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1.
Med. U.P.B ; 42(1): 49-56, ene.-jun. 2023. ilus, tab
Article Dans Espagnol | LILACS, COLNAL | ID: biblio-1416179

Résumé

Los alimentos de origen animal como la carne de pollo, res, pescado y cerdo poseen una amplia demanda en todo el mundo debido, entre otros aspectos, a su valor nutricional, asociado al alto contenido proteico. No obstante, este tipo de proteínas son susceptibles de sufrir reacciones de oxidación, las cuales pueden mediar procesos de fragmentación, agregación, pérdida de solubilidad, funcionalidad y digestibilidad proteica; eventos implicados en la pérdida de su valor nutricional. En este sentido, las proteínas agrega­das tienden a no ser digeridas en el tracto gastrointestinal y acumularse en el intestino (colon), donde la microbiota colónica las degrada a productos mutagénicos como fenol y p-cresol, lo que incrementa el riesgo de cáncer colorrectal. Por otra parte, los ami­noácidos o péptidos oxidados liberados en la digestión podrían incorporarse en las vías de señalización celular intestinal y favorecer o exacerbar procesos intestinales crónicos como colon irritable o enfermedad de Crohn. Debido al gran interés de esta temática en los últimos años, el objetivo de esta revisión es realizar una descripción general del impacto de proteínas oxidadas de origen animal sobre la salud intestinal.


Animal foods such as chicken, beef, fish and pork are in wide demand throughout the world due, among other things, to their nutritional value, associated with their high protein content. However, this type of protein is susceptible to oxidation reactions, which can mediate processes of fragmentation, aggregation, loss of solubility, functionality, and protein digestibility, which are events involved in the loss of their nutritional value. In this sense, aggregated proteins tend not to be digested in the gastrointestinal tract and accumulate in the intestine (colon), where the colonic microbiota degrades them into mutagenic products such as phenol and p-cresol, which increases the risk of colorectal cancer. On the other hand, the oxidized amino acids or peptides released in digestion could be incorporated into intestinal cell signaling pathways and favor or exacerbate chronic intestinal processes such as irritable bowel syndrome or Crohn's disease. Due to the great interest in this topic in recent years, the objective of this review is to provide a general overview of the impact of oxidized proteins of animal origin on intestinal health.


Alimentos de origem animal como frango, carne bovina, peixe e carne suína são muito procurados em todo o mundo devido, entre outros fatores, ao seu valor nutricional, associado ao seu alto teor de proteínas. No entanto, esse tipo de proteína é suscetível a reações de oxidação, que podem mediar processos de fragmentação, agregação, perda de solubilidade, funcionalidade e digestibilidade da proteína; eventos envolvidos na perda de seu valor nutritivo. Nesse sentido, as proteínas agregadas tendem a não ser digeridas no trato gastrointestinal e se acumulam no intestino (cólon), onde a microbiota colônica as degrada em produtos mutagênicos como fenol e p - cresol, aumentando o risco de câncer colorretal. Por outro lado, os aminoácidos ou peptídeos oxidados liberados na digestão poderiam ser incorporados às vias de sinalização das células intestinais e favorecer ou exacerbar processos intestinais crônicos, como a síndrome do intestino irritável ou a doença de Crohn. Devido ao grande interesse neste tema nos últimos anos, o objetivo desta revisão é fornecer uma descrição geral do impacto das proteínas oxidadas de origem animal na saúde intestinal.


Sujets)
Humains , Animaux , Aliments , Tumeurs colorectales , Protéines , Côlon , Phénol , Digestion , Alimentation d'Origine Animale , Microbiote , Viande rouge
2.
China Journal of Chinese Materia Medica ; (24): 958-965, 2023.
Article Dans Chinois | WPRIM | ID: wpr-970567

Résumé

This study was aimed at identifying the bioactive components of the crude and stir-baked hawthorn for invigorating spleen and promoting digestion, respectively, to clarify the processing mechanism of hawthorn by applying the partial least squares(PLS) algorithm to build the spectrum-effect relationship model. Firstly, different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions were prepared, respectively. Then, the contents of 24 chemical components were determined by ultra-high performance liquid chromatography-mass spectrometry. The effects of different polar fractions of crude hawthorn and stir-baked hawthorn aqueous extracts and combinations of different fractions were evaluated by measuring the gastric emptying rate and small intestinal propulsion rate. Finally, the PLS algorithm was used to establish the spectrum-effect relationship model. The results showed that there were significant differences in the contents of 24 chemical components for different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions, and the gastric emptying rate and small intestinal propulsion rate of model rats were improved by administration of different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions. The bioactive components of crude hawthorn identified by PLS models were vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, neochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, malic acid, quinic acid and fumaric acid, while neochlorogenic acid, cryptochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, quinic acid and fumaric acid were the bioactive components of stir-baked hawthorn. This study provided data support and scientific basis for identifying the bioactive components of crude and stir-baked hawthorn, and clarifying the processing mechanism of hawthorn.


Sujets)
Animaux , Rats , Rate , Crataegus , Acide quinique , Méthode des moindres carrés , Acide vanillique , Algorithmes , Digestion
3.
Braz. j. biol ; 83: e246568, 2023. tab, graf
Article Dans Anglais | LILACS, VETINDEX | ID: biblio-1278537

Résumé

Abstract Fishmeal; being a limited and costly feed ingredient is continuously been substituted with locally available plant proteins. However, the occurrence of anti-nutritional factors in plant meal suppresses its potential to be fully replaced. Therefore, in this study we aimed to study the synergistic effects of dietary additives like citric acid and phytase enzyme supplementation on growth performance and nutrient digestibility of Cirrhinus mrigala fingerlings. Canola meal (CM) was used as a test ingredient to replace fishmeal (FM) as; 0%, 25%, 50% and 75%. These four diets were further supplemented by varying levels of phytase (0 and 750 FTU kg-1) and citric acid (0% and 2.5%) to formulate total sixteen test diets as T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 and T16. Each treatment contained three replicates; applied to fish groups having 15 fingerlings each; following 3×3 factorial arrangement. 1% of chromic oxide was added as an inert marker. Maximum weight gain% (288%) and the lowest value of FCR (1.07) were recorded when fish was fed on diet T12 as compared to fish fed control diet (T1). Similarly, optimum nutrient digestibility values such as crude protein (77%), crude fat (84%) and gross energy (70%) were noted on same level. It was concluded that 50% canola meal can optimally replace fishmeal when supplemented with phytase and citric acid at the levels of 750 FTU kg-1 and 2.5%, respectively.


