ABSTRACT
Lactobacillus spp. isolated from different portions of chickens' gastrointestinal tract were evaluated concerning their ability to survive in a water-in-oil (W/0) emulsion containing sesame and sunflower oil. After sixty days of emulsion storage under refrigeration, three of five strains tested survived in number equal to or higher than 10(6)cfu/g. Lactobacillus reuteri 2M14C, which presented the highest survival in W/O emulsion (10(7)cfu/g), was tested for its capacity to resist throughout the passage through gnotobiotic mice gastrointestinal tract and for the ability to stimulate murine peritoneal macrophages phagocytosis. This strain remained at a number above 10(9)cfu/g feces during ten days of monoassociation, and monoassociated mice showed phagocytic activity significantly greater than the germ-free controls (P<0.05). The results suggest that the formulation can be used to incorporate viable Lactobacillus spp. cells in animal feed. Moreover, the results suggest that L. reuteri 2M14C is a strong candidate to be incorporated in probiotic formulations for use in chicken.
Lactobacillus spp. isolados de diferentes porções do trato gastrintestinal de frangos foram testados quanto à capacidade de se manterem viáveis em uma emulsão água/óleo (A/O) contendo óleos de gergelim e de girassol. Após sessenta dias de estocagem sob refrigeração, três de cinco linhagens testadas sobreviveram em concentração igual ou superior a 10 6 UFC/g. Lactobacillus reuteri 2M14C, que apresentou maior capacidade de sobrevivência na emulsão desenvolvida (10 7 UFC/g), foi testado quanto à sua capacidade de sobreviver às condições do trato gastrintestinal in vivo em camundongos gnotobióticos. Após dez dias de monoassociação com L. reuteri 2M14C, foi testada também a capacidade de estimulação da atividade fagocítica de macrófagos peritoneais. A linhagem permaneceu em número superior a 10 9 UFC/g de conteúdo fecal durante os dez dias de monoassociação, e os camundongos monoassociados apresentaram atividade fagocítica maior (P<0,05) que a do grupo controle isento de germe. Os resultados sugerem que a formulação proposta é capaz de manter a viabilidade de células de lactobacilos para adição em ração animal, necessitando, no entanto, de um acompanhamento dessa viabilidade por tempo maior de estocagem. Além disso, os resultados demonstram que Lactobacillus reuteri 2M14C é um forte candidato a ser adicionado em formulações probióticas para uso em frangos.
Subject(s)
Animals , Lactobacillus , Phagocytosis , Probiotics/analysis , Gastrointestinal Tract/anatomy & histology , Chickens/classificationABSTRACT
Puba or carimã is a Brazilian staple food obtained by spontaneous submerged fermentation of cassava roots. A total of 116 lactobacilli and three cocci isolates from 20 commercial puba samples were recovered on de Man, Rogosa and Sharpe agar (MRS); they were characterized for their antagonistic activity against foodborne pathogens and identified taxonomically by classical and molecular methods. In all samples, lactic acid bacteria were recovered as the dominant microbiota (7.86 ± 0.41 log10 CFU/g). 16S-23S rRNA ARDRA pattern assigned 116 isolates to the Lactobacillus genus, represented by the species Lactobacillus fermentum (59 isolates), Lactobacillus delbrueckii (18 isolates), Lactobacillus casei (9 isolates), Lactobacillus reuteri (6 isolates), Lactobacillus brevis (3 isolates), Lactobacillus gasseri (2 isolates), Lactobacillus nagelii (1 isolate), and Lactobacillus plantarum group (18 isolates). recA gene-multiplex PCR analysis revealed that L. plantarum group isolates belonged to Lactobacillus plantarum (15 isolates) and Lactobacillus paraplantarum (3 isolates). Genomic diversity was investigated by molecular typing with rep (repetitive sequence)-based PCR using the primer ERIC2 (enterobacterial repetitive intergenic consensus). The Lactobacillus isolates exhibited genetic heterogeneity and species-specific fingerprint patterns. All the isolates showed antagonistic activity against the foodborne pathogenic bacteria tested. This antibacterial effect was attributed to acid production, except in the cases of three isolates that apparently produced bacteriocin-like inhibitory substances. This study provides the first insight into the genetic diversity of Lactobacillus spp. of puba.
