Publicaciones científicas SCI 2009
Reflection paper on the use of third and fourth generation cephalosporins in food producing animals in the European Union: development of resistance and impact on human and animal health
Journal Of Veterinary Pharmacology And Therapeutics 32(6):515-33.
Resistance to third and fourth generation cephalosporins is rapidly increasing in bacteria infecting humans. Although many of these problems are linked to human to human transmission and to use of antimicrobials in human medicine, the potential role of community reservoirs such as food producing animals needs to be scrutinized. Resistance to third and fourth generation cephalosporins is emerging in enteric bacteria of food producing animals and also in food of animal origin. The genes encoding resistance to these cephalosporins are transferrable and often linked to other resistance genes. Systemic use of third and fourth cephalosporins selects for resistance, but co-selection by other antimicrobials is also likely to influence prevalence o… Leer más
Greko C., Badiola JI., Catry B., van Duijkeren E., Moreno MA., Matias-Ferreira MC., Pyorala S., Ruzauskas M., Sanders P., Threlfall EJ., Ungemach F., Torneke K., Torren-Edo J., Caprioli A., Mevius DJ. y Wallman J.
Novel genetic environment of qnrB2 associated with TEM-1 and SHV-12 on pB1004, an IncHI2 plasmid, in Salmonella Bredeney BB1047 from Spain
The Journal Of Antimicrobial Chemotherapy 64(6):1334-6.
Resistance to fluoroquinolones through plasmid-mediated qnr genes, especially when associated with resistance to cephalosporins by extended-spectrum b-lactamases (ESBLs), is an emerging phenomenon in enterobacteria. This is worrisome in the case of Salmonella, as these are two of the major antimicrobials used in invasive salmonellosis… Leer más
Gutierrez B., Herrera-Leon S., Escudero JA., Hidalgo L., Gonzalez-Sanz R., Arroyo M., San Millan A., Echeita MA. y Gonzalez-Zorn B..
Unilateral scrotal pyocele in ram caused by Staphylococcus capitis
Australian Veterinary Journal 87(12):484-6.
A massive unilateral scrotal pyocele caused by Staphylococcus capitis in a 6-year-old ram is reported. Ultrasound examination of the right hemiscrotum showed an irregular hyperechoic mass in an anechoic fluid. A dense exudate was collected from the scrotum for microbiological analysis. Grossly, there was an extensive greenish purulent exudate and a completely atrophied right testis. Coagulase-negative S. capitis was isolated in pure culture. To the authors` knowledge, this is the first report of genital infection by S. capitis in rams. This microorganism should be included in the differential diagnosis of ovine genital infections… Leer más
Lacasta D., Ferrer LM., Ramos JJ., Ortin A., Vela AI. y Latre MV.
First data on Eurasian wild boar response to oral immunization with BCG and challenge with a Mycobacterium bovis field strain
Vaccine 27(48):6662-8.
The Eurasian wild boar (Sus scrofa) is considered a reservoir for bovine tuberculosis (bTB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex in south-central Spain. The vaccination of wildlife with BCG offers an alternative to culling and to movement restriction for the control of bTB among wildlife reservoirs. In this study, we hypothesized that oral BCG immunization of wild boar would affect the expression of immunoregulatory genes and confer protection against M. bovis. Three groups were used to describe the infection, pathological findings and gene expression profiles in wild boar: BCG-vaccinated and M. bovis-challenged (vaccinated challenged group; N=6), non-vaccinated and M. bovis-cha… Leer más
Ballesteros C., Garrido JM., Vicente J., Romero B., Galindo RC., Minguijon E., Villar M., Martin-Hernando MP., Sevilla I., Juste R., Aranaz A., de la Fuente J. y Gortazar C.
One-step real-time quantitative PCR assays for the detection and field study of Sacbrood honeybee and Acute bee paralysis viruses
Journal Of Virological Methods 161(2):240-6.
