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Papers by Ana Vela

Journal of Wildlife Diseases, 2003

Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Al... more Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Although P. multocida is not ordinarily associated with disease in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), numerous isolates were cultured in high numbers from free-ranging bighorn sheep in the Hells Canyon area of Idaho, Washington, and Oregon (USA) during the winter of 1995-96. Animals captured in Hells Canyon and held in captivity, and their offspring, also harbored P. multocida. Biochemical utilization tests on 90 isolates identified three subspecies: P. multocida multocida a (nϭ54); P. multocida multocida b (nϭ13); and P. multocida gallicida (nϭ15); and a non-speciated biotype, U 6 (nϭ8). Genomic DNA digestion with restriction endonuclease Hha I separated the isolates into 62 unique restriction fragment length polymorphism profiles. Capsular type A was predominant (72% of isolates). Only one isolate type, which may have been transmitted from a feral goat, was capsular type D, possessed the structural gene, toxA, for dermonecrotoxin detected by polymerase chain reaction, and produced toxin as determined by monoclonal antibody immunoblot. In conclusion, bighorn sheep in this study carried diverse types of generally non-toxigenic P. multocida associated with epizootic pneumonia.

Anaporc: revista de …, 2007

... Caracterización de Salmonella. Autores: Ana Isabel Vela, José Francisco Fernández-Garayzábal,... more ... Caracterización de Salmonella. Autores: Ana Isabel Vela, José Francisco Fernández-Garayzábal, Lucas Domínguez Rodríguez; Localización: Anaporc: revista de la Asociación de Porcinocultura Científica, ISSN 1697-2147, Vol. 4, Nº. 36, 2007 , págs. 34-39. Fundación Dialnet. ...

Microorganisms

Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically sig... more Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the s...

Journal of Antimicrobial Chemotherapy, 2017

already evolved to form Tn6172 (pHypo in Figure 1c). Using the tni genes and the IRl of Tn6022 to... more already evolved to form Tn6172 (pHypo in Figure 1c). Using the tni genes and the IRl of Tn6022 together with the IRr of Tn6172, this segment has transposed as a single unit into the comM gene of the ancestor of the GC2 clone. Subsequently, AbGRI1 has evolved in situ into the large variety of forms seen in current GC2 isolates, some of which have acquired additional antibiotic resistance genes, such as tet(B), bla PER or oxa23 in Tn2006 or AbaR4.

International journal of systematic and evolutionary microbiology, Jan 16, 2016

Biochemical and molecular genetic studies were performed on two novel Gram-positive catalase-nega... more Biochemical and molecular genetic studies were performed on two novel Gram-positive catalase-negative coccus-shaped organisms isolated from liquid joint samples of two pigs. The microorganisms were not identified as a recognized species based on their cellular, morphological and biochemical tests. 16S rRNA gene sequence comparison studies allowed their identifications as members of the genus Jeotgalibaca, but the organisms were different to Jeotgalibaca dankookensis, the single species of the genus. The two microorganisms shared 96.3% and 96.9% 16S rRNA gene sequence similarity values with their nearest phylogenetic relative, J. dankookensis. The novel bacterial isolates were distinguished from J. dankookensis using biochemical tests. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacteria be classified as two new species of the genus Jeotgalibaca, Jeotgalibaca porci and Jeotgalibaca arthritidis sp. nov. The type strain of Jeotgalibaca porci is 1...

PloS one, 2017

The application of MALDI-TOF MS for identifying streptococcal isolates recovered from clinical sp... more The application of MALDI-TOF MS for identifying streptococcal isolates recovered from clinical specimens of diseased pigs was evaluated. For this proposal, the MALDI BDAL Database (Bruker Daltoniks, Germany) was supplemented with the main spectrum profiles (MSP) of the reference strains of S. porci, S. porcorum and S. plurextorum associated with pneumonia and septicemia. Although these three species showed similar MALDI profiles, several peaks were recognized that can be useful for their differentiation: S. porci (4113, 6133, 7975 and 8228 m/z Da), S. plurextorum (3979, 4078, 4665, 6164, 6491, 6812, 7959 and 9330 m/z Da) and S. porcorum (3385, 3954, 4190, 6772, 7908, and 8381 m/z Da). After adding these MSPs, an evaluation was conducted to determine the accuracy of MALDI-TOF MS for the identification of streptococci from diseased pigs using 74 field isolates. Isolates were identified as S. suis, S. porcinus, S. dysgalactiae, S. hyovaginalis, S. porcorum, S. alactolyticus, S. hyointe...

