RESUMO
Four Gram-stain-positive bacterial strains were isolated from the gut of honeybee (Apis mellifera) in China. These strains were characterized using a polyphasic taxonomic approach. The data demonstrated that three of the four strains represented two novel species of the genus Lactobacillus, strains F306-1T and F551-2T were designated as the type strains. Results of 16S rRNA gene sequence analysis indicated that strains F306-1T, F447 and F551-2T were phylogenetically related to the type strains of Lactobacillus kimbladii and Lactobacillus kullabergensis, having 99.1-99.7â% 16S rRNA gene sequence (about 1400 bp) similarities. The phylogenetic tree based on concatenated pheS, rpoA, gyrB, hsp60, recA, rpoB and tuf sequences (4114 bp) and the phylogenomic tree based on whole genome sequences indicated that strains F306-1T and F447 were most closely related to L. kullabergensis Biut2NT, and strain F551-2T was most closely related to L. kimbladii Hma2NT. Strains F306-1T and F447 shared 99.9â% average nucleotide identity (ANI), 99.7â% digital DNA-DNA hybridization (dDDH) and 99.9â% average amino acid identity (AAI) values, indicating that they belong to the same species. Strain F306-1T exhibited the highest ANI (94.4â%), dDDH (56.7â%) and AAI (94.7â%) values to L. kullabergensis Biut2NT. Strain F551-2T had the highest ANI (94.0â%), dDDH (54.3â%) and AAI (95.8â%) values with L. kimbladii Hma2NT. Acid production from amygdalin, maltose, starch, gentiobiose and turanose, activity of esterase (C4) and α-glucosidase, growth with 3â% NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth with 1-6% NaCl at 37 °C under aerobic condition (on mMRS agar plates supplemented with 0.05â% cysteine or with 1â% cysteine and 2â% fructose) could differentiate strains F306-1T and F447 from L. kullabergensis DSM 26262T. Acid production from d-glucose, arbutin and gentiobiose, growth with 3â% NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth at 45 °C under strict anaerobic condition (on mMRS agar plates) could differentiate strain F551-2T from L. kimbladii DSM 26263T. Based upon the data obtained in the present study, two novel species, Lactobacillus huangpiensis sp. nov. and Lactobacillus laiwuensis sp. nov., are proposed and the type strains are F306-1T (=LMG 32144T=JCM 34361T=CCTCC AB 2020300T) and F551-2T (=JCM 34502T=CCTCC AB 2021027T), respectively.
Assuntos
Abelhas/microbiologia , Lactobacillus/classificação , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Abstract This study assessed the inhibitory potential of the probiotics Lactobacillus (LB) exopolysaccharides (EPS) with or without extracts of Satureja calamintha on enteropathogenic Escherichia coli (EPEc) responsible for gastroenteritis. Methanolic and hydromethanolic extracts were prepared by cold maceration and subjected to phytochemical screening. The compounds of the extracts were determined with the colorimetric assays and identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Antioxidant activities of the extracts were also evaluated by using 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging. Antibacterial effect on EPEc was evaluated by using both agar disc diffusion and microdilution methods. The in vitro test of auto-aggregation was investigated. Microbiological analysis showed that 63% of the isolated LB were producing EPS, with the amount ranging from 8.21 to 43.13 mg/L. Chemical analysis of the extracts revealed the presence of polyphenols and flavonoids, more abundant in the hydromethanolic extract, which presented the highest content with 2.11 mg EGA/g of polyphenol and 1.64 mg EC/g of flavonoids and 1.71 mg EGA/g of polyphenol and 1.15 mg EC/g of flavonoids in the methanolic extract. Hydromethanolic extracts and EPS exhibited a more important activity than did the methanolic extract against EPEc. The combined action of EPS and extracts reduced the aggregation ability of EPEc and decreased the rate of their adhesion.