Two one-step real-time RT-PCR assays, based on SYBR Green (SG) chemistry, were developed or adapted respectively, for the detection, differentiation, and quantitation of two important honeybee viruses: Sacbrood virus (SBV) and Acute bee paralysis virus (ABPV). Both reactions were optimized to yield the highest sensitivity and specificity. The genome equivalent copies (GEC) detection limit per reaction was 389.3 for the ABPV RT-PCR. The GEC detection limit per reaction was 298.9 for the SBV RT-PCR. Viral detection and identification were confirmed by melting curve analysis and sequencing of the PCR products. Both techniques were used to evaluate Spanish field samples and establish the distribution of these viruses. Acute bee paralysis virus … Leer más
Kukielka D. y Sanchez-Vizcaino JM..
Occurrence of Mycobacterium avium subspecies paratuberculosis across host species and European countries with evidence for transmission between wildlife and domestic ruminants
BMC Microbiology 9:212.
Mycobacterium avium subspecies paratuberculosis (Map) causes an infectious chronic enteritis (paratuberculosis or Johne`s disease) principally of ruminants. The epidemiology of Map is poorly understood, particularly with respect to the role of wildlife reservoirs and the controversial issue of zoonotic potential (Crohn`s disease). Genotypic discrimination of Map isolates is pivotal to descriptive epidemiology and resolving these issues. This study was undertaken to determine the genetic diversity of Map, enhance our understanding of the host range and distribution and assess the potential for interspecies transmission. RESULTS: 164 Map isolates from seven European countries representing 19 different host species were genotyped by standardiz… Leer más
Stevenson K., Alvarez J., Bakker D., Biet F., de Juan L., Denham S., Dimareli Z., Dohmann K., Gerlach GF., Heron I., Kopecna M., May L., Pavlik I., Sharp JM., Thibault V., Willemsen P., Zadoks R. y Greig A.
Prevalence and diversity of extended-spectrum beta-lactamases in faecal Escherichia coli isolates from healthy humans in Spain
Clinical Microbiology And Infection 15(10):954-6.
Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates were detected in seven of 105 faecal samples from healthy humans, from two Spanish cities, during 2007. In these isolates, five ESBLs were detected, CTX-M-14 (n = 2), CTX-M-1 (n = 2), CTX-M-32 (n = 1), CTX-M-8 (n = 1) and TEM-52 (n = 1). Both bla(CTX-M-14a) (surrounded by ISEcp1-IS903) and bla(CTX-M-14b) variants (included in an integron structure) were identified in this study. This is the first time that the bla(CTX-M-8) gene and ESBLs of the CTX-M-8 group have been found in Europe and Spain, respectively. Faecal E. coli of healthy humans therefore constitute a reservoir of bla(CTX-M) genes with different surrounding genetic elements… Leer más
Vinue L., Saenz Y., Martinez S., Somalo S., Moreno MA., Torres C. y Zarazaga M.
Nervous signs associated with otitis and cranial osteomyelitis and with Ornithobacterium rhinotracheale infection in red-legged partridges (Alectoris rufa)
Avian Pathology: Journal Of The W.V.P.A 38(5):341-347.
A case of nervous signs in red-legged partridges (Alectoris rufa) associated with a severe otitis and osteomyelitis is reported. The outbreak was characterized by abnormal head position, torticollis and difficulty in standing, walking and flying. Pathological, microbiological and molecular genetic data supported an association with Ornithobacterium rhinotracheale (ORT) infection. Clinical signs persisted for several days and were accompanied by weight loss leading to death. Morbidity was approximately 20% and most birds died if untreated. Lesions were mainly characterized by a severe osteomyelitis of the cranial bones and purulent inflammation of the external, middle and inner ears. O. rhinotracheale was isolated from ear samples, skull and… Leer más
Moreno B., Chacon G., Villa A., Fernandez A., Vela AI., Fernandez-Garayzabal JF., Ferre S. y Gracia E.
Genetic and virulence-phenotype characterization of serotypes 2 and 9 of Streptococcus suis swine isolates
International Microbiology 12(3):161-6.
The aim of this study was to analyze the genetic characteristics and virulence phenotypes of Streptococcus suis, specifically, in clinical isolates of serotypes 2 and 9 (n = 195), obtained from diverse geographical areas across Spain. Pulsed-field gel electrophoresis (PFGE) typing identified 97 genetic profiles, 68% of which were represented by single isolates, indicative of a substantial genetic diversity among the S. suis isolates analyzed. Five PFGE profiles accounted for 33.3% of the isolates and were isolated from 38% of the herds in nine different provinces, indicative of the bacterium`s widespread distribution in the Spanish swine population. Representative isolates of the most prevalent PFGE profiles of both serotypes were subjected… Leer más
Blume V., Luque I., Vela AI., Borge C., Maldonado A., Dominguez L., Tarradas C. y Fernandez-Garayzabal JF..