Resumo A farinha de peixe, por ser um ingrediente alimentar limitado e caro, é continuamente substituída por proteínas vegetais disponíveis localmente. No entanto, a ocorrência de fatores antinutricionais na farinha de plantas suprime seu potencial de ser totalmente substituída. Portanto, neste estudo objetivamos estudar os efeitos sinérgicos de aditivos dietéticos como ácido cítrico e suplementação com enzima fitase sobre o desempenho de crescimento e digestibilidade de nutrientes de alevinos de Cirrhinus mrigala. A farinha de canola (CM) foi usada como ingrediente de teste para substituir a farinha de peixe (FM) como: 0%, 25%, 50% e 75%. Essas quatro dietas foram suplementadas por níveis variados de fitase (0 e 750 FTU kg-1) e ácido cítrico (0% e 2,5%) para formular um total de 16 dietas de teste como T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15 e T16. Cada tratamento continha três repetições; aplicado a grupos de peixes com 15 alevinos cada; seguindo o arranjo fatorial 3 × 3. 1% de óxido crômico foi adicionado como um marcador inerte. % de ganho de peso máximo (288%) e o valor mais baixo de FCR (1,07) foram registrados quando os peixes foram alimentados com dieta T12 em comparação com peixes alimentados com dieta controle (T1). Da mesma forma, valores ótimos de digestibilidade de nutrientes, como proteína bruta (77%), gordura bruta (84%) e energia bruta (70%) foram anotados no mesmo nível. Concluiu-se que 50% da farinha de canola pode substituir de forma ideal a farinha de peixe quando suplementada com fitase e ácido cítrico nos níveis de 750 FTU kg-1 e 2,5%, respectivamente.


Sujets)
Animaux , Phytase , Nutriments , Poulets , Acide citrique , Compléments alimentaires , Régime alimentaire/médecine vétérinaire , Digestion , Aliment pour animaux/analyse , Phénomènes physiologiques nutritionnels chez l'animal
4.
Arch. latinoam. nutr ; 72(1): 11-22, mar. 2022. tab, graf
Article Dans Espagnol | LILACS, LIVECS | ID: biblio-1368344

Résumé

El arándano (Vaccinium corymbosum L.) posee un alto contenido de compuestos fenólicos los cuales han sido estudiados principalmente por su actividad antioxidante, antiobesogénica, antiinflamatoria, entre otras. Objetivo. Evaluar el efecto de la digestión gastrointestinal in vitro sobre la bioaccesibilidad de compuestos fenólicos y actividad antioxidante de una formulación nutracéutica de arándano (cápsula), comparado con arándano fresco y polvo. Materiales y métodos. Se obtuvieron extractos metanólicos de muestras de arándano fresco y liofilizado y se determinó su contenido de fenoles, flavonoides y antocianinas totales, así como también actividad antioxidante. Se llevó a cabo un ensayo de simulación de digestión gastrointestinal para evaluar la bioaccesibilidad de los compuestos fenólicos presentes en las muestras. Resultados. Los resultados mostraron que la digestión gástrica de arándano en polvo y en cápsula promovió una mayor bioaccesibilidad de fenoles (42% y 40%), flavonoides (52% y 33%) y antocianinas (45% y 40%) comparado con digestos de arándano fresco. Posterior a la digestión intestinal, la bioaccesibilidad de fenoles (63%) y flavonoides (67%) fue mayor en la cápsula de arándano comparada con su contraparte arándano en polvo. Las condiciones de digestión intestinal afectaron negativamente la bioaccesibilidad de las antocianinas independientemente del tipo de muestra evaluada. Conclusión. Las condiciones de digestión gástrica promueven una mayor estabilidad de los compuestos fenólicos en arándano en polvo y en cápsula lo que pudiera ser relevante para el mantenimiento de un ambiente antioxidante a este nivel. Las condiciones de digestión intestinal afectaron de manera particular a los compuestos fenólicos de arándano fresco y polvo, pero no a la cápsula, lo que puede sugerir que el encapsulamiento protegió de las condiciones alcalinas a los fenoles presentes. Se sugieren estudios posteriores sobre absorción in vitro de los componentes remanentes en intestino y sus posibles efectos sobre biomarcadores de estrés oxidativo en modelos in vivo(AU)


Blueberry (Vaccinium corymbosum L.) has a high content of phenolic compounds which have been studied mainly for their antioxidant, antiobesogenic, anti-inflammatory activity, among others. Objetive. The objective of the present study was to evaluate the effect of in vitro gastrointestinal digestion on the bioaccessibility of phenolic compounds and antioxidant activity of a nutraceutical formulation of blueberry (capsule), compared to fresh and powder blueberry. Materials and methods. Methanolic extracts of fresh and lyophilized blueberry were obtained and determined its total phenols, flavonoids, anthocyanins content, as well as antioxidant activity. A gastrointestinal digestion simulation test also was carried out to assess the bioaccessibility of the phenolic compounds found in samples. Results. The results showed that gastric digestion of powder and capsule blueberry promoted greater bioaccessibility of phenols (42% and 40%), flavonoids (52% and 33%) and anthocyanins (45% and 40%), compared to fresh blueberry digests. After intestinal digestion, the bioaccessibility of phenols (63%) and flavonoids (67%) was higher in the blueberry capsule compared to its powdered blueberry counterpart. The intestinal digestion conditions negatively affected the bioaccessibility of anthocyanins regardless of the type of sample evaluated. Conclusion. Gastric digestion conditions promote greater stability of phenolic compounds in powdered and capsule blueberries, which could be relevant for the maintenance of an antioxidant environment at this level. The intestinal digestion conditions particularly affected the phenolic compounds of fresh and lyophilized blueberry, but not the capsule, which may suggest that encapsulation protected the phenols present from alkaline conditions. Further studies on in vitro absorption of the remaining components in the intestine and their possible effects on oxidative stress biomarkers in in vivo models are suggested(AU)


