physical properties (density and fluidity) and rehydration properties (wetting, dispersibility, and solubility) also were affected as the percentage of soy extract in the samples increased. When there was an addition of up to 20 % soy extract, the samples are still wettable. All dairy compounds showed solubility above 69 %. The use of soy extract in the polyphenolic and mineral enrichment of the dairy compounds is important to add nutritional value to powdered milk, we emphasize that this product has enormous potential to be used in diets that require mineral supplementation.

Block freeze concentration by centrifugation and vacuum increases the content of lactose-free milk macronutrients.

Lactose-free milk is rising in popularity among consumers due to its claim to be a better digestible product compared to regular fluid milk. For that reason, concentrating on this food is a good alternative for increasing its versatility and usability in different dairy industry segments. Block freeze concentration (BFC) is a simple technology used to concentrate liquid foods through ice crystal formation and subsequent removal of water. Thus, this work aimed to test two variants of the BFC technique on lactose-free milk concentration. In the first approach, it was investigated the centrifugation-assisted BFC of skim lactose-free milk by applying a factorial experimental design. Temperature, time, and rotation speed were the factors, and the response variables included the concentrate yield, concentration index, and efficiency of the process. Concentrate yield and concentration index were mainly affected by the centrifugation temperature. On the other hand, individual factors did not have a significant effect on the efficiency, only their interactions. In the case of centrifugation-assisted BFC in a single step, the condition at 40°C, 70 min, and 4500 rpm was considered the best, given the highest values of efficiency and concentrate yield (80.87 and 67.02, respectively), and still an excellent value for concentration index (2.05). Conversely, the condition at 30°C, 45 min, and 3500 rpm was chosen to integrate a freeze concentration process in two stage. Then, the ice obtained from the first cycle was subjected to the vacuum-assisted BFC, which consisted in the second cycle. The concentrate obtained from the vacuum-assisted BFC presented contents of total solids, carbohydrates, and protein 2.95, 3.00, and 2.91 times more than the initial lactose-free milk, respectively. Therefore, we believe that the concentrates obtained can be used for the development of innovative lactose-free dairy products. PRACTICAL APPLICATION: Using concentration processes in the dairy industry can significantly contribute to enhancing the overall efficiency of milk processing since huge quantities of water from milk can be reduced, increasing the total solids content. In turn, dairy products that provide a high amount of solids (especially protein) are gaining in popularity among consumers, with consequent interest from researchers. In addition, milk concentration shows advantages in terms of processing, packaging, transportation, and handling. Since most changes occur in an aqueous environment, the removal of some parts of water results in the preservation of milk. It is noteworthy that dairy industries are concerned principally with food preservation, green technologies, and the production of high-quality products. Thus, concentration processes could favor the development of milk products rich in proteins to meet certain demands on functional and nutritional properties, for example in beverages and formulated food.

Valorization of biowastes from sustainable viticulture with bioactive potential: application in functional yogurt.

Seed and peel flours of organic Bordeaux grapes (Vitis labrusca L.), containing phenolics and antioxidant capacity, influenced both the composition and properties of a yogurt. The total phenolic content (TPC) of the yogurts containing 3% of grape seed flour (GSFY) and 3% of the mixture of flours (MFY, containing 50% of seed and 50% of peel grape flours, w/w) were 18.800 ± 1.060 and 19.509 ± 1.216 mg/g of gallic acid equivalents (GAE), respectively, significantly higher than the content of the control formulation (CY, 3.199 ± 0.326 mg GAE/g). The GSFY, MFY and CY exhibited an antioxidant capacity (mean values), respectively, of 0.6100, 0.7833 and zero µmol TEAC/g by the FRAP method; and 3.6658, 2.9217 and 0.2468 µmol TEAC/g by the ABTS method. The yogurts presented typical coloration of each flour and the texture of the yogurts did not vary significantly compared to the CY. Principal Component Analysis (PCA) results distinguished the yogurts containing the grape flours and the control sample, regarding their composition and properties. The grape bioresidues were valorized by obtaining a functional and clean label yogurt.