Assuntos
Probióticos/efeitos adversos , Satureja/efeitos adversos , Escherichia coli Enteropatogênica/classificação , Lactobacillus/classificação , Extratos Vegetais/análise , Cromatografia Líquida de Alta Pressão/métodos , Nepeta/efeitos adversos , Compostos Fitoquímicos , Gastroenterite , Antioxidantes/farmacologiaRESUMO
BACKGROUND: Previous studies have demonstrated the acrylamide-removing properties of probiotic monocultures; however, potential advantages of consortia over monocultures in reducing the dietary exposure to acrylamide have not been proven. Hence this work aims to assess the acrylamide (AA)-binding properties of bacterial consortia, consisting of either probiotic strains and / or representative bacteria of duodenal microbiota, exposed to simulated gastrointestinal conditions (SGC). The AA binding capacity of ten probiotic strains (PS) and six duodenal strains (NDS) was evaluated under different conditions; then, three different consortia (PS, NDS, and PS + NDS) were assessed under SGC. RESULTS: Among individual PS, Bacillus coagulans GBI-30, Lactobacillus fermentum J23, L. pentosus J37 and J24, and L. casei Shirota, exhibited the highest AA-binding capacity (80-87%), while Bifidobacterium catenulatun ATCC27676, Streptococcus salivarius subsp. thermophilus ATCC19258, and S. gallolyticus ATCC9809 were the best (ca. 68%) NDS monocultures. Probiotic strain consortia showed higher (P < 0.05) AA binding capacity (> 90%) than monoculture bacteria. Conversely, individual NDS cultures displayed higher (P < 0.05) binding capacity than NDS consortia (60%). A significant reduction (P < 0.05) in AA removal capacity was observed when consortia were exposed to SGC, PS consortia being the most effective (> 60% removal). CONCLUSION: These results suggest that consortia of specific PS could play an important role in reducing the intestinal availability of acrylamide. © 2021 Society of Chemical Industry.
Assuntos
Acrilamida/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/microbiologia , Lactobacillus/metabolismo , Probióticos/farmacologia , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Avaliação Pré-Clínica de Medicamentos , Trato Gastrointestinal/metabolismo , Humanos , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Viabilidade Microbiana , Modelos BiológicosRESUMO
Human vagina is colonised by a diverse array of microorganisms that make up the normal microbiota and mycobiota. Lactobacillus is the most frequently isolated microorganism from the healthy human vagina, this includes Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners, and Lactobacillus jensenii. These vaginal lactobacilli have been touted to prevent invasion of pathogens by keeping their population in check. However, the disruption of vaginal ecosystem contributes to the overgrowth of pathogens which causes complicated vaginal infections such as bacterial vaginosis (BV), sexually transmitted infections (STIs), and vulvovaginal candidiasis (VVC). Predisposing factors such as menses, pregnancy, sexual practice, uncontrolled usage of antibiotics, and vaginal douching can alter the microbial community. Therefore, the composition of vaginal microbiota serves an important role in determining vagina health. Owing to their Generally Recognised as Safe (GRAS) status, lactobacilli have been widely utilised as one of the alternatives besides conventional antimicrobial treatment against vaginal pathogens for the prevention of chronic vaginitis and the restoration of vaginal ecosystem. In addition, the effectiveness of Lactobacillus as prophylaxis has also been well-founded in long-term administration. This review aimed to highlight the beneficial effects of lactobacilli derivatives (i.e. surface-active molecules) with anti-biofilm, antioxidant, pathogen-inhibition, and immunomodulation activities in developing remedies for vaginal infections. We also discuss the current challenges in the implementation of the use of lactobacilli derivatives in promotion of human health. In the current review, we intend to provide insights for the development of lactobacilli derivatives as a complementary or alternative medicine to conventional probiotic therapy in vaginal health.
Assuntos
Lactobacillus/metabolismo , Microbiota , Vagina/microbiologia , Vagina/fisiologia , Terapias Complementares/métodos , Feminino , Humanos , Lactobacillus/classificação , Probióticos/administração & dosagem , Vaginose Bacteriana/microbiologia , Vaginose Bacteriana/prevenção & controle , Vaginose Bacteriana/terapiaRESUMO
Maternal milk is an important source of essential nutrients for the optimal growth of infants. Breastfeeding provides a continuous supply of beneficial bacteria to colonize the infant gastrointestinal tract (GIT) and offers health benefits for disease prevention and immunity. The purpose of this study was to isolate novel probiotic strains from the breast milk of native Pakistani mothers and to evaluate their probiotic potential. We isolated 21 strains of bacteria from the colostrum and mature milk of 20 healthy mothers, who had vaginal deliveries and were not taking antibiotics. After phenotypic and genotypic characterization, these isolates were tested for survival in the GIT using in vitro acid and bile tests. Nine strains showing good acid tolerance were assessed for their growth rate, bile resistance and ability to hydrolyze bile salts. Out of the four Lactobacillus isolates adjudged to be most promising as probiotics, three were Lactobacillus fermentum strains and one was a strain of Lactobacillus oris. This study demonstrates that human milk is a viable source of commensal bacteria beneficial to both adults and babies.
