A novel sampling method for the investigation of gut mirobiota.

AIM: In order to characterize the qualitative and quantitative microorganisms in different sites of the lower digestive tract (LDT) in healthy volunteers, a specific technique was developed for collecting mucous of the distal ileum, colon and rectum. METHODS: A polyethylene tube was designed to go through the colonoscope channel with a No.8 French tube. In order to avoid internal contamination, the distal extremity was protected with a membrane of microfilm after being sterilized in ethilene oxid. To facilitate the aspiration of a precise volume, its interior was coated with silicone. One hundred microlliter (0.1 mL) sample of mucous was collected and transferred into an Eppenddorff tube containing nine hundred microlliter (0.9 mL) of VMGA-3 (viable medium of Goteborg). This procedure was repeated at each site of the LDT with a new sterilized catheter. RESULTS: All sites revealed the "non pathogenic" anaerobic bacteria Veillonella sp (average 10(5) colony forming units/mL-CFU/mL), allowing to conclude an environment of low oxidation-reduction potential (redox) in the LDT. It was also characterized the presence of Klebisiella sp with significant statistical predominance (SSP) in the ileum. Enterobacter sp was found with SSP in the sigmoid colon, Bacteroides sp non-pigmented (npg) and E.coli with SSP in the sigmoid colon and rectum, Enterococcus sp and Lactobacillus sp with SSP in the rectum, all in a mean concentration of 10(5) CFU/mL. CONCLUSION: This procedure is feasible and efficient and can point out a similar distribution of the aerobic and anaerobic bacteria with the presence of biological markers of normal microbiota in the LDT.

Digestive tract microbiota in healthy volunteers.

PURPOSE: The aim of this study was to standardize the methods of sample collection of mucus from the digestive tract and to determine the microbiota in healthy volunteers from Brazil, collecting samples from the mouth, esophagus, stomach, duodenum, jejunum, ileum, colon, and rectum. METHODS: Microbiota of selected healthy volunteers from the oral cavity (n=10), the esophagus (n=10), the upper digestive tract (n=20), and the lower digestive tract (n=24) were evaluated through distinct collection methods. Collection methods took into account the different sites, using basic scraping and swabbing techniques, stimulated saliva from the oral cavity, irrigation-aspiration with sterile catheters especially designed for the esophagus, a probe especially designed for upper digestive tract, and a special catheter for the lower digestive tract. RESULTS: (i) Mixed microbiota were identified in the oral cavity, predominantly Gram-positive aerobic and anaerobic cocci; (ii) transitional flora mainly in the esophagus; (iii) Veillonella sp, Lactobacillus sp, and Clostridium sp in the stomach and duodenum; (iv) in the jejunum and upper ileum, we observed Bacteroides sp, Proteus sp, and Staphylococcus sp, in addition to Veillonella sp; (v) in the colon, the presence of "nonpathogenic" anaerobic bacteria Veillonella sp (average 10(5) UFC) indicates the existence of a low oxidation-reduction potential environment, which suggests the possibility of adoption of these bacteria as biological markers of total digestive tract health. CONCLUSIONS: The collection methods were efficient in obtaining adequate samples from each segment of the total digestive tract to reveal the normal microbiota. These procedures are safe and easily reproducible for microbiological studies.

Evaluation of normal ocular bacterial flora with two different culture media.

BACKGROUND: The purpose of this study was to determine if the use of broth culture medium is efficient in investigating bacterial flora of the normal eyelid and conjunctiva. METHODS: Samples from the conjunctiva and eyelid of healthy patients of various ages who were undergoing ocular surgeries were obtained and cultured at 3 periods: before topical antibiotic prophylaxis, in the postoperative period during topical antibiotic treatment, and 15 days after discontinuation of antibiotic use. Samples were inoculated into both brain heart infusion broth and blood agar plate, and the growth results of both media were analyzed. RESULTS: Brain heart infusion broth medium showed a significantly higher bacterial growth of gram-positive cocci in most periods. The solid blood agar medium had a higher recovery of gram-positive bacilli before prophylaxis only in the older patients. INTERPRETATION: Our results show that a more complete analysis of eyelid and conjunctival flora can be obtained using both liquid and solid media to increase the chances of isolate recovery. The inclusion of liquid media in this analysis was even more relevant in the period of concomitant use of antibiotic treatment.