Freeze concentration techniques as alternative methods to thermal processing in dairy manufacturing: A review.

Freeze concentration technology is applied to concentrate liquid foods at low temperatures, thus separating pure ice crystals from the final concentrate solution. This method is an interesting alternative to concentrate food with high water levels and significant nutritional value such as dairy products, since several bioactive compounds are reduced when exposed to elevated temperatures. Considered that, this technique may be a great alternative to concentrating and maintaining both nutritional and sensory characteristics of liquid foods. The present review aims to introduce freeze concentration procedures as an eligible choice for conserving dairy products', also addressing its effects on the dairy matrix. PRACTICAL APPLICATION: This study reports the main techniques of freeze concentration applications in dairy products, to be used both on an industrial and laboratory scale, aiming to improve the nutritional quality of the products obtained.

Encapsulated Bifidobacterium BB-12 addition in a concentrated lactose-free yogurt: Its survival during storage and effects on the product's properties.

This work aims to manufacture a new concentrated lactose-free probiotic yogurt. For this purpose, the probiotic Bifidocaterium BB-12 was incorporated in a concentrated lactose-free yogurt, both in its free form and previously encapsulated. Previous cell encapsulation was performed using the spray-drying technique with the following wall materials: lactose-free milk, lactose-free milk and inulin, and lactose-free milk and oligofructose. Thus, three different probiotic powders were obtained and added separately to three fractions of concentrated lactose-free yogurt. The probiotic survival of both powders and yogurts was evaluated during refrigerated storage. Likewise, the viability of starter cultures in yogurt (Lactobacillus bulgaricus and Streptococcus thermophilus) was controlled. In addition, the physicochemical properties of the four yogurts were also measured (color, pH and acidity, and texture properties). All three powders showed good probiotic viability (>8 log CFU g-1) throughout 120 days of storage at 4 °C. In turn, yogurt formulations (with the addition of powders or free bifidobacteria) presented probiotic viability above 7 log CFU g-1 after storage; as well as the starter cultures (>8 log UFC g-1). Yogurt with probiotic powder from lactose-free milk showed a more yellowish color; however, these differences would not be detected by the human eye (ΔE < 3.00). The yogurt with bifidobacteria free cells showed a greater post-acidification process (pH 4.18 to 4.02 and titratable acidity 1.52 to 1.89). It was not observed differences for firmness values of yogurt with free cells addition and yogurt with lactose-free milk and oligofructose powder addition. A slight significant decrease in the cohesiveness was observed in the yogurt elaborated with bifidobacteria free cells. The gumminess showed fluctuating values between all concentrated lactose-free yogurts. At the end of this study, we conclude that these probiotic powders can be incorporated into innovative lactose-free yogurts.

Influence of guabiroba pulp (campomanesia xanthocarpa o. berg) added to fermented milk on probiotic survival under in vitro simulated gastrointestinal conditions.

In fermented milks inoculated with two thermophilic strains (Lactobacillus bulgaricus and Streptococcus thermophilus), guabiroba pulp (Campomanesia xanthocarpa O. Berg) was added in different concentrations: 5% (I5 sample) and 10% (I10 sample), compared to a control sample, with no pulp addition. In these fermented milks, Bifidobacterium BB-12 was added and the samples were submitted to a progressive gastrointestinal simulation in vitro. The cells count was performed, including the survival rates for all the progressive steps of the simulated digestion. Total phenolic content (TPC) and antioxidant activity analysis by FRAP (Ferric Reducing Antioxidant Power) and DPPH (2,2-diphenyl-1-picrylhydrazyl) were performed in all the gastrointestinal steps. Before and during the entire gastrointestinal tract, the Bifidobacterium BB-12 count was 8-9 log CFU g-1, above the recommended for a probiotic product, with a highlight in intestinal colon steps. The I10 sample showed the highest viable cell count, the highest total phenolic content and antioxidant activity throughout the entire gastric steps (p < 0.05). The fermented milk proved to be an effective matrix for the probiotic stability and incorporation of guabiroba components. Bioactive compounds present in the guabiroba pulp may have occasioned a prebiotic and protective effect on Bifidobacterium BB-12 after gastric conditions. The possible bioconversion of these compounds in more active forms can contribute to the absorption in epithelial cells, enhancing fermented milks with guabiroba pulp as important sources of dietary accessible bioactive compounds.

