Publicaciones científicas 2008

Persistence and molecular evolution of Mycobacterium bovis population from cattle and wildlife in Doñana National Park revealed by genotype variation

Veterinary Microbiology 132(1-2):87-95.

     The role of wildlife in tuberculosis epidemiology is being widely studied since it can affect the effectiveness of eradication campaigns in cattle. The health problem is enhanced when it concerns also wildlife welfare and biodiversity conservation. This study was performed to understand the epidemiology of Mycobacterium bovis population affecting livestock and wild animals in the Doñana National Park using bacteriology and molecular characterisation techniques. Tuberculosis research was performed on 1209 cattle and wild animals (artiodactyla and carnivore) collected over 6 years in the Park. One hundred and sixty-three animals were found to be infected with M. bovis, comprising 7.96% of the cattle and 20.53% of the wild animals tested. Spol… Leer más

Romero B., Aranaz A., Sandoval A., Alvarez J., de Juan L., Bezos J., Sanchez C., Galka M., Fernandez P., Mateos A. y Dominguez L..

Analytical sensitivity and specificity of a RT-PCR for the diagnosis and characterization of the spatial distribution of three Apis mellifera viral diseases in Spain

Apidologie 39(6):607-617.

     The occurrence and spatial distribution of deformed wing virus (DWV), black queen cell virus (BQCV), and Kashmir bee virus (KBV) were assessed in 294 honeybee colonies in Spain by employing a SYBR-Green based real time RT-PCR. 60% of them were positive for both DWV and BQCV, and those two viruses were detected in 84% and 68% of the samples, respectively. Conversely, KBV was detected in only 1.7% of the samples. Increments in the number of bee colonies per region, adjusted by the number of samples collected, were associated with increased risk of finding DWV, BQCV, and KBV, as estimated by mixed Bayesian regression models. The residual risk for DWV, BQCV, and KBV decreased northerly and westerly, suggesting that factors or forces that favour… Leer más

Kukielka D., Perez AM., Higes M., Bulboa MC. y Sanchez-Vizcaino JM..

Mycobacterium peregrinum infection in farmed European tench (Tinca tinca L.)

Veterinary Microbiology 131(3-4):393-9.

     This work is the first description of Mycobacterium peregrinum as an etiological agent for mycobacteriosis in farmed fishes. We report the mycobacterial infection in farmed European tench (Tinca tinca L.) which was confirmed by culture, molecular identification methods (PCRs aimed at 16S rRNA, rpobeta and hsp65 sequencing), and histopathology. Since M. peregrinum infection has been described in humans, their clinical significance in fishes should be considered of healthcare interest. With this case report, we also show that a multidisciplinary approach was needed to overcome difficulties associated to diagnosis of piscine mycobacteriosis… Leer más

Aranaz A., Gibello A., Alvarez J., Mata AI., Rodriguez-Bertos A., Fallola C., Fernandez-Garayzabal JF. y Dominguez L..

Prevalence and diversity of integrons and associated resistance genes in faecal Escherichia coli isolates of healthy humans in Spain

The Journal Of Antimicrobial Chemotherapy 62(5):934-7.

     OBJECTIVES: To analyse the prevalence and diversity of integrons in faecal Escherichia coli isolates from healthy humans in Spain. METHODS: One hundred E. coli isolates were obtained in Levine agar plates from faecal samples of 100 healthy humans during March to October 2007. Susceptibility to 16 antimicrobial agents was determined by the disc diffusion method. The presence and characterization of class 1, 2 and 3 integrons, as well as the presence of other antimicrobial resistance genes, were performed by PCR and DNA sequencing. RESULTS: Integrases associated with class 1 and/or class 2 integrons were identified in 29 E. coli isolates (intI1 gene in 26 isolates, intI2 in 1 isolate and intI1 + intI2 in 2 isolates), the remaining 71 isolates… Leer más

Vinue L., Saenz Y., Somalo S., Escudero E., Moreno MA., Ruiz-Larrea F. y Torres C.

Herpes simplex-like infection in a bottlenose dolphin stranded in the Canary Islands

Diseases Of Aquatic Organisms 81(1):73-6.

