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SCI scientific publications 2004

December 1st, 2004

Improving dioxygenase stability by gene chromosome insertion: implementation in immobilized-cell systems

Current Microbiology 49(6):390-5.

     The immobilization of recombinant cells by using the unstable 3,4-dihydroxyphenylacetate 2,3-dioxygenase was studied as a model. Dioxygenase activity and cell viability were compared in immobilized-cell systems and cells in suspension. Immobilization increased enzyme stability and the efficient degradation of 3,4-dihydroxyphenylacetate. The stability of the cloned enzyme and the viability of the immobilized recombinant cells were well maintained for at least 15 days. We used the strain Escherichia coli CC118-D in which the hpaB gene from Klebsiella pneumoniae, coding for the subunit of 3,4-dihydroxyphenylacetate 2,3-dioxygenase, was inserted into the chromosome. This study has demonstrated that the implementation of E. coli CC118-D in a pil… Leer más

Gibello A., Garbi C., Allende JL. and Martin M.

November 1st, 2004

Spoligotyping profile change caused by deletion of a direct variable repeat in a Mycobacterium tuberculosis isogenic laboratory strain

Journal Of Clinical Microbiology 42(11):5388-91.

     Spoligotyping is a major tool for molecular typing of Mycobacterium tuberculosis complex organisms. For epidemiological purposes, strains are considered clonal only when their spoligotyping patterns are identical. We report a change in the spoligotyping profiles of truly isogenic strains (a clinical isolate and a subculture derived in the laboratory) caused by deletion of a direct variable repeat. Without the information about the relationship between them, a link between these strains would have gone unnoticed. Evolutionary events should be taken into account in the interpretation of spoligotyping results and in the design of databases… Leer más

Aranaz A., Romero B., Montero N., Alvarez J., Bezos J., de Juan L., Mateos A. and Dominguez L..

November 1st, 2004

Corynebacterium ciconiae sp. nov., isolated from the trachea of black storks (Ciconia nigra)

International Journal Of Systematic And Evolutionary Microbiology 54(Pt 6):2191-5.

     Eight unidentified Gram-positive, rod-shaped organisms were recovered from the tracheas of apparently healthy black storks (Ciconia nigra) and subjected to a polyphasic taxonomic analysis. Based on cellular morphology and biochemical criteria the isolates were tentatively assigned to the genus Corynebacterium, although three of the organisms did not appear to correspond to any recognized species. Comparative 16S rRNA gene sequencing studies demonstrated that all of the isolates were phylogenetically members of the genus Corynebacterium. Five strains were genotypically identified as representing Corynebacterium falsenii, whereas the remaining three strains represented a hitherto unknown subline, associated with a small subcluster of species … Leer más

Fernandez-Garayzabal JF., Vela AI., Egido R., Hutson RA., Lanzarot MP., Fernandez-Garcia M. and Collins MD.

November 1st, 2004

Streptococcus equi subsp. ruminatorum subsp. nov., isolated from mastitis in small ruminants

International Journal Of Systematic And Evolutionary Microbiology 54(Pt 6):2291-6.

     Six isolates of an unknown Gram-positive, catalase-negative, chain-forming, coccus-shaped organism isolated from ovine and caprine mastitis were characterized by phenotypic and molecular taxonomic methods. On the basis of cellular morphology and the results of biochemical tests, the organism was tentatively identified as a streptococcal species. Comparative 16S rRNA gene sequencing studies confirmed that the organism is a member of the genus Streptococcus, with Streptococcus equi as its closest phylogenetic relative (98.8 % similarity). DNA-DNA pairing studies showed that the unidentified organism displayed more than 70 % relatedness to the type strains of S. equi subsp. equi and subsp. zooepidemicus. Despite the relatively high DNA-DNA rea… Leer más

Fernandez E., Blume V., Garrido P., Collins MD., Mateos A., Dominguez L. and Fernandez-Garayzabal JF..

September 1st, 2004

A highly sensitive and specific gel-based multiplex RT-PCR assay for the simultaneous and differential diagnosis of African swine fever and Classical swine

Veterinary Research 35(5):551-63.