Assuntos
Lactobacillus/fisiologia , Leite Humano/microbiologia , Probióticos , Ácidos/metabolismo , Adulto , Ácidos e Sais Biliares/metabolismo , Colostro/microbiologia , Feminino , Trato Gastrointestinal/microbiologia , Humanos , Lactente , Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Filogenia , Gravidez , Probióticos/classificação , Probióticos/isolamento & purificação , Adulto JovemRESUMO
The proper development of the early gastrointestinal tract (GIT) microbiota is critical for newborn ruminants. This microbiota is susceptible to modification by diverse external factors (such as diet) that can lead to long-lasting results when occurring in young ruminants. Dietary supplementation with prebiotics, ingredients nondigestible and nonabsorbable by the host that stimulate the growth of beneficial GIT bacteria, has been applied worldwide as a potential approach in order to improve ruminant health and production yields. However, how prebiotics affect the GIT microbiota during ruminants' early life is still poorly understood. We investigated the effect of milk supplementation with a combination of two well-known prebiotics, fructooligosaccharides (FOS) from sugar beet and garlic residues (all together named as "additive"), exerted on preweaned lamb growth and the composition of their fecal microbiota, by using 16S rRNA gene amplicon high-throughput sequencing. The results showed a significant increase in the mean daily weight gain of lambs fed with the additive. Lamb fecal microbiota was also influenced by the additive intake, as additive-diet lambs showed lower bacterial diversity and were significantly more abundant in Bifidobacterium, Enterococcus, Lactobacillus and Veillonella. These bacteria have been previously reported to confer beneficial properties to the ruminant, including promotion of growth and health status, and our results showed that they were strongly linked to the additive intake and the increased weight gain of lambs. This study points out the combination of FOS from sugar beet and garlic residues as a potential prebiotic to be used in young ruminants' nutrition in order to improve production yields.
Assuntos
Bactérias/classificação , Beta vulgaris/química , Peso Corporal/efeitos dos fármacos , Alho/química , Oligossacarídeos/administração & dosagem , Extratos Vegetais/administração & dosagem , Animais , Animais Recém-Nascidos , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , Bifidobacterium/classificação , Bifidobacterium/genética , Bifidobacterium/crescimento & desenvolvimento , Bifidobacterium/isolamento & purificação , Suplementos Nutricionais , Microbioma Gastrointestinal , Sequenciamento de Nucleotídeos em Larga Escala , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Leite/química , Oligossacarídeos/farmacologia , Extratos Vegetais/farmacologia , Prebióticos/administração & dosagem , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ovinos , Veillonella/classificação , Veillonella/genética , Veillonella/crescimento & desenvolvimento , Veillonella/isolamento & purificaçãoRESUMO
OBJECTIVES: Lactobacillus probiotics have been proposed as an antibiotic-sparing prevention strategy for urinary tract infections (UTIs). Our objective was to examine the relative ability of the 4 most common vaginal Lactobacillus species to inhibit the growth of Escherichia coli, the most common cause of UTIs. METHODS: Conditioned media (CM) was created from 4 laboratory strains of Lactobacillus species: Lactobacillus crispatus, Lactobacillus jensenii, Lactobacillus gasseri, and Lactobacillus iners, and 6 clinical strains of L. crispatus. One laboratory strain of E. coli was cocultured with each CM, as well as with various acidic solutions. Three clinical strains of E. coli from women with acute cystitis were cocultured with the CM from a laboratory strain of L. crispatus. Bacterial growth was compared between experimental variants and media control using analysis of variance. RESULTS: Growth of E. coli was inhibited by CM from L. crispatus, L. jensenii, and L. gasseri, but not L. iners. The magnitude of inhibition was correlated with the pH of the individual CM and the concentration of D-lactic acid. Different acids inhibited E. coli growth in proportion to the pH of the acid solution. Similar levels of inhibition were seen when L. crispatus was incubated with clinical E. coli strains as with laboratory E. coli. CONCLUSIONS: Three of the most common vaginal Lactobacillus species inhibit E. coli growth, likely through creating a low pH environment. However, L. iners, one of the most common species found after menopause, does not. These findings might be leveraged to more effectively manage UTIs.
