In vitro prebiotic activity of rhLf and galactooligosaccharideson infant intestinal microbiota / Actividad prebiótica in vitro de lactoferrina y galactooligosácridossobre la microbiota intestinal infantil

Objective: human lactoferrin (Lf) and human milk oligosaccharides possess a wide range of functions. So, the present study focusses on the role of Lf and/or galactooligosaccharides (GOS) in the modulation of gut microbiota composition.Methods: recombinant human lactoferrin (rhLf) was added to the first infant formula (0.10, 0.15, 0.20 %) alone or in combination with GOS (1 %) in vessels of a small-scale batch culture fermentation model. Short-chain fatty acids (SCFAs), microbial population groups, and pH were monitored through fermentation for 24 hours. Results: insignificant changes were observed in pH values and acetic acid accumulated during fermentation. Propionic acid content has been insignificantly increased while butyric acid has been insignificantly decreased. Moreover, increments in all bacterial groups except for Bacteroides were observed through the fermentation process. Lactobacillus and Bifidobacterium showed an increase in relation to initial time over thefermentation process, demonstrating the prebiotic effect of lactoferrin and GOS. After 24 hours of fermentation, all tested ingredients showed significant similarities in Enterococcus for controls except for 0.20 % rhLf + 1 % GOS, which provoked a diminution of Enterococci growth. Conclusion: despite the importance of the batch culture fermentation technique in uncovering the prebiotic activity of food ingredients, it is not useful for detecting the prebiotic nature of Lf due to its nature as a protein. Thus, Lf maybe shows its prebiotic activity on the gut microbiota through other mechanisms.(AU)

Objetivo: la lactoferrina humana (Lf) y los oligosacáridos de leche materna presentan un amplio rango de funciones. El presente estudio se centra en el papel de la Lf y/o galactooligosácridos (GOS) en la modulación de la composición de la microbiota intestinal. Métodos: se añadió lactoferrina humana recombinante (rhLf) a fórmula infantil (0,10, 0,15, 0,20 %), sola o en combinación con GOS (1 %) en botes de fermentación colónica. A lo largo de 24 horas de fermentación, se monitorizaron ácidos grasos de cadena corta, grupos de poblaciones microbianas y pH. Resultados: se observaron pequeños cambios en valores de pH y cantidad de ácido acético durante la fermentación. El contenido de ácido propiónico aumentó ligeramente, mientras que el butírico sufrió un ligero descenso. Todos los grupos bacterianos estudiados incrementaron, excepto los Bacteroides, durante la fermentación. Lactobacillus and Bifidobacterium mostraron un incremento respecto al valor inicial, demostrando el efecto prebiótico de la lactoferrina y los GOS. A las 24 horas de fermentación, todos los ingredientes estudiados mostraron similitud al control en cuanto a Enterococcus, excepto para 0,20 % rhLf + 1 % GOS, donde disminuyó el crecimiento de los enterococos. Conclusión: a pesar de la importancia de los estudios de fermentación in vitro para descubrir potenciales ingredientes prebióticos, no fue útil en el caso de lactoferrina debido a su naturaleza proteica. Por tanto, la lactoferrina podría mostrar su actividad prebiótica en la microbiota intestinal a través de otros mecanismos.(AU)

[Anti-inflammatory effect of milk whey from different species after in vitro digestion]. / Efecto antiinflamatorio del suero de leche de diferentes especies tras una digestión in vitro.

Introduction: Introduction: there is a close relationship between obesity, gut health and immune system. A low-grade of inflammation, which could precede obesity, may have implications for the development of metabolic syndrome and insulin resistance. Objective: analyzing the anti-inflammatory capacity of several types of whey (cow, sheep, goat and a mixture of them). Methods: an in vitro model of intestinal inflammation employing a cell co-culture (Caco-2 and RAW 264.7) was performed after an in vitro digestion and fermentation (simulating mouth-to-colon conditions). Inflammatory markers such as IL-8 and TNF-α, as well as the transepithelial electrical resistance (TEER) of Caco-2 monolayer, were determined. Results: digested and fermented whey had a protective effect on cell permeability, being lower in the case of fermented goat whey and mixture. The anti-inflammatory activity of whey was greater the more digestion progressed. Fermented whey showed the greatest anti-inflammatory effect, inhibiting IL-8 and TNF-α secretion, probably due to its composition (protein degradation products such as peptides and amino acids, and SCFA). However, fermented goat whey did not show this degree of inhibition, perhaps due to its low SCFA concentration. Conclusion: milk whey, especially after being fermented in the colon, can be useful nutritional strategy to preserve the intestinal barrier and mitigate the low-grade of inflammation that characterizes metabolic disorders and obesity.

