[Change in resistance to quinolones and betalactams in different serogroups of Salmonella during the last decade in a Madrid hospital]. / Evolución de la resistencia a quinolonas y betalactámicos en distintos serogrupos de Salmonella durante la última década en un centro hospitalario de Madrid.

The prevalence of antibiotic resistance was studied in 3230 strains of Salmonella enterica isolated in the Hospital Universitario La Paz in Madrid, Spain, from 1991 to 2001. Betalactam antibiotic resistance has been notorious in serogroup B4; the highest prevalence of ampicillin resistance (84%) was reached in 2000 and that of amoxicillin-clavulanic acid (45%) in 1996. Resistance to cephalosporins has been controlled, although in 2000 cefazolin resistance reached 37% in serogroup C2-8. An increase in first generation quinolone resistance was detected in every serogroup, especially in D9 and C2-8, which showed an increase from 6% and 15% in 1991, respectively, to 40% and 85% in 2001. Although important resistance to ciprofloxacin has not yet been detected, the activity of fluoroquinolones against Salmonella must be closely monitored.

Tendinitis cálcica del tendón infraespinoso / Infraspinatus calcific tendinitis

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Evolución de la resistencia a quinolonas y betalactámicos en distintos serogrupos de Salmonella durante la última década en un centro hospitalario de Madrid / Change in resistance to quinolones and betalactams in different serogroups of Salmonella during the last decade in a Madrid hospital

Se ha estudiado la resistencia a antimicrobianos en 3230 cepas de Salmonella enterica aisladas en el Hospital Universitario La Paz (Madrid) durante 1991-2001. Ha destacado la resistencia a los betalactámicos del serogrupo B4: del 84 por ciento a la ampicilina en 2000 y del 45 por ciento a amoxicilina-ácido clavulánico en 1996. La resistencia a las cefalosporinas se mantiene moderada, aunque en el año 2000 llegó al 37 por ciento para cefazolina en el serogrupo C2-8. El aumento de la resistencia a las quinolonas de primera generación se observa para todos los serogrupos, pero es especialmente llamativo para D9 y C2-8, que pasan de una resistencia del 6 por ciento y 15 por ciento, respectivamente, en el año 1991, al 40 por ciento y 85 por ciento en el año 2001. Aunque de momento no se han observado resistencias importantes al ciprofloxacino, consideramos necesario un seguimiento de la actividad de las fluoroquinolonas sobre este microorganismo (AU)

The virulence plasmids of Salmonella.

Certain Salmonella serovars belonging to subspecies I carry a large, low-copy-number plasmid that contains virulence genes. Virulence plasmids are required to trigger systemic disease; their involvement in the enteric stage of the infection is unclear. Salmonella virulence plasmids are heterogeneous in size (50-90 kb), but all share a 7.8 kb region, spv, required for bacterial multiplication in the reticuloendothelial system. Other loci of the plasmid, such as the fimbrial operon pef, the conjugal transfer gene traT and the enigmatic rck and rsk loci, may play a role in other stages of the infection process. The virulence plasmid of Salmonella typhimurium LT2 is self-transmissible; virulence plasmids from other serovars, such as Salmonella enteritidis and Salmonella choleraesuis, carry incomplete tra operons. The presence of virulence plasmids in host-adapted serovars suggests that virulence plasmid acquisition may have expanded the host range of Salmonella.

A ColE1-type plasmid from Salmonella enteritidis encodes a DNA cytosine methyltransferase.

The multicopy plasmid pFM366 was isolated from a virulent Salmonella enteritidis strain and was found to code for DNA methylase activity (Ibáñez and Rotger, 1993). The present work was aimed at characterizing the genetic organization and functional features of this 5.6 kb plasmid. We found pFM366 almost identical to the plasmid P4 isolated from Shigella sonnei, that encodes the SsoII restriction-modification system (Karyagina et al., 1993), and related to other ColE1-type plasmids. Examination of these plasmids revealed a common organization which suggests they were the result of similar recombinational events. The cytosine methylase of pFM366 is nearly identical to M. SsoII, whereas the gene encoding the restrictase homologous to R. SsoII is truncated and its product is inactive. The expression of the cytosine methylase encoded by pFM366 is strongly affected by deletion of regions located upstream and downstream of its ORF, and is negatively controlled by the rpoS gene in Escherichia coli. The methylase activity encoded by pFM366 induces the SOS response, which could be responsible for the observed delay in the growth of E. coli.