Sujets)
Tanins , Flavonoïdes , Myrtillier , Composés Phénoliques , Absorption gastro-intestinale , Techniques in vitro , Maladie chronique , Digestion , Lyophilisation
5.
Semina cienc. biol. saude ; 43(1): 101-118, jan./jun. 2022. tab, ilus
Article Dans Anglais | LILACS | ID: biblio-1354575

Résumé

Achyrocline satureioides is popularly known for its richness in phenolic compounds and medicinal properties (anti-inflammatory, analgesic, and hepatoprotective). The present study aimed at broadening the knowledge about the pharmacological potential exerted by the aqueous and ethanolic extracts of A. satureioides. These extracts were characterized by HPLC and tested for their modulatory action on phospholipases A2 and proteases of snake venoms. In addition, they were tested on the activities of digestive enzymes. Snake venoms were used as tools since they have enzymes with high functional and structural homology to human enzymes. The results demonstrate that the extracts of A. satureioides act as enzymatic inhibitors or potentiators, interfering in processes related to the hemostasis, such as coagulation and thrombus dissolution. In addition, the anti-genotoxic activity and inhibitions exerted on digestive enzymes suggests their potential use in the prevention and/or treatment of several pathologies. New studies could provide information on how the compounds present in the extracts and the different enzymes interact.


A Achyrocline satureioides é popularmente conhecida por sua riqueza em compostos fenólicos e por suas propriedades medicinais (anti-inflamatória, analgésica e hepatoprotetora). No presente estudo, com o objetivo de ampliar o conhecimento sobre o potencial farmacológico exercido por esses extratos, os extratos aquoso e etanólico de A. satureioides foram caracterizados por HPLC e testados quanto à sua ação modulatória sobre as fosfolipases A2 e proteases de peçonhas de serpentes. Além disso, também foram testados em atividades de enzimas digestivas. As peçonhas de serpentes foram usadas como ferramentas por apresentarem enzimas com alta homologia funcional e estrutural às humanas. Os resultados demonstram que os extratos de A. satureioides atuam como inibidores ou potencializadores enzimáticos, interferindo em processos relacionados à hemostasia, como coagulação e dissolução do trombo. Além do mais, destacam seu potencial antigenotóxico e as inibições exercidas sobre as enzimas digestivas direcionando seu potencial de uso na prevenção e/ou tratamento de diversas patologias. Novos estudos poderão fornecer informações sobre os mecanismos de interação entre os compostos presentes nos extratos e as diferentes enzimas.


Sujets)
Humains , Animaux , Serpents , Coagulation sanguine , Achyrocline , Digestion , Enzymes , Dissolution , Phospholipases A2 , Hémostase , Analgésiques , Inflammation
6.
Arq. bras. med. vet. zootec. (Online) ; 73(2): 469-476, Mar.-Apr. 2021. tab
Article Dans Anglais | LILACS, VETINDEX | ID: biblio-1248951

Résumé

This study was aimed to evaluate and compare the nutrient digestibility of grain-inclusive and grain-free commercial dry dog foods using the method of total fecal collection and also explore their effects on fecal consistency. 21 different foods, including 14 grain-inclusive (7 grain-chicken meat, 7 grain-lamb meat) and 7 grain-free were investigated. 12 adult Golden retriever dogs (age 3-4 years, body weight=22.5±1.7kg) were divided into 3 groups. The results of digestibility trials indicated that the grain-lamb meat foods showed the highest digestibility of dry matter, organic matter, and crude fiber. Overall, in the grain-inclusive and grain-free group evaluation, grain-inclusive foods showed higher digestibility of crude fibers while grain-free foods had higher ether extract digestibility whereas the differences in the scores of fecal consistencies between the groups were insignificant. Contrary to popular belief, grain-inclusive foods were more digestive than grain-free foods in terms of dry matter and organic matter. There are studies involving starch sources in the dog food formulations, but there is a need to study the digestibility of complete dog food to supply them with adequate nutrients. Also, each diet should be assessed based on its overall nutrient profile and digestibility rather than individual ingredients.(AU)


Este estudo teve como objetivo avaliar e comparar as digestibilidades de nutrientes de alimentos secos para cães comerciais, com grãos e livres de grãos, usando o método de coleta fecal total, bem como explorar seus efeitos na consistência fecal. Vinte e um alimentos diferentes, incluindo 14 grãos inclusivos (sete grãos de carne de frango, sete grãos de carne de cordeiro) e sete livres de grãos, foram investigados. Doze cães Golden Retriever adultos (idade de três-quatro anos, peso corporal = 22,5 ± 1,7kg) foram divididos em três grupos. Os resultados dos ensaios de digestibilidade indicaram que os alimentos cárneos de cordeiro apresentaram as maiores digestibilidades de matéria seca, matéria orgânica e fibra bruta. Em geral, na avaliação do grupo com grãos inclusivos e do grupo sem grãos, os alimentos com grãos inclusivos mostraram maior digestibilidade das fibras brutas, enquanto os alimentos sem grãos tiveram maior digestibilidade do extrato etéreo; já as diferenças nos escores de consistência fecal entre os grupos foram insignificantes. Ao contrário da crença popular, os alimentos com inclusão de grãos eram mais digestivos do que os sem grãos, em termos de matéria seca e matéria orgânica. Existem estudos envolvendo fontes de amido em formulações de rações, mas é necessário estudar a digestibilidade de rações completas para fornecer nutrientes adequados. Além disso, cada dieta deve ser avaliada com base em seu perfil geral de nutrientes e digestibilidade, em vez de ingredientes individuais.(AU)


Sujets)
Animaux , Chiens , Protéines alimentaires/administration et posologie , Nutriments/analyse , Grains comestibles , Digestion , Aliment pour animaux/analyse , Aliment pour animaux/classification
7.
Arq. bras. med. vet. zootec. (Online) ; 73(1): 82-90, Jan.-Feb. 2021. tab, graf
Article Dans Anglais | LILACS, VETINDEX | ID: biblio-1153042