Subject(s)
Humans , Fermentation , Genetic Variation , In Vitro Techniques , Lactobacillus plantarum/genetics , Lactobacillus plantarum/isolation & purification , Manihot/genetics , Polymerase Chain Reaction/methods , Food Samples , MethodsABSTRACT
Amostras de queijo de minas artesanal foram coletadas em 18 queijarias localizadas em propriedades rurais da região da Serra da Canastra, Minas Gerais, com o objetivo de avaliar a influência da altitude sobre a população de bactérias acidolácticas. As queijarias estavam distribuídas nas altitudes de 600 a 900m, 900 a 1000m e mais de 1000m. Observaram-se populações mais elevadas de bactérias acidolácticas nas amostras de queijo da altitude de 600 a 900m. Lactobacillus rhamnosus, Lactobacillus casei e Lactobacillus plantarum foram os principais microrganismos isolados e identificados por PCR ARDRA 16S-23S rDNA, além de Enterococcus spp., Lactococcus spp. e outras espécies de Lactobacillus. Sugere-se que estas espécies estejam adaptadas ao ambiente de produção do queijo de minas artesanal produzido na região, o que resultaria em características sensoriais próprias do produto.
Samples of minas artisanal cheese were collected in 18 small-scale producer properties located in the rural region of Serra da Canastra, Minas Gerais state, aiming to evaluate the influence of three altitudes, from 600 to 900m, 900 to 1000m, and higher than 1000m, on the lactic acid bacteria (LAB) population. High populations of LAB were observed in the cheese samples, mainly in the lowest altitude. Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus plantarum were the major LAB isolated from the cheese samples and identified according to PCR ARDRA 16S-23S rDNA. Enterococcus spp., Lactococcus spp., and other species of Lactobacillus genus were also found. It is suggested that these microorganisms are adapted to the production environment of the minas artisanal cheese which result in the unique sensorial properties of the product.
ABSTRACT
Lactobacilli isolated from the vaginal tract of women with and without bacterial vaginosis (BV) were identified and characterized for the production of antagonists. Bacterial samples were isolated from healthy women (N = 16), from patients with clinical complaints but without BV (N = 30), and from patients with BV (N = 32). Identification was performed using amplified ribosomal DNA restriction analysis. Production of antagonistic compounds was evaluated by the double-layer diffusion technique using Gram-positive (N = 9) and Gram-negative bacteria (N = 6) as well as yeast (N = 5) as indicator strains. Of a total of 147 isolates, 133 were identified as pertaining to the genus Lactobacillus. Lactobacillus crispatus was the species most frequently recovered, followed by L. johnsonii and L. jensenii. Statistical analysis showed that L. crispatus was more frequent in individuals without BV (P < 0.05). A higher production of antagonists was noted in L. crispatus isolates from healthy women (P < 0.05). More acidic local pH and higher H2O2 production by isolated lactobacilli from healthy women suggest these mechanisms as the possible cause of this antagonism. In conclusion, a significant correlation was detected between the presence and antagonistic properties of certain species of Lactobacillus and the clinical status of the patients.