Campomanesia spp. native fruits as potential source of health-promoting compounds

Campomanesia xanthocarpa var. littoralis, Campomanesia xanthocarpa (Berg), and Campomanesia eugenioides are native fruit plants found in Brazil. Due to the scarce number of controlled scientific studies comparing different native Campomanesia species, this study sought to determine their bioactive compounds and antioxidant properties. C. eugenioides proved to be a rich source of total phenolic compounds, also showing the best antioxidant capacity by the ABTS, DPPH and molybdenum reduction power methods. On the other hand, C. xanthocarpa var. littoralis showed the best results for total flavonoids content, and Iron(II) chelation power. The phenolic compounds contents present in C. eugenioides could be responsible for the best antioxidant activity. This study provides key scientific data regarding the use of valuable fruits from different edible Campomanesia species to produce bioactive ingredients, as well as natural preservatives for food products. Thus, our results contribute to the discovery of the potential application of these native Campomanesia Brazilian fruits, as a natural product with functional and antioxidant properties.

Bifidobacterium animalis ssp. lactis BB-12 enumeration by quantitative PCR assay in microcapsules with full-fat goat milk and inulin-type fructans.

The current study was conducted to develop a quantitative polymerase chain reaction (qPCR) assay for Bifidobacterium animalis ssp. lactis BB-12 quantification in microcapsules matrix with full-fat goat milk and inulin-type fructans. DNA was isolated from milk, feed solutions (before spray drying) and microcapsules (after spray drying) using DNAzol. Two primer pairs targeting Bal-23S or Tuf sequences were evaluated by qPCR. The qPCR efficiency was higher (89.5%) using the Tuf primers than Bal-23S primers (84.8%). Tuf primer pair was able to selectively detect B. animalis ssp. lactis BB-12. After, quantification of bifidobacteria in the microcapsules matrix by Tuf qPCR assay was compared to conventional enumeration by plate counting. The analysis of probiotic feed solutions and microcapsules showed higher (P < 0.05) bacterial enumeration determined by Tuf qPCR assay compared to those obtained by plate counting. This qPCR assay was considered a rapid and sensitive alternative for the quantification of B. animalis ssp. lactis BB-12 in probiotic microcapsules compared to plate counting.

Stability of bifidobacteria entrapped in goat's whey freeze concentrate and inulin as wall materials and powder properties.

Goat's whey was submitted to two cycles of block freeze concentration process, resulting in concentrate 1 and concentrate 2. Concentrate 1 was added with 5 g of inulin and both concentrates were inoculated with Bifidobacterium animalis ssp. lactis BB-12, the concentrates were then denoted as feed solutions 1 and 2, respectively. Feed solutions were spray-dried, resulting in powder 1 and 2. The stability of the bifidobacteria entrapped within the powders was evaluated for both spray-dried powders stored at 4 °C and 25 °C for 60 days. The spray-dried powders were also evaluated in relation to their physical and thermal properties. It was noted that Bifidobacteria displayed increased stability at refrigeration temperature. Analysis of physical properties indicated that the addition of inulin resulted in increased water solubility. However, both spray-dried powders displayed less flowability, as well as a yellow-greenish color. By evaluating the spray-dried powders thermal properties, it was possible to confirm that goat whey concentrates behave as excellent wall materials.