Introducción: Introducción: existe una estrecha relación entre obesidad, salud intestinal y sistema inmune. Un bajo grado de inflamación, que precedería a la obesidad, puede tener implicaciones en el desarrollo de síndrome metabólico y resistencia a la insulina. Objetivo: analizar el poder antiinflamatorio de varios tipos de lactosuero (vaca, oveja, cabra y mezcla de los anteriores). Metodología: se utilizó un modelo in vitro de inflamación intestinal, empleando un cocultivo celular (Caco-2 y RAW 264.7). Para ello, se realizó una digestión y fermentación in vitro (simulando las condiciones de boca a colon). Se estudiaron IL-8 y TNF-α como marcadores inflamatorios y la resistencia eléctrica transepitelial celular (RETE) de la monocapa celular Caco-2. Resultados: el suero digerido y fermentado tuvo un efecto protector sobre la permeabilidad celular que fue menor en el caso de lactosuero fermentado de cabra y mezcla. La actividad antiinflamatoria del suero fue mayor cuanto más progresaba la digestión. El lactosuero fermentado mostró el mayor efecto antiinflamatorio, inhibiendo la secreción de IL-8 y TNF-α, probablemente debido a su composición (productos de degradación proteica como péptidos y aminoácidos, y ácidos grasos de cadena corta [AGCC]). Sin embargo, el suero fermentado de cabra no mostró ese grado de inhibición, quizás debido a su baja concentración en AGCC. Conclusión: el lactosuero, sobre todo tras ser fermentado en colon, puede ser una estrategia nutricional útil para preservar la barrera intestinal y mitigar el bajo grado de inflamación que caracteriza a desordenes metabólicos y a la obesidad.

In vitro prebiotic activity of rhLf and galactooligosaccharides on infant intestinal microbiota.

Introduction: Objective: human lactoferrin (Lf) and human milk oligosaccharides possess a wide range of functions. So, the present study focusses on the role of Lf and/or galactooligosaccharides (GOS) in the modulation of gut microbiota composition. Methods: recombinant human lactoferrin (rhLf) was added to the first infant formula (0.10, 0.15, 0.20 %) alone or in combination with GOS (1 %) in vessels of a small-scale batch culture fermentation model. Short-chain fatty acids (SCFAs), microbial population groups, and pH were monitored through fermentation for 24 hours. Results: insignificant changes were observed in pH values and acetic acid accumulated during fermentation. Propionic acid content has been insignificantly increased while butyric acid has been insignificantly decreased. Moreover, increments in all bacterial groups except for Bacteroides were observed through the fermentation process. Lactobacillus and Bifidobacterium showed an increase in relation to initial time over the fermentation process, demonstrating the prebiotic effect of lactoferrin and GOS. After 24 hours of fermentation, all tested ingredients showed significant similarities in Enterococcus for controls except for 0.20 % rhLf + 1 % GOS, which provoked a diminution of Enterococci growth. Conclusion: despite the importance of the batch culture fermentation technique in uncovering the prebiotic activity of food ingredients, it is not useful for detecting the prebiotic nature of Lf due to its nature as a protein. Thus, Lf maybe shows its prebiotic activity on the gut microbiota through other mechanisms.

