Navegando por Autor "Colombo, Monique"
Agora exibindo 1 - 5 de 5
- Resultados por Página
- Opções de Ordenação
Item Beneficial properties and safety of lactic acid bacteria isolated from the dairy production environment(Universidade Federal de Viçosa, 2017-06-30) Colombo, Monique; Nero, Luis Augusto; http://lattes.cnpq.br/1188501382442265Lactic acid bacteria isolated from dairy environment were evaluated for beneficial potential. Preliminary screening and PCR analysis were applied to select and identified through 16s rRNA sequencing 15 LAB strains: Lactobacillus (n = 11; Lb. casei MSI1, Lb. casei MSI5, Lb. casei MRUV1, Lb. casei MRUV6, Lb. acidophilus MVA3, Lb. nagelli MSIV4, Lb. harbinensis MSI3, Lb. harbinensis MSIV2, Lb. fermentum SIVGL1, Lb. plantarum MLE5 and Lb. plantarum MSI2), Pediococcus (n = 2; P. pentosaceus MLEV8 and P. acidilactici MSI7) and Weissella (n = 2; W. paramesenteroides MRUV3 and W. paramesenteroides MSAV5). All selected strains showed resistance to acidic pH and to presence of bile salt. API ZYM test characterized enzymatic activity of the strains and high β-galactosidase activity was observed in 13 strains. All strains presented high values for survival rate to simulated gastric and intestinal conditions, ability to auto and co-aggregate with indicators microorganisms and high cell surface hydrophobicity. Most of the strains were positive for map and EFTu beneficial genes. Strong bile salts deconjugation was applied for all strains and all strains showed good results for assimilating lactose. After this first part of the study, the 15 BAL were evaluated for potential virulence and antimicrobial resistance. The production of virulence factors (hemolysis, gelatinase, lipase, deoxyribonuclease and biogenic amines: lysine, tyrosine, histidine and ornithine) was assessed by phenotypic methods at 25 °C and 37 °C, as well as the resistance to 17 antimicrobials. The isolates were also subjected to PCR to identify the presence of 49 genes associated with virulence factors. None of the strains presented hemolytic activity or the production of gelatinase, lipase, deoxyribonuclease and tested biogenic amines. Of the 15 selected cultures, for 12 types of antibiotics in the disc diffusion method, all strains were resistant for oxacillin and sulfa/trimethoprim, 14 were resistant to gentamicin, 11 were resistant to clindamycin, nine strains were resistant to vancomycin, eight strains to rifampicin, five were resistant to erythromycin, four were resistant to tetracycline, two strains were resistant to ampicillin, one strain was resistant to chloramphenicol and none was resistant for imipenem. For a quantitative test of the antibiogram, five antibiotics were selected in Etest ® strips (bioMérieux). All 15 strains were resistant to vancomycin, two for rifampicin, one for gentamicin and one for chloramphenicol. Regarding the virulence related genes, 19 genes from 49 tested were present in some strains. Results showed that five cultures showed the presence of the int gene, four cultures showed the presence of the ant(4')-Ia gene, three cultures were positive for vanC2, cpd and tdc, two cultures for vanA, tet(K), tet(S), ermA, bcrR, mur-2ed, asa1 and ccf, and one culture was positive for vanC1, ermB, aph(3')-IIIa, aac(6’)-le-aph(2”)-Ia, bcrB and hyl. After characterizing the virulent potential of the 15 BAL, these strains were evaluated for the technological potential for application in the dairy industry. All strains presented acidification capacity, reaching pH values between 0.73 and 2.11 in 24 hours: Lb. casei MRUV6 presented the highest acidification ability (pH 2.11 after 24 h). Ten strains were able to produce diacetyl at 37 °C, except by Lb. casei MSI1, Lb. harbinensis MSI3, Lb. fermentum SIVGL1, Lb. plantarum MLE5 and W. paramesenteroides MRUV3. All strains were able to produce exopolysaccharides, and only two strains presented proteolytic activity (Lb. casei MSI5 and W. paramesenteroides MSAV5). Based on this characterization, Lb. casei MRUV6 was selected for producing fermented milk, stored at 4 °C and 10 °C and monitored until 35 days of shelf life. Samples were subjected to phenotypical and molecular methods to quantify the presence of Lb. casei MRUV6 (conventional plating and RT-PCR, by checking the expression of gapdh, a housekeeping gene) and to verify the expression of bsh gene, related to resistance to bile salts (RT-PCR). Lb. casei MRUV6 population was stable during storage period at 4 and 10 °C at levels around 9.9 log CFU/g, and by monitoring