Assuntos
Escherichia coli , Concentração de Íons de Hidrogênio , Lactobacillus , Probióticos , Vagina , Adulto , Fatores Etários , Técnicas Bacteriológicas/métodos , Correlação de Dados , Suplementos Nutricionais , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Feminino , Humanos , Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Menopausa/fisiologia , Pessoa de Meia-Idade , Probióticos/administração & dosagem , Probióticos/efeitos adversos , Infecções Urinárias/prevenção & controle , Vagina/química , Vagina/microbiologiaRESUMO
Structure of lactic acid bacteria biota in ivy flowers, fresh bee-collected pollen (BCP), hive-stored bee bread, and honeybee gastrointestinal tract was investigated. Although a large microbial diversity characterized flowers and fresh BCP, most of lactic acid bacteria species disappeared throughout the bee bread maturation, giving way to Lactobacillus kunkeei and Fructobacillus fructosus to dominate long stored bee bread and honeybee crop. Adaptation of lactic acid bacteria was mainly related to species-specific, and, more in deep, to strain-specific features. Bee bread preservation seemed related to bacteria metabolites, produced especially by some L. kunkeei strains, which likely gave to lactic acid bacteria the capacity to outcompete other microbial groups. A protocol to ferment BCP was successfully set up, which included the mixed inoculum of selected L. kunkeei strains and Hanseniaspora uvarum AN8Y27B, almost emulating the spontaneous fermentation of bee bread. The strict relationship between lactic acid bacteria and yeasts during bee bread maturation was highlighted. The use of the selected starters increased the digestibility and bioavailability of nutrients and bioactive compounds naturally occurring in BCP. Our biotechnological protocol ensured a product microbiologically stable and safe. Conversely, raw BCP was more exposed to the uncontrolled growth of yeasts, moulds, and other bacterial groups.
Assuntos
Abelhas/microbiologia , Microbiologia de Alimentos , Pólen/metabolismo , Pólen/microbiologia , Própole/metabolismo , Animais , Anti-Infecciosos , Fermentação , Flores/microbiologia , Trato Gastrointestinal/microbiologia , Hanseniaspora/metabolismo , Hedera , Lactobacillales/classificação , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/isolamento & purificação , Lactobacillales/metabolismo , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Interações Microbianas , Microbiota , Pólen/química , Especificidade da EspécieRESUMO
BACKGROUND: Sagunja-tang (SGT) is widely used in traditional herbal medicine to treat immune system and gastrointestinal disorders and reportedly has protective effects against inflammation, cancer, and osteoporosis. In this study, we fermented SGT with different Latobacillus strains and investigated the change in phytochemical compositions in SGT and enhancement of it neuroprotective effects in SH-SY5Y human neuroblastoma. METHODS: Marker components, including ginsenoside Rg1, glycyrrhizin, liquiritin, liquiritigenin, atractylenolide I, atractylenolide II, atractylenolide III, and pachymic acid, in SGT, were qualitatively and quantitatively analyzed using high-performance liquid chromatography-diode array detection and liquid chromatography-mass spectrometry. SGT was fermented with eight different Lactobacillus strains to yield eight fermented SGTs (FSGTs). The conversion efficiencies of SGT marker components were determined in each FSGT. To detect the protective effect of SGT and FSGT, reactive oxygen species (ROS) assay and mitochondrial membrane potentials (MMPs) assay were performed in SH-SY5Y cells. RESULTS: Compared with the other FSGTs, SGT166, i.e., SGT fermented with L. plantarum 166, had high conversion efficiency, as indicated by increased amounts of glycyrrhizin, liquiritigenin, and atractylenolides I-III. In SH-SY5Y cells, protection against cell death induced by H2O2 and etoposide was high using SGT166 and very low using SGT. Furthermore, ROS production and mitochondrial membrane potential disruption in SH-SY5Y cells were markedly suppressed by SGT166 treatment, which demonstrated that inhibition of ROS generation may be one of the neuroprotective mechanisms of SGT166. CONCLUSIONS: This study demonstrated that fermentation of SGT with L. plantarum 166 enhanced suppression of oxidative stress and MMP loss. This enhanced neuroprotective effect was thought to be caused by the conversion of SGT phytochemicals by fermentation. SGT166 shows potential for treating neurological damage-related diseases.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Lactobacillus/metabolismo , Fármacos Neuroprotetores/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/metabolismo , Fermentação , Humanos , Lactobacillus/classificação , Espectrometria de Massas , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Nowadays, the strong relationship between diet and health is well known. Although the primary role of diet is to provide nutrients to fulfill metabolic requirements, the use of foods to improve health and the state of well-being is an idea increasingly accepted by society in the last three decades. During the last years, an important number of scientific advances have been achieved in this field and, although in some situations, it is difficult to establish a distinction between "harmful" and "good" bacteria, experts agree in classifying the genera Bifidobacterium and Lactobacillus as beneficial bacteria. Thus, several strategies can be used to stimulate the proliferation of these beneficial intestinal bacteria, being one of them the consumption of prebiotics. The development of new prebiotics, with added functionality, is one of the most serious challenges shared not only by the scientific community but also by the food industry. The objective of this work was to evaluate the potential prebiotic effect of red and white grape residues, both obtained during the winemaking process. For such purpose, an in vitro study with pure cultures of Lactobacillus and Bifidobacterium was first conducted. Secondly, a study with mixed cultures using human fecal inocula was carried out in a simulator of the distal part of the colon. The obtained results showed an increase in the Lactobacillus and Bifidobacterium population, indicating that these ingredients are serious candidates to be considered as prebiotics.