Introducción: Objetivo: la lactoferrina humana (Lf) y los oligosacáridos de leche materna presentan un amplio rango de funciones. El presente estudio se centra en el papel de la Lf y/o galactooligosácridos (GOS) en la modulación de la composición de la microbiota intestinal. Métodos: se añadió lactoferrina humana recombinante (rhLf) a fórmula infantil (0,10, 0,15, 0,20 %), sola o en combinación con GOS (1 %) en botes de fermentación colónica. A lo largo de 24 horas de fermentación, se monitorizaron ácidos grasos de cadena corta, grupos de poblaciones microbianas y pH. Resultados: se observaron pequeños cambios en valores de pH y cantidad de ácido acético durante la fermentación. El contenido de ácido propiónico aumentó ligeramente, mientras que el butírico sufrió un ligero descenso. Todos los grupos bacterianos estudiados incrementaron, excepto los Bacteroides, durante la fermentación. Lactobacillus and Bifidobacterium mostraron un incremento respecto al valor inicial, demostrando el efecto prebiótico de la lactoferrina y los GOS. A las 24 horas de fermentación, todos los ingredientes estudiados mostraron similitud al control en cuanto a Enterococcus, excepto para 0,20 % rhLf + 1 % GOS, donde disminuyó el crecimiento de los enterococos. Conclusión: a pesar de la importancia de los estudios de fermentación in vitro para descubrir potenciales ingredientes prebióticos, no fue útil en el caso de lactoferrina debido a su naturaleza proteica. Por tanto, la lactoferrina podría mostrar su actividad prebiótica en la microbiota intestinal a través de otros mecanismos.

In vitro digestion effect on CCK and GLP-1 release and antioxidant capacity of some plant-based milk substitutes.

Recently, plant-based milk substitutes, as an emerging industry, are receiving more attention. Despite that, these dairy alternatives have not been adequately studied for their functional properties. Thus, the current research was devoted to study the satiety potential through in vitro secretion of cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1), and the antioxidant capacity of these dairy alternatives after in vitro digestion. The enteroendocrine cell line, STC-1, was used to measure satiety hormones release (CCK and GLP-1) by enzyme-linked immunoassay (ELISA). Also, total phenolic and flavonoid contents and antioxidant capacity (using oxygen radical absorbance capacity [ORAC], ferric reducing antioxidant power [FRAP], and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid [ABTS] assays) were measured before and after in vitro digestion. The results demonstrated that CCK secretion was significantly (p < 0.05) higher for cow's milk (350.64 pg ml-1 ) as compared to plant-based milk substitutes. Among the plant-based milk substitutes, tiger nut milk showed the highest CCK stimulant (228.96 pg ml-1 ), followed by hazelnut milk (220.04 pg ml-1 ). Concerning GLP-1 release, the data exhibited that spelt milk was the food with the highest induction of GLP-1 hormone secretion, followed by cow's milk (910.17 and 876.59 pg ml-1 , respectively), but without any significant differences between them. total phenolic content (TPC) values strongly increased after in vitro digestion, cow's milk and soymilk being the samples with the highest TPC values after in vitro digestion (165.76 and 153.71 mg GAE/100 ml, respectively). In line with TPC values, soymilk had the highest ORAC, ABTS, and FRAP values after in vitro digestion (25.41, 8.17, and 2.51 µmol TE/ml, respectively). Thus, these dairy alternatives could be an adequate substitute for cow's milk, according to its satiety and antioxidant capacity. PRACTICAL APPLICATION: The first study deals with the efficacy of plant-based milk substitutes as satiating foods. Spelt milk was the highest stimulant for GLP-1 secretion. Tiger nut milk showed the highest CCK stimulant, followed by hazelnut milk. TPC and total antioxidant capacity increased after digestion.

Platos olvidados o de consumo ocasional en la gastronomía española: Murcia / Forgotten dishes or occasional consumption in Spanish gastronomy: Murcia

La diversidad geográfica murciana se traduce en una enorme variedad de recetas integradas por productos autóctonos, entre los que se puede encontrar los pescados y mariscos del Mar Menor y del Mediterráneo, las frutas y verduras de la huerta, las carnes y embutidos tradicionales y los excelentes vinos que ofrece la tierra. El paso de distintas civilizaciones por estas tierras ha dejado una influencia gastronómica perdurable, aún presente en las cocinas. Así, podemos encontrar claras semejanzas entre platos típicos actuales y comidas del antiguo mundo greco-romano. También la cultura árabe legó una tremenda herencia de sabores y aromas al recetario murciano. El tomate y el pimiento llegaron tras el descubrimiento de las Américas, del nuevo continente, y se han convertido en elementos emblemáticos de la cocina murciana. (AU)