PLOS ONE, 2015

Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycob... more Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycobacteria. In this work a histopathological and microbiological evaluation of TBL in pigs is shown. A total of 352 samples belonging to 171 pigs totally condemned at slaughterhouse due to generalized TBL were sampled and selected for analysis. Pyogranulomatous (56.2%) and granulomatous lesions (20.2%) were observed in all analysed organs. Most of the granulomas observed in both lymph nodes and lungs belonged to more advanced stages of development (stages III and IV) whereas in the liver and the spleen most of lesions belonged to intermediate stages (stages II and III). Different microorganisms were simultaneously detected from TBL in the 42.7% of the animals. Mycobacterium tuberculosis complex (MTC) (38%), coryneform bacteria (40.3%) and streptococci (28.1%) were the main groups of microorganisms detected after bacteriological analysis, with Trueperella pyogenes and Streptococcus suis as the most frequently isolated species. Mycobacteria belonging to MTC were the most frequently detected pathogens in granulomatous and pyogranulomatous lesions in submandibular lymph nodes (32.7%) and coryneform bacteria were the microorganisms more frequently isolated from lungs (25.9%) and spleen samples (37.2%). These results may provide new insights into the pathogenesis and diagnosis of this pathology. The importance of coryneform bacteria and streptococci in such processes must be evaluated in future studies.

International Journal of Systematic and Evolutionary Microbiology, 2015

Four isolates of an unknown Gram-stain-positive, catalase-negative coccus-shaped organism, isolat... more Four isolates of an unknown Gram-stain-positive, catalase-negative coccus-shaped organism, isolated from the pharynx of four wild rabbits, were characterized by phenotypic and molecular genetic methods. The micro-organisms were tentatively assigned to the genus Streptococcus based on cellular morphological and biochemical criteria, although the organisms did not appear to correspond to any species with a validly published name. Comparative 16S rRNA gene sequencing confirmed their identification as members of the genus Streptococcus, being most closely related phylogenetically to Streptococcus porcorum 682-03T (96.9 % 16S rRNA gene sequence similarity). Analysis of rpoB and sodA gene sequences showed divergence values between the novel species and S. porcorum 682-03T (the closest phylogenetic relative determined from 16S rRNA gene sequences) of 18.1 and 23.9 %, respectively. The novel bacterial isolate could be distinguished from the type strain of S. porcorum by several biochemica...

Archivos de medicina veterinaria, 2010

Determinación de los tejidos y medios de elección para la confirmación microbiológica de los resu... more Determinación de los tejidos y medios de elección para la confirmación microbiológica de los resultados serológicos de las campañas de control de Brucella melitensis en el ganado ovino Determination of best tissues and culture media for microbiological confirmation of the serological results of the control campaigns of Brucella melitensis in sheep

Microbial Ecology, 2014

In this work, we describe the biodiversity of cloacal and pharynx culture-based bacteria (commens... more In this work, we describe the biodiversity of cloacal and pharynx culture-based bacteria (commensal and pathogenic), in 75 Eurasian griffon vultures (Gyps fulvus) from two geographic areas. We address the question of whether the cultivable microbiota of vultures is organised into assemblages occurring by chance. In addition, we assess bacterial diversity in both anatomic regions and geographic areas. Bacterial diversity was represented by 26 Gram-negative and 20 Gram-positive genera. The most common genera were Escherichia, Enterococcus, Staphylococcus, Clostridium and Lactococcus. Escherichia coli and Enterococcus faecalis were the most common species in cloacal and pharyngeal samples. Staphylococcus and Erysipelothrix were isolated from the pharynx and Salmonella and Corynebacterium from the cloacae, and no Campylobacter was isolated from the cloacal swabs. Ten cloacal swabs were positive for Salmonella, of which five isolates were Salmonella enterica serotype 4,(5),12:i:-, one isolate was S. enterica serotype Derby, three isolates were S. enterica serotype 61:k:1,5,7 and one isolate was S. enterica serotype Infantis. The null modelling approach revealed that the commensal bacteria of vultures are not structured in assemblages. On the other hand, differences in bacterial genus and species richness between cloacal and pharyngeal samples or between geographic areas were clear, with the pharynx in vultures from both geographic areas being richer. The results of this study indicate also that vultures can serve as a reservoir of certain pathogenic zoonotic bacteria. The dissemination of these zoonotic pathogens in wildlife could be prevented by periodic sanitary surveys.