Assuntos
Microbioma Gastrointestinal , Modelos Biológicos , Pigmentos Biológicos/metabolismo , Componentes Aéreos da Planta/química , Extratos Vegetais/metabolismo , Prebióticos , Vitis/química , Adulto , Animais , Bifidobacterium/classificação , Bifidobacterium/crescimento & desenvolvimento , Bifidobacterium/isolamento & purificação , Bifidobacterium/metabolismo , Digestão , Fezes/microbiologia , Feminino , Fermentação , Indústria de Processamento de Alimentos/economia , Humanos , Resíduos Industriais/análise , Resíduos Industriais/economia , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Masculino , Viabilidade Microbiana , Tipagem Molecular , Componentes Aéreos da Planta/metabolismo , Extratos Vegetais/economia , Prebióticos/economia , Espanha , Vitis/metabolismoRESUMO
The aim of this study is to select and isolate autochthonous bacteria with probiotic potential for use in a supplemented diet for bullfrog tadpoles, Lithobates catesbeianus. A total of 20 strains of lactic acid bacteria were isolated. Nine out of these were used in the following in vitro assays: antagonism against pathogenic bacteria (ANT), antimicrobial activity from extracellular compounds (MIC), tolerance to bile salts (TBS), pH reduction, protease production, sensitivity to antimicrobial tetracycline, cell viability, growth rate and doubling time. Using these data was defined an ideotype (ideal strain) based on the best results. Distances were estimated with the Mahalanobis (D2) test, and the best candidates, presenting the shortest ideotype distances, were considered to be used. The best strain was found to be Lactobacillus plantarum because it presented 10.00 ± 0.50 mm of ANT against Aeromonas hydrophila, 3.99 ± 0.01 of MIC independent of pathogenic bacteria, 85.07 ± 0.01 of TBS, 4.20 ± 0.02 of final pH, 17.67 ± 1.15 of protease production, 13.50 ± 2.00 sensitivity to antimicrobial tetracycline, 9.36 ± 0.04 of cell viability, 0.20 ± 0.00 of growth rate and 3.46 ± 0.00 doubling time. Therefore this probiotic candidate was then supplemented (2.045 ± 1.07 × 107 colony forming unities. g-1) into the diets of bullfrog tadpoles for a period of 42 days. At the end of the trial, samples of blood and intestines were collected to verify the haematological alterations and the intestinal morphology using transmission and scanning electron microscopy. Tadpoles fed the supplemented diet showed successful lactic acid bacterium colonisation, an increased number of circulating thrombocytes, monocytes, eosinophil and LG-PAS+ and also an increase in the length and density of intestinal microvilli. This study shows the feasibility of using probiotics isolated from farmed bullfrogs as a supplement in the diets of tadpoles, providing a promising alternative for modulating the health of these animals.
Assuntos
Larva/metabolismo , Probióticos/administração & dosagem , Rana catesbeiana/microbiologia , Ração Animal/análise , Animais , Suplementos Nutricionais/análise , Hematologia , Intestinos/crescimento & desenvolvimento , Intestinos/microbiologia , Intestinos/ultraestrutura , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Larva/crescimento & desenvolvimento , Larva/microbiologia , Larva/ultraestrutura , Microscopia Eletrônica , Rana catesbeiana/sangue , Rana catesbeiana/crescimento & desenvolvimentoRESUMO
We present a 65-year-old diabetic patient with a complex liver abscess and bacteraemia from Lactobacillus paracasei The abscess resulted in a prolonged hospital stay due to ongoing sepsis despite ultrasound-guided drainage and broad-spectrum antibiotics. Furthermore, the patient developed several secondary complications including a right-sided pleural effusion, an inferior vena cava thrombus and septic lung emboli. The abscess was eventually managed successfully with a prolonged course of antibiotics and multiple ultrasound-guided drainage procedures.To our knowledge, this is the first reported case of probiotic consumption, confirmed by strain identification, as the likely source of a liver abscess. Probiotic products have been widely used for many years and are advocated to the general public for their health benefits with no warning of side effects. Lactobacilli are one group of bacteria commonly used in these products. Although rare, complications have been reported. Susceptible patients, such as those who are immunocompromised, should be advised against excessive consumption.
