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1.
Molecules ; 26(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34443333

RESUMO

The latest data link the chronic consumption of large amounts of fructose present in food with the generation of hypertension and disturbances in carbohydrate and lipid metabolism, which promote the development of obesity, non-alcoholic fatty liver disease, insulin resistance, and type 2 diabetes. This effect is possible after fructose is absorbed by the small intestine cells and, to a lesser extent, by hepatocytes. Fructose transport is dependent on proteins from the family of glucose transporters (GLUTs), among which GLUT5 selectively absorbs fructose from the intestine. In this study, we examined the effect of four phenolic-rich extracts obtained from A. graveolens, B. juncea, and M. chamomilla on fructose uptake by Caco-2 cells. Extracts from B. juncea and M. chamomilla most effectively reduced fluorescent fructose analogue (NBDF) accumulation in Caco-2, as well as downregulated GLUT5 protein levels. These preparations were able to decrease the mRNA level of genes encoding transcription factors regulating GLUT5 expression-thioredoxin-interacting protein (TXNIP) and carbohydrate-responsive element-binding protein (ChREBP). Active extracts contained large amounts of apigenin and flavonols. The molecular docking simulation suggested that some of identified phenolic constituents can play an important role in the inhibition of GLUT5-mediated fructose transport.


Assuntos
Dieta , Frutose/metabolismo , Transportador de Glucose Tipo 5/metabolismo , Fenóis/análise , Extratos Vegetais/farmacologia , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Transporte Biológico/efeitos dos fármacos , Células CACO-2 , Proteínas de Transporte/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , RNA Mensageiro/genética
2.
Cells ; 10(3)2021 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-33809924

RESUMO

Numerous honeybee (Apis mellifera) products, such as honey, propolis, and bee venom, are used in traditional medicine to prevent illness and promote healing. Therefore, this insect has a huge impact on humans' way of life and the environment. While the population of A. mellifera is large, there is concern that widespread commercialization of beekeeping, combined with environmental pollution and the action of bee pathogens, has caused significant problems for the health of honeybee populations. One of the strategies to preserve the welfare of honeybees is to better understand and protect their natural microbiota. This paper provides a unique overview of the latest research on the features and functioning of A. mellifera. Honeybee microbiome analysis focuses on both the function and numerous factors affecting it. In addition, we present the characteristics of lactic acid bacteria (LAB) as an important part of the gut community and their special beneficial activities for honeybee health. The idea of probiotics for honeybees as a promising tool to improve their health is widely discussed. Knowledge of the natural gut microbiota provides an opportunity to create a broad strategy for honeybee vitality, including the development of modern probiotic preparations to use instead of conventional antibiotics, environmentally friendly biocides, and biological control agents.


Assuntos
Abelhas/microbiologia , Microbioma Gastrointestinal , Trato Gastrointestinal/microbiologia , Mel , Lactobacillales/metabolismo , Animais , Criação de Abelhas , Abelhas/efeitos dos fármacos , Abelhas/metabolismo , Disbiose , Interações Hospedeiro-Patógeno , Humanos , Inseticidas/efeitos adversos , Controle Biológico de Vetores , Probióticos
3.
Int J Mol Sci ; 21(14)2020 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-32664580

RESUMO

Bone mass loss occurs with a decrease in osteoblast proliferation and differentiation, or the enhancement of bone resorption, which further leads to the impairment of bone mineral density and increase in bone fracture. Recent studies suggest that some phenolic compounds found in food play positive role in bone metabolism. High content of phenolic compounds with potential beneficial effects on bone metabolism have been identified in the Viburnum opulus fruit. The aim of the study was to determine the influence of V. opulus fresh juice (FJ) and juice purified by solid phase extraction (PJ) on osteogenesis processes with osteosarcoma Saos-2 cell lines. V. opulus purified juice revealed stronger potential as an inducer of Saos-2 osteogenic differentiation. Saos-2 cells matrix mineralization was evaluated with alkaline phosphatase (ALP) activity measurement and alizarin red S staining. Gene expression analysis showed the elevation of the mRNA levels of Runt-related transcription factor 2 (RUNX2), ALP, collagen type 1 and osteonectin, whereas the nuclear factor-κB ligand and osteoprotegerin ratio (RANKL/OPG) decreased. Furthermore, V. opulus was able to diminish the secretion of pro-inflammatory cytokines Il6 and TNFα, however had no effect on vascular endothelial growth factor (VEGF). It decreased intracellular oxidative stress and induced DNA repair, but had no effect on the growth inhibition of lactic acid beneficial microorganisms.