the expression of gapdh gene. However, bsh gene was not expressed during storage period. The study demonstrated the potential use of the beneficial strain Lb. casei MRUV6 isolated from a dairy environment for the production of a fermented milk product, and its stability during storage at 4 and 10 °C. All isolates from the study presented beneficial characteristics, safety for use in food and technological potential for use in the dairy industry. In addition, they may further be subjected to further studies for in vivo evaluations and characterization as probiotics.Item Beneficial properties of lactic acid bacteria naturally present in dairy production(BMC Microbiology, 2018-12) Colombo, Monique; Castilho, Nathália P. A.; Todorov, Svetoslav D.; Nero, Luís AugustoConsumers are increasingly demanding for natural and beneficial foods, in order to improve their health and well-being. Probiotics play an important role in such demand, and dairy foods are commonly used as vehicles for such bacteria, represented predominantly by lactic acid bacteria. Due to consumers demand, food industry is constantly looking for novel bacterial strains, leading to studies that aims the isolation and characterization of their beneficial features. This study aimed to characterize the naturally occurring lactic acid bacteria obtained from a dairy environment, in order to assess their potential use as probiotics.Preliminary screening and PCR analysis, based on 16S rRNA sequencing, were applied to select and identify 15 LAB strains from the genera Lactobacillus (n = 11), Pediococcus (n = 2) and Weissella (n = 2). All strains showed resistance to low pH and the evaluated bile salt concentrations in vitro. The API ZYM test characterized the enzymatic activity of the strains, and a high β-galactosidase activity was observed in 13 strains. All strains presented resistance to simulated gastric (3 h) and intestinal (4 h) conditions in vitro, the ability to auto- and co-aggregate with indicator microorganisms and a high cell surface hydrophobicity. Most of the strains were positive for map and EFTu beneficial genes. All strains exhibited strong deconjugation of bile salts in vitro and all assimilated lactose.The phenotypes exhibited in vitro and the presence of beneficial genes revealed the beneficial potential of the studied strains, demanding further analyses in a food matrix and in vivo to allow the development of a functional product, with health-related properties.Item Desenvolvimento de um meio de cultura alternativo para enumeração seletiva de Lactobacillus casei em leites fermentados(Universidade Federal de Viçosa, 2013-07-25) Colombo, Monique; Carvalho, Antônio Fernandes de; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781655T2; Nero, Luís Augusto; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763792E2; http://lattes.cnpq.br/1188501382442265; Beloti, Vanerli; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790396D2Micro-organismos da espécie Lactobacillus casei são os probióticos mais estudados, uma vez que são capazes de promover efeitos benéficos para a saúde humana. Além disso, os micro-organismos dessa espécie são ácido-tolerantes, sendo capazes de sobreviver ao ambiente gastrointestinal sem serem inativados. Quando adicionados em alimentos, devem se apresentar numa concentração mínima (maior que 106 UFC/mL ou g) para exercerem efetivamente a atividade probiótica. Por essas características, é necessário o monitoramento das populações dessas culturas quando adicionadas em alimentos, apesar da ausência de um protocolo oficial com esta finalidade. O sistema Petrifilm AC pode representar uma alternativa para enumeração de culturas starter e probióticas, desde que associado com meios de cultura seletivos. O presente estudo teve como objetivo a avaliação de substâncias químicas como agentes seletivos para Lb. casei, a fim de propor um meio de cultura seletivo para ser associado a placas Petrifilm AC, a ser avaliado como um protocolo alternativo de enumeração deste probiótico em leites fermentados. Vinte e seis culturas probióticas e starter (incluindo 6 cepas de Lb. casei) foram plaqueadas em ágar MRS com diferentes concentrações de ácido nalidíxico, bile, cloreto de lítio, metronidazol, propionato de sódio e vancomicina; somente vancomicina (10 mg/L) apresentou atividade seletiva para Lb. casei. Cloreto de 2,3,5-trifeniltetrazólio foi identificado como não inibitório para Lb. casei e placas de Petrifilm AC associadas a MRS adicionado de vancomicina a 10 mg/L (MRS-V) apresentaram contagens mais elevadas quando incubadas sob anaerobiose que em aerobiose. Leites fermentados acidificados e iogurtes foram preparados, adicionados de cepas de Lb. casei, estocados a 4 °C e as populações de Lb. casei foram monitoradas usando MRS-V e MRTLV por plaqueamento convencional e pelo sistema Petrifilm AC. Todos os índices de correlação entre as contagens obtidas por plaqueamento convencional e pelo sistema Petrifilm AC foram significativos (p < 0.05), mas o melhor desempenho foi observado para MRS-V. Os dados obtidos indicaram a adequação do uso do MRS-V associado às placas Petrifilm AC para enumeração de Lb. casei em leites fermentados.Item Lactobacillus curvatus UFV-NPAC1 and other lactic acid bacteria isolated from calabresa, a fermented meat product, present high bacteriocinogenic activity against Listeria monocytogenes(BMC Microbiology, 2019) Castilho, Nathália Parma Augusto; Colombo, Monique; Oliveira, Leandro Licursi de; Todorov, Svetoslav Dimitrov; Nero, Luís AugustoBacteriocins produced by lactic acid bacteria (LAB) can be considered as viable alternatives for food safety and quality, once these peptides present antimicrobial activity against foodborne pathogens and spoilage bacteria. Fermented foods, such as artisanal sausages and cured meats, are relevant sources of LAB strains capable of producing novel bacteriocins, with particular interest by the food industry.Three LAB strains (firstly named as Lactobacillus curvatus 12, L. curvatus 36 and Weissella viridescens 23) were obtained from calabresa by presenting promising bacteriocinogenic activity, distinct genetic profiles (rep-PCR, RAPD, bacteriocin-related genes) and wide inhibitory spectrum. Among these strains, L. curvatus 12 presented higher bacteriocin production, reaching 25,000 AU/mL after incubation at 25, 30 and 37 °C and 6, 9 and 12 h. Partially purified bacteriocins from L. curvatus 12 kept their inhibitory activity after elution with isopropanol at 60% (v/v). Bacteriocins produced by this strain were purified by HPLC and sequenced, resulting in four peptides with 3102.79, 2631.40, 1967.06 and 2588.31 Da, without homology to known bacteriocins.LAB isolates obtained from calabresa presented high inhibitory activity. Among these isolates, bacteriocins produced by L. curvatus 12, now named as L. curvatus UFV-NPAC1, presented the highest inhibitory performance and the purification procedures revealed four peptides with sequences not described for bacteriocins to date.Item Risky consumption habits and safety of fluid milk available in retail sales outlets in Viçosa, Minas Gerais State, Brazil(Foodborne Pathogens and Disease, 2014-06-02) Pieri, Fabio Alessandro; Colombo, Monique; Merhi, Carolina Milner; Juliati, Vinı́cius Augusto; Ferreira, Marcello Sebe; Nero, Marcelo Antônio; Nero, Luis AugustoThis study aimed to assess raw milk consumption habits in the urban population of Viçosa, Minas Gerais, Brazil, and the microbiological safety and quality of the fluid milk available in retail sales outlets in the same region. A simplified questionnaire regarding raw milk consumption was applied to the persons responsible for food acquisition in 411 residences. The regular consumption of raw milk was observed by 18.5% of the interviewers, and lack of knowledge of possible risks related to this food product. Microbiological safety and quality were assessed for raw (n = 69), pasteurized (n = 80), and ultra-high-temperature (UHT)-treated milk (n = 80) by analyzing the counts of mesophilic aerobes, coliforms, and Escherichia coli, and detection of Listeria monocytogenes and Salmonella spp.; raw milk samples were also subjected to enumeration of coagulase- positive Staphylococcus. Concerning raw milk, 59.4% of the samples were considered as produced in inadequate hygienic conditions, 5.8% of the samples presented counts of coagulase-positive Staphylococcus lower than 100 colony-forming units (CFU)/mL, and no samples presented with positive results for L. monocytogenes or Salmonella spp. All pasteurized and UHT milk samples presented with low counts of mesophilic aerobes and coliforms, while L. monocytogenes and Salmonella spp. were absent. The data demonstrated that raw milk was consumed by the population studied. Despite the absence of potential hazards, raw milk was of poor hygienic quality, in contrast with the processed fluid milk available in retail sales outlets that was safe and of good hygienic quality, highlighting the suitability of pasteurized and UHT milk for human consumption.