Veterinary Microbiology, 2005

The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from disease... more The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from diseased pigs in Spain was determined by a microdilution method. Isolates were mostly susceptible to beta-lactam antimicrobials, aminoglycosides, enrofloxacin, novobiocin and spectinomycin. More than 87% of the S. suis isolates were resistant to tetracyclines, sulphonamides, macrolides and clindamycin. Strains of serotype 9 were significantly more resistant than strains of serotype 2 (P<0.05) to tylosin (94% versus 77%) and clindamycin (94% versus 64%). Eighty-seven percent of the S. suis isolates were resistant to at least four antimicrobials and nine isolates (6%) were resistant to at least six antimicrobials. The most frequently identified multidrug pattern involved resistance against tetracyclines, sulphonamides, macrolides and lincosamides, with 69% of the isolates exhibiting this resistotype. Fifteen out of the 22 strains of serotype 2 (68.2%), and 84 out of the 98 of the strains of serotype 9 (85.7%) exhibited this resistotype, indicating its widespread distribution among the strains of the two most frequently isolated serotypes.

Veterinary Microbiology, 2013

Veterinary Microbiology, 2012

Staphylococcus aureus is an important pathogen in humans and many animal species. The prevalence ... more Staphylococcus aureus is an important pathogen in humans and many animal species. The prevalence of different clonal types in animal species remains largely unknown. We analyzed 267 S. aureus from intramammary infections in goats (47) and sheep (220) by spa typing, multi-locus sequence typing (MLST) and antimicrobial susceptibility. The most frequent spa types in goats were t337 (N=9), t759 (N=6) and t1534 (N=5). Sheep isolates mainly belonged to spa types t1534 (N=72), t2678 (N=29) and t3576 (N=20). Eighteen novel spa-types were observed; two from goat strains, 13 from sheep and three in both species. The majority of the goat strains grouped in MLST CC133 (N=10) and ST522 (N=10), followed by CC9 (N=9), while the majority of the sheep strains were of ST522 (N=108) followed by CC133 (N=86) and CC130 (N=11). Nine new MLST types were detected; three in goat and sheep isolates (ST1739, ST1758 and ST1780), two identified in goats only (ST1740 and ST2061) and four in sheep only (ST1742, ST1743, ST1781 and ST2011). Strains showed resistance below 20% against penicillin and tetracycline; a strong association between CC-types and penicillin resistance was observed. No resistance was detected to cefoxitin, quinupristin-dalfopristin, rifampicin and vancomycin. This study suggests that ST522 is the most common S. aureus clone associated with small ruminants followed by CC133.

Vaccine, 2006

Three outbreaks of clostridial disease in ruminants were reported in Spain. Out of 202,525 animal... more Three outbreaks of clostridial disease in ruminants were reported in Spain. Out of 202,525 animals in affected herds, 41,767 were infected and 22,189 died. Epidemiological investigation linked these outbreaks with vaccination with three different commercial anti-clostridial vaccines contaminated with Clostridium sordellii. Vaccines were produced by the same manufacturer. Microbiological and molecular genetic analyses confirmed this association, and isolates of C. sordellii with identical biochemical and pulsed-field gel electrophoresis (PFGE) macrorestriction patterns were isolated from both diseased animals and the epidemiologically related vaccines. Contamination of the commercial vaccines was not detected by the sterility test proposed by the European (EU) Pharmacopoeia. However, growth was obtained when the commercial vaccines were inoculated into specific culture media for Clostridium and incubated for up to 60 days.