Assuntos
Conservadores da Densidade Óssea/farmacologia , Neoplasias Ósseas/patologia , Sucos de Frutas e Vegetais/análise , Osteogênese/efeitos dos fármacos , Osteossarcoma/patologia , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Viburnum/química , Bactérias/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Reparo do DNA/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Microbioma Gastrointestinal/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-6/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/enzimologia , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Osteogênese/genética , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
4.
Biomolecules ; 10(5)2020 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-32384772

RESUMO

An effective and ecological method for liberation of pectin-derived oligosaccharides (POS) from sugar beet pulp (SBP) was developed using enzymatic and microorganism-mediated biomass conversion. The POS may be applied in the production of prebiotic feed additives. Various yeast strains were screened for their capacity for protein synthesis and monosaccharide assimilation. Combined yeast cultivation and pectin hydrolysis were found to be an effective method of producing prebiotics. Separate enzymatic hydrolysis and fermentation of SBP resulted in the release of 3.6 g of POS per 100 g d.w., whereas the yield of POS acquired after the combined process was 17.9% higher, giving 4.2 g of POS per 100 g d.w. Introducing the yeast into the process improved hydrolysis performance due to lower enzyme inhibition by mono- and disaccharides. The prebiotic effect of the POS was assessed by in vitro fermentation using individual cultures of gastrointestinal bacteria. The POS in the SBP hydrolysate effectively promoted the growth of lactobacilli and bifidobacteria. A large increase in adherence to Caco-2 cells in the presence of POS was noted for beneficial Lactobacillus brevis strains, whereas pathogenic bacteria and yeast (C. albicans, C. lusitanie, C. pelliculosa), responsible for infections in breeding animals, showed much weaker adhesion.


Assuntos
Ração Animal , Beta vulgaris/química , Candida/metabolismo , Fermentação , Pectinas/química , Prebióticos , Biomassa , Hidrólise , Microbiologia Industrial/métodos , Pectinas/metabolismo
5.
Molecules ; 25(9)2020 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-32403328

RESUMO

American ginseng, Panax quinquefolium (L.), is traditionally used in folk medicine. It exhibits a range of anti-inflammatory, hepatoprotective, anti-diabetic, anti-obesity, anti-hyperlipidemic and anti-carcinogenic effects. Its main components are ginsenosides, also known as panaxosides or triterpene saponins. In order to obtain high yields of ginsenosides, different methods of controlled production are involved, i.e., with hairy root cultures. However, they are still employed under in vitro conditions. Our studies revealed that hairy root cultures subjected to an elicitation process can be considered as a potent source of ginsenosides. The present study examines the biological activity of ginseng hairy root cultures against the Caco-2 human adenocarcinoma cell line. Among our six different clones of P. quinquefolium hairy roots, extracts B and Be (treated with elicitor) were the strongest inhibitors of the cellular metabolic activity. While all extracts induced DNA damage, B and Be also generated reactive oxygen species (ROS) in a concentration-dependent manner, which was correlated with the depletion of the mitochondrial membrane potential and induction of apoptosis. These findings indicate that further research concerning P. quinquefolium hairy root cultures should focus on the activity of rare ginsenosides and other biologically active compound profiles (i.e., phenolic compounds).


Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Dano ao DNA/efeitos dos fármacos , Ginsenosídeos/farmacologia , Panax/química , Raízes de Plantas/química , Adenocarcinoma/metabolismo , Trifosfato de Adenosina/metabolismo , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Células Clonais/química , Neoplasias do Colo/metabolismo , Humanos , Concentração Inibidora 50 , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Panax/metabolismo , Espécies Reativas de Oxigênio/metabolismo
6.
Antioxidants (Basel) ; 8(8)2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31374918