Murcia’s geographical diversity translates into anenormous variety of recipes made up of native products, among which you can find fish and shellfish from the Mar Menor and the Mediterranean, fruit and vegetables from the garden, traditional meats and sausages and excellent wines. The passage of different civilizations through these lands left a long-lasting gastronomic hallmark. Thus, we can find clear similarities between current typical dishes and foods from the ancient Greco-Romanworld. The Arab culture also bequeathed a tremendous heritage of flavors and aromas to the Murcian recipe repertoire. Tomatoes and peppers arrived after the discovery ofthe Americas and became emblematic elements of Murcian cuisine. (AU)

Valorization of Citrus Co-Products: Recovery of Bioactive Compounds and Application in Meat and Meat Products.

Citrus fruits (orange, lemon, mandarin, and grapefruit) are one of the most extensively cultivated crops. Actually, fresh consumption far exceeds the demand and, subsequently, a great volume of the production is destined for the citrus-processing industries, which produce a huge quantity of co-products. These co-products, without proper treatment and disposal, might cause severe environmental problems. The co-products obtained from the citrus industry may be considered a very important source of high-added-value bioactive compounds that could be used in the pharmaceutical, cosmetic, and dietetic industries, and mainly in the food industry. Due to consumer demands, the food industry is exploring a new and economical source of bioactive compounds to develop novel foods with healthy properties. Thus, the aim of this review is to describe the possible benefits of citrus co-products as a source of bioactive compounds and their applications in the development of healthier meat and meat products.

Evaluación cuantitativa y cualitativa del desayuno en estudiantes universitarios y su relación con parámetros antropométricos / Quantitative and qualitative evaluation of breakfast in university students and its relationship with anthropometric parameters

Fundamentos: La etapa universitaria conforma un periodo de cambio en el entorno que repercute en los estilos de vida saludables de los estudiantes. El objetivo es Analizar el hábito del desayuno y determinar su relación con parámetros antropométricos y el riesgo de enfermedad cardiovascular. Métodos: Estudio transversal con 123 estudiantes de la Universidad de Murcia analizando la ingesta calórica, variedad de alimentos y datos antropométricos (peso, talla, pliegues cutáneos y circunferencias de cintura (PCi) y cadera).Resultados: Los valores de índice de masa corporal (IMC),PCi, índice cintura-cadera (ICC) e índice cintura-talla (ICT)fueron mayores en hombres. El porcentaje de grasa corporal fue mayor en las mujeres. El 50% de los hombres presentó IMC > 25, y un 27,3% superó el valor del ICT. El 50% de los hombres y el 39,4% de las mujeres consumieron un único grupo de alimentos. La energía cubierta fue del 9,62% para hombres y 12,59% para mujeres. Sólo el 21,8% de las mujeres alcanzó el 20% de la energía diaria en el desayuno, en los varones ninguno (p < 0,05).Conclusiones: El desayuno es una ingesta que los estudiantes universitarios realizan de forma inadecuada, tanto en cantidad como en variedad. (AU)

Background: The university stage implies a change in theenvironment that affects the life style, such as the omissionof breakfast. The aim is to analyze the breakfast habit anddetermine its relationship with anthropometric parametersand the risk of cardiovascular disease.Methods: The study was carried out with 123 students ofthe University of Murcia. The energy intake, variety of foodand and anthropometric parameters (weight, height, skinfolds and waist circumferences (WC) and hip) was analyzed.Results: The values of body mass index (BMI), WC, waisthip index (WHI) and waist-height index (WHtI) were higherin men. The percentage of body fat was higher in women.50% of men presented BMI > 25, and 27.3% exceeded thevalue of WHtI, associated with an increased risk ofcardiovascular disease. 50% of men and 39.4% of womenconsumed a single group of foods, mainly dairy and lessfruit. The energy covered was 9.62% for men and 12.59%for women. None of the men reached 20% of the dailyenergy at breakfast while 21.8% of the women did.Conclusions: Breakfast is a meal that university studentsmake inadequately, both in quantity and variety of foods. (AU)

Effect of Gazpacho, Hummus and Ajoblanco on Satiety and Appetite in Adult Humans: A Randomised Crossover Study.