     The development and standardisation of a novel, highly sensitive and specific one-step hot start multiplex RT-PCR assay is presented for the simultaneous and differential diagnosis of African swine fever (ASF) and Classical swine fever (CSF). The method uses two primer sets, each one specific for the corresponding virus, amplifying DNA fragments different in length, allowing a gel-based differential detection of the PCR products. Universal detection of ASF and CSF virus strains was achieved through selection of primers in conserved viral genome regions. The detection range was confirmed by analysis of a large collection of isolates of the two viruses. The high specificity of the assay was proven by testing related viruses, uninfected cell l… Leer más

Aguero M., Fernandez J., Romero LJ., Zamora MJ., Sanchez C., Belak S., Arias M. and Sanchez-Vizcaino JM..

August 1st, 2004

Salmonella diversity associated with wild reptiles and amphibians in Spain

Environmental Microbiology 6(8):868-71.

     During the spring and summer of 2001, faeces from 166 wild reptiles (94 individuals) and amphibians (72 individuals) from 21 different species found in central Spain were examined for the presence of Salmonella. Thirty-nine reptiles (41.5%) yielded 48 Salmonella isolates, whereas all the amphibians examined were negative. Subspecies Salmonella enterica enterica (I) accounted for up to 50% of isolates. Fourteen isolates (29.2%) belonged to subspecies diarizonae (IIIb), six isolates (12.5%) to subspecies salamae (II), and four isolates (8.3%) to subspecies arizonae (IIIa). Twenty-seven different serotypes were identified. Serotypes Anatum (12.5%), Herzliya (8.3%), Abony, 18:l,v:z, 9,12:z29:1,5 and 38:z10:z53 (6.2%/each) were the most frequent… Leer más

Briones V., Tellez S., Goyache J., Ballesteros C., Lanzarot MP., Dominguez L. and Fernandez-Garayzabal JF..

July 8th, 2004

Beta-lactamase characterization in Escherichia coli isolates with diminished susceptibility or resistance to extended-spectrum cephalosporins recovered from sick animals in Spain

Microbial Drug Resistance (Larchmont, N.Y.) 9(2):201-9.

     A total of 1439 Escherichia coli isolates from sick animals were received from the Spanish Network of Veterinary Antimicrobial Resistance Surveillance (VAV) from 1997 to 2001. Antimicrobial susceptibility tests were performed and diminished susceptibility to cefotaxime and ceftazidime was identified in 2.5% and 2.8% of the isolates, respectively. Beta-lactamase characterization was carried out in the group of 20 E. coli isolates with both characteristics. The MIC ranges of different beta-lactams showed by these 20 isolates were as follows (in microg/ml): ampicillin (64-->256), amoxicillin-clavulanic acid (4-64), ticarcillin (8-->128), cefazolin (32-->256), cefoxitin (4-->128), cefotaxime (1-64), ceftazidime (2-->64), ceftriaxone (0.5-64), i… Leer más

Brinas L., Moreno MA., Teshager T., Zarazaga M., Saenz Y., Porrero MC., Dominguez L. and Torres C.

June 21st, 2004

Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value

Veterinary Microbiology 101(2):109-16.

     Streptococcus iniae is a well-known pathogen of both fish and humans that is difficult to identify by conventional biochemical tests. A PCR assay based on the lactate oxidase (lctO) gene of S. iniae was developed for the rapid and specific detection and identification of this pathogen from different sources. The PCR assay had a detection limit of 62–31 cells, and 25 pg of DNA per PCR reaction mixture. The PCR was also effective in detecting the bacterium from inoculated tissue homogenates, suggesting its potential use for a rapid and accurate diagnosis of S. iniae infections… Leer más

Mata AI., Blanco MM., Dominguez L., Fernandez-Garayzabal JF. and Gibello A.

June 1st, 2004

Bovine tuberculosis (Mycobacterium bovis) in wildlife in Spain

Journal Of Clinical Microbiology 42(6):2602-8.