Genetic map of the virulence plasmid of Salmonella enteritidis and nucleotide sequence of its replicons.

Partial sequencing of a genomic library of the virulence plasmid of Salmonella enteritidis has been used to localize in the restriction map of this plasmid the genetic loci already described in other Salmonella plasmids. The comparison of the vestigial tra region with the corresponding genes in the F plasmid allowed us to define the extent of the deletions that the S. enteritidis plasmid should have suffered. The putative replicons of the plasmid, repB and repC, were isolated and both proved to be functional in Escherichia coli, but repC was segregationally unstable. The nucleotide sequence of repB showed the typical organization of RepFIIA replicons, although the similarity was lower than usual in this group of replicons. The highest homology was found with the replicon of the virulence plasmid pYVe439-80 from Yersinia enterocolitica (72.5%). Replicon repC also showed a maximum identity of 72.6% with known replicons, namely the RepFIB of pColV-K30 and P307, both virulence plasmids isolated from E. coli. We conclude that the S. enteritidis plasmid could arise from the S. typhimurium plasmid through deletions, and that they are evolutionary distant from other IncFI and IncFII plasmids.

Analysis of the leaching and toxicity of new amine activators for the curing of acrylic bone cements and composites.

The comparative reactivity of new tertiary amine activators with the basic chemical structure of N,N-dimethyl-4-toluidine, but reduced toxicity, is analysed. The leaching of the amine compounds from cured cements was studied by analysis of the concentration of the corresponding amine in a physiological saline solution after 3 months of immersion, giving lower values for the new amine compounds as compared to N,N-dimethyl-4-toluidine. The acute toxicity was determined by intravenous injection of saline solutions of the corresponding chlorhydrates in mice and the cytotoxicity by the evolution of specific culture media. The results obtained demonstrate a lower acute toxicity and cytotoxicity of the new activators, even with a noticeable antiseptic action, which makes these materials very interesting from a practical point of view as activators of the curing process of acrylic bone cements for orthopaedic surgery and dentistry.

Enterotoxin and cytotoxin production by Salmonella enteritidis strains isolated from gastroenteritis outbreaks.

Seventy-six Salmonella enteritidis, three Salmonella virchow and one Salmonella bradenrup strains were screened for enterotoxigenicity by using the Chinese hamster ovary (CHO), Y1 adrenal, Vero and HeLa cell tests. All the strains gave positive reactions for enterotoxin production, except one, and the relative sensitivity to the toxin exhibited by the different cell lines was evaluated. An enterotoxic activity has been identified in sonicated extracts of Salm. enteritidis. This enterotoxin was purified on Agarose A-5m (Bio-Rad) and Superose 12 HR 10/30 column. The enterotoxic activity was eluted from the Superose column in the first peak. Like Vibrio cholerae toxin CT and Escherichia coli enterotoxin LT, it was blocked by GM1 ganglioside, but at a higher concentration. In addition, a cytotoxic factor has been partially identified. The procedure for isolating the cytotoxin included ammonium sulphate precipitation, size-exclusion chromatography and anion exchange chromatography. This cytotoxin factor caused inhibition of protein synthesis in cultured cells, as determined by flow cytometry and [3H]-leucine incorporation. Flow cytometry analysis also showed an activation of CHO cells when exposed to this cytotoxic factor resulting in a state of active growth. Cytotoxic activity was not blocked by gangliosides.

Restriction map of the Salmonella enteritidis virulence plasmid and its homology with the plasmid of Salmonella typhimurium.

A restriction map of the 61 kb virulence plasmid of Salmonella enteritidis was constructed and compared with the Salmonella typhimurium virulence plasmid map. Polymerase chain reaction was used to detect a region homologous to rck gene in the S. enteritidis plasmid which was localized in the map by hybridization. Regions homologous to traT and vagD/vagC loci were not detected in this plasmid by this method.

