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1.
Euro Surveill ; 28(14)2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-37022212

RESUMO

BackgroundAntimicrobial resistance (AMR) is caused by AMR determinants, mainly genes (ARGs) in the bacterial genome. Bacteriophages, integrative mobile genetic elements (iMGEs) or plasmids can allow ARGs to be exchanged among bacteria by horizontal gene transfer (HGT). Bacteria, including bacteria with ARGs, can be found in food. Thus, it is conceivable that in the gastrointestinal tract, bacteria from the gut flora could take up ARGs from food.AimThe study objective was to gain insight into the ARG set carried by commonly used probiotic bacteria that may enter the human body with non-fermented foods, fermented foods, or probiotic dietary supplements (FFPs) and to assess ARG mobility.MethodsNext generation sequencing whole genome data from 579 isolates of 12 commonly employed probiotic bacterial species were collected from a public repository. Using bioinformatical tools, ARGs were analysed and linkage with mobile genetic elements assessed.ResultsResistance genes were found in eight bacterial species. The ratios of ARG positive/negative samples per species were: Bifidobacterium animalis (65/0), Lactiplantibacillus plantarum (18/194), Lactobacillus delbrueckii (1/40), Lactobacillus helveticus (2/64), Lactococcus lactis (74/5), Leucoconstoc mesenteroides (4/8), Levilactobacillus brevis (1/46), Streptococcus thermophilus (4/19). In 66% (112/169) of the ARG-positive samples, at least one ARG could be linked to plasmids or iMGEs. No bacteriophage-linked ARGs were found.ConclusionThe finding of potentially mobile ARGs in probiotic strains for human consumption raises awareness of a possibility of ARG HGT in the gastrointestinal tract. In addition to existing recommendations, screening FFP bacterial strains for ARG content and mobility characteristics might be considered.


Assuntos
Farmacorresistência Bacteriana , Genes Bacterianos , Bactérias Gram-Positivas , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/genética , Farmacorresistência Bacteriana/genética , Probióticos , Bifidobacterium animalis/efeitos dos fármacos , Bifidobacterium animalis/genética , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Genoma Bacteriano
2.
World J Microbiol Biotechnol ; 37(12): 205, 2021 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-34698975

RESUMO

n-Butanol is an essential chemical intermediate produced through microbial fermentation. However, its toxicity to microbial cells has limited its production to a great extent. The anaerobe lactic acid bacteria (LAB) are the most resistant to n-butanol, so it should be the first choice for improving n-butanol production. The present article aims to review the following aspects of n-butanol production by LAB: (1) the tolerance of LAB to n-butanol, including its tolerance level and potential tolerance mechanisms; (2) genome editing tools in the n-butanol-resistant LAB; (3) methods of LAB modification for n-butanol production and the production levels after modification. This review will provide a theoretical basis for further research on n-butanol production by LAB.


Assuntos
1-Butanol/metabolismo , Lactobacillales/genética , Lactobacillales/metabolismo , Engenharia Metabólica , 1-Butanol/farmacologia , Anaerobiose , Butanóis , Tolerância a Medicamentos , Fermentação , Edição de Genes , Microbiologia Industrial , Lactobacillales/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Estresse Fisiológico
3.
Int J Mol Sci ; 22(15)2021 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-34360567

RESUMO

Resistance to antimicrobials is a growing problem of worldwide concern. Plasmids are thought to be major drivers of antibiotic resistance spread. The present work reports a simple way to recover replicative plasmids conferring antibiotic resistance from the bacteria in cheese. Purified plasmid DNA from colonies grown in the presence of tetracycline and erythromycin was introduced into plasmid-free strains of Lactococcus lactis, Lactiplantibacillus plantarum and Lacticaseibacillus casei. Following antibiotic selection, the plasmids from resistant transformants were isolated, analyzed by restriction enzyme digestion, and sequenced. Seven patterns were obtained for the tetracycline-resistant colonies, five from L. lactis, and one each from the lactobacilli strains, as well as a single digestion profile for the erythromycin-resistant transformants obtained in L. lactis. Sequence analysis respectively identified tet(S) and ermB in the tetracycline- and erythromycin-resistance plasmids from L. lactis. No dedicated resistance genes were detected in plasmids conferring tetracycline resistance to L. casei and L. plantarum. The present results highlight the usefulness of the proposed methodology for isolating functional plasmids that confer antibiotic resistance to LAB species, widen our knowledge of antibiotic resistance in the bacteria that inhabit cheese, and emphasize the leading role of plasmids in the spread of resistance genes via the food chain.