Evaluation of the interaction between microencapsulated Bifidobacterium BB-12 added in goat's milk Frozen Yogurt and Escherichia coli in the large intestine.

Goat milk and goat milk and inulin were used as encapsulating agents of Bifidobacterium BB-12 and applied in Frozen Yogurt (GF2 and GF3, respectively) in order to evaluate the antagonistic effect against Escherichia coli. GF1 is a control containing only Escherichia coli. Simulation of gastrointestinal digestion occurred sequentially. Quantification of Bifidobacterium BB-12 was performed using plate counting while E. coli count was compared with quantification by qPCR with viable and nonviable cell differentiation. The Bifidobacterium BB-12 count was <1.0, 9.23 and 9.11 log CFU g-1 for GF1, GF2 and GF3, respectively. In the ascending colon, all samples showed E. coli counts of approximately 5 log CFU g-1 by plate counting and by qPCR. Throughout the transverse (24 h) and descending colon (48 h) samples GF2 and GF3 showed decrease in E. coli number. GF3 showed higher decrease of E. coli in the descending colon because of inulin bifidogenic characteristic. The production of organic acids by bifidobacteria was directly related to the decrease in the E. coli count. In plate counts, E. coli was not detected for the GF3 sample in the descending colon. However, when quantified by qPCR the sample presented amplification that corresponded to 3 log CFU g-1. In this way, it was possible to observe the phenomenon of the viable but not-culturable cells of E. coli. Finally, it is recommended the microcapsule produced with goat milk and the inulin for application in goat milk products, due to the better antagonist effect against E. coli.

Dairy foods and positive impact on the consumer's health.

The objective of the present chapter was to demonstrate the state of the art in the recent advances in nutritional and functional components of dairy products research. In this chapter, the main mechanisms responsible and essential for a better understanding of nutritional and functional values of the components of milk and dairy products are highlighted. It also includes a discussion about the positive impacts of fermented milk, cheese, butter, ice cream, and dairy desserts components on the consumer's health.

Effect of full-fat goat's milk and prebiotics use on Bifidobacterium BB-12 survival and on the physical properties of spray-dried powders under storage conditions.

The effects of full-fat goat's milk and/or inulin and/or oligofructose, as carrier agents, were investigated to improve the survival rates of Bifidobacterium BB-12, and the physical properties of the microcapsules under storage conditions. On the day of their manufacture, the microcapsules were evaluated for morphology, particle size, and distribution of fat and bifidobacteria. The viability of the bifidobacteria, moisture and fat content, water activity, solubility, bulk and tapped density, flowability, cohesiveness and color properties were evaluated for 120 days at 4 °C and 25 °C. The full-fat goat's milk powder with or without inulin as encapsulating agents showed the highest survival rates of Bifidobacterium BB-12 after spray drying and storage. Considering the bifidobacteria survival, both of these spray-dried powders showed the most desirable physical properties, i.e., lowest water activity and solubility, respectively. Both properties are highlighted for better stability of spray-dried powders, with microcapsules, during storage time. These results are credited to full-fat goat's milk (200 g L-1) and the presence of inulin (100 g L-1), besides the fat content showing a high correlation with the solubility values. The lowest volume occupied by the spray-dried powders was noted when oligofructose was used as the carrier agent. The samples that showed the presence of cracks influenced negatively on the bifidobacteria viability. These cracks were responsible by the greater water escape, resulting in powders with more desirable lower water activity. In relation to the color parameters, lower stability was noted when oligofructose was used, while the best stability was also noted for the powders with full-fat goat's milk and/or inulin. During storage time, the best performance was achieved by the microencapsulation process that used only full-fat goat's milk and/or inulin and storage at 4 °C.

Progressive freeze concentration of skimmed milk in an agitated vessel: Effect of the coolant temperature and stirring rate on process performance.