Phage typing of Salmonella enteritidis from different sources in Brazil.

The occurrence of Salmonella Enteritidis (SE) phage types (PTs) in samples collected from healthy and diseased chickens, in outbreaks of human gastroenteritis related to the consumption of egg products, in samples of poultry meat, in pipped embryos of broiler chickens, in meat meal, in poultry-rearing environments, and in many foods (cheese, mayonnaise, cake, and bacon) is described for strains isolated from 1995 to 1997 in Brazil. SE strains were isolated, and the most common PT was found to be PT 4, followed by PTs 7, 21, 35, 6, 4a, 8, 30, 6a, 5a, 1, and 1b. Fourteen strains were classified as react-but-do-not-conform strains, and one strain was not typeable. The results of this study demonstrate that PT 4 has a wider distribution among the sources studied than do any other SE phage types and is the most important phage type in human salmonellosis.

Tuberculosis survey of free-ranging marsh deer (Blastocerus dichotomus) in Brazil.

Esophageal-pharyngeal fluids from 53 free-ranging marsh deer (Blastocerus dichotomus) captured for a research program in the state of Mato Grosso do Sul, Brazil, were assayed for tuberculosis. Total DNA was extracted. amplified by polymerase chain reaction using specific primers for Mycobacterium tuberculosis complex (M. tuberculosis, M. bovis, M. microti, and M. africanum), and observed by agarose gel electrophoresis stained with ethidium bromide. All samples were negative. This, along with necropsy and histopathology data, suggests that these animals are not shedding and probably do not have active disease.

Single-use trocar: is it possible to reprocess it after the first use?

The aim of this study was to verify and describe the presence of microorganisms in the single-use trocar after its use in surgical procedures, and after this device was submitted to cleaning, conditioning, and sterilization by physicochemical processes (formaldehyde, ethylene oxide, and hydrogen peroxide plasma). Twenty-eight trocars of the Ethicon, Auto-suture, and Aesculap brands, were randomly selected and analyzed after laparoscopic cholecystectomy. The results have shown that cultures grown of the material collected from the trocars, immediately after its use and before its sterilization process, showed the presence of bacteria and fungi in 46.5% (13). In 53.5% (15) of the trocars, the presence of microorganisms was not detected, very likely due to niche's scarcity. In the cultures grown of the 28 trocars after being submitted to sterilization processes, the presence of microorganisms was not verified. We can therefore conclude that although trocars possess compartments not easily accessed for cleaning, these devices can be adequately cleaned and effectively sterilized, when well manipulated, in the institution where the study was carried out by the processes of steam sterilization at low temperature and formaldehyde, ethylene oxide, and hydrogen peroxide plasma.

