Comparación de medios de cultivo selectivos para el aislamiento de especies del Taxón K perteneciente al complejo Burkholderia cepacia / Comparison of selective culture media for the isolation of Taxon K species belonging to the Burkholderia cepacia complex

Objetivo: Dentro del complejo Burkholderia cepacia (cBc), el Taxón K, integrado por B. contaminans y B. lata, es frecuentemente aislado de muestras clínicas e industriales. Los métodos para aislar bacterias del cBc están consensuados en el ámbito clínico pero no para muestras de origen industrial y tampoco hay información documentada sobre la capacidad de recuperación de los medios de cultivo frente a especies del Taxón K. Dada la importancia del uso correcto de medios selectivos para la recuperación de microorganismos, el objetivo de este trabajo fue comparar el agar Trypan Blue-Tetraciclina (TB-T), el agar selectivo para Burkholderia cepacia (BCSA) y el BCSA comercial modificado (BCSAm) en el aislamiento de cepas del Taxón K. Métodos: empleamos el método ecométrico utilizado en el control de calidad de medios de cultivo. Analizamos criterios de productividad, selectividad y especificidad frente a cepas de referencia del cBc, aislamientos clínicos e industriales del Taxón K y cepas de otras especies. Resultados: no se observaron diferencias de productividad y selectividad entre los medios BCSA. Con ambos se obtuvo adecuada productividad y selectividad parcial por permitir el crecimiento de especies taxonómicamente cercanas al cBc. El medio TB-T presentó menor productividad (especialmente con B. contaminans) y menor selectividad. Por otra parte, no se observaron diferencias atribuibles al origen clínico o industrial de los aislamientos. Conclusión: los resultados permiten sugerir al BCSA o BCSAm como los medios selectivos de elección para el aislamiento del Taxón K, tanto en muestras de origen clínico como industrial

Objective: Within the Burkholderia cepacia complex (Bcc), the so called Taxon K, integrated by B. contaminans and B. lata, is frequently isolated from clinical and industrial samples. There is consensus in the use of culture media for the isolation of Bcc from clinical origin but not for industrial samples. By the other side there is no documented information about the performance of culture media recovering Taxon K species. Regarding the importance of the proper use of selective media in the recovery of microorganisms from clinical and industrial samples, the objective of this work was to compare Trypan Blue-Tetracycline agar (TB-T), Burkholderia cepacia selective agar (BCSA) and commercial modified Burkholderia cepacia selective agar (BCSAm) in the isolation of Taxon K strains. Methods: we employed the ecometric method for culture media quality control. Productivity, selectivity and specificity criteria were analyzed by testing Bcc reference strains, clinical and industrial Taxon K isolates and non Bcc strains. Results: no differences in terms of productivity and selectivity were observed between BCSA and BCSAm. Both medium, displayed adequate productivity and partial selectivity since the growth of non Bcc isolates was observed. Productivity (especially with B. contaminans isolates) and selectivity in TB-T was lower than BCSA medium. No differences were observed related to the clinical or industrial origin of isolates. Conclusion: results allow us to suggest BCSA or BCSAm selective media for the isolation of Taxon K strains in clinical or industrial samples

Comparison of ChromID C. difficile agar and cycloserine-cefoxitin-fructose agar for the recovery of Clostridium difficile.

AIM: The rapidly changing epidemiology of Clostridium difficile infection highlights the need for improved and continuing surveillance involving stool culturing to enable molecular tracking. Culture of C. difficile can be difficult and time consuming. In this report ChromID C. difficile agar (CDIF) was compared to cycloserine-cefoxitin-fructose-egg-yolk agar which contained 0.1% sodium taurocholate (TCCFA) as a germinant. RESULTS: All ribotypes of C. difficile tested (n=90) grew well on CDIF within 24 h and most gave characteristic small irregular black colonies with a raised umbonate profile. Counts from standard suspensions of C. difficile at 24 h (p<0.005) and 48 h (p=0.01) were significantly higher on CDIF than on TCCFA. Similar results were achieved after alcohol shock. When temperature shock was used to differentiate vegetative cells and spores, the total number of culturable and vegetative cells on CDIF was significantly higher than on TCCFA (culturable cells, p=0.003 at 24 h and p=0.002 at 48 h; vegetative cells, p=0.0003 at 24 h and p=0.0002 at 48 h). CONCLUSIONS: These data suggest that CDIF is a better medium for the recovery of vegetative C. difficile than TCCFA and equal to TCCFA for spore recovery.

Recovery of mutans streptococci on MSB, SB-20 and SB-20M agar media.

OBJECTIVE: The recovery of mutans streptococci in saliva and dental biofilm samples depends, in part, on the culture medium used. In this study, we compared (i) the culture media Sucrose-Bacitracin agar (SB-20), Modified SB-20 (SB-20M) and Mitis Salivarius Bacitracin agar (MSB) in the count of colony forming units (cfu) of mutans streptococci and (ii) in the morphological and biochemical differentiation between Streptococcus mutans and Streptococcus sobrinus. DESIGN: Samples of non-stimulated saliva from 20 children were plated on SB-20, SB-20M and MSB, and incubated in microaerophilia at 37°C for 72h. Identification of microorganisms was based on analysis of colony morphology under stereomicroscopy. The biochemical identification of colonies was done by biochemical tests using sugar fermentation, resistance to bacitracin and hydrogen peroxide production. RESULTS: There was no significant difference (p>0.05) in the number of cfu of mutans streptococci recovered on SB-20 and SB-20M agar. Comparing the media, SB-20 and SB-20M yielded a larger number of mutans streptococci colonies (p<0.05) and were more effective than MSB in the identification of S. sobrinus (p<0.05), but not of S. mutans (p>0.05). CONCLUSION: There was no significant difference between SB-20 and SB-20M culture media in the count of mutans streptococci, demonstrating that the replacement of sucrose by coarse granular cane sugar did not alter the efficacy of the medium. Compared with MSB, SB-20 and SB-20M allowed counting a larger number of mutans streptococci colonies and a more effective morphological identification of S. sobrinus.