Nowadays, overweight and obesity has reached an epidemic level around the world. With the aim to tackle them, an interesting strategy is the study of food and ingredients with satiety properties. In addition to reducing food and/or calorie intake, this type of foods must be included as part of a healthy diet. With regard to this, it is well known that the Mediterranean Diet (MD) is a feeding pattern that helps us to maintain good health, providing an adequate intake of micronutrients and active compounds. With this background, the main aim of this research was to identify MD foods with a high satiating potential capacity. For this purpose, three typical foods of the Mediterranean region, mainly based on vegetables, were selected: hummus, ajoblanco and gazpacho. As a control, white bread was used. Twenty-four human healthy volunteers consumed a standard breakfast followed by the different typical Mediterranean foods, and then the subjective sensation of hunger and satiety for each food was assessed by visual analogue scales (VAS) during 3 h. Subsequently, volunteers had ad libitum access to a standard meal. The results indicate that gazpacho showed the highest satiating scores, despite the fact that it was not the food that provided the highest protein or fibre amount. More studies of this type are needed to determine the proportion and/or combination of ingredients from these classical Mediterranean recipes that could enhance human satiety.

Occurrence of Clostridioides (Clostridium) difficile in Poultry Giblets at Slaughter and in Retail Pork and Poultry Meat in Southeastern Spain.

ABSTRACT: Raw meat and meat products contaminated with Clostridioides difficile could be a vehicle for spreading community-associated C. difficile infection. This study was conducted to determine the occurrence of C. difficile in pork and poultry meat samples (n = 325) from retail establishments and in edible giblet samples (n = 36) from a poultry processing plant in Murcia (southeastern Spain). C. difficile was isolated after selective enrichment from 2% (6 of 361) of the samples, all of which were from the poultry processing plant. These isolates were recovered from 17% (6 of 36) of the edible chicken giblets, i.e., 28% (5 of 18) of the gizzard samples and 6% (1 of 18) of the liver samples. All six C. difficile isolates were negative for toxin A and B genes by PCR assay. These findings indicate that C. difficile can survive in the gastric acid of the chicken gizzard and could be transmitted to other meat products. However, the very low prevalence of C. difficile in the tested samples indicates that retail meat may not be an important source for transmission of C. difficile to humans.

Anti-Inflammatory and Antioxidant Effects of Regular Consumption of Cooked Ham Enriched with Dietary Phenolics in Diet-Induced Obese Mice.

Oxidative damage and chronic inflammation have been proven as one of the major factors associated with obesity, which increases the incidence of non-communicable chronic diseases. In this sense, the development of new functional products aiming at the palliation of oxidative stress and inflammatory disruption can be a determining factor for public health as seen in previous researches. In this study, a blend of potentially bioavailable dietary phenolics was added to low sodium and low-fat cooked ham. A diet-induced obesity model in C57/BL6J mice has been used for testing the effectiveness of the phenolic blend and the new functionalized product, which bioavailability was tested by UPLC-ESI-QTOF-MS. After obesity induction, different oxidative and inflammatory biomarkers were evaluated. Results in the murine induced obesity model, demonstrate a robust statistically significant improvement in key parameters related with obesity risk in the groups feed with a phenolic-enriched diets (P) + high-fat diet (HFD) and phenolic enriched cooked ham (PECH) + HFD. In both groups there was an improvement in body composition parameters, inflammatory biomarkers and antioxidant enzymes levels. Specifically in the group feed with the phenolic enriched cooked ham (PECH + HFD) there was an improvement of total fat volume (23.08% reduction), spleen index (22.04% of reduction), plasmatic MCP-1 (18% reduction), IL-6 (38.94% reduction), IL-10 (13.28% reduction), TNF-α (21.32% reduction), gut IL-1ß (10.86% reduction), gut IL-6 (13.63% reduction) and GPx (60.15% increase) and catalase (91.37% increase) enzymes. Thus, the functionalized ham could be considered an appropriate dietary polyphenol source, which might improve the oxidative and inflammatory status and could finally result in the potential decrease of the risk of certain non-communicable chronic diseases.