     A bottlenose dolphin, stranded in the Canary Islands in 2001 exhibited non-suppurative encephalitis. No molecular detection of cetacean morbillivirus (CeMV) was found, but a herpesviral-specific band of 250 bp was detected in the lung and brain. The sequenced herpesviral PCR product was compared with GenBank sequences, obtaining 98% homology (p-distance of 0.02) with Human herpesvirus 1 (herpes simplex virus 1 or HSV-1). This is the first report of a herpes simplex-like infection in a stranded dolphin… Leer más

Esperon F., Fernandez A. y Sanchez-Vizcaino JM..

Prevalence of antimicrobial resistance among bacterial pathogens isolated from cattle in different European countries: 2002-2004

Acta Veterinaria Scandinavica 50:28.

     BACKGROUND:
The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-2005, with the aim to establish a continuous monitoring of antimicrobial susceptibility among veterinary laboratories in European countries based on validated and harmonised methodologies. Available summary data of the susceptibility testing of the bacterial pathogens from the different laboratories were collected.
METHOD:
Antimicrobial susceptibility data for several bovine pathogens were obtained over a three year period (2002-2004). Each year the participating laboratories were requested to fill in excel-file templates with national summary data on t… Leer más

Hendriksen RS., Mevius DJ., Schroeter A., Teale C., Meunier D., Butaye P., Franco A., Utinane A., Amado A., Moreno MA., Greko C., Stark K., Berghold C., Myllyniemi AL., Wasyl D., Sunde M. y Aarestrup FM.

Orbiviruses in the Mediterranean basin: updated epidemiological situation of Bluetongue and new methods for the detection of BTV serotype 4

Transboundary And Emerging Diseases 55(5-6):205-14.

     Bluetongue virus (BTV), a member of the Orbivirus genus, has contributed to great economical losses in countries across the Mediterranean basin. Although BTV has an African origin, it has been reported in the South of Europe since 1924 when it was first detected in Cyprus. After this first Bluetongue (BT) outbreak, many others followed in most Mediterranean countries, resulting in seven BTV serotypes detected in 17 countries in Central-, South-Europe and North Africa in the last 10 years. Currently, six BTV serotypes (1, 2, 4, 8, 9 and 16) are circulating throughout Central- and Western Europe. The unexpected occurrence of BTV serotype 8 in Central Europe in 2006 and its spread and persistence have evidenced changes in the BTV scenario: new… Leer más

Rodriguez B., Iglesias I., Martinez-Aviles M. y Sanchez-Vizcaino JM..

Improved diagnosis for nine viral diseases considered as notifiable by the world organization for animal health

Transboundary And Emerging Diseases 55(5-6):215-25.

     Nine viral diseases included in the World Organization for Animal Health list of notifiable diseases (former list A) were chosen for their contagiousness and high capacity of spreading to improve their diagnosis using new and emerging technologies. All the selected diseases--foot-and-mouth disease, swine vesicular disease, vesicular stomatitis, classical swine fever, African swine fever, bluetongue, African horse sickness, Newcastle disease and highly pathogenic avian influenza--are considered as transboundary diseases, which detection causes the prohibition of livestock exportation, and, thus, it leads to high economical losses. The applied diagnostic techniques can fall into two categories: (i) nucleic-acid detection, including padlock pr… Leer más

Rodriguez B., Sanchez-Vizcaino JM., Uttenthal A., Rasmussen TB., Hakhverdyan M., King DP., Ferris NP., Ebert K., Reid SM., Kiss I., Brocchi E., Cordioli P., Hjerner B., McMenamy M., McKillen J., Ahmed JS. y Belak S.

Quantitative risk assessment of foot-and-mouth disease introduction into Spain via importation of live animals

Preventive Veterinary Medicine 86(1-2):43-56.

     Spain has been a foot-and-mouth disease (FMD)-free country since 1986. However, the FMD epidemics that recently affected several European Union (EU) member countries demonstrated that the continent is still at high risk for FMD virus (FMDV) introduction, and that the potential consequences of those epidemics are socially and financially devastating. This paper presents a quantitative assessment of the risk of FMDV introduction into Spain. Results suggest that provinces in north-eastern Spain are at higher risk for FMDV introduction, that an FMD epidemic in Spain is more likely to occur via the import of pigs than through the import of cattle, sheep, or goats, and that a sixfold increase in the proportion of premises that quarantine pigs pri… Leer más

Martinez B., Perez AM., de la Torre A. y Sanchez-Vizcaino JM..