Assuntos
Hospedeiro Imunocomprometido/imunologia , Lactobacillus/patogenicidade , Abscesso Hepático/microbiologia , Probióticos/efeitos adversos , Sepse/microbiologia , Idoso , Antibacterianos/uso terapêutico , Contagem de Colônia Microbiana , Drenagem , Feminino , Fluconazol/uso terapêutico , Humanos , Lactobacillus/classificação , Lactobacillus/imunologia , Lactobacillus/isolamento & purificação , Letargia , Linezolida/uso terapêutico , Abscesso Hepático/etiologia , Metronidazol/uso terapêutico , Testes de Sensibilidade Microbiana , Derrame Pleural/microbiologia , Sepse/etiologia , Resultado do Tratamento , Trombose Venosa/microbiologiaRESUMO
The aim of this study was to investigate the effects of dietary supplementation with two alternatives to antibiotics (Candida tropicalis and mulberry leaf flavonoids) on intestinal microbiota of preweaned calves challenged with Escherichia coli K99. Sixty Holstein calves were randomly assigned to 5 treatments: fed a basal diet (N-CON); fed a basal diet and challenged with E.coli K99 (P-CON); fed a basal diet supplemented with C.tropicalis (CT), mulberry leaf flavonoids (MLF), and the combination of the two additives (CM), respectively, and challenged with E.coli K99. The MLF and CM groups had significantly higher average daily grain and feed efficiency, and significantly lower fecal scores compared with the P-CON group after E. coli K99 challenge. The supplementation groups increased the relative abundance, at the phylum level, of Bacteroidetes and Proteobacteria, whereas at the genus level, they increased the relative abundance of Prevotella, Lactobacillus, and Enterococcus. Quantitative PCR revealed that the CT, MLF, and CM groups had significantly lower copy numbers of E.coli K99 compared with the P-CON group. The CT, MLF, and CM treatments reduce days of diarrhea, improve intestinal health, and beneficially manipulate the intestinal microbiota in preweaned calves.
Assuntos
Candida tropicalis/fisiologia , Doenças dos Bovinos/dietoterapia , Diarreia/veterinária , Suplementos Nutricionais , Infecções por Escherichia coli/veterinária , Flavonoides/administração & dosagem , Morus/química , Ração Animal/análise , Animais , Antibiose/genética , Técnicas de Tipagem Bacteriana , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/patologia , Diarreia/dietoterapia , Diarreia/microbiologia , Diarreia/patologia , Enterococcus/classificação , Enterococcus/genética , Enterococcus/isolamento & purificação , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/patogenicidade , Infecções por Escherichia coli/dietoterapia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Fezes/microbiologia , Microbioma Gastrointestinal/genética , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Masculino , Folhas de Planta/química , Reação em Cadeia da Polimerase , Prevotella/classificação , Prevotella/genética , Prevotella/isolamento & purificação , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , DesmameRESUMO
The aim of this study was to characterise the gut microbiota composition of piglets fed bovine colostrum (BC), milk replacer (MR) or sow milk (SM) in the post-weaning period. Piglets (n 36), 23-d old, were randomly allocated to the three diets. Faecal samples were collected at 23, 25, 27 and 30 d of age. Digesta from the stomach, ileum, caecum and mid-colon was collected at 30 d of age. Bacterial DNA from all samples was subjected to amplicon sequencing of the 16S rRNA gene. Bacterial enumerations by culture and SCFA analysis were conducted as well. BC-piglets had the highest abundance of Lactococcus in the stomach (P<0·0001) and ileal (P<0·0001) digesta, whereas SM-piglets had the highest abundance of Lactobacillus in the stomach digesta (P<0·0001). MR-piglets had a high abundance of Enterobacteriaceae in the ileal digesta (P<0·0001) and a higher number of haemolytic bacteria in ileal (P=0·0002) and mid-colon (P=0·001) digesta than SM-piglets. BC-piglets showed the highest colonic concentration of iso-butyric and iso-valeric acid (P=0·02). Sequencing and culture showed that MR-piglets were colonised by a higher number of Enterobacteriaceae, whereas the gut microbiota of BC-piglets was characterised by a change in lactic acid bacteria genera when compared with SM-piglets. We conclude that especially the ileal microbiota of BC-piglets had a closer resemblance to that of SM-piglets in regard to the abundance of potential enteric pathogens than did MR-piglets. The results indicate that BC may be a useful substitute for regular milk replacers, and as a feeding supplement in the immediate post-weaning period.
