RESUMO
Alterations of the microbiota-gut-brain axis has been associated with intestinal and neuronal inflammation in Parkinson's disease (PD). The aim of this work was to study some mechanisms associated with the neuroprotective effect of a combination (MIX) of lactic acid bacteria (LAB) composed by Lactiplantibacillus plantarum CRL2130 (riboflavin overproducing strain), Streptococcus thermophilus CRL808 (folate producer strain), and CRL807 (immunomodulatory strain) in cell cultures and in a chronic model of parkinsonism induced with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in aged mice, and under levodopa-benserazide treatment. In vitro, N2a differentiated neurons were exposed to the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or with conditioned media from BV-2 cells exposed to the bacterial extracts. In vivo, motor skills, tyrosine hydrolase (TH) in brain and cytokine concentrations in serum and in brain were evaluated. The study of the faecal microbiota and the histology of the small intestine was also performed. The results showed that the neuroprotective effect associated with LAB MIX administration did not interfere with levodopa-benserazide treatment. This effect could be associated with the antioxidant and immunomodulatory potential of the LAB selected in the MIX, and was associated with the significant improvement in the motor tests and a higher number of TH + cells in the brain. In addition, LAB MIX administration was associated with modulation of the immune response. LAB administration decreased intestinal damage with an increase in the villus length /crypt depth ratio. Finally, the administration of the LAB MIX in combination with levodopa-benserazide treatment was able to partially revert the intestinal dysbiosis observed in the model, showing greater similarity to the profiles of healthy controls, and highlighting the increase in the Lactobacillaceae family. Different mechanisms of action would be related to the protective effect of the selected LAB combination which has the potential to be evaluated as an adjuvant for conventional PD therapies.
Assuntos
Benserazida , Levodopa , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores , Transtornos Parkinsonianos , Animais , Levodopa/farmacologia , Benserazida/farmacologia , Benserazida/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Fármacos Neuroprotetores/farmacologia , Transtornos Parkinsonianos/tratamento farmacológico , Transtornos Parkinsonianos/metabolismo , Masculino , Camundongos , Combinação de Medicamentos , Microbioma Gastrointestinal/efeitos dos fármacos , Modelos Animais de Doenças , Lactobacillales , Probióticos/uso terapêutico , Antiparkinsonianos/farmacologia , Antiparkinsonianos/uso terapêutico , Streptococcus thermophilus/efeitos dos fármacosRESUMO
In addition to their use in human medicine, antimicrobials are also used in food animals and aquaculture, and their use can be categorized as therapeutic against bacterial infections. The use of antimicrobials in aquaculture may involve a broad environmental application that affects a wide variety of bacteria, promoting the spread of bacterial resistance genes. Probiotics and bacteriocins, antimicrobial peptides produced by some types of lactic acid bacteria (LAB), have been successfully tested in aquatic animals as alternatives to control bacterial infections. Supplementation might have beneficial impacts on the intestinal microbiota, immune response, development, and/or weight gain, without the issues associated with antibiotic use. Thus, probiotics and bacteriocins represent feasible alternatives to antibiotics. Here, we provide an update with respect to the relevance of aquaculture in the animal protein production sector, as well as the present and future challenges generated by outbreaks and antimicrobial resistance, while highlighting the potential role of probiotics and bacteriocins to address these challenges. In addition, we conducted data analysis using a simple linear regression model to determine whether a linear relationship exists between probiotic dose added to feed and three variables of interest selected, including specific growth rate, feed conversion ratio, and lysozyme activity.
RESUMO
There is now strong evidence to support the interest in using lactic acid bacteria (LAB)in particular, strains of lactococci and lactobacilli, as well as bifidobacteria, for the development of new live vectors for human and animal health purposes. LAB are Gram-positive bacteria that have been used for millennia in the production of fermented foods. In addition, numerous studies have shown that genetically modified LAB and bifodobacteria can induce a systemic and mucosal immune response against certain antigens when administered mucosally. They are therefore good candidates for the development of new mucosal delivery strategies and are attractive alternatives to vaccines based on attenuated pathogenic bacteria whose use presents health risks. This article reviews the most recent research and advances in the use of LAB and bifidobacteria as live delivery vectors for human and animal health.
