Marco Antônio Lemos Miguel

Microbiological, technological and therapeutic properties of kefir: a natural probiotic beverage

Kefir is a fermented milk beverage produced by the action of bacteria and yeasts that exist in symbiotic association in kefir grains. The artisanal production of the kefir is based on the tradition of the peoples of Caucasus, which has spread to other parts of the world, from the late 19th century, and nowadays integrates its nutritional and therapeutic indications to the everyday food choices of several populations. The large number of microorganisms present in kefir and their microbial interactions, the possible bioactive compounds resulting of microbial metabolism, and the benefits associated with the use this beverage confers kefir the status of a natural probiotic, designated as the 21th century yoghurt. Several studies have shown that kefir and its constituents have antimicrobial, antitumor, anticarcinogenic and immunomodulatory activity and also improve lactose digestion, among others. This review includes data on the technological aspects, the main beneficial effects on human health of kefir and its microbiological composition. Generally, kefir grains contain a relatively stable and specific microbiota enclosed in a matrix of polysaccharides and proteins. Microbial interactions in kefir are complex due to the composition of kefir grains, which seems to differ among different studies, although some predominant Lactobacillus species are always present. Besides, the specific populations of individual grains seem to contribute to the particular sensory characteristics present in fermented beverages. This review also includes new electron microscopy data on the distribution of microorganisms within different Brazilian kefir grains, which showed a relative change in its distribution according to grain origin.

Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains

A total of 34 lactic acid bacteria isolates from 4 different Brazilian kefir grains were identified and characterized among a group of 150 isolates, using the ability to tolerate acidic pH and resistance to bile salts as restrictive criteria for probiotic potential. All isolates were identified by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of representative amplicons. Eighteen isolates belonged to the species Leuconostoc mesenteroides, 11 to Lactococcus lactis (of which 8 belonged to subspecies cremoris and 3 to subspecies lactis), and 5 to Lactobacillus paracasei. To exclude replicates, a molecular typing analysis was performed by combining repetitive extragenic palindromic-PCR and random amplification of polymorphic DNA techniques. Considering a threshold of 90% similarity, 32 different strains were considered. All strains showed some antagonistic activity against 4 model food pathogens. In addition, 3 Lc. lactis strains and 1 Lb. paracasei produced bacteriocin-like inhibitory substances against at least 2 indicator organisms. Moreover, 1 Lc. lactis and 2 Lb. paracasei presented good total antioxidative activity. None of these strains showed undesirable enzymatic or hemolytic activities, while proving susceptible or intrinsically resistant to a series of clinically relevant antibiotics. The Lb. paracasei strain MRS59 showed a level of adhesion to human Caco-2 epithelial cells comparable with that observed for Lactobacillus rhamnosus GG. Taken together, these properties allow the MRS59 strain to be considered a promising probiotic candidate.

Tapered plastic optical fiber-based biosensor – Tests and application

Cells detection is crucial in microbiological analysis of clinical, food, water or environmental samples. However, currently employed methods are time consuming. Plastic optical fiber (POF) biosensors consist in a viable alternative for rapid and inexpensive scheme for detection. In order to study the sensitivity of tapers for microbiological detection, geometric parameters are studied, such as the taper waist diameter since the formation of taper regions are the key sensing element in this particular type of sensors. In this study, a series of POF taper sensors were prepared using a specially developed tapering machine, and the dispersion of geometric dimensions is evaluated, aiming to achieve the best tapering characteristics which will provide a better sensitivity on the sensor response. The fiber tapers that presented the finest results were those constructed in U-shaped (bended) configurations, with taper waist diameters ranging from 0.40 mm up to 0.50 mm. These fiber tapers were used as the main section of the monitoring device, and when chemically treated as immunosensors for the detection of bacteria, yeast and erythrocytes.