Combined application of social network and cluster detection analyses for temporal-spatial characterization of animal movements in Salamanca, Spain
Preventive Veterinary Medicine 91(1):29-38.
Social network analysis was used in combination with techniques for detection of temporal-spatial clusters to identify operations at high risk of receiving or dispatching pigs, from January through December 2005, in the Spanish province of Salamanca. The temporal-spatial structure of the network was explicitly analyzed to estimate the statistical significance of observed clusters. Significant (P<0.01) temporal-spatial clusters identified were grouped into two compartments based on the nature and extent of the contacts among operations within the clusters. One of the compartments was identified from January through April, included a high proportion of extensive farms (0.39), and was likely to be related with the production and trade of Iberi… Leer más
Martinez-Lopez B., Perez AM. y Sanchez-Vizcaino JM..
Multiresistance in Pasteurella multocida is mediated by coexistence of small plasmids
Antimicrobial Agents And Chemotherapy 53(8):3399-404.
In most gram-negative bacteria, acquired multiresistance is conferred by large plasmids compiling numerous antimicrobial resistance genes. Here, we show an evolutionary alternative strategy used by Pasteurella multocida to become resistant to multiple clinically relevant antibiotics. Thirteen beta-lactam-resistant clinical isolates, concomitantly resistant to tetracyclines and/or streptomycin as well as to sulfonamides, were studied. Pulsed-field gel electrophoresis analysis revealed different profiles among the isolates, showing that clonal dissemination was not the sole event responsible for the spread of multiresistance. Each P. multocida strain carried two or three small plasmids between 4 and 6 kb in size. A direct association between … Leer más
San Millan A., Escudero JA., Gutierrez B., Hidalgo L., Garcia N., Llagostera M., Dominguez L. y Gonzalez-Zorn B..
Reclassification of the members of the genus Tetrathiobacter Ghosh et al. 2005 to the genus Advenella Coenye et al. 2005
International Journal Of Systematic And Evolutionary Microbiology 59(8):1914-8.
The taxonomic position of the genera Advenella and Tetrathiobacter was examined. 16S rRNA gene sequence analysis revealed that the two genera are closely related, representing a monophyletic cluster with high sequence similarity (98.1-99.7 %) within the family Alcaligenaceae. The phenotypic characteristics of the type strains of Advenella incenata, Tetrathiobacter kashmirensis and Tetrathiobacter mimigardefordensis were re-examined using the API 20NE, API ZYM and API 50CH systems. Phylogenetic data together with similarities in phenotypic characteristics, G+C content and cellular acid composition suggest that they should be classified in the same genus. On the basis of the data presented, the two species of the genus Tetrathiobacter should … Leer más
Gibello A., Vela AI., Martin M., Barra-Caracciolo A., Grenni P. y Fernandez-Garayzabal JF..
Single nucleotide polymorphisms in the IS900 sequence of Mycobacterium avium subsp. paratuberculosis are strain type specific
Journal Of Clinical Microbiology 47(7):2260-4.
Insertion sequence IS900 is used as a target for the identification of Mycobacterium avium subsp. paratuberculosis. Previous reports have revealed single nucleotide polymorphisms within IS900. This study, which analyzed the IS900 sequences of a panel of isolates representing M. avium subsp. paratuberculosis strain types I, II, and III, revealed conserved type-specific polymorphisms that could be utilized as a tool for diagnostic and epidemiological purposes… Leer más
Castellanos E., Aranaz A., de Juan L., Alvarez J., Rodriguez-Campos S., Romero B., Bezos J., Stevenson K., Mateos A. y Dominguez L..
Defining output-based standards to achieve and maintain tuberculosis freedom in farmed deer, with reference to member states of the European Union
Preventive Veterinary Medicine 90(3-4):254-67.