Assuntos
Antibacterianos/farmacologia , Queijo/microbiologia , Resistência Microbiana a Medicamentos , Eritromicina/farmacologia , Lactobacillales/crescimento & desenvolvimento , Plasmídeos/genética , Animais , Lactobacillales/efeitos dos fármacos , Lactobacillales/isolamento & purificação
4.
Eur J Clin Pharmacol ; 77(11): 1639-1648, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34085115

RESUMO

BACKGROUND AND AIM: Low-dose aspirin (LDA) administration prevents cerebral infarction and myocardial infarction, but many studies found an association with mucosal injury. Proton-pump inhibitors (PPIs) can prevent gastric and duodenal mucosal damage, but they may exacerbate small-intestinal mucosal injury by altering the microbiota. We aimed to assess the effect of PPIs on the intestinal flora of LDA users. METHODS: Thirty-two recruited patients, who received LDA (100 mg/day) but did not take PPIs, were divided into 15 patients additionally receiving esomeprazole (20 mg/day) and 17 patients additionally receiving vonoprazan (10 mg/day). On days 0, 30, 90, and 180, the microbiota of each patient was examined by terminal restriction fragment length polymorphism analysis, and the serum gastrin, hemoglobin, and hematocrit levels were measured. RESULTS: Additional PPI administration increased the proportion of Lactobacillales in the microbiota of LDA users. This trend was more prevalent in the vonoprazan group (p < 0.0001) than in the esomeprazole group (p = 0.0024). The Lactobacillales proportion was positively correlated with the gastrin level (r = 0.5354). No significant hemoglobin or hematocrit level reduction was observed in subjects receiving LDA with additional PPI. CONCLUSIONS: Additional PPI administration increased the Lactobacillales proportion in the microbiota of LDA users. The positive correlation between the gastrin level and the proportion of Lactobacillales suggested that the change in the intestinal flora was associated with the degree of suppression of gastric acid secretion. Additional oral PPI did not significantly promote anemia, but the risk of causing PPI-induced small-intestinal mucosal injury in LDA users should be considered.


Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Aspirina/administração & dosagem , Microbioma Gastrointestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Inibidores da Bomba de Prótons/farmacologia , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Comorbidade , Relação Dose-Resposta a Droga , Esomeprazol/farmacologia , Feminino , Gastrinas/sangue , Hemorragia Gastrointestinal/induzido quimicamente , Hematócrito , Hemoglobinas , Humanos , Lactobacillales/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Fragmento de Restrição , Estudos Prospectivos , Pirróis/farmacologia , Sulfonamidas/farmacologia
5.
Braz J Microbiol ; 52(2): 869-881, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33694058

RESUMO

The aim of this research was to identify the key lactic acid bacteria associated with the fermentation of dairy traditional fermented products for developing starter cultures for controlled fermentation. A total of 100 lactic acid bacteria (LAB) were isolated from dairy traditional fermented products. Samples were obtained from eight producers in the South East of Nigeria. Isolates were identified by phenotypic and genotypic techniques including rep-PCR genotyping and sequencing of the 16S rRNA, pheS and rpoA genes. Isolates were characterised for antimicrobial activity against foodborne pathogens, exopolysaccharide (EPS) production and survival at low pH and in the presence of bile salts. All isolates clustered into 11 distinct rep-PCR groups and were identified as Lactobacillus fermentum (40%), Lactobacillus delbrueckii (23%), Streptococcus thermophilus (22%), Streptococcus infantarius (10%), Lactobacillus senioris (2%), Leuconostoc pseudomesenteriodes (2%) and Enterococcus thailandicus (1%). Lactobacillus fermentum showed a broad spectrum antimicrobial activity and survival at low pH, while Lactobacillus delbrueckii was able to tolerate low pH and produce EPS. All isolates survived in vitro exposure to 1% (w/v) bile salts over a 3-h period. L. fermentum, L. delbrueckii and S. thermophilus could be used to simulate the fermentation of dairy traditional fermented products.


Assuntos
Produtos Fermentados do Leite/microbiologia , Lactobacillales/isolamento & purificação , Lactobacillales/fisiologia , Antibiose , Ácidos e Sais Biliares/farmacologia , Fermentação , Genes Bacterianos/genética , Genótipo , Concentração de Íons de Hidrogênio , Lactobacillales/classificação , Lactobacillales/efeitos dos fármacos , Viabilidade Microbiana/efeitos dos fármacos , Nigéria , Polissacarídeos Bacterianos/metabolismo , RNA Ribossômico 16S/genética
6.
Meat Sci ; 176: 108467, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33640646

RESUMO

Zinc protoporphyrin IX (ZnPP)-forming food-grade lactic acid bacteria (LAB) were screened from various sources for their ability to improve the color of meat products. The effects of salt and nitrite on the ZnPP-forming ability of these bacteria were also investigated. Finally, these bacteria were applied in salt-added minced meat to assess their ability to improve the color. Twenty-five LAB were screened for their ZnPP-forming ability in pork. Most of the strains exhibited maximum growth anaerobically in 3% salt at 30 °C and grew well at pH 5.5 and 6.5. Moreover, 3% salt slightly retarded ZnPP formation; however, nitrite completely inhibited ZnPP formation in all the ZnPP-forming LAB. Thirteen LAB (avoiding duplication and non-food-grade) could form ZnPP in salt-added minced meat, resulting in improvement of the bright red color, high ZnPP autofluorescence, and increased fluorescence intensity. Finally, considering the safety, Lactobacillus plantarum, Lactococcus lactis subsp. cremoris, and Leuconostoc lactis were suggested as promising candidates to improve the color of meat products.