Assuntos
Ração Animal/efeitos adversos , Colostro , Dieta/veterinária , Disbiose/veterinária , Microbioma Gastrointestinal , Sus scrofa/microbiologia , Doenças dos Suínos/prevenção & controle , Animais , Bovinos , Cruzamentos Genéticos , Dinamarca , Dieta/efeitos adversos , Disbiose/etiologia , Disbiose/microbiologia , Disbiose/prevenção & controle , Enterobacteriaceae/classificação , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/metabolismo , Ácidos Graxos Voláteis/metabolismo , Fezes/microbiologia , Conteúdo Gastrointestinal/química , Conteúdo Gastrointestinal/microbiologia , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Lactococcus/classificação , Lactococcus/crescimento & desenvolvimento , Lactococcus/isolamento & purificação , Lactococcus/metabolismo , Tipagem Molecular , Especificidade de Órgãos , Distribuição Aleatória , Sus scrofa/crescimento & desenvolvimento , Sus scrofa/imunologia , Suínos , Doenças dos Suínos/etiologia , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , DesmameRESUMO
The aim of this study was to perform the microbiological analysis of quality of 25 probiotic products, available on the Polish market. Analysis of bacterial viability in probiotic products showed that not all of these preparations possess a suitable number of bacteria. Moreover, some of the tested probiotic products contained bacterial strains other than those declared by the manufacturer. All tested strains recovered from probiotic products were found to be resistant to metronidazole and susceptible to nitrofurantoin. The susceptibility to other antibiotics was strain specific. Probiotic products should be subject to regular and thorough inspection by appropriate institutions.
Assuntos
Bifidobacterium/isolamento & purificação , Lactobacillus/isolamento & purificação , Probióticos , Carga Bacteriana , Técnicas de Tipagem Bacteriana , Bifidobacterium/classificação , Suplementos Nutricionais , Humanos , Lactobacillus/classificação , PolôniaRESUMO
Vitamin B12 (B12) production is a strain specific, rare and hidden functional attribute of lactobacilli and a cogent protocol for selection of such isolates from the herd of lactobacilli is required. The present study included isolation of lactobacilli from human samples (milk and fecal), screening them by a polyphasic (three-phase) methodology for probable B12 production potential and validating the screening protocol by exploring selected strains for in vitro vitamin production (two-phase fermentation) and quantification [micro-assay and ultra fast liquid chromatography (UFLC)]. Fifty-nine Lactobacillus strains were recovered from tested biological samples. Contrary to screening inapplicabilities of first [growth potential (GP) in B12-free medium] and second phases (GP in B12-free and cobalt chloride-supplemented conditions), third phase (cbiK gene detection on genomic DNA) alone was revealed as a validated strategy for selection of two probable B12-producing lactobacilli. Microbiological assay confirmed production and bioavailability of produced vitamin, while UFLC testing validated the results by precisely quantifying the cyanocobalamin (industrially produced bio-available form of B12) in cell extracts of both possible B12 producers [BHM10 (10.91 ± 1.55 µg/l) and BCF20 (23.90 ± 1.73 µg/l)] and positive standard [Lactobacillus reuteri DSM20016 (20.03 ± 4.17 µg/l)]. Moreover, this study generates a novel report for genomic detection, partial amplification and sequencing of cbiK gene in Lactobacillus plantarum species (both BHM10 and BCF20). In conclusion, contrary to first two phases, cbiK gene detection strategy successfully selects B12-producing strains from a group of human-originated lactobacilli and can be used in the future for similar screening studies.
Assuntos
Fezes/microbiologia , Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Leite Humano/microbiologia , Vitamina B 12/biossíntese , Humanos , Lactobacillus/genética , Tipagem Molecular , RNA Ribossômico 16S/genéticaRESUMO
Riboflavin has an important role in various cellular metabolic activities through its participation in oxidation-reduction reactions. In this study, as many as 60 lactobacilli were screened for the presence or absence of riboflavin biosynthesis genes and riboflavin production. Of these, only 14 strains were able to grow in a commercial riboflavin-free medium. We observed that the presence of riboflavin biosynthesis genes is strain-specific across different species of lactobacilli. The microbiological assay was found to be appreciably reproducible, sensitive, rapid, and inexpensive and, hence, can be employed for screening the riboflavin-producing strains. The study thus represents a convenient and efficient method for selection of novel riboflavin producers. These riboflavin(+) strains thus identified and characterized could be explored as potent candidates for the development of a wide range of dairy- and cereal-based foods for the delivery of in situ riboflavin to consumers.
