RESUMO
In order to investigate the microflora of Slovakian bryndza cheese (a cheese containing unpasteurized or pasteurized ewes' milk component) by a culture-independent method, DNA was extracted directly from 7 bryndza samples and analysed by an innovative method. Using the universal prokaryotic and fungal primers, ribosomal DNA internal transcribed spacer (ITS) regions with variable length were amplified. The standard universal reverse primer L1 aligning to bacterial 23s rDNA was found unsuitable for some lactic acid bacteria and other species based on in silico analysis. Therefore, L1 primer was replaced by a combination of novel primers GplusR and GminusR aligning to the adjacent, more conserved DNA region. The amplification profiles were visualised by both standard electrophoresis and by fluorescent capillary gel electrophoresis. From representative samples, major amplicons were excised from the gel, cloned and sequenced. Sequencing revealed that the samples contained Lactobacillus delbrueckii, Lactobacillus brevis, Streptococcus thermophilus, Lactococcus lactis, Lactococcus raffinolactis, Streptococcus macedonicus, Leuconostoc pseudomesenteroides, Debaromyces hansenii, Mucor fragilis, Yarrowia lipolytica and Galactomyces geotrichum. These results represent an extension of the knowledge on the microflora of Slovakian bryndza cheese. The introduced automated ribosomal DNA intergenic spacer analysis of the bacterial and fungal genomes proved to be very effective in the application of studying microflora of cheese.
Assuntos
Queijo/microbiologia , Microbiologia de Alimentos , Animais , Bactérias/classificação , Bactérias/genética , Contagem de Colônia Microbiana , DNA Bacteriano/análise , DNA Bacteriano/genética , Feminino , Humanos , Lactobacillus/classificação , Lactobacillus/genética , Lactococcus/classificação , Lactococcus/genética , Lactococcus lactis/classificação , Lactococcus lactis/genética , Leuconostoc/classificação , Leuconostoc/genética , Leite/química , Leite/microbiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Eslováquia , Streptococcus/classificação , Streptococcus/genética , Streptococcus thermophilus/classificação , Streptococcus thermophilus/genéticaRESUMO
The 16S rRNA gene sequence of the type strain of Ruminococcus flavefaciens, the type species of the genus Ruminococcus, was determined by PCR direct sequencing. A comparative sequence analysis showed that R. flavefaciens is phylogenetically related to a small cluster (cluster IV of Collins et al. [M. D. Collins, P. A. Lawson, A. Willems, J. J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994]) of organisms which includes several Clostridium and Eubacterium species. R. flavefaciens was found to be phylogenetically only remotely related to Ruminococcus gnavus, Ruminococcus torques, Peptostreptococcus productus, and Streptococcus hansenii. These findings demonstrate that the genus Ruminococcus is not a monophyletic group, and the proposed transfer of P. productus and S. hansenii to this genus (T. Ezaki, N. Li, Y. Hashimoto, H. Miura, and H. Yamamoto, Int. J. Syst. Bacteriol. 44:130-136, 1994) is not supported.
Assuntos
Clostridium/classificação , Eubacterium/classificação , Cocos Gram-Positivos/classificação , Sequência de Bases , Clostridium/genética , DNA Ribossômico/genética , Eubacterium/genética , Cocos Gram-Positivos/genética , Dados de Sequência Molecular , Peptostreptococcus/classificação , Peptostreptococcus/genética , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Homologia de Sequência do Ácido Nucleico , Streptococcus/classificação , Streptococcus/genéticaRESUMO
Streptococci that belong to groups other than A are involved in a variety of human infections. In the present study, we report the pyogenic infections associated with group G streptococci (GGS), observed at the Christian Medical College Hospital, Vellore, Tamil Nadu, India. Of 2314 beta-haemolytic streptococci isolated from 31,857 pus specimens processed during the period 1975-1986, GGS accounted for 338 (14.6%). Twelve of these were grown in pure culture. The clinical conditions associated with them included pyogenic arthritis, osteomyelitis, paravertebral sinus and an abdominal burst. Clinical analysis of 99 of 125 patients with GGS, grown in mixture with another organism or skin contaminants, showed that 59% of them were from surgical wound infections, diabetic lesions or wound infections in leprosy patients. Surgical management and antibiotic treatment resulted in full recovery of these clinical conditions.