Assuntos
Microbioma Gastrointestinal , Lactobacillales , Vacinas , Animais , Bifidobacterium/genética , Vetores Genéticos/genética , Humanos , Lactobacillales/genéticaRESUMO
Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by deterioration and loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc), resulting in motor deficits. Many studies have revealed an inverse relationship between thiamine consumption and susceptibility to PD. Previously, Lactiplantibacillus (L.) plantarum CRL 1905 was selected as thiamine-producing lactic acid bacteria (LAB), and its amprolium-resistant clone, L. plantarum CRL 1905*, was able to produce higher levels of this vitamin and inhibited neuronal death in an in vitro model. The present work aimed to evaluate the neuroprotective effect of these thiamine-producing LAB in an in vivo parkinsonian mouse model. Male C57BL/6 mice injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were orally administered with one of the LAB strains or commercial thiamine for 1 month. The administration of either thiamine-producing LAB prevented the motor deficits of mice similar to the effects of the commercial vitamin. These benefits were associated with increased number of tyrosine hydroxylase positive (TH+) neurons in the SNpc. The evaluation of the inflammatory response caused by the neurotoxin showed that both LAB decreased pro-inflammatory cytokines in serum; moreover, the strain selected as the higher thiamine producer showed the best anti-inflammatory effect locally in the brain and significantly decreased the levels of IL-6, TNF-α, IFN-γ and MCP-1, which remained similar to the levels of healthy control animals. These results demonstrated that thiamine-producing L. plantatum CRL 1905* has the potential to be used as part of a strategy to prevent or to complement the treatments of neurodegenerative diseases such as PD. A limitation of this study is that we cannot guarantee whether LAB are capable of producing thiamine in the intestinal tract or release the vitamin after lysis; however, the results show that thiamine production by L. plantarum CRL 1905 is implicated in the observed effect, in addition to other benefits associated with the LAB strain that are also involved and are currently under study.
Assuntos
Lactobacillales , Fármacos Neuroprotetores , Doença de Parkinson , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina , Animais , Modelos Animais de Doenças , Neurônios Dopaminérgicos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/farmacologia , Doença de Parkinson/tratamento farmacológico , Substância Negra , Tiamina , Vitaminas/farmacologiaRESUMO
Oxidative stress and inflammatory processes might contribute to the cascade of events leading Parkinson disease (PD); and vitamins such as riboflavin can exert protection on vulnerable neurons in neurodegenerative conditions. Previously, it was demonstrated that a mixture of lactic acid bacteria (including a riboflavin-producing strain) improved motor skills in a parkinsonian model. The aim of the present work was to investigate the neuroprotective potential of Lactiplantibacillus (L.) plantarum CRL2130, a riboflavin-producing strain in PD models. In vitro, N2a differentiated neurons were exposed the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or commercial riboflavin. In vivo, adult male C57BL/6 mice were injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and probenecid, and received orally L. plantarum CRL2130, L. plantarum CRL725 (parent strain that produces low levels of riboflavin) or commercial vitamin. Results showed that when N2a cells were incubated with intracellular extract from L. plantarum CRL2130 maintained the viability, and significantly decreased the release of IL-6 and the formation of reactive oxygen species (ROS), all affected by MPP+. In vivo, the administration of L. plantarum CRL2130 attenuated motor deficits and prevented dopaminergic neuronal death. Decrease of pro-inflammatory cytokines and increase of IL-10 in both serum and brain were observed in samples from mice that received L. plantarum CRL2130 compared to MPTP control group (without treatment). In addition, these beneficial effects were similar or improved when compared with animals that received commercial riboflavin. In conclusion, L. plantarum CRL2130 showed a neuroprotective effect in both PD models through anti-oxidant/anti-inflammatory mechanisms.
