A total of 54 rapidly growing mycobacteria (RGM) isolated from patients attended in the two hospitals of Cádiz Bay (Spain) were selected during a seven-year-period (2000-2006) in order to evaluate the INNO-LiPA Mycobacteria v2 assay for mycobacterial identification, based on the reverse hybridization principle. The strains were cultured in Löwenstein-Jensen and Middlebrook 7H9 media and identified to the species level by sequencing of the 16S rRNA, PCR-restriction enzyme analysis of the hsp65 gene, conventional tests and INNO-LiPA Mycobacteria v2 assay. By the molecular methods we identified a total of 12 different species: 23 Mycobacterium fortuitum, 11 M. chelonae, 10 M. abscessus, 2 M. senegalense, 1 M. alvei, 1 M. brumae, 1 M. mageritense, 1 M. mucogenicum, 1 M. neoaurum, 1 M. peregrinum, 1 M. septicum and 1 M. smegmatis. Fifty two strains (96.3%) were correctly identified by conventional techniques and 47 strains (87.0%) by INNO-LiPA Mycobacteria v2 assay. We find INNO-LiPA Mycobacteria v2 assay simple to perform but it provides few advantages in comparison with conventional methods and sometimes needs complementary tests to identify Mycobacterium fortuitum complex, M. chelonae complex and specific species due to the great heterogeneity in the RGM group.

Background: Tuberculous spondylodiscitis is relatively uncommon but represents the most common location of osseous tuberculosis. Aim: To describe clinical features, imaging studies and laboratory tests to establish the diagnosis in a group of patients living in Cádiz (Spain). Material and Methods: Retrospective analysis of medical records of patients with tuberculous spondylodiscitis diagnosed between 2000 and 2009. The diagnosis was based on microorganism recovery from vertebral samples obtained by imaging guided biopsies. Results: Six patients with positive Mycobac-terium tuberculosis cultures from vertebral samples, were identified (10% of extra-pulmonary tuberculosis). In only 2 patients the Ziehl-Nielsen stain was positive, and histology was compatible in 4 cases. Four patients were females, their mean age was 54.3 years and the mean duration of symptoms was 7.3 months. Three patients had lumbar location and a positive Mantoux test. A soft tissue abscess was present in 4 cases. None of these patients had neurological complications. The treatment with four tuberculostatic agents (isoniazid, rifampicin, pyrazinamide and ethambutol) was effective in 5 patients. Conclusions: Tuberculous spondylodiscitis may become a serious disease due to diagnostic and treatment delays. The main examinations to establish diagnosis are magnetic resonance imaging and biopsy with microbiological culture. Generally, antituberculous therapy is effective in this clinical situation.

Background: Rapidly growing mycobacteria (RGM) are considered opportunistic pathogens. An increasing number of post traumatic or surgical infections are caused by these microorganisms. Aim: To determine the antimicrobial susceptibility of RGM using the E-test method. Material and methods: A total of 54 isolates of RGM was obtained from several clinical samples and selected for this study Strains were identified to the species level by phenotypic and biochemical characteristics, PCR-restriction enzyme analysis of the hsp65 gene (PRA) and sequencing of the 16S rRNA. Susceptibility was investigated by E-test to amikacin, cefoxitin, ciprofioxacin, clarithromycin, imipenem, quinupristin/dalfopristin, linezolid and tigecycline. Results: Twelve different species of RGM were identified: Mycobacterium fortuitum (23 strains), M chelonae (11), M abscessus (10), Msenegalense (2), Malvei (1), Mbrumae (1), Mmageritense (1), mucogenicum (1), M neoaurum (1), Mperegrinum (1), M septicum (1) y M smegmatis (1). All the strains were inhibited by low concentrations of amikacin and tigecycline. Susceptibility to cefoxitin, fluoroquinolones, clarithromycin, imipenem and linezolid was variable. All but two strains were resistant to quinupristin/ dalfopristin. Conclusions: Due to the uneven antimicrobial susceptibility of different species of RGM, an antimicrobial susceptibility test is mandatory for these microorganisms. The E-test method is well suited to determine minimum inhibitory concentrations.

Background : Tuberculosis in the elderly is an increasing public health problem. Aim: To describe the epidemiological, clinical and microbiological characteristics of tuberculosis in patients over 65 years old, as compared to younger patients. Material and methods: We reviewed the case histories of tuberculosis, microbiologically diagnosed by Ziehl-Neelsen/auramine smear and Löwenstein-Jensen culture, in patients over 65 years old, in the sanitary area of Cadiz (Spain). Results: We detected 49 new cases (37 males) of tuberculosis in elderly subjects. The infection rate in this age group (16.3/100.000 inhabitants) is lower than in people younger than 65 years old (21.1/100.000 inhabitants). The most common symptoms were cough (44.9%), fever (40.8%) and dyspnea (44.9%). Neither of them were infected by the human immunodeficiency virus nor were parenteral drug abusers. Thirty seven percent of cases had a history of previous tuberculosis, and in 75.5% the disease location was pulmonary. The Ziehl-Neelsen/auramine smear was negative in 53.1% of the samples. Conclusions: The number of patients older than 65 years old with tuberculosis has increased in the sanitary area of Cadiz (Spain). This group of patients shows different epidemiological and clinical characteristics than younger patients (Rev Méd Chile 2004; 132: 325-30).

The efficiency of a new chromogenic differential medium CHROMagar Candida was evaluated against two media of efficiency proved. We tested 34 different yeast species, pertaining to eight genera: Candida, Cryptococcus, Trichosporon, Blastoschizomyces, Rhodotorula, Kloeckera, Pichia and Saccharomyces, previously identified. Isolates were cultured in each plates of chromogenic media. The colony colour, morphology and growth were noted. Colonies in new medium developed small size and high intensity colour. Several yeasts showed same morphotype. New CHROMagar medium has not demonstrated similar efficiency for identification of yeast species that another media. It presents less sensibility, longer incubation time for colony growth, and the differentiation of some clinical interest yeasts as C. tropicalis, C. dubliniensis and C. parapsilosis was difficult.

Se evaluó la utilidad de un nuevo medio cromogénico diferencial CHROMagar Candida frente a dos medios de eficacia probada. Ensayamos 34 levaduras de diferentes especies, pertenecientes a ocho géneros: Candida, Cryptococcus, Trichosporon, Blastoschizomyces, Rhodotorula, Kloeckera, Pichia y Saccharomyces. Tras su correcta identificación se cultivaron en los tres medios a estudiar, apreciando la morfología colonial, color y capacidad de crecimiento. En el nuevo medio las colonias presentaron un tamaño menor y más intensidad de color. Muchas especies ofrecieron el mismo morfotipo. El nuevo medio CHROMagar Candida no muestra la misma eficacia que los otros medios para la identificación de especies de levaduras, al presentar menor sensibilidad, requerir un mayor tiempo de incubación para que las colonias se desarrollen y dificultar la diferenciación de algunas especies de interés clínico como C. tropicalis, C. dubliniensis y C. parapsilosis.