Serratia rubidaea biotype 1 was isolated from the bile and blood of a patient with a bile tract carcinoma obstructing the common bile duct and who underwent invasive procedures. The infection was cleared after adequate treatment with antibiotics.

The usefulness of single-enzyme amplified-fragment length polymorphism (AFLP) analysis for the subtyping of Mycobacterium kansasii type I isolates was evaluated. This simplified technique classified 253 type I strains into 12 distinct clusters. The discriminating power of this technique was high, and the technique easily distinguished between the epidemiologically unrelated control strains and our clinical isolates. Overall, the technique was relatively rapid and technically simple, yet it gave reproducible and discriminatory results. This technique provides a powerful typing tool which may be helpful in solving many questions concerning the reservoirs, pathogenicities, and modes of transmission of these isolates.Mycobacterium kansasii is the most common cause of pulmonary nontuberculous mycobacterial infection in the nonhuman immunodeficiency virus-infected population in many parts of the world (4,6,9,12,16,19,25). Of all nontuberculous mycobacterial diseases, the clinical course of M. kansasii lung disease most closely parallels that caused by M. tuberculosis infection (12,36). Although it has seldom been recovered from soil (10, 22), M. kansasii has frequently been isolated from tap water and is thought to be acquired from the environment rather than by case-to-case transmission (8,17,18,20,24,29,31,37).The first typing method developed for M. kansasii was phage typing. Other features, especially catalase activity, have been used to type M. kansasii isolates. Isolates with high catalase activities were considered more virulent (23). Analysis of the 16S rRNA sequence (27), amplification of the 16S-23S rRNA spacer region (1), PCR-restriction fragment length polymorphism (RFLP) analysis of the hsp65 gene (7, 33), and detection of insertion sequence element IS1652 (38) showed that M. kansasii contains a subspecies genetically distinct from the typical M. kansasii isolates.M. kansasii has been classified into five subspecies or types (types I to V) on the basis of PCR-RFLP analysis of the hsp65 gene (7, 33). These results have been confirmed by differences in the sequences of the 16S-23S rRNA spacer region (2) and by RFLP analysis with the major polymorphic tandem repeat probe (23). Recently, two new types (types VI and VII) have been described (26,32). Of the seven types identified, M. kansasii type I is the most prevalent type from human sources worldwide. Moreover, large restriction fragment-pulsed-field gel electrophoresis (LRF-PFGE) (2, 14, 23), amplified-fragment length polymorphism (AFLP) analysis (23), and randomly amplified polymorphic DNA analysis (2) have produced polymorphic patterns within each type.By use of these typing methods, minimal genetic polymorphism was noted among type I strains. These results gave the impression that type I shows substantial clonality (21). This apparent clonality may have resulted from the insufficient discriminatory powers of the techniques used. Solutions to questions about the reservoirs, pathogenicities, and modes of transmission of these isolates require more discrim...

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Molecular epidemiology of circulating clinical isolates is crucial to improve prevention strategies. The Spanish Working Group on multidrug resistant tuberculosis (MDR-TB) is a network that monitors the MDR-TB isolates in Spain since 1998. The aim of this study was to present the study of the MDR-TB and extensively drug-resistant tuberculosis (XDR-TB) patterns in Spain using the different recommended genotyping methods over time by a national coordinated system. Based on the proposed genotyping methods in the European Union until 2018, the preservation of one method, MIRU-VNTR, applied to selected clustered strains permitted to maintain our study open for 20 years. The distribution of demographic, clinical and epidemiological characteristics of clustered and non-clustered cases of MDR/XDR tuberculosis with proportion differences as assessed by Pearson’s chi-squared or Fisher’s exact test was compared. The differences in the quantitative variables using the Student's-t test and the Mann–Whitney U test were evaluated. The results obtained showed a total of 48.4% of the cases grouped in 77 clusters. Younger age groups, having a known TB case contact (10.2% vs 4.7%) and XDR-TB (16.5% vs 1.8%) were significantly associated with clustering. The largest cluster corresponded to a Mycobacterium bovis strain mainly spread during the nineties. A total of 68.4% of the clusters detected were distributed among the different Spanish regions and six clusters involving 104 cases were grouped in 17 and 18 years. Comparison of the genotypes obtained with those European genotypes included in The European Surveillance System (TESSy) showed that 87 cases had become part of 20 European clusters. The continuity of MDR strain genotyping in time has offered a widespread picture of the situation that allows better management of this public health problem. It also shows the advantage of maintaining one genotyping method over time, which allowed the comparison between ancient, present and future samples.

publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.Poster Presentations S49 quantitative detection is difficult from environments in which mycobacteria co-exist with usual bacteria. Aim/Objective: To evaluate a method for the selective and quantitative detection of mycobacteria from water in hospital environments, and to detect usual bacteria and mycobacteria quantitatively using with the new method in endoscope reprocessors. Methods: We have developed a modified oxalic-acid method (OAM) anew. In the OAM, samples are treated with oxalic-acid solutions for 30 min. After neutralization with a phosphate buffer, samples are filtrated with filters (pore size: 0.45 mm). The filters are placed onto 7H11 agar which contained nalidixic acid and amphotericin B for culture of mycobacteria. Results: On the concentration of oxalic acid in OAM, a 0.05% solution killed most of gram-negative rods, whereas it did not decrease CFU of M. fortuitum, a rapid grower. Some of methylobacteria survived after the treatment, whose red or orange colonies were easily distinguished from those of other bacteria. A 0.5% solution reduced CFU of M. fortuitum, whereas most of M. avium were still alive after the treatment. Bacteriological surveillance was performed with OAM at 9 water samples in 9 endoscope reprocessors. In contaminated bacteria, range at usual bacteria distributed widely (from 0 to 258 CFU/ ml). Mycobacterial contamination surpassed that of usual bacteria (range from 6.8 to 635 CFU/ml, average 251 CFU/ml). The contamination of mycobacteria did not correlate with that of usual bacteria. In contaminated mycobacteria, rapid growers, especially M. fortuitum, were mainly detected. M. avium was also detected to some extent at 4 samples. Discussion/Conclusion: The new method of ours disclosed that water and supply water pipes in endoscope reprocessors were contaminated with mycobacteria more than with usual bacteria. P9.09Background: The usefulness of bacterial and fungal cultures in operating rooms has been questioned. In our country air cultures are mandatory because of local regulations. Objectives: The aim of this study is to assess the efficacy of bacterial and fungal cultures in air samples in operating rooms. Methods: Operating rooms for vascular, heart, ocular and ortophedic surgery with prosthesis are classified as high risk. Cultures for bacteria and fungi early in the morning with an empty operating room and a second culture on the same day during surgical activity are made every month. The MAS 100 (MERCK) microbial air sampler is used. Samples for bacteria are cultured in blood agar and incubated 48 hours at 35ºC. Fungal cultures are made in Saboureaud + Cloranfenicol and incubated 7 days. Standard for high risk: fungal cultures 0 cfu/1000 liters of air and bacterial cultures <10...