Assuntos
Lactobacillales/metabolismo , Produtos da Carne/microbiologia , Protoporfirinas/biossíntese , Animais , Cor , Microbiologia de Alimentos , Lactobacillales/efeitos dos fármacos , Lactobacillales/crescimento & desenvolvimento , Produtos da Carne/análise , Nitritos/química , Cloreto de Sódio/química , Suínos
7.
J Infect Dev Ctries ; 15(1): 102-112, 2021 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-33571152

RESUMO

INTRODUCTION: Probiotic and postbiotic potential of thirty-two strains of lactic acid bacteria (LAB), obtained earlier from artisanal dairy sources in Pakistan, have been investigated against major multi-drug resistant (MDR) and food borne pathogenic bacteria. METHODOLOGY: LAB strains were identified by 16S rRNA gene sequencing and their antibacterial activity was assessed by the microdilution method. Four LAB isolates, Weissella confusa PL6, Enterococcus faecium PL7, and Lactobacillus delbrueckii PL11 and PL13 were shortlisted. Their ability to degrade lactose and safety for human consumption in terms of hemolysis and antibiotic susceptibility were assessed in vitro. The antibacterial components in the cell-free supernatants (CFSs) of isolate cultures were characterized biochemically by HPLC. RESULTS: Acid neutralization but not protease treatment abolished the antibacterial activity of CFSs. Lactic, acetic and propionic acids were the main acids in the CFSs, and acid production peaked in the stationary phase of growth. The antibacterial activity of the LAB cultures resulted from secretion of organic acids that lowered the pH. The strains exhibited variable ability to degrade lactose and were non-hemolytic and susceptible to the most common antibiotics. CONCLUSIONS: These LAB strains are probiotic candidates for further investigation of their postbiotic role in naturally preserving processed foods and for attenuation of lactose intolerance.


Assuntos
Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Lactobacillales/metabolismo , Probióticos/metabolismo , Probióticos/farmacologia , Antibacterianos/farmacologia , Laticínios/microbiologia , Farmacorresistência Bacteriana Múltipla , Escherichia coli/efeitos dos fármacos , Humanos , Lactobacillales/química , Lactose/metabolismo , Interações Microbianas , Paquistão , Filogenia , Probióticos/química , Pseudomonas aeruginosa/efeitos dos fármacos , RNA Ribossômico 16S , Salmonella typhi/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
8.
World J Microbiol Biotechnol ; 37(3): 52, 2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33594545

RESUMO

Fermented milk products are a major source of health-promoting microorganisms known as probiotics. To characterize the probiotic properties of lactic acid bacteria isolated from Ghanaian traditionally fermented milk, thirty (30) isolates comprising Enterococcus faecium (1), Lactobacillus fermentum (14), Lb. plantarum (2) and Pediococcus acidilactici (13) identified by 16S rRNA gene sequencing, were tested for survival at low pH (2.5) and bile salts (0.3% (w/v)), hydrophobicity, co-aggregation, auto-aggregation and antimicrobial activities against selected pathogens. Safety of potential probiotic bacteria was assessed by hemolytic activity on blood agar and susceptibility to nine different antibiotics. Majority (90%) of the strains showed survival rates above 80% at pH (2.5) and in bile salts (0.3% (w/v)). Hydrophobicity ranged from 5 to 61% while cell auto-aggregation ranged from 41 to 80% after 24 h. Co-aggregation with E. coli (3.7-43.9%) and S. Typhimurium (1.3-49.5%) were similar for the LAB strains at 24 h. Cell- free supernatants of all LAB strains inhibited E. coli while S. Typhimurium was not sensitive to cell-free supernatants of five Pd. acidilactici strains: OS24h20, OS18h3, OY9h19, OS9h8 and 24NL38. None of the LAB strains showed ß-hemolysis but 38% of strains showed α-hemolysis. Susceptibilities to antibiotics were strain-specific; only four strains, two Lb. fermentum and two Pd. acidilactici were susceptible to all nine antibiotics tested. Based on high survival rates in bile salts, low pH and generally good hydrophobicity, auto-aggregation, co-aggregation and inhibitory activities, 15 out of 30 strains tested were considered qualified candidates for development of probiotic cultures for fermented milk products in sub-Saharan Africa.