Within the European Union (EU), detailed legislation has been developed for cattle, but not deer, to minimise disease risks associated with trade in animals and animal products. This legislation is expressed as input-based standards, providing a detailed outline of the activity required (for example, testing of animals and application of defined control measures), on the expectation that an adequate output (for example, confidence in freedom) will be achieved. Input-based standards are at odds with the increasing shift towards output-based standards, particularly in OIE rules governing international trade. In this paper, we define output-based standards to achieve and maintain freedom from tuberculosis (TB) in farmed deer, with reference to… Leer más
More JS., Cameron AR., Greiner M., Clifton-Hadley RS., Rodeia SC., Bakker D., Salman MD., Sharp JM., De Massis F., Aranaz A., Boniotti MB., Gaffuri A., Have P., Verloo D., Woodford N. y Wierup M.
The SOS Response Controls Integron Recombination
Science 324(5930):1034.
Integrons are found in the genome of hundreds of environmental bacteria but are mainly known for their role in the capture and spread of antibiotic resistance determinants among Gram-negative pathogens. We report a direct link between this system and the ubiquitous SOS response. We found that LexA controlled expression of most integron integrases and consequently regulated cassette recombination. This regulatory coupling enhanced the potential for cassette swapping and capture in cells under stress, while minimizing cassette rearrangements or loss in constant environments. This finding exposes integrons as integrated adaptive systems and has implications for antibiotic treatment policies… Leer más
Guerin E., Cambray G., Sanchez-Alberola N., Campoy S., Erill I., Da Re S., Gonzalez-Zorn B., Barbe J., Ploy MC. y Mazel D.
VanB-type Enterococcus faecium clinical isolate successively inducibly resistant to, dependent on, and constitutively resistant to vancomycin
Antimicrobial Agents And Chemotherapy 53(5):1974-82.
Three Enterococcus faecium strains isolated successively from the same patient, vancomycin-resistant strain BM4659, vancomycin-dependent strain BM4660, and vancomycin-revertant strain BM4661, were indistinguishable by pulsed-field gel electrophoresis and harbored plasmid pIP846, which confers VanB-type resistance. The vancomycin dependence of strain BM4660 was due to mutation P(175)L, which suppressed the activity of the host Ddl D-Ala:D-Ala ligase. Reversion to resistance in strain BM4661 was due to a G-to-C transversion in the transcription terminator of the vanRS(B) operon that lowered the free energy of pairing from -13.08 to -6.65 kcal/mol, leading to low-level constitutive expression of the resistance genes from the P(RB) promoter, as… Leer más
San Millan A., Depardieu F., Godreuil S. y Courvalin P.
Social network analysis. Review of general concepts and use in preventive veterinary medicine
Transboundary And Emerging Diseases 56(4):109-20.
Social network analysis (SNA) and graph theory have been used widely in sociology, psychology, anthropology, biology and medicine. Social network analysis and graph theory provide a conceptual framework to study contact patterns and to identify units of analysis that are frequently or intensely connected within the network. Social network analysis has been used in human epidemiology as a tool to explore the potential transmission of infectious agents such as HIV, tuberculosis, hepatitis B and syphilis. In preventive veterinary medicine, SNA is an approach that offers benefits for exploring the nature and extent of the contacts between animals or farms, which ultimately leads to a better understanding of the potential risk for disease spread… Leer más
Martinez-Lopez B., Perez AM. y Sanchez-Vizcaino JM..
Risk assessment and cost-effectiveness analysis of Aujeszky´s disease virus introduction through breeding and fattening pig movements into Spain
Preventive Veterinary Medicine 90(1-2):10-6.
Movement of infected animals is considered the most likely route of Aujeszky´s disease virus (ADV) introduction into free areas and the main obstacle to eradicating Aujeszky´s disease (AD) in those areas, which have achieved a low prevalence (>0 and Leer más
Martinez-Lopez B., Carpenter TE. y Sanchez-Vizcaino JM..
Risk of introduction of H5N1 HPAI from Europe to Spain by wild water birds in autumn
Transboundary And Emerging Diseases 56(3):86-98.