Résumé

Essential oils (EO) such as carvacrol represent a wide range of mainly volatile aromatic plant compounds which hold antioxidant, antibacterial and antifungal potential, in addition to other properties of interest to animal health, such as the ability to modulate the microbiome. Current horse care commonly involves an intensive management system with an excessive use of concentrated feed, which can lead to severe digestive and metabolic disorders. Studies with EO in horses are limited, but the use of carvacrol essential oil (CEO) can promote benefits in microbial fermentation. The objective was to investigate the effect of different quantities of CEO on the apparent total digestibility of nutrients, microbial profile in the feces and postprandial blood glucose and insulin response when added to the equine diet. Eight Mini-Horse geldings were used (42±6 months; 135±15 kg BW) and fed with a proportion of 60% concentrate and 40% grass hay. The treatments were: 0, 100, 200 and 300 ppm of CEO. The addition of CEO up to 300 ppm did not influence the apparent digestibility of nutrients or the postprandial plasma glucose and insulin response. The use of CEO maintained the fermentative digestive health of horses fed with concentrate diets.(AU)


Os óleos essenciais (EO), como o carvacrol, são descritos por representarem ampla gama de compostos principalmente voláteis de plantas aromáticas, com potencial antioxidante, antibacteriano, antifúngico, entre outras propriedades de interesse para a saúde animal, como a modulação do microbioma. Atualmente, os cavalos são submetidos a manejo intensivo, com uso excessivo de ração concentrada, o que pode causar graves distúrbios digestivos e metabólicos. Em cavalos, estudos com EO são limitados, mas o uso de óleo essencial de carvacrol (CEO) poderia promover benefícios na fermentação microbiana. O objetivo da presente pesquisa foi investigar o efeito de diferentes quantidades de óleo essencial de carvacrol, adicionadas à dieta de equinos, sobre a digestibilidade aparente total de nutrientes, o perfil microbiano por meio das fezes e a resposta sanguínea pós-prandial de glicose e insulina. Foram utilizados oito cavalos castrados, da raça Mini-Horse (42±6 meses), 135±15kg PV, alimentados na proporção de 60% concentrado e 40% feno de capim. Os tratamentos foram: 0, 100, 200 e 300ppm de CEO. A adição de CEO até 300ppm não influencia a digestibilidade aparente dos nutrientes e a resposta de glicose e insulina plasmática pós-prandial. O uso de EO demonstra manter a saúde digestiva fermentativa quando os cavalos são alimentados com dieta rica em concentrado.(AU)


Sujets)
Animaux , Huile essentielle/usage thérapeutique , Digestion/effets des médicaments et des substances chimiques , Glucose , Equus caballus/sang , Insuline/sang , Compléments alimentaires/analyse , Monoterpènes
8.
Braz. j. biol ; 81(1): 44-52, Feb. 2021. tab
Article Dans Anglais | LILACS | ID: biblio-1153322

Résumé

Abstract Three trials were executed to examine the nutritive profile, metabolisable energy and digestible amino acid (AA) contents of four indigenous feed ingredients including wheat (W), wheat middling (WM), canola meal (CM) and rapeseed meal (RSM) in Nile tilapia. Three samples of each test ingredient were collected from three different locations of Multan (MUL) and Sukkar (SKR), of Pakistan. The collected three samples were pooled thereafter to make a homogenous/ representative sample of each test ingredient from a particular study site. Nutrients composition, AA and energy digestibility of these indigenous ingredients were evaluated by using laboratory analyses and fish studies. Proximate analysis indicated variations in some of the nutrients due to location (p < 0.05). Differences were also observed in some AA including arginine, lysine, serine, cysteine, glutamic and aspartic acids, histidine, valine and glycine contents of these ingredients (p < 0.05). Digestibility of leucine, glycine and glutamic acid was higher (p < 0.05) in RSM from MUL. Among W samples from MUL, AA digestibility for lysine, threonine, and aspartic acid was higher (p < 0.05). Crude protein, arginine, alanine, serine, and aspartic acid had higher digestibility (p < 0.05), whereas digestibility was lower (p < 0.05) for threonine, valine and tyrosine in RSM from MUL. Metaboliseable energy contents did not differ among W, WM, CM and RSM regarding their origin (p > 0.05). The results indicated that nutritional profiles and their digestibility indices vary with the location for Nile tilapia.


Resumo Três experimentos foram executados para examinar o perfil nutritivo, a energia metabolizável e o conteúdo de aminoácidos digestíveis (AA) de quatro ingredientes alimentícios, incluindo trigo (W), farelo de trigo (WM), farelo de canola (CM) e farelo de colza (RSM) em tilápia do Nilo. Três amostras de cada ingrediente do teste foram coletadas de dois locais diferentes (Multan (MUL) e Sukkar (SKR), do Paquistão) e assim agrupadas. A composição nutricional, AA e digestibilidade energética desses ingredientes indígenas foram avaliadas por meio de análises laboratoriais e estudos de peixes. A análise imediata indicou variações (p <0,05) em alguns dos nutrientes devido à localização. Variações (p <0,05) também foram observadas em alguns teores de AA desses ingredientes. A digestibilidade da leucina, glicina e ácido glutâmico foi maior (p <0,05) em RSM de MUL. Entre as amostras de W da MUL, a digestibilidade de AA para Lys, Thr e Asp foi maior (p <0,05). Proteína Crud, arginina, alanina, serina e ácido aspártico apresentaram maior digestibilidade (p <0,05), e menor (p <0,05) para treonina, valina e tirosina em MRS. Nenhuma diferença (p> 0,05) relacionada a energia metabolizável foi observada entre esses ingredientes em relação à sua origem. Os resultados indicaram que os perfis nutricionais e sua digestibilidade variam com a localização.