Characterization of a small cryptic plasmid from Salmonella enteritidis that affects the growth of Escherichia coli.

We examined the plasmid content of 25 clinical isolates of Salmonella enteritidis, and detected the presence of small plasmids (3-5.3 kb) in 9 of them, alone, or in addition to the large, so-called virulence plasmid. A 5.3-kb plasmid isolated as unique extrachromosomal DNA from a strain responsible for a high-mortality outbreak was characterized by restriction mapping and cloning. The plasmid replicon was localized in a 1.7-kb fragment, that hybridized with three of the small plasmids detected in S. enteritidis, and with another small plasmid from Salmonella typhimurium. A strain of Escherichia coli carrying this plasmid, or a cloned 3.7-kb PvuII restriction fragment, showed a slower growth rate, especially in minimal medium, as well as a noticeable increase in DNA methyltransferase activity.

New naphthalene-degrading marine Pseudomonas strains.

Over 100 strains that utilized naphthalene as the only carbon and energy source were isolated from samples of marine sediments taken from a heavily polluted area. The isolates were characterized taxonomically and physiologically. Most of these strains belonged to the genus Pseudomonas, and seven of them did not fit any previous taxonomic description. They differed from type strains in a few biochemical characteristics and in the utilization of aromatic compounds. None had catechol 1,2-dioxygenase activity, and catechol 2,3-dioxygenase was responsible for the aromatic ring cleavage. DNA hybridization demonstrated a close relationship between two isolates and the Pseudomonas stutzeri type strain, and between five isolates and the Pseudomonas testosteroni type strain. On the basis of nutritional and enzymatic characteristics, it was assumed that the seven isolates represent new biovars belonging to the species P. testosteroni and P. stutzeri that are able to degrade aromatic hydrocarbons.

Characterization of a beta-lactamase-specifying plasmid isolated from Eikenella corrodens and its relationship to a commensal Neisseria plasmid.

A 9.4-kilobase plasmid encoding penicillin, streptomycin, and sulfonamide resistance was isolated from a beta-lactamase-producing Eikenella corrodens strain. This plasmid appears to be identical to a resistance plasmid common to saprophytic Neisseria strains.

A multi-resistance plasmid isolated from commensal Neisseria species is closely related to the enterobacterial plasmid RSF1010.

pFM739, an R plasmid from Neisseria sicca that encodes penicillin, streptomycin and sulphonamide resistance, and the enterobacterial IncQ(P-4) plasmid RSF1010, which encodes streptomycin and sulphonamide resistance, were incompatible, and were mobilized by the same conjugative plasmids. Restriction mapping confirmed a high degree of similarity between both R plasmids; pFM739 carried DNA fragments corresponding to the known replication and resistance regions of RSF1010. pFM739 also carried an extra segment with the same restriction map as that described for the beta-lactamase-coding region of transposon Tn3. It is suggested that the R plasmids isolated from commensal Neisseria sp. could have resulted from transposition of a Tn3-like genetic element to an RSF1010-like plasmid, and that they contain deletion derivatives of transposon Tn3.

Multiresistance plasmid from commensal Neisseria strains.

Antibiotic-resistant commensal strains of Neisseria spp. and Branhamella catarrhalis were isolated from throat cultures, on the basis of their capacity to grow in the presence of penicillin, streptomycin, or sulfamethoxazole-trimethoprim. Several strains, which belonged to different species of Neisseria, were resistant to beta-lactams, streptomycin, sulfamethoxazole, and trimethoprim, harbored a 6.0-megadalton plasmid with identical HinfI restriction patterns, and produced beta-lactamase and streptomycin phosphotransferase. The resistance determinants for beta-lactams, streptomycin, and sulfamethoxazole, but not for trimethoprim, were transferred from all these strains to Escherichia coli by conjugation or transformation. The resulting transconjugants or transformants acquired the plasmid and the capacity to produce beta-lactamase and streptomycin phosphotransferase. The 6.0-megadalton plasmid complemented a mutation which determines production of thermosensitive dihydropteroate synthetase in E. coli. We conclude that an R plasmid coding for beta-lactamase, streptomycin phosphotransferase, and a sulfonamide-resistant dihydropteroate synthetase is common to these strains.