Assuntos
Lactobacillales , Fármacos Neuroprotetores , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Animais , Modelos Animais de Doenças , Neurônios Dopaminérgicos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Riboflavina/farmacologia , Riboflavina/uso terapêuticoRESUMO
AIMS: To evaluate the capacity of autochthonous lactic acid bacteria (LAB) (43) from Andean grains to increase the antioxidant activity (AOA) and total phenolic compounds (TPCs) in quinoa sourdough to select best performing strains to be used as starter cultures in the elaboration of biscuits. METHODS AND RESULTS: Microbial growth (CFU per g) and pH were evaluated during quinoa dough fermentation. Counts were increased in a range of 0.61-2.97 log CFU per g and pH values between 3.95 and 4.54 were determined after 24 h at 30°C of fermentation. Methanolic (ME) and aqueous (AE) extracts were obtained at the end of fermentation, and free radical scavenging capacity was performed by the DPPH and ABTS methods. ME was selected for further analysis using other methods and TPC quantification. Principal component analysis showed the highest scores of growth, acidification capacity, AOA and TPC for the strains Lc. mesenteroides subsp. mesenteroides CRL 2131 and L. plantarum CRL 1964 and CRL 1973. AOA and TPC in biscuits made with sourdough from these LAB were higher than the acidified and uninoculated controls. CONCLUSIONS: Autochthonous LAB strains (3) increased the AOA of quinoa-based biscuits. SIGNIFICANCE AND IMPACT OF THE STUDY: Quinoa sourdough obtained with selected LAB is suitable as an ingredient for bakery foods with improved antioxidant status.
Assuntos
Chenopodium quinoa , Lactobacillales , Antioxidantes/análise , Pão/microbiologia , Chenopodium quinoa/química , Chenopodium quinoa/microbiologia , Fermentação , Microbiologia de AlimentosRESUMO
Thiamine or vitamin B1, an essential micronutrient mainly involved in energy production, has a beneficial impact on the nervous system, and its deficiency can be associated with the development and progression of neurodegenerative diseases. The aim of this work was to select thiamine-producing lactic acid bacteria (LAB) and study their physiological effects using neuron cell cultures. In this study, 23 LAB able to produce thiamine were identified by growth in thiamine-free synthetic medium. Intra- and extracellular thiamine concentrations were determined using a microbiological method and results confirmed by HPLC techniques. A wide variation in vitamin production was found showing that this property was not only species specific but also a strain-dependent trait. Five of these strains were pre-selected for their capacity to produce higher concentrations of thiamine. Only the pre-treatment with the intracellular extract of Lactiplantibacillus (L.) plantarum CRL 1905 increased significantly neuronal survival in N2a cells' model of neurotoxicity (MPP+) with thiamine deficiency conditions (amprolium). Furthermore, amprolium-resistant variants of CRL 1905 were isolated by exposition of the strain to increasing concentrations of this toxic thiamine analogue. The variant A9 was able to increase more than 2 times the intracellular thiamine production of the original strain. A9 bacterial extract significantly prevented neuronal cell death and the increase of IL-6. The amprolium-resistant strain A9 showed a modulating and neuroprotective effect in an in vitro model of neurotoxicity constituting a potential bio-strategy to counteract thiamine deficiencies and thus prevent or treat neurodegenerative diseases. KEY POINTS: ⢠LAB can produce thiamine in a species- and strain-dependant manner. ⢠L. plantarum CRL 1905 significantly reduce MPP+-induced neurotoxicity in N2a cells. ⢠Amprolium-resistant strain A9 has neuroprotective effect and prevents IL-6 increase.
Assuntos
Lactobacillales , Doenças Neurodegenerativas , Amprólio , Morte Celular , Humanos , Doenças Neurodegenerativas/prevenção & controle , TiaminaRESUMO
OBJECTIVES: Parkinson's disease (PD) is a neurodegenerative process that affects the motor function and involves an inflammatory response and B vitamin deficiencies. The aim of this study was to evaluate the effect of B-group vitamin-producing and immunomodulatory lactic acid bacteria (LAB) in a murine model of PD. METHODS: The effect of Lactobacillus plantarum CRL 2130 (a riboflavin producer), Streptococcus thermophilus CRL 807 (an immunomodulatory strain), and Streptococcus thermophilus CRL 808 (a folate producer) were evaluated individually and as a mixture in mice injected with 1-methyl-4-fenil-1,2,3,6-tetrahidropiridina. Motor capacity, tyrosine hydrolase in the brain, and cytokine concentrations in serum and brain tissues were evaluated in 1-methyl-4-fenil-1,2,3,6-tetrahidropiridina-treated mice after bacterial supplementation. RESULTS: The mice receiving the selected LAB showed significantly improved motor skills compared with those that did not receive bacterial supplementation. When given the mixture of all 3 strains together, the animals had higher brain tyrosine hydrolase-positive cell counts, decreased inflammatory cytokines interleukin 6 and tumor necrosis factor alpha in serum, and increased antiinflammatory cytokine interleukin 10 in serum and brain tissues compared with animals that did not receive LAB supplementation. CONCLUSIONS: The results showed the potential of a selected LAB mixture to improve motor behavior and neuroinflammation in PD. This probiotic mixture could be used as an adjunct treatment in the control of PD.