Assuntos
Lactobacillus/metabolismo , Reação em Cadeia da Polimerase/métodos , Riboflavina/biossíntese , Bioensaio , Laticínios , Grão Comestível , Fermentação , Alimentos Fortificados , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Oxirredução , Riboflavina/genética , Especificidade da EspécieRESUMO
AIM: To analyze the changes of different Lactobacillus species in ulcerative colitis patients and to further assess the therapeutic effects of selected Lactobacillus strains on dextran sulfate sodium (DSS)-induced experimental colitis in BALB/c mice. METHODS: Forty-five active ulcerative colitis (UC) patients and 45 population-based healthy controls were enrolled. Polymerase chain reaction (PCR) amplification and real-time PCR were performed for qualitative and quantitative analyses, respectively, of the Lactobacillus species in UC patients. Three Lactobacillus strains from three species were selected to assess the therapeutic effects on experimental colitis. Sixty 8-week-old BALB/c mice were divided into six groups. The five groups that had received DSS were administered normal saline, mesalazine, L. fermentum CCTCC M206110 strain, L. crispatus CCTCC M206119 strain, or L. plantarum NCIMB8826 strain. We assessed the severity of colitis based on disease activity index (DAI), body weight loss, colon length, and histologic damage. RESULTS: The detection rate of four of the 11 Lactobacillus species decreased significantly (P < 0.05), and the detection rate of two of the 11 Lactobacillus species increased significantly (P < 0.05) in UC patients. Relative quantitative analysis revealed that eight Lactobacillus species declined significantly in UC patients (P < 0.05), while three Lactobacillus species increased significantly (P < 0.05). The CCTCC M206110 treatment group had less weight loss and colon length shortening, lower DAI scores, and lower histologic scores (P < 0.05), while the CCTCC M206119 treatment group had greater weight loss and colon length shortening, higher histologic scores, and more severe inflammatory infiltration (P < 0.05). NCIMB8826 improved weight loss and colon length shortening (P < 0.05) with no significant influence on DAI and histologic damage in the colitis model. CONCLUSIONS: Administration of an L. crispatus CCTCC M206119 supplement aggravated DSS-induced colitis. L. fermentum CCTCC M206110 proved to be effective at attenuating DSS-induced colitis. The potential probiotic effect of L. plantarum NCIMB8826 on UC has yet to be assessed.
Assuntos
Colite Ulcerativa/terapia , Modelos Animais de Doenças , Microbioma Gastrointestinal/efeitos dos fármacos , Lactobacillus/classificação , Probióticos/uso terapêutico , Adulto , Animais , Colite Ulcerativa/induzido quimicamente , Colo/microbiologia , Colo/patologia , Sulfato de Dextrana , Suplementos Nutricionais , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de Doença , Redução de Peso/efeitos dos fármacosRESUMO
In many vertebrates, social interactions and nutrition can affect the colonization of gut symbionts across generations. In the highly social honey bee, it is unknown to what extent the hive environment and older worker individuals contribute to the generational transmission of core gut bacteria. We used high-throughput sequencing to investigate the effect of nest materials and social contact on the colonization and succession of core hindgut microbiota in workers. With only brief exposure to hive materials following natural eclosion, gut bacterial communities at 3 and 7 days contained phylotypes typically found in the guts of mature adults regardless of treatment. Continuous exposure to nest materials or direct social interactions with mature adults did not affect the diversity or abundance of gut bacterial communities at the scale examined. Similarly, a common pollen supplement fed by beekeepers during pollen dearth had no effect. A consideration of unique OTUs revealed extensive microbial succession independent of treatment. The dominant Lactobacillus strain at 3 days was largely replaced by a different strain at day 7, revealing the colonization signature of a pioneer species. Similar but less pronounced patterns were evident in less abundant OTU's, many of which may influence community succession via alteration of the gut environment. Our results indicate that the process of bacterial community colonization in the hindgut is resilient to changes in the nutritional, hive, and social environment. Greater taxonomic resolution is needed to accurately resolve questions of ecological succession and typical proportional variation within and between core members of the gut bacterial community.
Assuntos
Abelhas/microbiologia , Ecologia , Lactobacillus/classificação , Animais , Carga Bacteriana , Abelhas/crescimento & desenvolvimento , Biodiversidade , Microbioma Gastrointestinal , Relações Interpessoais , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Microbiota , Filogenia , Pólen/microbiologia , SimbioseRESUMO
Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 2(4) factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R (2) = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L(-1) and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL(-1)) occurred in the MRS broth supplemented with 5.5 g L(-1) glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL(-1).