Screening ultrafiltration membranes to separate lactose and protein from sheep whey: application of simplified model.

Several studies demonstrated that protein from whey milk could be a new strategy to reduce energy intake and increase satiety. Sheep whey has high protein content, but it is also rich in lactose. The aim of this study was to screening different ultrafiltration membranes to separate protein and lactose from sheep whey in one step. Protein was recovered in the concentrate feed, and lactose passed through three membranes and was recovered in the permeate feed. Membranes with different chemical composition and molecular weight cut-offs were assayed, and the influence of operating pressure and lactose concentration feed in the permeate flux and lactose rejection coefficients were studied. Lactose separation was not affected by pressure in GR60PP or GR90PP, and 85% and 80%, respectively of the lactose was separated into permeate feed. When the feed concentration increased, lactose separation remained stable in all three membranes, being GR60PP the most efficient, as 90% of the disaccharides were separated. In all cases 100% of the protein was recovered. Finally, the Spiegler-Kedem-Katchalsky model perfectly fitted the results obtained about lactose rejection coefficients.

In Vitro Modulation of Gut Microbiota and Metabolism by Cooked Cowpea and Black Bean.

Legumes are a rich source of a wide range of compounds that may represent an important tool to overcome gut dysbiosis. In this work, the prebiotic potential of two cooked legumes (cowpea and black bean) was investigated in comparison with potato:beef mixture, as substrates in batch faecal culture fermentation. Prior to the fermentation, all the samples were in vitro digested, passing through three phases, namely mouth, gastric and small intestine simulation, and then in vitro fermented for 6, 24 and 48 h. The shift of pH, production of gas and short-chain fatty acids (SCFAs) and changes in gut microbiota were evaluated along the fermentation time. The pH decreased significantly over time in all media with fermentable sources when compared with the negative control. Gas production was higher in the media containing fermentable source than in the negative control and decreased with fermentation time. The concentration of SCFAs increased over time and it was significantly higher for both legumes than in inulin (positive control) and potato:beef meal. Acetate was the major SCFAs produced during fermentation, particularly in media containing legumes. Both legumes presented a strong prebiotic effect on gut microbiota, showing a significant increase in Bifidobacterium and Lactobacillus. These results suggest that consumption of cooked cowpea and black bean, used alone or as an ingredient of novel functional foods, may contribute to improving intestinal health and therefore human health promotion.

Nondigestible Carbohydrates Affect Metabolic Health and Gut Microbiota in Overweight Adults after Weight Loss.

BACKGROUND: The composition of diets consumed following weight loss (WL) can have a significant impact on satiety and metabolic health. OBJECTIVE: This study was designed to test the effects of including a nondigestible carbohydrate to achieve weight maintenance (WM) following a period of WL. METHODS: Nineteen volunteers [11 females and 8 males, aged 20-62 y; BMI (kg/m2): 27-42] consumed a 3-d maintenance diet (15%:30%:55%), followed by a 21-d WL diet (WL; 30%:30%:40%), followed by 2 randomized 10-d WM diets (20%:30%:50% of energy from protein:fat:carbohydrate) containing either resistant starch type 3 (RS-WM; 22 or 26 g/d for females and males, respectively) or no RS (C-WM) in a within-subject crossover design without washout periods. The primary outcome, WM after WL, was analyzed by body weight. Secondary outcomes of fecal microbiota composition and microbial metabolite concentrations and gut hormones were analyzed in fecal samples and blood plasma, respectively. All outcomes were assessed at the end of each dietary period. RESULTS: Body weight was similar after the RS-WM and C-WM diets (90.7 and 90.8 kg, respectively), with no difference in subjectively rated appetite. During the WL diet period plasma ghrelin increased by 36% (P < 0.001), glucose-dependent insulinotropic polypeptide (GIP) decreased by 33% (P < 0.001), and insulin decreased by 46% (P < 0.001), but no significant differences were observed during the RS-WM and C-WM diet periods. Fasting blood glucose was lower after the RS-WM diet (5.59 ± 0.31 mmol/L) than after the C-WM diet [5.75 ± 0.49 mmol/L; P = 0.015; standard error of the difference between the means (SED): 0.09]. Dietary treatments influenced the fecal microbiota composition (R2 = 0.054, P = 0.031) but not diversity. CONCLUSIONS: The metabolic benefits, for overweight adults, from WL were maintained through a subsequent WM diet with higher total carbohydrate intake. Inclusion of resistant starch in the WM diet altered gut microbiota composition positively and resulted in lower fasting glucose compared with the control, with no apparent change in appetite. This trial was registered at clinicaltrials.gov as NCT01724411.