Digestive tract microbiota in healthy volunteers

PURPOSE: The aim of this study was to standardize the methods of sample collection of mucus from the digestive tract and to determine the microbiota in healthy volunteers from Brazil, collecting samples from the mouth, esophagus, stomach, duodenum, jejunum, ileum, colon, and rectum. METHODS: Microbiota of selected healthy volunteers from the oral cavity (n=10), the esophagus (n=10), the upper digestive tract (n=20), and the lower digestive tract (n=24) were evaluated through distinct collection methods. Collection methods took into account the different sites, using basic scraping and swabbing techniques, stimulated saliva from the oral cavity, irrigation-aspiration with sterile catheters especially designed for the esophagus, a probe especially designed for upper digestive tract, and a special catheter for the lower digestive tract. RESULTS: (i) Mixed microbiota were identified in the oral cavity, predominantly Gram-positive aerobic and anaerobic cocci; (ii) transitional flora mainly in the esophagus; (iii) Veillonella sp, Lactobacillus sp, and Clostridium sp in the stomach and duodenum; (iv) in the jejunum and upper ileum, we observed Bacteroides sp, Proteus sp, and Staphylococcus sp, in addition to Veillonella sp; (v) in the colon, the presence of "nonpathogenic" anaerobic bacteria Veillonella sp (average 10(5) UFC) indicates the existence of a low oxidation-reduction potential environment, which suggests the possibility of adoption of these bacteria as biological markers of total digestive tract health. CONCLUSIONS: The collection methods were efficient in obtaining adequate samples from each segment of the total digestive tract to reveal the normal microbiota. These procedures are safe and easily reproducible for microbiological studies.

OBJETIVO: Padronizar os métodos de coleta do muco do trato digestivo e determinar a microbiota, em voluntários saudáveis no Brasil, coletando amostras da boca, esôfago, estômago, duodeno, jejunos e íleo, cólons e reto. MÉTODOS: A microbiota de voluntários saudáveis foi avaliada através de diferentes métodos de coleta: cavidade oral (n=10 voluntários), do esôfago (n=10), do trato digestivo alto (n=20) e do trato digestivo baixo (n=24). Métodos de coleta foram adotados em cada sítio restrito, usando derramar saliva, técnica de esfregar a mucosa e saliva estimulada da cavidade oral, irrigação-aspiração, cateteres específicos designados para o esôfago, sonda especial para o trato digestivo alto e cateteres especiais para o trato digestivo baixo. RESULTADOS: Identificados: (i) na cavidade oral, microbiota mista, predominando cocos aeróbios e anaeróbios Gram positivos; (ii) no esôfago, flora transitória; (iii) no estômago e duodeno, Veillonella sp, Lactobacillus sp and Clostridium sp; (iv) no jejuno e íleo proximal, Bacteróides sp, Proteus sp and Staphilococcus sp, além da Veillonella sp ; (v) no colon, foi revelada a presença "não patogênica" da bactéria anaeróbica Veillonella sp numa concentração média de 10(5) unidades formadoras de colônia, indicando um meio de baixo potencial de oxido-redução e a possibilidade de se conceituar esta bactéria como um marcador biológico do trato digestivo total em sadios. CONCLUSÃO: Estes métodos de coleta foram considerados eficientes para obtenção adequada de amostra em cada segmento do trato digestivo total para caracterizar a microbiota normal. Estes procedimentos são seguros e facilmente reprodutível para estudo microbiológico.

Molecular epidemiology of Lyme disease spirochetes based on a probe complementary to ribosomal RNA

Lyme disease is caused by the spirochete, Borrelia burgdorferi, a bacteria which infects many vertebrates including humans. Borrelia have been isolated from many parts of the world, and there is interest to identify commun genetic markers to improve molecular methods of diagnosis, and to aid in understanding varied manifestations of the disease. A total of 48 Borrelia burgdorferi strains, including: 38 isolated from ticks (Ixodes dammini, I. persulcatus, I. ricinus and I. pacificus), 3 from animals (dog, bird and hamster), and 7 from human clinical cases (skin, CSF, plasma and blood) from different geographic areas, were studied by DNA/DNA hybridization and rRNA gene restriction patterns by using a biotinylated pKK3535 probe (Altewegg M., Mayer L.W., 1989). The migration patterns of rRNA generestriction fragments after clevage by Hind III separate these strains into 5 ribotypes of Borrelia burgdorferi: Type I (38 American, 2 European strains); Type II (13 American strains); Type III (3 Asian and 1 European strains); Type IV (1 European and 2 Asian strains) and Type V (1 Asian strain). The use of ribotyping has provided and additional tool to investigate the diferences or commun patterns which cause various Lyme disease syndromes.