Subject(s)
Female , Humans , Hydrogen Peroxide/metabolism , Lactobacillus/metabolism , Vagina/microbiology , Vaginosis, Bacterial/microbiology , Bacterial Typing Techniques/methods , DNA, Ribosomal/analysis , DNA, Ribosomal/genetics , Lactobacillus/classification , Lactobacillus/isolation & purification , Restriction MappingABSTRACT
Em amostras de S. flexneri, isoladas no período de 1989 a 1993, foi estudado o mecanismo molecular que mediava a multirresistência. Foram utilizadas no estudo 26 amostras de S. flexneri. Estas amostras foram submetidas a teste de sensibilidade a antimicrobianos, experimentos de conjugação e extração de plasmídios. Com relação ao padrão de resistência a antimicrobianos, observou-se que todas as amostras de S. flexneri eram resistentes a pelo menos três antimicrobianos. Das 26 amostras de S. flexneri doadoras submetidas ao processo de conjugação, 34,6% (9 amostras) resultaram em uma freqüência variável de transconjugantes. Das amostras que conjugaram, 100% transferiram o fator de resistência relacionado à ampicilina; sendo que em todas as transconjugantes foi evidenciado apenas um plasmídio de 23,1Kb. Este plasmídio, encontrado em todas as amostras de Shigellas, caracterizou-se como o portador de marca de resistência para ampicilina.
In Shigella strains were studied the molecular mechanism that mediated the multiply antibiotic-resistance. Twenty-six strains of Shigella flexneri were utilised in this investigation. These strains were submitted to disk diffusion test, mating experiments and plasmid isolation. In relation to antibiotics resistance standard it was observed that all Shigella flexneri strains were resistant to at least, three antibiotics tested. From twenty-six Shigella flexneri strains donors submitted to conjugation process, 34.6% (nine strains) resulted in variable frequency of transconjugants. From strains that conjugated, 100%, transferred the resistance factor acquainted with ampicillin. Being that, in all transconjugants which were observed, just one plasmid with 23.1 Kb was evidenced. This plasmid found in all strains was characterised as the cause of resistance to ampicillin.
Subject(s)
R Factors , Drug Resistance, Microbial , Drug Resistance, Multiple , Shigella flexneri/drug effects , Ampicillin Resistance , Brazil , Microbial Sensitivity TestsABSTRACT
Foram submetidos a exame clínico laboratorial de dermatofitose e inqueridos sobre o possível contato com cães e gatos 158 pacientes residentes na área urbana de Fortaleza, que apresentavam lesões suspeitas de dermatofitose. Esta busca associada aos dados obtidos em questionário permitiram identificar a freqüência de surtos domiciliares. Dentre os 83 dermatófitos isolados de infecções humanas, predominaram as espécies antropofílicas sobre as zoofílicas, tendo sido observado uma confluência de diagnóstico humano e animal em 100% dos casos de dermatofitoses zoofílicas humanas, onde foram identificadas as mesmas espécies no homem e nos animais contactantes: Microsporum canis e Trichophyton mentagrophytes. Já os pacientes portadores de dermatófitos antropofílicos variaram quanto ao contato com animais domésticos, não tendo sido isolado estes fungos de nenhum animal contactante. Diante da baixa freqüência de dermatofitoses zoofílicas, considerou-se que o convívio do homem com cães e gatos domésticos foi pouco representativo como fator condicionante da ocorrência de dermatofitoses no meio urbano.
There have been submitted to clinical exam in laboratory of dermatophytosis and inquired about possible contact with domestic animals (dogs and cats) 158 patients living in the urban area of Fortaleza, that showed lesions suspected of dermatophytosis. This search associated to the obtained data in questionnaires permitted us to identify the frequency of domicile outbreaks. Within the 83 people with dermatosis isolated of human infections, prevailed the anthropophilic species over the zoophilic ones, and that it was observed a confluence of human and animal diagnosis in 100% of the human dermatophytosis zoophilic cases, where the same species were identified in men and contacting animals: M. canis and T. mentagrophytes. As the patients carrying anthropophilic dermatophytosis varied as to the contact with animals, not having been these fungus isolated from none of the contacting animals. Before the low frequency of zoophilic dermatophytosis, it was considered that the intimacy of men with domestic dogs and cats represented was little as a conditional factor of occurrence of dermatophytosis in the urban environment.