Assuntos
Produtos Fermentados do Leite/microbiologia , Lactobacillales/classificação , Probióticos/farmacologia , Animais , Antibacterianos/farmacologia , Ácidos e Sais Biliares , Tolerância a Medicamentos , Escherichia coli/genética , Fermentação , Gana , Concentração de Íons de Hidrogênio , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Lactobacillales/isolamento & purificação , Leite/microbiologia , RNA Ribossômico 16S/genética
9.
Arch Microbiol ; 203(2): 597-607, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32995979

RESUMO

In the present study, lactic acid bacteria were isolated from table olive in Morocco. Random Amplified Polymorphic DNA fingerprinting with (GTG)'(5) primer revealed a remarquable variability within isolates. According to the molecular identification, Enterococcus faecium was the most dominant species isolated with 32 strains (84.21%), followed by 4 strains of Weissella paramesenteroides (10.52%), 1 strain of Leuconostoc mesenteroides (2.63%) and Lactobacillus plantarum (2.63%). All of the strains that were identified showed occurrence of more than one bacteriocin-encoding gene. Based on the results obtained, L. plantarum 11 showed a mosaic of loci coding for nine bacteriocins (pln A, pln D, pln K, pln G, pln B, pln C, pln N, pln J, ent P). A phenotypic and genotypic antibiotic resistance was also examined. L. plantarum 11, L. mesenteroides 62, W. paramesenteroides 9 and W. paramesenteroides 36 as well as all the strains of E. faecium were susceptible to ampicillin, clindamycin and teicoplanin; however, isolates showed a resistance profile against tetracycline and erythromycin. Except E. faecium 114, E. faecium 130 and L. plantarum 11, no antibiotic resistance genes were detected in all of the strains, which might be due to resistances resulting from non-transferable or non-acquired resistance determinants (intrinsic mechanism).


Assuntos
Antibacterianos/farmacologia , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Olea/microbiologia , Proteínas de Bactérias/genética , Bacteriocinas/genética , Resistência Microbiana a Medicamentos/genética , Lactobacillales/classificação , Lactobacillales/isolamento & purificação , Testes de Sensibilidade Microbiana , Especificidade da Espécie
10.
Meat Sci ; 172: 108367, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33243446

RESUMO

Nitrite (0.25-2%) incorporated thermoplastic starch (TPS) and linear low-density polyethylene (LLDPE) blend films were produced by the conventional blown-film extrusion process. Films were characterized and determined for efficacy as active packaging for pork steak. Scanning electron micrographs showed enhanced starch granule disruption due to melting. Increased nitrite contents enhanced the starch network and improved mechanical properties. Water vapor and barrier properties of blend films were improved by nitrite incorporation, with increased compatibility between TPS and LLDPE networks. Films containing 0.5% nitrite effectively improved redness and corresponded to 0.06 ppm residual nitrite and 75 ppm nitrosomyoglobin in packaged pork. Nitrite addition modified protein secondary conformation involving CO stretching bonds via H-bonding, while films with 1% and 2% nitrite significantly inhibited growth of lactic acid bacteria, yeast and mold, and retained softer texture during storage. Nitrite incorporated films were demonstrated as efficient active packaging to improve the quality of red meat products.


Assuntos
Embalagem de Alimentos , Carne de Porco/análise , Animais , Fungos/efeitos dos fármacos , Lactobacillales/efeitos dos fármacos , Mioglobina/química , Nitritos/química , Polietileno/química , Carne de Porco/microbiologia , Amido/química , Vapor , Sus scrofa
11.
J Dairy Sci ; 103(11): 9980-9991, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32952010

RESUMO

Human milk is the main source of nutrition for infants and the transmission of various microorganisms. The lactic acid bacteria (LAB) in breast milk allow for the establishment of the gut microflora of infants. In this study, we aimed to assess the probiotic potential of LAB strains isolated from breast milk of healthy Chinese women. Two strains, Lacticaseibacillus rhamnosus (formerly Lactobacillus rhamnosus) LHL6 and LHL7, were selected and identified through morphology observation, Gram staining, and 16S rDNA phylogenetic analysis. Using Limosilactobacillus fermentum (formerly Lactobacillus fermentum) CECT5716 as the standard reference strain, the screened strains were characterized for aspects of growth, production of lactic acid and H2O2, antibiotic susceptibility, survival under simulated gastrointestinal conditions, and tolerance to cadmium (Cd). In de Man, Rogosa, and Sharpe (MRS) broth, LHL6 and LHL7 showed longer lag phases than CECT5716 but higher specific growth rates. For the production of lactic acid and H2O2, LHL7 performed better than LHL6 and CECT5716, indicating better antimicrobial ability. Strain LHL7 generated 9.99 mg/L H2O2, considerably higher than 1.25 mg/L for LHL6 and 2.33 mg/L for CECT5716. According to European Food Safety Authority minimum inhibitory concentrations, all of the investigated strains were resistant to chloramphenicol, streptomycin, and kanamycin. However, unlike LHL6 and CECT5716, LHL7 was susceptible to ampicillin and resistant to tetracycline. Resistance to azithromycin, cephalexin, and penicillin G were similar for all 3 strains, whereas CECT5716 was resistant to a higher concentration of roxithromycin. All 3 strains were able to survive in a simulated gastric-like solution, but a low percentage survived in the presence of 0.4% bile salt and 7% pancreatin. Encapsulation with protectants may enhance the survival rate. All 3 strains were tolerant to 500 mg/L Cd in MRS broth and to 1,000 mg/L Cd on MRS agar medium. In summary, 2 novel strains of LAB were obtained that have similar characteristics to the reference strain CECT5716. This work identified potential probiotic candidates for application in the food and pharmaceutical industries and facilitated identification of further probiotics.