Sujets)
Animaux , Cichlides , Acides aminés , Pakistan , Régime alimentaire , Digestion , Iléum , Aliment pour animaux/analyse , Phénomènes physiologiques nutritionnels chez l'animal
9.
Rev. argent. microbiol ; 52(4): 41-50, dic. 2020. graf
Article Dans Anglais | LILACS | ID: biblio-1340919

Résumé

Abstract The health benefits attributed to probiotics generate interest in the search of competent strains adapted to several ecological niches, especially those related to traditional beverages and foods of each country. Pineapple tepache, a traditional Mexican fermented beverage, was used for the isolation of lactic acid bacteria with probiotic potential, one of which withstood the in vitro tests. The isolated strain AB-05, which exhibited the tested probiotic functional properties, was designated as Lactobacillus pentosus ABHEAU-05. The sequence was registered in GenBank under access code MK587617. This study is the first report of a lactic acid bacterium with in vitro digestion resistance isolated from pineapple tepache. The survival of L. pentosus ABHEAU-05 in a symbiotic medium was proven using fermented milk enriched with inulin. The in vitro digestion-resistant probiotic activity of lactobacilli was measured through analysis of pH and proteolysis. Results showed that L. pentosus grew properly in fermented milk; therefore, this microorganism could be used in the manufacture of this kind of products. The concentration of L. pentosus reached up to 8.5 logCFU/ml after 40 h of fermentation. In addition, the production of peptides and the decrease in pH indicated the vigorous and active metabolic state of the lactic acid bacterium tested. The activity and the concentration of this microorganism were maintained during refrigeration. The results of this research conclude that L. plantarum ABHEAU-05 is an in vitro digestion-resistant microorganism that can be used as a starter culture for the production of functional foods of dairy origin.


Resumen Los beneficios a la salud atribuidos a los probióticos generan interés en la búsqueda de cepas competentes adaptadas a varios nichos ecológicos, especialmente los relacionados con bebidas y alimentos tradicionales de cada país. En este estudio, se aisló del tepache de pina, una bebida fermentada tradicional mexicana, una bacteria láctica resistente a la digestión in vitro. Entre 5 bacterias aisladas, una de ellas soportó las pruebas simuladas de digestión gastrointestinal. Se analizó la resistencia a sales biliares, a condiciones ácidas y al ataque enz-imático con pepsina. La bacteria aislada, que exhibió las propiedades funcionales probióticas referidas, fue identificada como Lactobacillus pentosus y designada como L. pentosus ABHEAU-05. La secuencia fue depositada en GenBank (acceso MK587617). Se comprobó la supervivencia de L. pentosus ABHEAU-05 en una leche fermentada adicionada con inulina y su resistencia a la digestión in vitro mediante el análisis del pH y la proteólisis. Los resultados muestran que la leche fermentada es una matriz adecuada, donde L. pentosus ABHEAU-05 se desarrolla sin inconvenientes, alcanzando un título de 8,5 logufc/ml después de 40 h de fermentación. Además, la producción de péptidos y el descenso del pH indicaron el estado metabólico vigoroso y activo del microorganismo probiótico. Se concluye que L. pentosus ABHEAU-05 es un microorganismo resistente a la digestión in vitro, que puede servir como cultivo iniciador para la producción de alimentos de origen lácteo. Este es el primer informe acerca del aislamiento de una bacteria ácido láctica resistente a la digestión in vitro a partir del tepache de piña.


Sujets)
Probiotiques , Lactobacillus pentosus , Aliments fermentés , Boissons , Acide lactique , Digestion , Fermentation , Microbiologie alimentaire
10.
Electron. j. biotechnol ; 48: 29-35, nov. 2020. ilus, tab, graf
Article Dans Anglais | LILACS | ID: biblio-1254696

Résumé

BACKGROUND: Cellulose as a potential feed resource hinders its utilization because of its complex structure, and cellulase is the key to its biological effective utilization. Animal endogenous probiotics are more susceptible to colonization in the intestinal tract, and their digestive enzymes are more conducive to the digestion and absorption of feed in young animals. Min pigs are potential sources of cellulase probiotics because of the high proportion of dietary fiber in their feed. In this study, the cellulolytic bacteria in the feces of Min pigs were isolated and screened. The characteristics of enzymes and cellulase production were studied, which provided a theoretical basis for the rational utilization of cellulase and high-fiber food in animal production. RESULTS: In our study, 10 strains of cellulase producing strains were isolated from Min pig manure, among which the M2 strain had the best enzyme producing ability and was identified as Bacillus velezensis. The optimum production conditions of cellulase from strain M2 were: 2% inoculum, the temperature of 35°C, the pH of 5.0, and the liquid loading volume of 50 mL. The optimum temperature, pH and time for the reaction of cellulase produced by strain M2 were 55°C, 4.5 and 5 min, respectively. CONCLUSIONS: Min pigs can be used as a source of cellulase producing strains. The M2 strain isolated from feces was identified as Bacillus velezensis. The cellulase from M2 strain had a good activity and the potential to be used as feed additive for piglets.


Sujets)
Animaux , Porc miniature , Bactéries/enzymologie , Cellulase/biosynthèse , Bacillus , Fibre alimentaire , Probiotiques , Digestion , Fèces , Aliment pour animaux
11.
Electron. j. biotechnol ; 48: 62-71, nov. 2020. tab, graf
Article Dans Anglais | LILACS | ID: biblio-1254807

Résumé

BACKGROUND: A key challenge for manufacturers of pro-health food containing active probiotic microorganisms is to develop a product with attractive sensory features along with maintenance of declared number of microorganisms during storage and transfer by alimentary tract. RESULTS: The highest concentration of polyphenols was observed in snacks without an additive of probiotics as well as those with an additive of L. rhamnosus and B. animalis bacteria and concentration of these compounds increased by 9.5% during six months of storage. None of the products distinguished itself in the sensorial assessment although each was assessed positively. The number of microorganisms was stable and comparatively high during six months of storage at a room temperature and in cooling conditions (108 cfu/g). In the digestion model, an influence of aggressive digestion conditions was examined in the alimentary tract on the number of microorganisms, which allowed to arrange strains from the most resistant (S. boulardii) to the most sensitive (B. breve). It must be noted that currently on the market there is no available snack containing probiotic yeast as well as there is no literature data on works on such formulation of food. CONCLUSIONS: In the newly developed snack made of chocolate, in which sugar has been replaced with maltitol, a raw material was added in the form of raspberry, prebiotic in the form of inulin and a strain of probiotic bacteria, including the unprecedented so far S. boulardii, which stands a high chance to occupy a good place on the market of functional food.