The aim of this study was to investigate the freeze-concentration of skimmed milk by a progressive freeze concentration process. The progressive freeze concentration procedure was performed at three different temperatures (-5, -10, and -15 ℃) and stirring rates (0, 500, and 1000 r/min). The solids concentration was determined and used for calculations of the efficiency of the process, concentrated yield, and experimental results validation. A general linear model was applied to determine the influence of the two factors studied, namely coolant temperature and agitation speed. In all tests, it was possible to concentrated skimmed milk with total solids content higher (P < 0.05) than ultra-high temperature skimmed milk. The highest concentration (P < 0.05) was achieved at low coolant temperature (-15 ℃) and high agitation speed (1000 r/min). The coolant temperature and the stirring rate both had a significant effect (P < 0.05) on the results of efficiency of the process and concentrated yield. Nevertheless, the parameter that showed the most significant effect in our study was the stirring rate. The tests presented a good fit since the root mean square values were below 25%. The freezing point temperatures of the concentrated milk fractions were lower than that of skimmed milk. Finally, the best-operating conditions in our study were achieved using a high coolant temperature (-5 ℃) and high mechanical stirring (1000 r/min), which was also the variable with the lowest (P < 0.05) retention of solids in the ice fraction. In our study, the progressive freeze concentration technique showed promise as an alternative for the dairy industry since it makes the development of new dairy products possible.

Quantification of Lactobacillus paracasei viable cells in probiotic yoghurt by propidium monoazide combined with quantitative PCR.

Propidium monoazide (PMA) coupled with qPCR has been successfully used for specific quantification of viable bacteria cells in diverse matrices food. The present study aimed to develop PMA-qPCR assay for quantification of Lactobacillus paracasei viable cells in probiotic yoghurt. L. paracasei grown in culture medium was submitted to heat treatment at 60°C for different periods of time and probiotic yoghurt containing L. paracasei were prepared and stored at 4°C for 30days. The viable cells were quantified using qPCR and PMA-qPCR assays targeting tuf gene and also by plate counting. Standard curves were prepared and mean efficiency obtained was 94% and 96% (R2>0.98) to L. paracasei in culture medium and probiotic yoghurt stored one day, respectively. The limit of detection (LOD) for both samples was 104 genome copies, corresponding to 32.1pg of DNA. For viable cells quantification, standard curves Cq versus log CFU were plotted using mean CFU by plate counting of L. paracasei grown in culture medium and probiotic yoghurt. Results obtained for L. paracasei heat-treated cells were concordant by PMA-qPCR and plate count, CFU decreased as the heat treatment time increased, while qPCR count remained constant. L. paracasei enumerations obtained by qPCR for probiotic yoghurt stored one day and 30days were higher than enumerations by PMA-qPCR for the same samples. The plate count values were similar to CFU values obtained by PMA-qPCR. These results showed that PMA-qPCR is a powerful approach compared with culture-dependent methods for quantification of L. paracasei viable cells in yoghurt. PMA-qPCR allowed reliable obtained results much faster than plate counting.

Concentration of biologically active compounds extracted from Ilex paraguariensis St. Hil. by nanofiltration.

The aim of this study was to characterise the bioactive compounds in mate (Ilex paraguariensis St. Hil) extract and in concentrated mate extract obtained by nanofiltration (NF). Also, the impact of NF on the antioxidant activity of both mate extracts was evaluated in vitro and using eukaryotic cells of Saccharomyces cerevisiae (yeast assay). The results showed a significant increase in the contents of total phenolics (338%), chlorogenic acid (483%), theobromine (323%), caffeine (251%), chlorophyll (321%), condensed tannins (278%) and saponins (211%) in the concentrated mate extract. The concentrated mate extract showed higher in vitro antioxidant activity than the mate extract. According to the results obtained, it can be stated that the use of nanofiltration membrane is a valid approach for the concentration of biologically active compounds in aqueous extract of mate.