[Evaluation of Agar Candida ID2 (bioMerieux) a chromogenic medium for yeasts differentiation]. / Ocena przydatnosci chromogennych podlozy Agar Candida ID 2 do identyfikacji grzybów rodzaju Candida.

Invasive diagnostic and therapeutic methods, widespread antibiotic therapy and rising percent of the immunocompromised patients cause incrementation of frequency of occurrence of the yeast infection. C. albicans is the most commonly isolated species of Candida from clinical samples. However, recently growth of frequency of isolation Candida non - albicans from clinical specimens have been observed. Yeast-like fungi different from C. albicans have become serious clinical problem. Conventional methods of identification of the yeast-like fungi carry away a lot time enough. Employment of chromogenic agar shortens latency on result. We decided to examine the usefulness ofAgar Candida ID2 (CAN2) (bioMérieux) in the identification of Candida species. The subjects within the study were 146 of Candida spp strains which were isolated from the clinical specimens of patients hospitalized at the University Hospital in Bydgoszcz. Germ tube test. Api 20C AUX test (bioMérieux) and Agar Candida ID2 (bioMérieux) were used. We have ascertained correspondence of identifying species amounted to 82.2% of analyzed Candida species between API 20C AUX test and kind of growth on CAN2 medium. Divergence of results received between CAN2 medium and API 20C AUX test suggests necessity of conducting of verification data with other methods. In conclusion, our study shows that Agar Candida ID2 is an effective medium for the isolation yeast-like fungi and in preliminary identification of Candida species direct from clinical materials.

Comparison of selective agars for the isolation and identification of Klebsiella oxytoca and Escherichia coli from environmental drinking water samples.

Various selective media were assessed for their ability to detect and differentiate Klebsiella oxytoca and Escherichia coli in environmental water samples. Only two, Membrane Lauryl Sulphate agar and Deoxycholate Agar, could differentiate the two coliforms from each other and from the 'background' heterotrophs in water and this was a consequence of E. coli's ability to grow at 44 degrees C and 37 degrees C whereas Kl. oxytoca could only grow at 37 degrees C. Modified M-FC medium effectively differentiated Kl. oxytoca but not E. coli in environmental samples. Other media characterized the different coliforms in pure culture but failed to do likewise in environmental samples. For example, pure cultures of E. coli fluoresced when MUG was added to the medium but single colonies on a mixed species plate failed to do so. MT7 agar distinguished the two coliforms from water heterotrophs but not from each other.

Enumeration of intestinal enterococci and interfering organisms with Slanetz-Bartley agar, KF streptococcus agar and the MUST method.

The recovery of intestinal species of enterococci and streptococci and potentially interfering nonfaecal species was measured on KF streptococcus agar, Slanetz-Bartley agar and in a medium based on 4-methylumbelliferyl-beta-D-glucoside (MUST method) using pure cultures. Both of the solid media yielded high recoveries of the target species. Their selectivity was better at elevated incubation temperature but nonfaecal Enterococcus and Staphylococcus species were not eliminated even at the elevated temperature. The MUST method tended to give slightly lower recoveries than the agar cultivation methods with some target species at 44 degrees C but recoveries were better at 41 degrees C.

Evaluation of two new chromogenic media for detection of Salmonella in stools.

A clinical collaborative study was conducted to compare two new chromogenic agar media, Rambach agar and the Salmonella Detection and Identification Medium (SMID) (bioMérieux, France), with two conventional media, Salmonella-Shigella agar and Hektoen agar. Thirty-nine Salmonella strains involving 14 serotypes were isolated from 1,454 stool specimens. After enrichment in a selective broth, 100% sensitivity was obtained with each medium. The SMID and Rambach agars are considerably more specific than the conventional media. Although SMID agar detects all Salmonella serotypes, it is not as specific as Rambach agar, which requires a complementary test (C8 esterase test) to detect all serotypes.

Bacteriological and serological study of pertussis in Abeokuta, Nigeria.

Charcoal horse blood agar is the medium of choice for isolation of Bordetella pertussis from patients with early whooping cough. Since sterile animal blood often is not available in developing countries, a field study in Nigeria was undertaken to evaluate donated human blood as supplement to charcoal agar. Out of 209 children with suspected early pertussis, 33 were culture-positive (isolation rate 16%). Out of 188 children studied serologically by enzyme immunoassay, 36 (19%) were seropositive. The satisfactory isolation rate of 16% shows that culturing for B. pertussis on charcoal human blood agar can be tried in countries, where there is no regular supply of bacteriological media with animal blood.

Contaminaciòn ambiental en el Servicio de Ginecologia Hospital de Clìnicas: La Paz-Bolivia / Eviromental pollution in the Gynocoloy Service of Hospital de Clinicas La Paz - Bolivia

En el servicio de Ginecologìa del Hospital de Clìnicas de La Paz, se investiga la contaminaciòn ambiental existente. Se utilizaron 4 cajas de Petri que se ubicaron en diferentes lugares del servicio; cada una de ellas contenia agar-sangre como medio de cultivo. Luego de la incubaciòn respectiva y el desarollo bacteriano observado, se identificaron cinco variedades de gèrmenes, de los cuales 3 son patògenos y 2 no patògenos. Se concluye en que la sala y el consultorio presentan un 60 porciento de contaminaciòn. Se sugiere tomar algunas medidas para disminuir esa contaminaciòn y asi disminuir tambièn la mortalidad existente en el servicio