Sujets)
Probiotiques , Aliment fonctionnel , Chocolat/microbiologie , Polyols , Température , Aliments totaux , Digestion , Stockage des aliments , Prébiotiques , Synbiotiques , Polyphénols , Casse-croute , Rubus , Maltose/analogues et dérivés
12.
Arq. bras. med. vet. zootec. (Online) ; 72(5): 1971-1979, Sept.-Oct. 2020. tab
Article Dans Anglais | LILACS, VETINDEX | ID: biblio-1131529

Résumé

The objective was to evaluate the effects of cranberry on blood and urinary parameters of dogs (experiment I), digestibility of nutrients (experiment II), palatability of diet (experiment III) and the influence of cranberry on E. coli UPEC-MRHA fimbriae in vitro (experiment IV). For experiment I and II, ten dogs were fed with diets containing 0% or 0.4% cranberry for 30 days. Experiment III compared the diets containing 0% and 0.4% cranberry using 16 adult dogs. There were no statistical differences (P>0.05) in the blood parameters evaluated. Dogs consuming cranberry presented lighter color and appearance of urine, compared to the control group (P<0.05). The diet containing cranberry showed higher digestibility of dry matter, organic matter, ether extract, higher metabolizable energy (P<0.05) and reduced fecal sialic acid concentration (P<0.05) compared to the control diet. There was no influence of cranberry on the formation of fimbriae of E. coli UPEC-MRHA. There was a lower intake ratio of the diet containing cranberry (P<0.05). The inclusion of 0.4% cranberry increases the digestibility of nutrients and influences the color and appearance of urine of dogs. However, it reduces diet palatability and does not alter the adhesion of E. coli UPEC-MRHA in vitro.(AU)


O objetivo foi avaliar os efeitos do cranberry nos parâmetros sanguíneos e urinários de cães (experimento I), na digestibilidade dos nutrientes (experimento II), na palatabilidade da dieta (experimento III) e a influência do cranberry sobre E. coli UPEC-MRHA fimbriae in vitro (experimento IV). Para os experimentos I e II, 10 cães foram alimentados com dietas contendo 0% ou 0,4% de cranberry por 30 dias. O experimento III comparou as dietas contendo 0% e 0,4% de cranberry usando 16 cães adultos. Não houve diferenças estatísticas (P>0,05) nos parâmetros sanguíneos avaliados. Cães que consumiram cranberry apresentaram cor e aparência mais claras da urina, em comparação com o grupo controle (P<0,05). A dieta contendo cranberry apresentou maior digestibilidade da matéria seca, extrato etéreo, matéria orgânica, maior energia metabolizável (P<0,05) e menor concentração de ácido siálico fecal (P<0,05) comparada à dieta controle. Não houve influência do cranberry na formação de fímbrias de E. coli UPEC-MRHA. Houve uma menor taxa de ingestão da dieta contendo cranberry (P<0,05). A inclusão de 0,4% de cranberry aumenta a digestibilidade dos nutrientes, influencia a cor e a aparência da urina dos cães. No entanto, reduz a palatabilidade da dieta e não altera a adesão de E. coli UPEC-MRHA in vitro.(AU)


Sujets)
Animaux , Chiens , Compléments alimentaires , Cystite , Vaccinium macrocarpon/métabolisme , Digestion , Infections urinaires/médecine vétérinaire , Phénomènes physiologiques nutritionnels chez l'animal
13.
Chinese Journal of Biotechnology ; (12): 2435-2442, 2020.
Article Dans Chinois | WPRIM | ID: wpr-878499

Résumé

In recent years, mass spectrometry has been widely used to study membrane protein structure and function. However, the application of mass spectrometry to study integral membrane protein is limited because there are many hydrophobic amino acids in the trans-membrane domain of integral membrane protein to cause low sequence coverage detected by LC-MS/MS. Therefore, we used vitamin K epoxide reductase (VKORC1), a human integral membrane protein, as a model to optimize the digestion conditions of chymotrypsin, and developed an in-gel digestion method of chymotrypsin to improve sequence coverage of membrane protein by mass spectrometry. By exploring the effects of calcium concentration, pH value and buffer system on the percentage of sequence coverage, number of total detected and types of unique peptide, and the size of unique peptide, sequence coverage and peptide diversity could be considered under condition of Tris-HCl buffer with 5-10 mmol/L calcium ion concentration and pH value 8.0-8.5. This method could make the sequence coverage of membrane protein to reach more than 80%. It could be widely used in the study of membrane protein structure and function, identification of interaction site between membrane proteins, and identification of binding site between membrane protein and small molecular drug.


Sujets)
Humains , Séquence d'acides aminés , Chromatographie en phase liquide , Chymotrypsine/métabolisme , Digestion , Protéines membranaires , Spectrométrie de masse en tandem , Trypsine , Vitamin K epoxide reductases
14.
Afro-Egypt. j. infect. enem. Dis ; 1(3): 199-208, 2020. ilus
Article Dans Anglais | AIM | ID: biblio-1258725

Résumé

Background: The aim of Trichinella inspection is to reliably detect larvae in meat at levels which are capable of causing human trichinellosis. Trichinoscopy is a rapid, cost effective but is less sensitive in detection of low muscle larvae (ML)numbers. Artificial digestion is more convenient, and flexible, but may destroy immature larvae. Baermann technique may enhance the efficiency of diagnosis. Giemsa and Leishman stains provide identical contrasting coloration as Hematoxylin and Eosin (H-E) with the both advantages of that they can be used for staining non-histological sectionsand rapid preparation.The primary aim of the study was to compare the efficiency of digestion and Baermann techniques for detection of ML in muscles. Secondary aim was detection of ML by Geimsa and Leishman stain and their comparison with routinely used stain H-E. Methods: Muscles from 36 mice orally infected with Trichinella spiralis encysted larvae were examined. Larvae from 1 g samples of different muscles were recovered by digestion and Baermann techniques after different dose of infection. Muscle samples were prepared for staining with Giemsa, Leishman and H-E. Results: Number of larvae recovered by Baermann was higher recovered by digestion method (P ˃0.5). Leishman stain was the best stain for rapid detection of ML after 30 min followed in validity by Geimsa then H-E. Conclusion: Baermann concentration technique showed higher sensitivity than digestion method. Leishman stain is superior to Giemsa stain as it takes less time. Both stains are considered good alternative to H-E for rapid and easy diagnosis of trichinellosis in post-mortem animals