Presence of toxigenic Clostridioides (Clostridium) difficile in edible bivalve mollusks in Spain.

Clostridioides difficile reservoirs other than humans are becoming increasingly recognized, and the occurrence of the pathogen in shellfish raises concern because spores can survive cooking temperature and edible bivalve mollusks are often consumed raw or poorly cooked. This study was conducted to determine the occurrence of pathogenic C. difficile in retail bivalve mollusks. The microbiological quality of samples was also checked through the isolation of Salmonella spp. and Escherichia coli. We analyzed 129 mollusk samples from different fishmongers and grocery stores in Murcia. C. difficile was isolated from 8.53% (11/129) of the mollusks investigated. Four C. difficile isolates harbored genes for the production of toxin A and B. Salmonella spp. were not isolated from any sample and E. coli was isolated from 1.55% (2/129) of the samples, in both cases in accordance with the current legal requirements for consumption. Our findings indicate that the intake of raw or poorly cooked contaminated bivalve mollusks could be a potential source of C. difficile, leading to a risk for human health.

Effect of Specimen Type and Processing on the Detection of Clostridioides [Clostridium] difficile in Piglet Fecal Samples.

Subclinical Clostridioides difficile colonization in piglets could be a potential source of this bacterium for community-acquired C. difficile infection. The purposes of this study were to assess the effect of specimen type and processing on C. difficile isolation, culture, and detection by polymerase chain reaction (PCR), and to determine the occurrence of C. difficile in piglets of different ages. We compared different culture procedures-direct plating, ethanol shock, and an enrichment step-to isolate C. difficile from swine feces and rectal swabs. DNA was isolated directly from feces, processed feces, and bacterial isolates to detect the triose phosphate isomerase (tpi) gene and identify the toxins A and B genes. The results show that ethanol shock increased the C. difficile isolation from feces, while it decreased it for rectal swabs, in comparison with direct plating. The use of the enrichment broth gave the highest C. difficile recovery from both types of specimen. Our findings show low sensitivity for tpi gene detection after the DNA extraction directly from feces and an increase in PCR-positive samples when feces were processed before the DNA extraction. The overall prevalence of C. difficile was 16.9% (22/130), of which 100% were found to be toxigenic as assessed by the enrichment culture of fecal samples. The rate of isolation of positive samples decreased with the animal age, regardless of the presence or absence of diarrhea. Our results demonstrate the persistent reservoir of toxigenic C. difficile in fecal samples of piglets and support the impact of specimen processing on its isolation.

In vitro effectiveness of recombinant human lactoferrin and its hydrolysate in alleviating LPS-induced inflammatory response.

This study aimed to evaluate the potential anti-inflammatory role of the most produced form of lactoferrin expressed in various expression systems (Fe-saturated recombinant human Lf, rhLf) and its hydrolysate in concentrations resembles that found in mature human milk. Co-culture model consisted of CaCo-2 and RAW 246.7 cell lines was used to evaluate the potential anti-inflammatory activity of rhLf and its hydrolysate. During this experiment, CaCo-2 monolayer permeability and integrity was assayed through the measurement of transepithelial electrical resistance (TEER values). Also, the production of reactive oxygen species (ROS), nitric oxide (NO) and different cytokines (IL-8, IL-1ß, IL-6, IL-10, IL-12p70, and TNF-α) were measured. The treatment with rhLf and its hydrolysate protected the monolayer integrity against LPS effect and reduced IL-8 and ROS production. This effect was dependent on the dose and 2mgmL-1 of rhLf hydrolysate was more effective. The addition of rhLf and its hydrolysate to infant formula is a prominent step towards improving both infant formula functionality and newborn health. Thus, these functional ingredients could be incorporated in infant foods. In this context, ongoing researches are conducted to clarify this effect whether by using synthetic peptides or by using LPS-sepsis animal.