Assuntos
Lactobacillales , Fármacos Neuroprotetores , Doença de Parkinson , Probióticos , Animais , Camundongos , Fármacos Neuroprotetores/farmacologia , Doença de Parkinson/tratamento farmacológico , Streptococcus thermophilusRESUMO
Folate plays a specific role as methyl donor for nucleotide synthesis and genomic methylation patterns, which in turn are important epigenetic determinants in gene expression. Previous studies have revealed the presence of folate in bovine oviductal fluid as well as the existence of a fine-tuned regulation of the gene expression of folate receptors and transporters in bovine oviduct epithelial cells (BOECs). However, the functional implications of folate in the oviduct remain unknown. The present study aimed to assess the effect of folic acid (FA) on expression levels of selected genes that potentially respond to the folate status in in vitro BOECs. To obtain an insight into the optimization of a culture system for assays, gene expression of folate receptors and transporters was compared between BOECs grown in monolayers and in suspension. The results showed that BOECs from isthmus and ampulla in suspension culture better preserved the region-dependent gene expression profile than in monolayers. Subsequently, BOECs from both anatomical regions were separately cultured in suspension for 24 h assaying different FA concentrations: I) TCM-199 (control); II) TCM-199 + 1 µM FA (similar to the oviduct concentration); III) TCM-199 + 10 µM FA and IV) TCM-199 + 100 µM FA. Expression analysis of genes related to important cellular processes including folate transport, DNA methylation, cell-cell interaction, antioxidant activity and signaling pathways was performed in BOECs using RT-qPCR. Our data demonstrated that addition of 1 µM FA did not affect mRNA levels of most genes analyzed. In contrast, BOECs cultured with 10 µM FA exhibited increased mRNA expression levels of genes involved in folate intake, DNA methylation and antioxidant protection. It is worth noting that at 100 µM FA, transcriptional response in BOECs mainly resulted in decreased mRNA levels of the majority of the genes assayed. Interestingly, cytotoxicity analysis showed a similar LDH activity in the culture media of the experimental groups, indicating that cell integrity was not affected by the FA concentrations assayed. In conclusion, our findings suggest that folate can affect BOECs, promoting changes in gene activity in a framework of functional readjustments in response to environmental conditions.
Assuntos
Tubas Uterinas , Oviductos , Animais , Bovinos , Células Epiteliais , Feminino , Ácido Fólico/farmacologia , RNA MensageiroRESUMO
The demand for animal protein for human consumption has been risen exponentially. Modern animal production practices are associated with the regular use of antibiotics, potentially increasing the emerging multi-resistant bacteria, which may have a negative impact on public health. In poultry production, substances capable of maximizing the animals' performance and displaying an antimicrobial activity against pathogens are very well desirable features. Probiotic can be an efficient solution for such a task. In the present work, lactic acid bacteria (LAB) were isolated from chicken cecum and screened for their antagonistic effect towards many pathogens. Their capacity of producing the B-complex vitamins folate and riboflavin were also evaluated. From 314 isolates, three (C43, C175 and C195) produced Bacteriocin-Like Inhibitory Substances (BLIS) against Staphylococcus aureus (inhibition zones of 18.9, 21.5, 19.5 mm, respectively) and also inhibited the growth of Salmonella Heidelberg. The isolate C43 was identified as Enterococcus faecium, while C173 and C195 were both identified as Lactococcus lactis subsp. lactis. Moreover, the isolates L. lactis subsp. lactis strains C173 and C195 demonstrated high potential to be used as probiotic in poultry feed, in addition to their advantage of producing folate (58.0 and 595.5 ng/mL, respectively) and riboflavin (223.3 and 175.0 ng/mL, respectively).