Sujets)
Arthrite expérimentale , Colorants azurés , Digestion , Égypte
15.
Arq. bras. med. vet. zootec. (Online) ; 71(3): 1015-1026, May-June 2019. tab, ilus
Article Dans Portugais | VETINDEX, LILACS | ID: biblio-1011301

Résumé

O objetivo do presente trabalho foi avaliar a conversão alimentar, a digestibilidade do amido, o comportamento ingestivo e o escore de sobras da dieta e de fezes de novilhos confinados, suplementados com doses do complexo enzimático (0; 2,5; 5,0 e 7,5g animal-1 dia-1) e alimentados com dieta constituída por 85% de grão de milho e 15% de núcleo proteico, vitamínico e mineral, na base seca, isenta de forragem. O delineamento experimental foi o de blocos ao acaso contendo quatro tratamentos e quatro repetições. Trinta e dois novilhos inteiros, ½ sangue Angus Nelore, com idade média de 12 meses e peso vivo médio inicial de 422kg, foram confinados por um período de 77 dias. Cada grama de inclusão de complexo enzimático melhorou a conversão alimentar em 0,1652%, reduziu a matéria seca das fezes em 0,4648% e o tempo de ingestão de água em 0,0068 horas dia-1. A máxima digestibilidade do amido foi alcançada na dose de 5,08g animal-1 dia-1. A inclusão progressiva do complexo enzimático à dieta de alta densidade energética promoveu melhoria na conversão alimentar, redução na matéria seca das fezes e diminuição do tempo de ingestão de água. A dose de 5g animal-1 dia-1 do complexo enzimático aumentou a digestibilidade do amido.(AU)


The objective was to evaluate the feed conversion, starch digestibility, ingestive behavior, diet leftover score and feces score of steers supplemented with doses of enzyme complex (0; 2.5; 5.0 and 7 .5g animal -1 day -1 ) fed with roughage-free diet composed of a mixture of 85% whole corn grain and 15% protein-mineral-vitamin mix, on a dry matter basis. A completely randomized block design was adopted, consisting of four treatments and four replicates. Thirty-two ½ Angus ½ Nellore crossbred steers at an average age of 12 months, with an average initial weight of 422kg, were kept in a feedlot for 77 days. Each gram of enzyme complex inclusion improved feed conversion in 0.1652%, decreased feces dry matter in 0,4648% and time of water intake in 0.0068 hours day -1 . The maximum starch digestibility was reached in the dose of 5,08g animal -1 day -1 . The gradual inclusion of enzyme complex promoted improvement in feed conversion, reduction in the dry matter of feces and redution in the time of water intake. The enzyme complex dose of 5.0g animal -1 day -1 increased the starch digestibility.(AU)


Sujets)
Animaux , Bovins , Amidon/métabolisme , Régime alimentaire/médecine vétérinaire , Digestion , Enzymes/administration et posologie , Amidon et Fécule
16.
Arq. bras. med. vet. zootec. (Online) ; 71(3): 990-996, May-June 2019. tab
Article Dans Portugais | VETINDEX, LILACS | ID: biblio-1011324

Résumé

Objetivou-se avaliar o efeito de complexos enzimáticos sobre a energia metabolizável e o coeficiente de digestibilidade de nutrientes do milheto para frangos de corte. Quinhentos e setenta e seis frangos machos foram distribuídos em 36 gaiolas, com três tratamentos: T1 - composição de milheto sem complexo enzimático; T2 - composição de milheto com complexo enzimático (CES) e T3 - composição de milheto com complexo enzimático (CEV). Os tratamentos foram definidos com base em seis dietas (três dietas referências e três dietas testes). As dietas testes foram obtidas pela substituição de 40% da dieta referência por milheto inteiro, e a adição de enzimas consistiu de dois complexos enzimáticos: CES, constituído pelas enzimas fitase, protease, xilanase, ß-glucanase, celulase, amilase e pectinase; e CEV. constituído pelas enzimas protease, celulase e amilase. Na fase de 11 a 20 dias, a suplementação com o CEV reduziu os valores de EMA, EMAn e CDPB. A suplementação com CES melhorou o CDPB, e não houve efeito significativo para CDMS e CDEB. Na fase de 21 a 30 dias, houve menor aproveitamento da energia e dos nutrientes com as suplementações CES e CEV. Na fase de 31 a 40 dias, as suplementações reduziram os valores de EMA, EMAn, e o complexo CEV foi efetivo em aumentar o valor de CDPB. A inclusão dos complexos enzimáticos CES (fitase, protease, xilanase, ß-glucanase, celulase, amilase e pectinase) e CEV (protease, celulase e amilase) não favoreceu a utilização da energia do milheto, no entanto melhorou o coeficiente de digestibilidade da proteína do milheto nos períodos de 11 a 20 e de 31 a 40 dias de idade.(AU)


The objective of this study was to evaluate the effect of enzymatic complexes on metabolizable energy and nutrient digestibility coefficient of millet for broilers chickens. 576 male chickens, were distributed in 36 cages with three treatments: T1 - millet composition without enzymatic complex; T2 - millet composition with enzymatic complex (ECS); and T3 - millet composition with enzymatic complex (ECV). The treatments were defined from six diets (3 reference diets and 3 test diets). The test diets were obtained from the substitution of 40% for reference diet by whole millet, and the enzyme addition consisted of two enzymatic complex, ECS constituted by phytase, protease, xylanase, ß-glucanase, cellulase, amylase and pectinase enzymes, and ECV constituted by protease, cellulase and amylase enzymes. In the 11 to 20 days phase, a supplementation with the ECV reduced the AME, AMEn and CDPB values, a ECS supplementation improved the CDPB, and there was no significant effect for CDMS and CDEB. In the 21 to 30 days phase, there were less profit of the energy and nutrients with ECS and ECV supplements. In the 31 to 40 days phase as supplements reduced the values of AME, AMEn, and the ECV complex was effective in increasing the value of CDPB. The inclusion of ECS enzymatic complexes, (phytase, protease, xylanase, ß-glucanase, cellulase, amylase and pectinase) and ECV (protease, cellulase and amylase), did not favor millet's energy utilization, however, favored the millet's protein digestibility coefficient on 11 to 20 and 31 to 40 periods.(AU)