Novel gel-based and real-time PCR assays for the improved detection of African horse sickness virus

Journal Of Virological Methods 151(1):87-94.

     In order to improve, ensure and accelerate the diagnosis of African horse sickness, a highly devastating, transboundary animal disease listed by the World Animal Health Organisation, (OIE) three novel diagnostic PCR assays were developed and tested in this study. The reverse transcription-PCR (RT-PCR) tests were the following: (a) a conventional, gel-based RT-PCR, (b) a real-time PCR with SYBR-Green-named rRT-PCR SYBR-Green-, and (c) a real-time PCR rRT-PCR with TaqMan probe (termed rRT-PCR TaqMan). The same pair of primers-directed against African Horse Sickness Virus (AHSV) segment 5, encoding the non-structural protein NS1, is used in the three tests listed above. The three PCR assays detected similarly the nine AHSV serotypes from culti… Leer más

Rodriguez B., Fernandez J., Sailleau C., Zientara S., Belak S., Arias M. y Sanchez-Vizcaino JM..

Bluetongue control - A new challenge for Europe

Berliner Und Münchener Tierärztliche Wochenschrift 121(7-8):306-13.

Makoschey B., Beer M., Zientara S., Haubruge E., Rinaldi L., Dercksen D., Millemann Y., von Rijn P., de Clercq K., Oura C., Saegerman C., Domingo M. y Sanchez-Vizcaino JM..

Occurrence of antimicrobial resistance among bacterial pathogens and indicator bacteria in pigs in different European countries from year 2002 - 2004: the ARBAO-II study

Acta Veterinaria Scandinavica 50:19.

     BACKGROUND:
The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-05. The aim of this project was to establish a program for the continuous monitoring of antimicrobial susceptibility of pathogenic and indicator bacteria from food animals using validated and harmonised methodologies. In this report the first data on the occurrence of antimicrobial resistance among bacteria causing infections in pigs are reported.
METHODS:
Susceptibility data from 17,642 isolates of pathogens and indicator bacteria including Actinobacillus pleuropneumoniae, Streptococcus suis and Escherichia coli isolated from pigs were collected from f… Leer más

Hendriksen RS., Mevius DJ., Schroeter A., Teale C., Jouy E., Butaye P., Franco A., Utinane A., Amado A., Moreno MA., Greko C., Stark K., Berghold C., Myllyniemi AL., Hoszowski A., Sunde M. y Aarestrup FM.

Morbillivirus and pilot whale deaths, Mediterranean Sea

Emerging Infectious Diseases 14(5):792-4.

     An outbreak of a lethal morbillivirus infection of long-finned pilot whales occurred in the Mediterranean Sea from the end of October 2006 through April 2007. Sequence analysis of a 426-bp conserved fragment of the morbillivirus phosphoprotein gene indicates that the virus is more closely related to dolphin morbillivirus than to pilot whale morbillivirus… Leer más

Fernandez A., Esperon F., Herraez P., Espinosa A., Clavel C., Bernabe A., Sanchez-Vizcaino JM., Verborgh P., DeSephanis R., Toledano F. y Bayon A.

Interference of paratuberculosis with the diagnosis of tuberculosis in a goat flock with a natural mixed infection

Veterinary Microbiology 128(1-2):72-80.

     Detection of infected animals is a key step in eradication programs of tuberculosis. Paratuberculosis infection has been demonstrated to compromise the specificity of the diagnostic tests. However, its effect on their sensitivity has not been clarified. In the present study, skin tests and the interferon-gamma (IFN-gamma) assay were evaluated in a goat flock (n=177) with a mixed tuberculosis-paratuberculosis infection in order to assess the possible effect of paratuberculosis on their sensitivity. Culture of mycobacteria was performed as the gold standard to determine the true infection status. All techniques showed lower sensitivities than previously described; the single intradermal tuberculin (SIT) test and the IFN-gamma assay detected 7… Leer más

Alvarez J., de Juan L., Bezos J., Romero B., Saez-Llorente JL., Reviriego FJ., Briones V., Moreno MA., Mateos A., Dominguez L. y Aranaz A.