RESUMO

In recent years, there has been increasing interest in studying food-originated phytocompounds with beneficial influences for humans. Amongst the most active natural substances are polyphenols, for which high content has been identified in the Viburnum opulus berry, and which are unused in Western Europe. Due to its strong antioxidant activity we explored the potential of V. opulus as a preventive agent against diet-related chronic diseases, such as obesity and type 2 diabetes. Among the causes of these ailments is oxidative stress, as well as impaired glucose and free fatty acids (FFA) uptake. Thus, the purpose of this study was to determine biological activity of V. opulus phenolic extracts as cytoprotective agents able to decrease induced oxidative stress, lower lipid accumulation and attenuate glucose and FFA uptake by Caco-2 cells via GLUT2 and CD36/FAT transporters. To determine the source of the most biologically active phenolic compounds, we obtained four phenolic compounds extracts as crude juice, phenolics isolated from juice and two preparations of phenolics obtained with different extraction agents from fruit pomace. Among the studied extracts, the phenolic rich fraction obtained from fruit juice revealed the strongest activity to decrease uptake of glucose, FFA and accumulation of lipid droplets in Caco-2 cells without affecting their viability (IC0 50 µg/mL). Observed uptake attenuation was followed by decrease of the CD36/FAT gene expression, without influence on the GLUT2 and PPARα levels. We suspect that V. opulus phenolics were able to modulate the cellular membrane dynamic, although that hypothesis requires further, more detailed studies. Extracts revealed strong chemo-preventive activity against oxidative stress induced chemically by tert-butylhydroperoxide (t-BOOH), as well as against DNA damage through the induction of DNA repair after cell exposition to methylnitronitrosoguanidine (MNNG) and H2O2. Our findings suggest Viburnum opulus fruit as a dietary source of phytocompounds, which could be considered as a tailored design food supplement components for the prevention and treatment of postprandial elevation of glucose and fatty acids through delaying the rate of glucose and fatty acid absorption by intestinal cells.

7.
Nutrients ; 11(5)2019 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-31075951

RESUMO

Panax quinquefolium L. (American Ginseng, AG) is an herb characteristic for regions of North America and Asia. Due to its beneficial properties it has been extensively investigated for decades. Nowadays, it is one of the most commonly applied medical herbs worldwide. Active compounds of AG are ginsenosides, saponins of the glycosides group that are abundant in roots, leaves, stem, and fruits of the plant. Ginsenosides are suggested to be primarily responsible for health-beneficial effects of AG. AG acts on the nervous system; it was reported to improve the cognitive function in a mouse model of Alzheimer's disease, display anxiolytic activity, and neuroprotective effects against neuronal damage resulting from ischemic stroke in animals, demonstrate anxiolytic activity, and induce neuroprotective effects against neuronal damage in ischemic stroke in animals. Administration of AG leads to inhibition of hypertrophy in heart failure by regulation of reactive oxygen species (ROS) in mice as well as depletion of cardiac contractile function in rats. It also has an anti-diabetic and anti-obesity potential as it increases insulin sensitivity and inhibits formation of adipose tissue. AG displays anti-cancer effect by induction of apoptosis of cancer cells and reducing local inflammation. It exerts antimicrobial effects against several pathogenic strains of bacteria. Therefore, AG presents a high potential to induce beneficial health effects in humans and should be further explored to formulate precise nutritional recommendations, as well as to assess its value in prevention and therapy of some disorders, including cancer.


Assuntos
Ginsenosídeos/uso terapêutico , Panax/química , Compostos Fitoquímicos/uso terapêutico , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Anti-Infecciosos/farmacologia , Anti-Infecciosos/uso terapêutico , Doenças Cardiovasculares/tratamento farmacológico , Ginsenosídeos/farmacologia , Humanos , Doenças Metabólicas/tratamento farmacológico , Neoplasias/tratamento farmacológico , Doenças do Sistema Nervoso/tratamento farmacológico , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia
8.
Nutrients ; 11(2)2019 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-30720717

RESUMO

Schisandra chinensis (Turcz.) Baill. (SCE) is a plant with high potential for beneficial health effects, confirmed by molecular studies. Its constituents exert anti-cancer effects through the induction of cell cycle arrest and apoptosis, as well as inhibition of invasion and metastasis in cancer cell lines and experimental animals. SCE displays antimicrobial effects against several pathogenic strains. It has anti-diabetic potential, supported by hypoglycemic activity. A diet rich in SCE improves pancreatic functions, stimulates insulin secretion, and reduces complications in diabetic animals. SCE prevents lipid accumulation and differentiation of preadipocytes, indicating its anti-obesity potential. SCE exerts a protective effect against skin photoaging, osteoarthritis, sarcopenia, senescence, and mitochondrial dysfunction, and improves physical endurance and cognitive/behavioural functions, which can be linked with its general anti-aging potency. In food technology, SCE is applied as a preservative, and as an additive to increase the flavour, taste, and nutritional value of food. In summary, SCE displays a variety of beneficial health effects, with no side effects. Further research is needed to determine the molecular mechanisms of SCE action. First, the constituents responsible for its beneficial effects should be isolated and identified, and recommended as preventative nutritional additives, or considered as therapeutics.