Plastic optical fiber-based biosensor platform for rapid cell detection

This work presents a novel, fast response time, plastic optic fiber (POF) biosensor to detect Escherichia coli. It discloses the technique for the development, calibration and measurement of this robust and simple-to-construct POF biosensor. The probes in U-shaped format were manufactured with a specially developed device. The calibration process led to the evaluation of the sensitivity, accuracy and repeatability by using solutions of sucrose for obtaining refractive indices (RI) in the range 1.33-1.39 IR equivalent of water and bacteria, respectively. The POF probes were functionalized with antibody anti-E. coli serotype O55 and tested firstly with saline and then with bacterial concentrations of 10(4), 10(6), and 10(8) colony forming units/ml (CFU/ml). The optoelectronic setup consists of an 880 nm LED connected to the U-shaped probe driven by a sine waveform generated by the Simulink (from Matlab(®)). On the other side of the probe a photodetector generates a photocurrent which is amplified by a transconductance amplifier. The output voltage signal is read by the analog-to-digital (A/D) input of the microcontroller. In all tested concentrations, the results presented a tendency of a decrease in the output signal with time, due to the attachment of the bacteria to the POF probe and consequent increase in the RI close to the sensitive area of the fiber surface. It has been shown that the system is capable of providing positive response to the bacterial concentration in less than 10 min, demonstrating good possibilities to be commercially developed as a portable field sensor.

A cluster of Listeria monocytogenes infections in hospitalized adults

BACKGROUND Listeriosis occurs mainly in persons at extremes of age and with immunocompromising conditions. It is believed that most cases of listeriosis are acquired in the community. A cluster of listeriosis in hospitalized patients prompted the present investigation. METHODS We conducted a case series study of listeriosis from August 21, 2006, to June 1, 2007, in a hospital in the city of Rio de Janeiro, Brazil. RESULTS Six patients with Listeria monocytogenes infection were identified: 5 during hospitalization and 1 at a day clinic. By the time the infection was diagnosed, 5 patients had been in the hospital for a mean of 9 days. All patients were elderly (median age, 80 years) and had immunocompromising conditions. Five (83%) patients died. Four patients developed bloodstream infections, 3 caused by serotype 1/2b. Two patients had peritonitis: one caused by serotype 3b and another by serotype 1/2b. Four L monocytogenes isolates belonged to a single pulse-field gel electrophoresis genotype, suggesting a common source. An epidemiologic investigation pointed to the hospital kitchen as the possible contamination. CONCLUSION Data suggest a health care-associated outbreak of listeriosis and highlight the importance of developing guidelines for prevention and treatment of health care-associated foodborne diseases, especially in hospitals with immunocompromised adult patients.

Microbiological and chemical characteristics of Brazilian kefir during fermentation and storage processes.

The microbial community composition and chemical characteristics of a Brazilian milk kefir sample produced during its manufacturing and refrigerated storage were investigated by culture-dependent and -independent methods and HPLC. Lactococcus lactis ssp. cremoris and ssp. lactis, Leuconostoc mesenteroides, Acetobacter lovaniensis, and Saccharomyces cerevisiae were isolated, whereas the detected bands on denaturing gel gradient electrophoresis corresponded to Lactobacillus kefiranofaciens, Lactobacillus kefiri, Lactobacillus parakefiri, and S. cerevisiae. After fermentation, lactic acid bacteria were present at levels of 10 log units, whereas acetic acid bacteria and yeast were present at levels of 7.8 and 6 log units, respectively. The lactic acid bacteria and yeast counts remained constant, whereas acetic acid bacteria counts decreased to 7.2 log units during storage. From fermentation to final storage, the pH, lactose content and citric acid of the kefir beverage decreased, followed by an increase in the concentrations of glucose, galactose, ethanol, and lactic, acetic, butyric, and propionic acids. These microbiological and chemical characteristics contribute to the unique taste and aroma of kefir. This research may serve as a basis for the future industrial production of this beverage in Brazil.

Estudo da estabilidade quÍmica e microbiológica do suco de caju in natura armazenado em diferentes condições de estocagem

O caju, rico em nutrientes, apresenta alta perecibilidade. Este trabalho teve como objetivo avaliar a estabilidade quimica, fisico-quimica e microbiologica do suco de caju in natura mantido em temperatura ambiente por 24 h, estocado sob refrigeracao por sete dias e sob congelamento por 120 dias. O teor de acido ascorbico no suco recem-extraido foi de 147,29 ± 0,41 mg/100 mL e reduziu em 6,57% em temperatura ambiente. Durante a estocagem sob refrigeracao e congelamento, as taxas de reducao dessa vitamina foram 1,16% ao dia e 0,05% ao dia, respectivamente. Foi observado aumento na contagem de bacterias mesofilas totais e fungos filamentosos e leveduras no suco mantido em temperatura ambiente. Nos sucos estocados sob refrigeracao durante sete dias houve reducao da contagem de bacterias mesofilas totais e aumento na contagem de fungos filamentosos e leveduras. Nos sucos congelados, a contagem de fungos filamentosos e leveduras permaneceu inferior a inicial, enquanto a de bacterias mesofilas totais apresentou variacao ate o trigesimo dia. A partir deste periodo, esta permaneceu estavel em menos de um ciclo logaritmico acima da contagem inicial. Nos periodos estudados, refrigeracao e congelamento mostraram-se eficazes na preservacao do acido ascorbico e da qualidade microbiologica do suco de caju in natura.