Assuntos
Antibacterianos/farmacologia , Lacticaseibacillus rhamnosus/isolamento & purificação , Lactobacillales/isolamento & purificação , Limosilactobacillus fermentum/isolamento & purificação , Leite Humano/microbiologia , Probióticos/farmacologia , Animais , Inocuidade dos Alimentos , Humanos , Peróxido de Hidrogênio/metabolismo , Ácido Láctico/metabolismo , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Lactobacillales/crescimento & desenvolvimento , Limosilactobacillus fermentum/efeitos dos fármacos , Limosilactobacillus fermentum/genética , Lacticaseibacillus rhamnosus/efeitos dos fármacos , Lacticaseibacillus rhamnosus/genética , Lacticaseibacillus rhamnosus/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Filogenia
12.
Int J Food Microbiol ; 334: 108837, 2020 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-32890943

RESUMO

This study investigated physiological alterations involved in the inactivation of Levilactobacillus (L.) brevis and Leuconostoc (Lc.) mesenteroides in orange juice caused by Citrus lemon essential oil (CLEO) and C. reticulata essential oil (CREO) alone and combined with mild heat treatment (MHT). Damage in DNA, membrane integrity, membrane potential, metabolic and efflux activity of bacterial cells were measured after exposure (6 and 12 min) to CLEO or CREO (0.5 µL/mL) and/or MHT (54 °C) using flow cytometry. Limonene was the major constituent in CLEO (66.4%) and CREO (89.4%). The size of the damaged cell subpopulations increased (p < 0.05) after longer exposure time and varied with the tested essential oil and/or bacterial isolate. After exposure to CLEO and CREO alone, the cell subpopulations with damage in measured physiological functions were in a range of 19.6-66.8% and 23.8-75.9%, respectively. Exposure to CREO resulted in larger Lc. mesenteroides cell subpopulations (35.4-68.7%) with damaged DNA, permeabilized and depolarized membrane and compromised metabolic or efflux activity compared to L. brevis (23.8-58.0%). In contrast, exposure to CLEO led to higher damaged L. brevis cell subpopulations (35.1-77%) compared to Lc. mesenteroides (25.3-36.6%). Exposure to combined treatments (CLEO or CREO and MHT) affected the measured physiological functions in almost the entire L. brevis and Lc. mesenteroides cell population (up to 99%), although the damage extension on each isolate varied with tested essential oil. Results show that inactivation of L. brevis and Lc. mesenteroides cells caused by CLEO and CREO alone and combined with MHT in orange juice involves a multi-target action, which causes DNA damage, altered permeability and depolarization of membrane and compromised metabolic and efflux activities.


Assuntos
Citrus/química , Sucos de Frutas e Vegetais/microbiologia , Temperatura Alta , Lactobacillales/fisiologia , Óleos Voláteis/farmacologia , Pasteurização/métodos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Membrana Celular/efeitos da radiação , Dano ao DNA , Microbiologia de Alimentos , Lactobacillales/classificação , Lactobacillales/efeitos dos fármacos , Lactobacillales/efeitos da radiação , Óleos Voláteis/química , Fatores de Tempo
13.
BMC Microbiol ; 20(1): 239, 2020 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-32753060

RESUMO

BACKGROUND: Probiotics have been reported to reduce total cholesterol levels in vitro, but more evidence is needed to determine the clinical relevance of this activity. Chinese traditional fermented pickles are a good source of lactic acid bacteria. Therefore, pickle samples were collected for screening lactic acid bacteria based on their ability to survive stresses encountered during gastrointestinal passage and cholesterol reducing potency. RESULTS: Seventy five lactic acid bacteria strains were isolated from 22 fermented pickles. From these bacteria, Lactobacillus plantarum E680, showed the highest acid (85.25%) and bile tolerance (80.79%). It was sensitive to five of the eight antibiotics tested, inhibited the growth of four pathogenic bacteria, and reduced the total cholesterol level by 66.84% in broth culture. In vivo testing using hypercholesterolemic mice fed high-fat emulsion, independent of food intake, found that L. plantarum E680 suppressed body weight gain and reduced total cholesterol and low-density lipoprotein cholesterol levels, with no effect on high-density lipoprotein cholesterol. CONCLUSIONS: Chinese traditional fermented pickles are a good source for probiotics. L. plantarum E680, isolated from pickles, was acid and bile tolerant, sensitive to antibiotics, and reduced cholesterol levels both in vitro and in vivo. Based on these results, L. plantarum E680 may have potential as a novel probiotic for the development of cholesterol-lowering functional food.