     Mycobacterium bovis infection in wildlife and feral species is a potential source of infection for livestock and a threat to protected and endangered species. The aim of this study was to identify Spanish wild animal species infected with M. bovis through bacteriological culture and spacer oligonucleotide typing (spoligotyping) of isolates for epidemiological purposes. This study included samples from red deer (Cervus elaphus), fallow deer (Dama dama), wild boar (Sus scrofa), Iberian lynx (Lynx pardina), hare (Lepus europaeus), and cattle (Bos taurus). They were collected in several geographical areas that were selected for their unique ecological value and/or known relationships between wildlife and livestock. In the areas included in this… Leer más

Aranaz A., de Juan L., Montero N., Sanchez C., Galka M., Delso C., Alvarez J., Romero B., Bezos J., Vela AI., Briones V., Mateos A. and Dominguez L..

May 26th, 2004

Subclinical bovine spongiform encephalopathy infection in transgenic mice expressing porcine prion protein

The Journal Of Neuroscience : The Official Journal Of The Society For Neuroscience 24(21):5063-9.

     The bovine-porcine species barrier to bovine spongiform encephalopathy (BSE) infection was explored by generating transgenic mouse lines expressing the porcine prion protein (PrP) gene. All of the porcine transgenic (poTg) mice showed clinical signs of BSE after intracerebral inoculation with a high-titer BSE inoculum. The protease-resistant PrP (PrP(res)) was detected in 14% (3 of 22) of the BSE-infected poTg mice by immunohistochemical or immunoblot analysis. Despite being able to infect 42% (5 of 12) of control mice, a low-dose BSE inoculum failed to penetrate the species barrier in our poTg mouse model. The findings of these infectivity studies suggest that there is a strong species barrier between cows and pigs. However, after second-p… Leer más

Castilla J., Gutierrez-Adan A., Brun A., Doyle D., Pintado B., Ramirez MA., Salguero FJ., Parra B., Segundo FD., Sanchez-Vizcaino JM., Rogers M. and Torres JM.

May 22nd, 2004

Distribution of serotypes of Streptococcus suis isolated from diseased pigs in Spain

The Veterinary Record 154(21):665-6.

     Streptococcus suis is well known as a major cause of meningitis, septicaemia, endocarditis, reproductive disorders, pneumonia and arthritis in pigs (Higgins and Gottschalk 1999).
It is also considered an important zoonotic agent (Arends and Zanen 1988), able to induce meningitis and septicaemia in human beings, and mainly affects pig handlers (Tarradas and others 2001). At present, 35 different serotypes, based on capsular antigens, are recognised (Higgins and others 1995); serotype 2 is the most frequently isolated capsular type in Italy, France and Denmark, while serotype 9 is more frequently isolated in Germany, the Netherlands and Belgium
(Gogolewski and others 1990, Aarestrup and others 1998, Sihvonen and others 1998, Be… Leer más

Tarradas C., Perea JA., Vela AI., Goyache J., Dominguez L., Fernandez-Garayzabal JF., Borge C., Huerta B. and Luque I.

May 1st, 2004

Multiplex PCR assay for detection of bacterial pathogens associated with warm-water Streptococcosis in fish

Applied And Environmental Microbiology 70(5):3183-7.

     A multiplex PCR-based method was designed for the simultaneous detection of the main pathogens involved in warm-water streptococcosis in fish (Streptococcus iniae, Streptococcus difficilis, Streptococcus parauberis, and Lactococcus garvieae). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. The sensitivity of the multiplex PCR using purified DNA was 25 pg for S. iniae, 12.5 pg for S. difficilis, 50 pg for S. parauberis, and 30 pg for L. garvieae. The multiplex PCR assay was useful for the specific detection of the four species of bacteria not only in pure culture but also in inoculated fish tissue homogenates and naturally infected fish. Therefore, this method could be … Leer más

Mata AI., Gibello A., Casamayor A., Blanco MM., Dominguez L. and Fernandez-Garayzabal JF..

March 1st, 2004

Dogs should be included in surveillance programs for vancomycin-resistant enterococci

Journal Of Clinical Microbiology 42(3):1384-5.