Genetic diversity of Mycobacterium avium isolates recovered from clinical samples and from the environment: molecular characterization for diagnostic purposes

Journal Of Clinical Microbiology 46(4):1246-51.

     Isolation of Mycobacterium avium complex (MAC) organisms from clinical samples may occur in patients without clinical disease, making the interpretation of results difficult. The clinical relevance of MAC isolates from different types of clinical samples (n = 47) from 39 patients in different sections of a hospital was assessed by comparison with environmental isolates (n = 17) from the hospital. Various methods for identification and typing (commercial probes, phenotypic characteristics, PCR for detection of IS1245 and IS901, sequencing of the hsp65 gene, and pulsed-field gel electrophoresis) were evaluated. The same strain was found in all the environmental isolates, 21 out of 23 (91.3%) of the isolates cultured from urine samples, and 5 … Leer más

Alvarez J., Garcia IG., Aranaz A., Bezos J., Romero B., de Juan L., Mateos A., Gomez-Mampaso E. y Dominguez L..

Lactobacillus ceti sp. nov., isolated from beaked whales (Ziphius cavirostris)

International Journal Of Systematic And Evolutionary Microbiology 58(Pt 4):891-4.

     Biochemical and molecular genetic studies were performed on three isolates of an unknown Gram-positive, catalase-negative and rod-shaped organism isolated from the lungs and liver of two beaked whales. The organisms were tentatively identified as Lactobacillus spp. based on cellular morphology and biochemical tests. 16S rRNA gene sequencing studies confirmed the provisional identification of the novel isolates as members of the genus Lactobacillus, but the isolates did not correspond to any recognized species of this genus. The novel strains shared the same phenotypic characteristics and exhibited 100 % 16S rRNA gene sequence similarity. The nearest phylogenetic relatives of the novel isolates were Lactobacillus satsumensis DSM 16230T (94.2… Leer más

Vela AI., Fernandez A., Espinosa A., Goyache J., Herraez P., Tames B., Cruz F., Dominguez L. y Fernandez-Garayzabal JF..

PCR amplification of species specific sequences of 16S rDNA and 16S-23S rDNA intergenic spacer region for identification of Streptococcus phocae

Microbiological Research 163(2):132-5.

     Streptococcus phocae, a bacterial pathogen of seals, could reliably be identified by PCR amplification using oligonucleotide primers designed according to species specific segments of the previously sequenced 16S rRNA gene and the 16S-23S rDNA intergenic spacer region of this species. The PCR mediated assay allowed an identification of S. phocae isolated from harbor and gray seals and from Atlantic salmons. No cross-reaction could be observed with 13 different other streptococcal species and subspecies and with Lactococcus garvieae strains investigated for control purposes… Leer más

Hassan AA., Vossen A., Lammler C., Siebert U. y Fernandez-Garayzabal JF..

Vaccines against bluetongue in Europe

Comparative Immunology, Microbiology And Infectious Diseases 31(2-3):101-20.

     After the incursion of bluetongue virus (BTV) into European Mediterranean countries in 1998, vaccination was used in an effort to minimize direct economic losses to animal production, reduce virus circulation and allow safe movements of animals from endemic areas. Vaccination strategies in different countries were developed according to their individual policies, the geographic distribution of the incurring serotypes of BTV and the availability of appropriate vaccines. Four monovalent modified live virus (MLV) vaccines were imported from South Africa and subsequently used extensively in both cattle and sheep. MLVs were found to be immunogenic and capable of generating strong protective immunity in vaccinated ruminants. Adverse side effects … Leer más

Savini G., MacLachlan NJ., Sanchez-Vizcaino JM. y Zientara S.

Role of hepatic macrophages during the viral haemorrhagic fever induced by African Swine Fever Virus

Histology And Histopathology 23(6):683-91.