Assuntos
Hipercolesterolemia/tratamento farmacológico , Lactobacillus plantarum/fisiologia , Probióticos , Ácidos/metabolismo , Animais , Antibacterianos/farmacologia , Antibiose , Anticolesterolemiantes/farmacologia , Anticolesterolemiantes/uso terapêutico , Ácidos e Sais Biliares/metabolismo , Peso Corporal/efeitos dos fármacos , Cucumis sativus , Alimentos Fermentados/microbiologia , Hipercolesterolemia/sangue , Hipercolesterolemia/patologia , Lactobacillales/efeitos dos fármacos , Lactobacillales/isolamento & purificação , Lactobacillales/fisiologia , Lactobacillus plantarum/efeitos dos fármacos , Lactobacillus plantarum/isolamento & purificação , Lipídeos/sangue , Camundongos , Probióticos/administração & dosagem , Probióticos/farmacologia
14.
J Dairy Sci ; 103(9): 7908-7926, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32684468

RESUMO

In this study a total of 220 isolates of lactic acid bacteria (LAB) recovered from 10 types of Brazilian artisanal cheeses marketed in 4 main regions of Brazil were evaluated regarding their safety and ability to produce diacetyl (a precursor of aromatic compounds), exopolysaccharides (EPS; from different sugar sources), and antagonistic activity against Listeria monocytogenes and Staphylococcus aureus. The results indicated that 131 isolates (59.6%) were classified as strong (40.5%) and moderate (19.1%) diacetyl producers; 28 isolates (12.7%) stood out due to their remarkable production of EPS from different sugars, including sucrose (3.2%), fructose (2.3%), lactose (2.3%), and glucose (6%). Furthermore, 94.1% and 95.9% of isolates presented antagonistic activity against S. aureus and L. monocytogenes, respectively, even though only 27 isolates (12.3%) exhibited positive results in the bacteriocin production test. None of the isolates tested presented hemolytic activity, and 117 were classified as safe, due to their intrinsic resistance to a maximum of 4 different antibiotics. The data obtained for assessment of antibiogram profile and technological potential (moderate and high production of diacetyl, EPS, and bacteriocins) were submitted to a multiple correspondence analysis to correlate them with the cheese of isolation. Regarding the antimicrobial profile of LAB strains, it was possible to verify an association between isolates from Minas artisanal cheeses from Araxá and resistance to tetracycline; Minas artisanal cheeses from Serro and resistance to erythromycin; Coalho and Minas artisanal cheese from Cerrado and resistance to penicillin; and isolates from Serrano and Colonial cheeses with clindamycin and ceftazidime resistance. Although the susceptibility of strains to these antibiotics was considered high (71.8-80.5%), these data may be related to the horizontal transfer of genes in the production chain of these cheeses. Results of multiple correspondence analysis also showed that isolates with antagonistic activity were mostly isolated from Manteiga, Colonial, and Coalho cheeses. The isolates with high or moderate EPS-producer ability from sucrose, glucose, and fructose were mainly associated with Minas artisanal cheeses from Cerrado. In contrast, isolates with high or moderate EPS-producer ability from lactose were isolated from Serrano, Minas artisanal cheeses from Canastra, and Campo das Vertentes microregions. Finally, isolates from Minas artisanal cheeses (from Araxá microregion), Coalho, and Caipira cheeses were associated with moderate/high diacetyl production. To the best of the authors' knowledge, this study provides, for the first time, data indicating that the dominant technological, biopreservative, and safety properties of LAB isolates can be correlated with the type of Brazilian artisanal cheeses, which denotes its singularity. This knowledge is of utmost relevance for the development of starter or adjunct cultures with tailored properties.


Assuntos
Queijo/microbiologia , Microbiologia de Alimentos , Lactobacillales/isolamento & purificação , Animais , Antibacterianos/farmacologia , Bacteriocinas , Brasil , Queijo/análise , Inocuidade dos Alimentos , Lactobacillales/efeitos dos fármacos , Listeria monocytogenes/fisiologia , Testes de Sensibilidade Microbiana , Análise Multivariada , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/fisiologia
15.
Food Microbiol ; 91: 103540, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32539956

RESUMO

Lactic acid bacteria (LAB) are commonly used in soymilk fermentation to improve health-related functionality, but their contribution to sensory qualities is less valued. We characterized Lactobacillus harbinensis M1, Lactobacillus mucosae M2, Lactobacillus fermentum M4, Lactobacillus casei M8 and Lactobacillus rhamnosus C1 from naturally-fermented tofu whey, along with Streptococcus thermophilus ST3 from kefir XPL-1 fermented soymilk, to investigate their potential as starter cultures of fermented soymilk. They were characterized for antibiotic susceptibility, probiotic potential and their performance as starter cultures. All the LABs showed sensitivity to the tested antibiotics. L. casei M8 had strongest tolerance to synthetic gastrointestinal juice (<1.0 log CFU/mL loss), as well as antagonistic effects towards five food-borne pathogens. GC/MS analysis showed that L. harbinensis M1 produced significantly higher abundance (P < 0.05) of 2,3-butanedione (2.45 ppm) and acetoin (44.30 ppm), thus improving the overall sensory acceptability of fermented soymilk. The coding genes for the synthesis of 2,3-butanedione/acetoin (alsS, alsD, butA) were predicted from the whole-genome. A co-culture of L. harbinensis M1 and L. casei M8 produced a fermented soymilk product with both markedly improved flavor and good probiotic potential. It appears that L. harbinensis M1 has much potential for improving the organoleptic properties of fermented soymilk.