Sujets)
Animaux , Mâle , Poulets , Enzymes , Millets , Aliment pour animaux/analyse , Digestion
17.
Gut and Liver ; : 569-575, 2019.
Article Dans Anglais | WPRIM | ID: wpr-763869

Résumé

BACKGROUND/AIMS: Cholangiocytes are capable of reabsorbing bile salts from bile, but the pathophysiological significance of this process is unclear. To this end, we detected the expression and distribution of bile acid transport proteins in cholangiocytes from normal rat liver and analyzed the possible pathophysiological significance. METHODS: Bile duct tissues of Sprague-Dawley rats were isolated by enzymatic digestion and mechanical isolation, and then divided into large and small bile duct tissues. Immunohistochemistry, real-time polymerase chain reaction and Western blotting were used to determine the expression of the apical sodium-dependent bile acid transporter (ASBT), ileal bile acid binding protein (IBABP), and basolateral organic solute transporter α (Ostα) in the biliary tract system of rats. Differences in the expression and distribution of these proteins were analyzed. RESULTS: In cholangiocytes, ASBT and IBABP were mainly expressed in cholangiocytes of the large bile ducts, in which the expression of both was significantly higher than that in the small ducts (p0.05). CONCLUSIONS: Bile acid transporters are expressed and heterogeneously distributed in rat bile ducts, indicating that bile acid reabsorption by cholangiocytes might mainly occur in the large bile ducts. These findings may help explore the physiology of bile ducts and the pathogenesis of various cholangiopathies.


Sujets)
Animaux , Rats , Acides et sels biliaires , Conduits biliaires , Bile , Voies biliaires , Technique de Western , Protéines de transport , Digestion , Immunohistochimie , Foie , Physiologie , Caractéristiques de la population , Rat Sprague-Dawley , Réaction de polymérisation en chaine en temps réel
18.
Mycobiology ; : 59-65, 2019.
Article Dans Anglais | WPRIM | ID: wpr-760526

Résumé

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.


Sujets)
Agrobacterium , Technique de Southern , Digestion , Génome , Génomique , Méthodes , Microscopie de fluorescence , Oxidoreductases , Réaction de polymérisation en chaîne , Réaction de polymérisation en chaine en temps réel , Plaies et blessures
19.
Journal of Veterinary Science ; : e67-2019.
Article Dans Anglais | WPRIM | ID: wpr-758954

Résumé

The intra-articular use of hyaluronic acid (HA) for the treatment of synovitis and osteoarthritis is still controversial. As a consequence, corticosteroids remain the most frequently employed therapeutic agents, despite their potential systemic and local deleterious effects. This study examined the anti-inflammatory, antioxidant, and chondroprotective activities of low and high molecular weight hyaluronic acid (LMW-HA and HMW-HA) on lipopolysaccharide (LPS)-induced synovitis in horses compared to triamcinolone acetonide (TA). LPS was injected in the metacarpophalangeal joints, which were treated intra-articularly with either TA (as control) or LMW-HA or HMW-HA. Joint clinical evaluation and synovial fluid (SF) analysis were performed at 0, 8, 24, and 48 h. The white blood cell counts (WBC), prostaglandin E2 (PGE2), interleukin (IL)-1, IL-6, IL-10, tumor necrosis factor-α, chondroitin sulfate (CS) and HA concentrations, oxidative burst, and HA molecular weights were measured. TA reduced the lameness, swelling, and PGE2 release but increased the SF CS concentrations enormously at 24h and 48h, and decreased the SF HA modal molecular weight. These results indicate the breakdown of articular cartilage aggrecan and SF HA. In contrast, LMW-HA and HMW-HA were less effective in reducing the inflammation symptoms, but preserved the joints because only a modest increase in CS occurred at 24 h, decreasing at 48 h, and the SF HA was maintained. The HA-treatment also had anti-inflammatory actions, and LMW-HA was the most effective in reducing the release of cytokine. In summary, the HA treatment inhibited efficiently the digestion of cartilage proteoglycans and SF HA breakdown.


Sujets)
Hormones corticosurrénaliennes , Agrécanes , Cartilage , Cartilage articulaire , Chondroïtines sulfate , Digestion , Dinoprostone , Equus caballus , Acide hyaluronique , Inflammation , Interleukine-10 , Interleukine-6 , Interleukines , Articulations , Numération des leucocytes , Articulation métacarpophalangienne , Masse moléculaire , Nécrose , Arthrose , Protéoglycanes , Stimulation du métabolisme oxydatif , Synovie , Synovite , Triamcinolone , Triamcinolone acétonide
20.
Korean Journal of Community Nutrition ; : 24-37, 2019.
Article Dans Coréen | WPRIM | ID: wpr-741039

Résumé

OBJECTIVES: The study purpose is to develop a content system for a healthy dietary education program for after-school care in lower grade in elementary school. METHODS: The contents of healthy dietary education in the 2015 revised curriculum and textbooks and the major education programs related to dietary life that are currently used in elementary school education were analyzed. Focus group interviews were held with field experts related to lower grade in elementary care class. Accordingly, the structuring of the education area and the detailed education contents were systematized. RESULTS: From the analysis results, the contents of curriculum, textbook, and administrative department were classified as hygiene safety, health, and culture. The goal of the educational content system was divided into three areas: nutritional dietary life, food hygiene and health, and food culture. The subjects consisted of dietary balance, healthy body weight, digestion and absorption, food hygiene, Korean agricultural products, traditional food, and table manners. The curriculum was composed of 12 content elements. CONCLUSIONS: In order to ensure that after-school care students can grow into healthy, growth-oriented and creative talents, the role of the caring guide is important, and associated guidelines are needed in the future.


Sujets)
Humains , Absorption , Aptitude , Poids , Programme d'études , Digestion , Éducation , Groupes de discussion , Hygiène
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