     To ascertain the role played by the various liver monocyte-macrophage populations in the course of a viral hemorrhagic fever, fifteen pigs were inoculated intramuscularly with the highly virulent isolate of African Swine Fever Virus (ASFV) España-70 and slaughtered at 1-7 days post-inoculation (dpi). Samples of liver were fixed in different solutions and routinely processed for morphological, immunohistochemical and ultrastructural studies. Viral antigen (vp73) was detected from 3 dpi onward, mainly in circulating monocytes of sinusoid and Kupffer`s cells (KC), as well as in portal macrophages and hepatocytes from 5 dpi. Anti-SWC3 immunolabelled cells were increased from 1 dpi, peaking between 3 and 5 dpi, thereafter declining until the end… Leer más

Sanchez-Cordon PJ., Romero-Trevejo JL., Pedrera M., Sanchez-Vizcaino JM., Bautista MJ. y Gomez-Villamandos JC.

Determination of the immunotoxic potential of heavy metals on the functional activity of bottlenose dolphin leukocytes in vitro

Veterinary Immunology And Immunopathology 121(3-4):189-98.

     Heavy metals may affect the immune system of cetaceans. But no information exists on their effects on the bottlenose dolphin (Tursiops truncatus) immune system, although this species is a coastal top predator which can bioaccumulate high concentrations of them. This work studies the effects of Hg (1, 5 and 10mg/L), Al (2,5, 25 and 50mg/L), Cd (1, 10, 20 and 40mg/L), Pb (1, 10, 20 and 50mg/L) and Cr (1 and 10mg/L), on the function of phagocytes and lymphocytes isolated from the peripheral blood of bottlenose dolphins under in vitro conditions. Cell viability, apoptosis, lymphocyte proliferation and phagocytosis were evaluated. Viability and lymphoproliferation were measured with Alamar Blue assay, and apoptosis and phagocytosis were evaluate… Leer más

Camara S., Munoz MJ., Carballo M. y Sanchez-Vizcaino JM..

Rapid and differential diagnosis of foot-and-mouth disease, swine vesicular disease, and vesicular stomatitis by a new multiplex RT-PCR assay

Journal Of Virological Methods 147(2):301-11.

     A highly sensitive and specific one-step multiplex RT-PCR assay has been developed and standardised for the simultaneous and differential detection of the most important vesicular viruses affecting livestock: foot-and-mouth disease virus (FMDV), swine vesicular disease virus (SVDV), and vesicular stomatitis virus (VSV). The method uses three primer sets, each one specific for the corresponding virus, selected to detect of all serotypes of FMD and VS. The detection range was confirmed by examination of a collection of 31 isolates of the three target viruses. The specificity of the assay was also demonstrated by testing other related viruses, uninfected cell line cultures and healthy pig tissues. The testing of blood and serum samples from an… Leer más

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

A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera

Journal Of Virological Methods 147(2):275-81.

     A one-step real-time RT-PCR based on SYBR Green (SG) chemistry was developed for the detection, differentiation and quantification of two of the most common viruses on the honeybee Apis mellifera L., deformed wing virus and black queen cell virus. Two sets of primers specific for each virus, were designed in conserved regions of the viral genome for their use in the one-step real-time RT-PCR. Both reactions were optimized for highest sensitivity and specificity and SG-based real-time was used to achieve quantitative detection. All samples evaluated in this study were from Spanish honeybee colonies. Viral detection and identification was confirmed by sequencing of the PCR products. The described one-step real-time SG RT-PCR proved to be a fa… Leer más

Kukielka D., Esperon F., Higes M. y Sanchez-Vizcaino JM..

Single-nucleotide polymorphism in two representative multidrug-resistant Mycobacterium bovis isolates collected from patients in a Spanish hospital harboring a human infection outbreak

Journal Of Clinical Microbiology 46(2):826-7.

     Mycobacterium bovis is the etiological agent of tuberculosis in domestic and wild animals. Its involvement as a human pathogen has been highlighted again with the recent descriptions of transmission through dairy products (18), reactivation or primary infection in human immunodeficiency virus-infected patients (5), and association with meat industry workers, animal keepers, or hunters (3). Strains resistant to antituberculous drugs (M. bovis is naturally resistant to pyrazinamide) pose an additional risk (2). Several studies have demonstrated that mutations in target genes are associated with resistance to antituberculous drugs (4, 7, 10, 11, 16). However, most of them have been developed in Mycobacterium tuberculosis strains and limited da… Leer más

Romero B., Aranaz A., Bezos J., Alvarez J., de Juan L., Mateos A., Gomez-Mampaso E. y Dominguez L..