Assuntos
Acetoína/metabolismo , Diacetil/metabolismo , Alimentos Fermentados/microbiologia , Lactobacillus/metabolismo , Leite de Soja , Antibacterianos/farmacologia , Antibiose , Aderência Bacteriana , Células CACO-2 , Fermentação , Alimentos Fermentados/análise , Microbiologia de Alimentos , Suco Gástrico/metabolismo , Humanos , Lactobacillales/classificação , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Lactobacillales/metabolismo , Lactobacillus/efeitos dos fármacos , Lactobacillus/genética , Lacticaseibacillus casei/efeitos dos fármacos , Lacticaseibacillus casei/genética , Lacticaseibacillus casei/metabolismo , Testes de Sensibilidade Microbiana , Probióticos , Paladar
16.
Appl Microbiol Biotechnol ; 104(14): 6375-6383, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32488313

RESUMO

The emergence of lactic acid bacteria (LABs) resistant to existing antimicrobial drugs is a growing health crisis. To decrease the overuse of antibiotics, molecular diagnostic systems that can rapidly determine the presence of antibiotic resistance (AR) genes in LABs from yogurt samples are needed. This paper describes a fully integrated, miniaturized plastic chip and closed-tube detection chemistry that performs multiplex nucleic acid amplification. High-throughput identification of AR genes was achieved through this approach, and six AR genes were analyzed simultaneously in < 2 h. This time-to-result included the time required for the extraction of DNA. The detection limit of the chip was 103 CFU mL-1, which was consistent with that of tube LAMP. We detected and identified multiple DNAs, including streptomycin, tetracycline, and vancomycin resistance-associated genes, with complete concordance to the Kirby-Bauer disk diffusion method.Key Points• A miniaturized chip was presented, and multiplex nucleic acid amplification was performed.• The device can be integrated with LAMP for rapid detection of antibiotic resistance genes.• The approach had a high throughput of AR gene analysis in lactic acid bacteria.


Assuntos
Farmacorresistência Bacteriana/genética , Genes Bacterianos , Lactobacillales/genética , Microfluídica/métodos , Antibacterianos/farmacologia , Microbiologia de Alimentos , Lactobacillales/efeitos dos fármacos , Lactobacillales/isolamento & purificação , Testes de Sensibilidade Microbiana , Microfluídica/instrumentação , Técnicas de Amplificação de Ácido Nucleico , Polimetil Metacrilato , Sensibilidade e Especificidade , Iogurte/microbiologia
17.
Molecules ; 25(11)2020 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-32545249

RESUMO

Soluble fibers, including pectins from apple and lemon, are commonly used as prebiotic and to prepare functional foods. The present study aimed to investigate the physicochemical and functional properties of pectins extracted from jujubes (Ziziphus jujuba Mill.). Pectins were extracted from jujubes at three stages of harvesting and characterized by FTIR and SEM analyses. Whole milk inoculated with kefir grains was supplemented by 0.25 mg·mL-1 of pectins. The pH value and vitamin C content were evaluated after 24 and 48 h of fermentation. Pectins from jujubes at the first harvesting stage (PJ1K) showed the lowest methoxylation degree. The addition of pectins enhanced the production of vitamin C during heterolactic process. This result was found to depend on jujube harvesting stage as PJ1K stimulated the growth of yeasts in kefir grains yielding to the highest amount of vitamin C (0.83 ± 0.01 µg·mL-1) compared to other samples (0.53-0.60 µg·mL-1) at 24 h. Lactic acid bacteria diminish pH rapidly with respect to control (4.13 ± 0.05), according to the stage of maturation, reducing its initial value by 38.3% in PJ1K. Besides being an excellent prebiotic, pectins from jujubes could be used to enrich kefir with vitamin C.