     Enterococci are commensal bacteria of the intestinal microbiota in humans and animals. Multidrug-resistant enterococci are among the most important pathogens responsible for nosocomial infections in humans (8). During the last decade vancomycin-resistant enterococci (VRE) have emerged as a worldwide health problem, i.e., as reservoirs of genes coding for antimicrobial resistance and as possessors of the ability to spread these resistance genes to other bacterial species (17). There are numerous reports on the presence of VRE in farm animals (2, 3). Some epidemiological studies suggest that animals carrying VRE in their gastrointestinal tract could be the source of VRE infections of humans (19). These VRE of animal origin can colonize humans… Leer más

Herrero IA., Fernandez-Garayzabal JF., Moreno MA. and Dominguez L..

January 1st, 2004

Proteinase K enhanced immunoreactivity of the prion protein-specific monoclonal antibody 2A11

Neuroscience Research 48(1):75-83.

     Here, we report the development and further characterisation of a novel PrP-specific monoclonal antibody: 2A11. By Western blot analysis, 2A11 reacts with PrPC from a variety of species including cow, sheep, pig, hamster, rabbit, cat, dog, deer and mouse but fails to react with human, chicken and turtle PrP. Reactivity to PrPC in Western blot was found to be dependent on the redox state of the protein since binding of mAb 2A11 to its epitope was more effective in reducing conditions. 2A11 binding site was mapped within a region comprised by residues 171-179 (six octarepeats bovine PrP notation; 163-171 for the ovine PrP notation). Interestingly, in immunohistochemistry (IHC) analysis, immunoreactivity was greatly enhanced after proteinase K… Leer más

Brun A., Castilla J., Ramirez MA., Praguer K., Parra B., Salguero FJ., Shiveral D., Sanchez C., Sanchez-Vizcaino JM., Douglas A. and Torres JM.

January 1st, 2004

Analysis of the gyrA gene of clinical Yersinia ruckeri isolates with reduced susceptibility to quinolones

Applied And Environmental Microbiology 70(1):599-602.

     Antimicrobial susceptibility of seven clinical strains of Yersinia ruckeri representative of those isolated between 1994 and 2002 from a fish farm with endemic enteric redmouth disease was studied. All isolates displayed indistinguishable pulsed-field gel electrophoresis restriction patterns, indicating that they represented a single strain. However, considering both inhibition zone diameters (IZD) and MICs, the isolates recovered in 2001-2002 formed a separate cluster with lower levels of susceptibility to all the quinolones tested, especially nalidixic acid (NA) and oxolinic acid (OA), compared with the isolates recovered between 1994 and 1998. Analysis of the PCR product of the quinolone resistance-determining region of the gyrA gene fro… Leer más

Gibello A., Porrero MC., Blanco MM., Vela AI., Liebana P., Moreno MA., Fernandez-Garayzabal JF. and Dominguez L..

Science Publication
ISI Scientific Publications
Total Last 60 mo. Last 12 mo.
1 ZTA 249 SUAT 61 SUAT 15
2 ICM 237 ZTA 52 MYC 11
3 MYC 227 MYC 48 SAP 11
4 SUAT 219 ICM 40 ZTA 5
5 NED 75 SAP 31 ICM 4
  VISAVET 1123 316 54

Impact Factor Average
Total Last 60 mo. Last 12 mo.
1 ZTA 3.903 ZTA 5.366 ZTA 6.42
2 SAP 3.818 SUAT 4.227 SAP 6.06
3 SUAT 2.987 SAP 4.095 NED 4.3
4 MYC 2.8 NED 3.858 SUAT 4.287
5 DICM 2.776 ICM 3.798 MYC 3.118
VISAVET 3.282 4.869 6.827

ISI Scientific Publications
Total Last 60 mo.
1 Transbound Emerg Dis 93 Front Vet Sci 35
2 Vet Microbiol 88 Transbound Emerg Dis 32
3 Prev Vet Med 77 Animals 16
4 Front Vet Sci 61 Res Vet Sci 14
5 PLoS ONE 56 Sci Rep 14
  All journals 354   All journals 38