Highly pathogenic avian influenza (HPAI) subtype H5N1 continues to circulate across Eurasia and Africa since its unprecedented rapid spread in 2005. Diffusion by wild bird movements has been evidenced in the European Union in 2006 and 2007. Spain is an important wintering quarter for aquatic birds from northern latitudes, so identifying the critical areas and species where an outbreak is prone to happen is necessary. This work presents an assessment of the risk of introduction of H5N1 HPAI in Spain by aquatic wild birds estimating a relative risk value per province. For this purpose, an assessment of the release and exposure to the risk of infection with H5N1 HPAI of 25 selected water bird species has been carried out. Parameters considered… Leer más
Martinez-Aviles M., Munoz MJ., de la Torre A., Iglesias I., Peris S., Infante O. y Sanchez-Vizcaino JM..
Rapid and sensitive detection of African horse sickness virus by real-time PCR
Research In Veterinary Science 86(2):353-8.
A highly sensitive and specific TaqMan-MGB real-time RT-PCR assay has been developed and standardised for the detection of African horse sickness virus (AHSV). Primers and MGB probe specific for AHSV were selected within a highly conserved region of genome segment 7. The robustness and general application of the diagnostic method were verified by the detection of 12 AHSV isolates from all of the nine serotypes. The analytical sensitivity ranged from 0.001 to 0.15 TCID(50) per reaction, depending on the viral serotype. Real-time PCR performance was preliminarily assessed by analysing a panel of field equine samples. The same primer pair was used to standardise a conventional RT-PCR as an affordable, useful and simple alternative method in la… Leer más
Fernandez J., Fernandez-Pacheco P., Rodriguez B., Sotelo E., Robles A., Arias M. y Sanchez-Vizcaino JM..
Moraxella pluranimalium sp. nov., isolated from animal specimens
International Journal Of Systematic And Evolutionary Microbiology 59(Pt 4):671-4.
Four unusual Gram-negative, catalase-positive, oxidase-positive, coccus-shaped bacteria isolated from one sheep and three pigs were characterized using phenotypic and molecular genetic methods. On the basis of cellular morphology and biochemical criteria, the isolates were tentatively assigned to the genus Moraxella, although the organisms did not appear to correspond to any recognized species. Comparative 16S rRNA gene sequencing studies demonstrated that the isolates represent a novel subline within the genus Moraxella. The most closely related species in phylogenetic terms was Moraxella cuniculi, with 16S rRNA gene sequence similarity of 97.9 % to the type strain CCUG 2154(T), although the DNA-DNA relatedness value was only 29 %. The nov… Leer más
Vela AI., Arroyo E., Aragon V., Sanchez-Porro C., Latre MV., Cerda-Cuellar M., Ventosa A., Dominguez L. y Fernandez-Garayzabal JF..
Effect of paratuberculosis on the diagnosis of bovine tuberculosis in a cattle herd with a mixed infection using interferon-gamma detection assay
Veterinary Microbiology 135:389-393.
Interferon-gamma (IFN-gamma) detection assay is being applied as an ancillary test to tuberculin tests in the diagnosis of bovine tuberculosis to detect the maximum number of infected animals. Among possible factors influencing the performance of tuberculosis-diagnostic tests, paratuberculosis, a widespread disease in Spain and other European countries, has been pointed out as a cause of false positive reactions. Still, its effect on the sensitivity of these tests in cattle has yet to be fully characterized. The impact of paratuberculosis in the apparent sensitivity of IFN-gamma assay was studied in a bullfighting cattle herd with a mixed tuberculosis-paratuberculosis infection, using culture of Mycobacterium bovis and Mycobacterium avium p… Leer más
Alvarez J., de Juan L., Bezos J., Romero B., Saez-Llorente JL., Marques S., Dominguez C., Minguez O., Fernandez-Mardomingo B., Mateos A., Dominguez L. y Aranaz A.
Occurrence of extended-spectrum beta-lactamase-producing Salmonella enterica in northern Spain with evidence of CTX-M-9 clonal spread among animals and humans
Clinical Microbiology And Infection 15(3):292-5.
Among the 1233 Salmonella enterica isolates obtained in two Spanish hospitals, five isolates (0.4%) (serovars: Virchow, four; Livingstone, one) had the phenotype of an extended-spectrum beta-lactamase (ESBL) producer. The genetic characterization of the ESBL of S. enterica Livingstone revealed a bla(SHV-2) gene. The bla(CTX-M-10) gene in a phage-related genetic environment was found in one S. enterica Virchow isolate, and the bla(CTX-M-9) gene within the In60 integron was found in the three remaining Virchow isolates. These three isolates presented indistinguishable or closely related pulsed-field gel electrophoresis patterns among themselves and also as compared with the two other bla(CTX-M-9)-containing isolates previously obtained from a… Leer más
Riano I., Garcia-Campello M., Saenz Y., Alvarez P., Vinue L., Lantero M., Moreno MA., Zarazaga M. y Torres C.