Assuntos
Ácido Ascórbico/análise , Kefir/microbiologia , Pectinas/farmacologia , Ziziphus/química , Ácido Ascórbico/metabolismo , Fermentação , Concentração de Íons de Hidrogênio , Kefir/análise , Lactobacillales/efeitos dos fármacos , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/metabolismo , Microscopia Eletrônica de Varredura , Pectinas/isolamento & purificação , Prebióticos/análise , Espectroscopia de Infravermelho com Transformada de Fourier , Leveduras/efeitos dos fármacos , Leveduras/crescimento & desenvolvimento , Leveduras/metabolismo
18.
Biotechnol Lett ; 42(10): 1927-1937, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32419046

RESUMO

OBJECTIVES: This research paper was to investigate the influence of soybean peptides addition on viable count of lactic acid bacteria, physicochemical parameters, flavor, and sensory evaluation of yoghurt. RESULTS: The number of fermenting strains (Streptococcus thermophilus + Lactobacillus delbrueckii subsp. bulgaricus) cells in yoghurt (stored at 4 °C for 19 days) added with 0.2% (w/v) of soybean peptides (808.34 Da) reached 1.4 times higher bacterial number than in the control group. A total of 34 volatile substances were detected in this study, while there were 22 volatiles occurred in the control group yoghurt, 30 volatiles were detected in yoghurt added with 0.2% soybean peptides. There was no significant difference in sensory evaluation (p > 0.05) between the yoghurt with and without soybean peptides. CONCLUSIONS: In our study, the addition of soybean peptides (0.2%) can be effective both in maintaining the viable bacterial count and yoghurt quality.


Assuntos
Proteínas de Soja , Iogurte , Carga Bacteriana/efeitos dos fármacos , Feminino , Fermentação/efeitos dos fármacos , Fermentação/fisiologia , Aromatizantes , Humanos , Lactobacillales/efeitos dos fármacos , Lactobacillales/metabolismo , Masculino , Peptídeos/metabolismo , Peptídeos/farmacologia , Proteínas de Soja/metabolismo , Proteínas de Soja/farmacologia , Iogurte/análise , Iogurte/microbiologia , Iogurte/normas
19.
Int J Mol Sci ; 21(7)2020 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-32276519

RESUMO

Lactic acid bacteria can act as reservoirs of antibiotic resistance genes that can be ultimately transferred to pathogens. The present work reports on the minimum inhibitory concentration (MIC) of 16 antibiotics to 25 LAB isolates of five Lactobacillus and one Bifidobacterium species from the human vagina. Acquired resistances were detected to kanamycin, streptomycin, chloramphenicol, gentamicin, and ampicillin. A PCR analysis of lactobacilli failed to identify genetic determinants involved in any of these resistances. Surprisingly, a tet(W) gene was detected by PCR in two Bifidobacterium bifidum strains, although they proved to be tetracycline-susceptible. In agreement with the PCR results, no acquired genes were identified in the genome of any of the Lactobacillus spp. strains sequenced. A genome analysis of B. bifidum VA07-1AN showed an insertion of two guanines in the middle of tet(W) interrupting the open reading frame. By growing the strain in the presence of tetracycline, stable tetracycline-resistant variants were obtained. An amino acid substitution in the ribosomal protein S12 (K43R) was further identified as the most likely cause of VA07-1AN being streptomycin resistance. The results of this work expand our knowledge of the resistance profiles of vaginal LAB and provide evidence for the genetic basis of some acquired resistances.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Lactobacillales/fisiologia , Polimorfismo Genético , Vagina/microbiologia , Bifidobacterium bifidum/efeitos dos fármacos , Bifidobacterium bifidum/genética , Bifidobacterium bifidum/fisiologia , Feminino , Humanos , Lactobacillales/efeitos dos fármacos , Lactobacillales/genética , Lactobacillus/efeitos dos fármacos , Lactobacillus/genética , Lactobacillus/fisiologia , Tetraciclina/farmacologia
20.
FEMS Microbiol Lett ; 367(9)2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32319520

RESUMO

The main hypothesis of this work was to evaluate the presence of lactic acid bacteria (LAB) intrinsically resistant to plant essential oils in sage (Salvia officinalis L.) and laurel (Laurus nobilis), for future applications in functional cheese production by addition of aromatic herbs. The effect of the drying process on the viability of LAB was evaluated with three biomass densities (3, 4 and 5 kg/m2). The drying densities did not affect weight loss, but influenced the levels of LAB of sage and laurel. A total of 10 different strains of Enterococcus faecium, Enterococcus mundtii, Enterococcus raffinosus and Leuconostoc mesenteroides were identified from laurel, while sage did not host any LAB species. In particular, L. mesenteroides was the only species sensitive to the heat treatment. Only five strains, all enterococci, were resistant to at least one antibiotic, even though no strain showed gelatinase or haemolytic activity. The investigation on the technological traits useful in cheese making demonstrated that all LAB can be considered non starter LAB, because they were characterized by a slow acidification capacity (the pH was still above 6.00 after 3 d) and a very limited autolysis (the maximum decrease of the optical density at 599 nm was barely 0.2).


Assuntos
Antibacterianos/farmacologia , Biodiversidade , Lactobacillales/classificação , Lactobacillales/crescimento & desenvolvimento , Queijo/microbiologia , DNA Bacteriano/genética , Dessecação , Microbiologia de Alimentos , Temperatura Alta , Lactobacillales/efeitos dos fármacos , Lauraceae/microbiologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana , Filogenia , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Salvia officinalis/microbiologia
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