Características microbiológicas do suco de laranja in natura

Fresh orange juices are very popular as a source of vitamin C, minerals and carbohydrates. However, for the same reasons, they are a suitable environment for the microorganism growth, including pathogens that are able to survive in such conditions, as E. coli, Salmonella and Listeria monocytogenes. To assure the quality of these products, microbiological safety monitoring is needed, through measuring lactic acid bacteria (LAB), moulds and yeasts, total and fecal coliforms and total plate counts. These parameters were monitored for 50 fresh juice samples commercialized in Rio de Janeiro, Brazil. The samples were acquired and maintained under refrigerated conditions. Tests were also made for the detection of E. coli and Salmonella sp. The LAB and total plate counts were, for the most of the samples, higher than the expected. The fecal coliforms levels of 15% of the samples were above those permitted by law. The susceptibility patterns for fecal coliforms isolates were as the expected to environmental or fecal samples that were not under selective pressure.

Inhibition of Vancomycin and High-Level Aminoglycoside-Resistant Enterococci Strains and Listeria monocytogenes by Bacteriocin-Like Substance Produced by Enterococcus faecium E86

Three hundred and thirty nine lactic bacteria strains isolated from food samples were screened for antimicrobial activity. Only one strain isolated from meat pie and identified as Enterococcus faecium produced a bacteriocin-like inhibitory substance (BLIS) showing activity against Enterococcus, Leuconostoc, Lactobacillus, Listeria, Corynebacterium and Staphylococcus aureus. The BLIS produced was resistant to acid and alkali treatment and 121ºC for 15 min. The addition of BLIS in BHI contaminated with Listeria monocytogenes decreased the contamination in 4.8 log cycles in 24 h. The inhibition of listeria was also obtained in milk. Forty multiresistant enterococci strains were inhibited in the well-diffusion test. Two vancomycin resistant strains tested in liquid with BLIS were also inhibited. The BLIS producer showed no pathogenicity marker.Three hundred and thirty nine lactic bacteria strains isolated from food samples were screened for antimicrobial activity. Only one strain isolated from meat pie and identified as Enterococcus faecium produced a bacteriocin-like inhibitory substance (BLIS) showing activity against Enterococcus, Leuconostoc, Lactobacillus, Listeria, Corynebacterium and Staphylococcus aureus. The BLIS produced was resistant to acid and alkali treatment and 121ºC for 15 min. The addition of BLIS in BHI contaminated with Listeria monocytogenes decreased the contamination in 4.8 log cycles in 24 h. The inhibition of listeria was also obtained in milk. Forty multiresistant enterococci strains were inhibited in the well-diffusion test. Two vancomycin resistant strains tested in liquid with BLIS were also inhibited. The BLIS producer showed no pathogenicity marker.

Caracterização microbiológica de lixiviados gerados por resíduos sólidos domiciliares e de serviços de saúde da cidade do Rio de Janeiro

The present paper consisted on the microbiological characterization of leachate generated from solid waste from health services and municipal solid waste. This study aimed to support the discussion about the need for differentiated treatment and disposal for the health services solid waste. The following parameters were selected: total coliforms, thermotolerant coliforms, Escherichia coli, Enterococci, trials for Pseudomonas aeruginosa, and Salmonella and other Enterobacteria, analysis of susceptibility to antimicrobials and research on antimicrobial activity of the leachate. Expressive densities were observed in microbial populations in health services and municipal solid waste, with values ranging from 108 to 109. In both wastes, resistant and susceptible strains to antimicrobials occurred and none were resistant to all antibiotics tested. Results showed similarities for both types of waste and reinforce the recommendation for health services and municipal solid waste co-disposal in sanitary landfills.