Expression of immunoregulatory genes in peripheral blood mononuclear cells of European wild boar immunized with BCG
Veterinary Microbiology 134(3-4):334-9.
The objective of this study was to analyze the expression of immunoregulatory genes in European wild boar (Sus scrofa) immunized with BCG. Eighteen immunoregulatory genes were selected for expression analysis based on their role in host immune response during tuberculosis and/or for their association with resistance to bovine tuberculosis in European wild boar populations. Initially, mRNA levels were analyzed by quantitative real-time reverse transcription PCR (qRT-PCR) in spleen samples from Mycobacterium bovis-infected (N=18) and uninfected (N=22) European wild boar. Statistical analysis of qRT-PCR data revealed that four genes, complement component C3, IFN-gamma, IL-4 and RANTES were downregulated in infected animals (P<0.05). These gene… Leer más
Lastra JM., Galindo RC., Gortazar C., Ruiz-Fons F., Aranaz A. y de la Fuente J.
Streptococcus plurextorum sp. nov., isolated from pigs
International Journal Of Systematic And Evolutionary Microbiology 59(Pt 3):504-8.
Biochemical and molecular genetic studies were performed on an unknown Gram-positive, catalase-negative, coccus-shaped organism isolated from clinical samples from pigs. On the basis of the results of cellular morphological and biochemical tests, the organism was identified as a streptococcal species. 16S rRNA gene sequence comparisons confirmed its identification as a member of the genus Streptococcus, but the organism was distinct from any recognized species of this genus. The closest phylogenetic relative of the unknown organism corresponded to Streptococcus suis NCTC 10234(T) (97.2 % 16S rRNA gene sequence similarity) and this phylogenetic position was confirmed by analysis of rpoB and sodA sequences. DNA-DNA hybridization studies showe… Leer más
Vela AI., Casamayor A., Sanchez V., Dominguez L. y Fernandez-Garayzabal JF..
A stochastic model to quantify the risk of introduction of classical swine fever virus through import of domestic and wild boars
Epidemiology And Infection 137(10):1505-15.
Classical swine fever (CSF) is a disease of pigs that imposes major hardship on the industry of infected regions. The recent history of CSF epidemics suggests that animal movements remain the main source of CSF virus (CSFV) infection for susceptible populations in Europe. This study presents an assessment of the risk of introducing CSFV into Spain through the importation of live susceptible animals. Results suggest that, if prevailing conditions persist, introduction of CSFV into Spain is likely to occur on average every 9 years and that introduction is almost three times more likely to occur via domestic pigs than through wild boars. The highest risk was concentrated in March and in the Northeastern provinces of Spain. Results were consist… Leer más
Martinez-Lopez B., Perez AM. y Sanchez-Vizcaino JM..
Discovery of Stable and Variable Differences in the Mycobacterium avium subsp. paratuberculosis Type I, II, and III Genomes by Pan-Genome Microarray Analysis
Applied And Environmental Microbiology 75(3):676-86.
Mycobacterium avium subsp. paratuberculosis is an important animal pathogen widely disseminated in the environment that has also been associated with Crohn`s disease in humans. Three M. avium subsp. paratuberculosis genomotypes are recognized, but genomic differences have not been fully described. To further investigate these potential differences, a 60-mer oligonucleotide microarray (designated the MAPAC array), based on the combined genomes of M. avium subsp. paratuberculosis (strain K-10) and Mycobacterium avium subsp. hominissuis (strain 104), was designed and validated. By use of a test panel of defined M. avium subsp. paratuberculosis strains, the MAPAC array was able to identify a set of large sequence polymorphisms (LSPs) diagnostic… Leer más
Castellanos E., Aranaz A., Gould KA., Linedale R., Stevenson K., Alvarez J., Dominguez L., de Juan L., Hinds J. y Bull TJ.