Assuntos
Preparações de Plantas/farmacologia , Schisandra , Animais , Anti-Infecciosos/farmacologia , Fármacos Antiobesidade/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Conservantes de Alimentos , Humanos , Fitoterapia/métodos , Substâncias Protetoras/farmacologia
9.
Molecules ; 21(7)2016 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-27420041

RESUMO

The paper presents the chemical characteristics of ellagitannins isolated from raspberry (Rubus idaeus L.) fruit and their in vitro and in situ antifungal activity against Geotrichum candidum LOCK 0511. The study investigated a complex preparation containing various raspberry ellagitannins at a concentration of 86% w/w, as well as pure lambertianin C and sanguiin H-6. The ellagitannin preparation was obtained by extracting raspberry press cake and purifying the extract using Amberlite XAD resin, while individual compounds were isolated by means of preparative HPLC. The complex preparation was analyzed for the content of ellagitannins, anthocyanins, and flavan-3-ols using HPLC and LC-MS. The antifungal activity of the complex ellagitannin preparation and the isolated ellagitannins was determined for the strain Geotrichum candidum. The MIC and MFC values (10.0 mg/mL and 30.0 mg/mL, respectively) were found to be the same for lambertianin C, sanguiin H-6, and the complex ellagitannin preparation. The fungistatic activity of the studied ellagitannin preparation at a concentration of 10 mg/mL, as determined by the poisoned medium method, was 65.2% following 6 day incubation of Geotrichum candidum, with the linear growth rate of only 16.2 mm/day. The corresponding parameters for the control sample were 0% and 56 mm/day, respectively. The study demonstrated both in vitro and in situ antifungal activity of raspberry ellagitannins against Geotrichum candidum.


Assuntos
Antifúngicos/farmacologia , Produtos Biológicos/farmacologia , Frutas/química , Geotrichum/efeitos dos fármacos , Taninos Hidrolisáveis/farmacologia , Extratos Vegetais/farmacologia , Rubus/química , Antifúngicos/química , Produtos Biológicos/química , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Taninos Hidrolisáveis/química , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Extratos Vegetais/química
10.
J Environ Manage ; 170: 50-9, 2016 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-26797046

RESUMO

The aim of this study was to determine the effectiveness of microbial preparation and Yucca schidigera in the removal of odorous volatile compounds from poultry manure as well as to evaluate antimicrobial properties of these amendments. It was demonstrated that the combined treatment of poultry manure (PM) with the microbial preparation and Y. schidigera extract can reduce the concentration of odorants by 58%-73%, depending on the tested compound. When Y. schidigera extract and the microbial preparation were applied at a time interval of 48 h, the deodorization efficiency was improved by 6-24%. Furthermore, Y. schidigera extract has antimicrobial properties, which affect poultry manure hygienization. It was found that when the microbial preparation was enriched with Lactobacillus plantarum, it became insensitive to the antimicrobial properties of Y. schidigera.


Assuntos
Bactérias/metabolismo , Esterco , Odorantes , Microbiologia do Solo , Yucca/metabolismo , Animais , Biodegradação Ambiental , Contagem de Colônia Microbiana , Aves Domésticas
11.
Environ Toxicol Pharmacol ; 34(3): 895-904, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22995401

RESUMO

The aim of the study was to investigate the effects of beetroot juice fermented by Lactobacillus brevis 0944 and Lactobacillus paracasei 0920 (FBJ) on carcinogen induction of aberrant crypt foci (ACF) in rat colon. 2-Amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP) was used as carcinogen, which was administrated intragastrically at a dose of 10 µg/day, every day of the experiment. Additionally, we investigated the cytotoxicity and genotoxicity of fecal water from experimental animals in the Caco-2 cell line, evaluated by MTT test and the comet assay, respectively, as well as by the count of bacteria adhered to colon epithelium assessed by fluorescence in situ hybridization. Oxidative stress in rats was expressed by measuring serum antioxidant status and the level of malondialdehyde in the kidneys and liver. The experimental rats were divided into four groups based on diet type: basal diet, basal diet supplemented with FBJ, basal diet and PhIP treatment, and basal diet supplemented with FBJ and PhIP treatment. FBJ significantly reduced the number of ACF in PhIP-treated rats (from 59 ± 18 to 26 ± 4). Moreover, the number of extensive aberrations (more than 4 crypts in a focus) decreased from 52 ± 18 to 18 ± 4. Fecal water obtained from rats fed with a PhIP-containing diet induced pronounced cytotoxic and genotoxic effects in Caco-2 cells, but FBJ supplementation of the diet abolished these effects. In groups fed dietary PhP and FBJ the latter was found to increase the antioxidant status of serum from 40% to 66% depending on the fraction. Reduced concentration of malondialdehyde was found only in the kidneys of rats fed with PhIP and FBJ. FBJ present in the diet of rats causes a reduction of MDA in the kidneys from 118.7 nmol/g tissue to 100 nmol/g tissue. The presence of FBJ in the diet of rats significantly increased the count of bacteria, including Lactobacillus/Enterococcus and Bacteroides-Prevotella group adhered to colonic epithelium. In conclusion, supplementation of the diet with lactofermented beetroot juice may provide protection against precancerous aberrant crypt formation and reduce the cytotoxic and genotoxic effects of fecal water and improve the oxidative status of the organism.