A review of Clostridioides [Clostridium] difficile occurrence through the food chain.

The epidemiology and transmission of Clostridioides difficile, particularly for community-associated infections, are not completely understood. Although there have been no confirmed cases of any foodborne disease caused by C. difficile, its occurrence in livestock and foods suggests that contaminated food products with spores could be a vehicle to spread C. difficile infection. This review proposes potential sources of C. difficile infection in the community and contamination routes of food products. Based on European research, it also summarizes the occurrence and organism characterization of C. difficile in animals at slaughterhouses and in human foods. Most of the analyzed literature reported prevalence in retail foods of less than 8%, including microorganism belonging to the ribotype 078, an important hypervirulent strain involved in disease in humans. This prevalence in Europe is underestimated, being lower that reported in North America (rates up to 42%), probably due of the lack of an ISO procedure for the detection of C. difficile in food products that preclude the comparison of prevalence data from different studies. The survival and growth of vegetative C. difficile cells and the resistance of its spores in foods are discussed as well as the risk factors of acquisition CDI from food products.

Effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf on the morphology and glass transition temperature of spray-dried lemon juice powder.

The aim of this research was to evaluate the effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf as a drying aid, on morphology, particle size and glass transition temperature of spray-dried lemon juice powder. Five concentrations of grapefruit mesocarp (0.4%, 0.8%, 1.2%, 1.6% and 2.0% (w/w)) and maltodextrin DE 10 (1.2%, w/w) were evaluated as encapsulant agents. The obtained data were evaluated by one-way ANOVA using Statistix version 8.0. For the means separation, the Tukey's test was applied with a 95% reliability. The morphology of the particles was described. According to the results, by applying different levels of coating agent in lemon juices, powder particles with different sizes (from 3.07 to 6.20 µm) and shapes (spherical, irregular and shrunken or reduced) are obtained; however, their glass transition temperature is not modified, finding values between 37.43 and 38.64°C.

In vitro effect of green tea and turmeric extracts on GLP-1 and CCK secretion: the effect of gastrointestinal digestion.

Plant extract activity can be modified by the digestion process. In order to assess the satiety effect of green tea and turmeric extracts, an in vitro gastrointestinal digestion process was performed and the STC-1 cell line was used. The enteroendocrine cell line was incubated (for 30, 60 and 120 minutes) with the digested and non-digested extracts measuring the secretion of cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) by enzyme-linked immunosorbent assay (ELISA). The release of satiety hormones by the STC-1 cells showed similar or higher results for plant extracts compared to the positive controls reported as satiatogenic (proteins and short chain fatty acids). For the non-digested samples, the largest amount of CCK and GLP-1 was found for turmeric, with 379 ± 46 pg ml-1 (60 min) and 347 ± 125 pg ml-1 (120 min) respectively. For the digested samples, the highest level of CCK was found for turmeric at 30 minutes (43.3 ± 11) and for GLP-1 by green tea at 120 minutes (165.7 ± 52). The in vitro gastrointestinal digestion process affects the satiety of plant extracts, increasing for green tea and decreasing for turmeric. Green tea showed the most stable satiating capacity after the in vitro digestion process and this may be useful for using it as a stable food supplement to reduce hunger associated with overweight and obesity.

Effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf. on the bromatological characteristics of spray-dried lemon juice powder.

The aim of this research was to evaluate the effect of different concentrations of pulverized mesocarp of Citrus paradisi Macf. as a drying aid on the bromatological characteristics and yield of spray-dried lemon juice powder. Five concentrations of grapefruit mesocarp encapsulant (0.4, 0.8, 1.2, 1.6, and 2.0% (w / w)) and maltodextrin DE 10 (1.2%, w / w) were evaluated as encapsulant agents. The highest yield (46.76%) was obtained with 1.2% of grapefruit encapsulant. Water activity and ash content were inversely proportional to the added encapsulant concentration. The highest moisture value was obtained with 0.4% and the highest soluble solids with 2.0%. For all treatments, the pH level did not change, except at 0.4% (it was lower). The concentrations of the encapsulants significantly affected the evaluated characteristics, except for the proteins.