Assuntos
Focos de Criptas Aberrantes/metabolismo , Antioxidantes/farmacologia , Beta vulgaris , Carcinógenos/toxicidade , Extratos Vegetais/farmacologia , Animais , Células CACO-2 , Ensaio Cometa , Fezes , Fermentação , Humanos , Imidazóis/toxicidade , Masculino , Mutagênicos/toxicidade , Estresse Oxidativo , Ratos
12.
Exp Toxicol Pathol ; 64(6): 599-604, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21185162

RESUMO

The aim of the study was to investigate the effects of beetroot juice fermented by Lactobacillus brevis 0944 and Lactobacillus paracasei 0920 (FBJ) on carcinogen induction of aberrant crypt foci (ACF) in rat colon. N-Nitroso-N-methylurea (MNU) was used as carcinogen, which was administrated intragastrically at a dose of 50 mg/kg on the 23rd and 26th day of the experiment. Additionally, we investigated the cytotoxicity and genotoxicity of fecal water from experimental animals in the Caco 2 cell line, evaluated by MTT/NRU tests and the comet assay, respectively, as well as by the count of bacteria adhered to colon epithelium assessed by fluorescence in situ hybridization and DAPI staining. The experimental rats were divided into four groups based on diet type: basal diet, basal diet supplemented with FBJ, basal diet and MNU treatment, and basal diet supplemented with FBJ and MNU treatment. FBJ significantly reduced the number of ACF in MNU-treated rats (from 55±18 to 21±6). Moreover, the number of extensive aberrations (more than 4 crypts in a focus) decreased from 45±21 to 7±4. Fecal water obtained from rats fed with an MNU-containing diet induced pronounced cytotoxic and genotoxic effects in Caco 2 cells, but FBJ supplementation of the diet abolished these effects. The presence of FBJ in the diet significantly increased the count of bacteria, including Lactobacillus/Enterococcus, adhered to colonic epithelium. In conclusion, supplementation of the diet with lactofermented beetroot juice may provide protection against precancerous aberrant crypt formation and reduce the cytotoxic and genotoxic effects of fecal water.


Assuntos
Focos de Criptas Aberrantes/prevenção & controle , Beta vulgaris , Neoplasias Colorretais/prevenção & controle , Lactobacillus , Extratos Vegetais , Probióticos/farmacologia , Focos de Criptas Aberrantes/induzido quimicamente , Focos de Criptas Aberrantes/microbiologia , Animais , Carcinógenos/toxicidade , Colo/efeitos dos fármacos , Colo/microbiologia , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/microbiologia , Ensaio Cometa , Modelos Animais de Doenças , Fezes/química , Fermentação , Hibridização in Situ Fluorescente , Masculino , Metilnitrosoureia/toxicidade , Testes de Mutagenicidade , Raízes de Plantas , Ratos , Ratos Wistar
13.
Res Vet Sci ; 89(3): 391-5, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20435327

RESUMO

The aim of the study was to evaluate the influence of a probiotic preparation on the genotoxicity of faecal water of broiler chickens fed with a fodder contaminated with aflatoxin B(1) (AFB(1)) at 1 or 5mg per kg. Human blood lymphocytes were exposed to chicken's faecal water samples and DNA damage was measured using the comet assay. Genotoxicity of faecal water did not depend on the AFB(1) concentration in the fodder. The mean DNA damage, measured as the percentage of DNA in the tail of the comets, for chickens fed with fodder with AFB(1) at 1 mg/kg was 16.80±0.66, at 5 mg/kg - 16.73±1.51 and in the controls - 12.79±0.66. The supplementation of fodder with the probiotic preparation decreased the extent of DNA damage to 10.02±0.39 for 1 mg/kg AFB(1) and to 11.89±0.72 for 5 mg/kg.


Assuntos
Aflatoxina B1/toxicidade , Ração Animal/efeitos adversos , Galinhas/fisiologia , Doenças das Aves Domésticas/prevenção & controle , Probióticos/uso terapêutico , Ração Animal/microbiologia , Criação de Animais Domésticos/métodos , Animais , Ensaio Cometa/veterinária , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/genética , Suplementos Nutricionais , Fezes/microbiologia , Feminino , Humanos , Linfócitos/efeitos dos fármacos , Testes de Mutagenicidade/veterinária
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