Veterinary Record, 2002

nosa are also responsible for a significant proportion of cases of mastitis in dairy sheep (Lafi ... more nosa are also responsible for a significant proportion of cases of mastitis in dairy sheep (Lafi and others 1998, Scott and Jones 1998, Rapoport and others 1999). P aeruginosa represents less than 10 per cent of the isolates from subclinical mastitis (Lafi and others 1998), but it is responsible for outbreaks of clinical mastitis with significant sanitary and economic repercussions (Las Heras and others 1999b, Rapoport and others 1999). Molecular characterisation studies provide relevant information towards greater knowledge of the epidemiology of the different bacterial pathogens which cause mastitis, as well as for the establishment of rational control measures. These studies have generally been carried out in dairy cows with bacteria which are of higher clinical significance, mainly with S aureus (Fitzgerald and others 1997, Lange and others 1999, Buzzola and others 2001, Joo and oth-

Veterinary Record, 2011

Two hundred and five isolates of Pasteurella multocida from pigs were phenotypically and genetica... more Two hundred and five isolates of Pasteurella multocida from pigs were phenotypically and genetically characterised by determining their biovar, capsular type, virulence-associated genes and pulsed-field gel electrophoresis (PFGE) profiles. All isolates were identified as P multocida subspecies multocida and most were assigned to biovar 3 (58 per cent) and biovar 2 (39.5 per cent). Biovar 1 represented 2.4 per cent of the isolates. According to the capsular type, the great majority of the isolates (79.0 per cent) belonged to capsular type A, 18.5 per cent belonged to capsular type D and 2.4 per cent were of capsular type F. All isolates harboured ompH, psl, oma87, ptfA, nanB, nanH, tonB, hgbA, sodA and sodC genes, while none of them possessed the transferrin-binding protein gene tbpA. The prevalence of toxA, pfhaA and hgbB genes was variable (7.8, 40.5 and 60.5 per cent of the isolates, respectively). After PFGE typing, isolates of biovar 2 and 3 were grouped in two different clusters (A and B) at a level of 45 per cent similarity. In addition, isolates of biovar 2 and 3 exhibited statistically significant differences (P<0.05) in the virulence-associated hgbB and pfhA genes (biovar 3 was hgbB(+) pfhA(-), while biovar 2 was hgbB- pfhA(+)).

The Veterinary Journal, 2010

This work aims to investigate the presence of Streptococcus suis in wild rabbits. A total of 65 S... more This work aims to investigate the presence of Streptococcus suis in wild rabbits. A total of 65 S. suis isolates were recovered from 33.3% of the wild rabbits examined. Most isolates (86.2%) belong to genotype cps9. These isolates were further characterized by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and virulence genotyping. Overall, S. suis exhibited a low genetic diversity. Only 5 genetic profiles were obtained by PFGE and most isolates (71.4%) were included in two pulsotypes that were also widely distributed among the wild rabbit population. MLST analysis assigned all cps9 isolates into three new singlestones (ST216, ST217 and ST284), which were not genetically related to the European ST87 and Spanish ST61 widespread swine clones, indicating a different genetic background for the S. suis isolates from wild rabbits and pigs. Wild rabbit isolates exhibited the genotype mrpÀ/epfÀ/slyÀ, different from those showed by most of the swine S. suis isolates of the ST87 and ST61 clones. None of the S. suis isolated from wild rabbits exhibited the genotype cps2/mrp+/epf+/sly+ associated with human infections. These results indicate that S. suis isolates from wild rabbits are not genetically related with prevalent clones usually associated with infections in pigs or humans in Europe and do not exhibit either their virulence genotypes. Therefore, although wild rabbits could represent an unknown reservoir of this pathogen, they could not represent a potential risk for pigs or humans.

Systematic and Applied Microbiology, 2012

Genotypic and phenotypic analyses were performed on five Gram-negative, catalase and oxidaseposit... more Genotypic and phenotypic analyses were performed on five Gram-negative, catalase and oxidasepositive, rod-shaped bacteria isolated from the gill and liver of four rainbow trout. Studies based on comparative 16S rRNA gene sequence analysis showed that the five new isolates shared 99.8-100% sequence similarity and that they belong to the genus Chryseobacterium. The nearest phylogenetic neighbours of the strain 701B-08 T were Chryseobacterium ureilyticum F-Fue-04IIIaaaa T (99.1% 16S rRNA gene sequence similarity) and Chryseobacterium joosteii LMG 18212 T (98.6%). DNA-DNA hybridization values between the five isolates were 91-99% and ranged from 2 to 53% between strain 701B-08 T and the type strains of phylogenetically closely related species of Chryseobacterium. Strain 701B-08 T had a DNA G + C content of 36.3 mol%, the major fatty acids were iso-C 15:0 , iso-C 17:1 ω9c, C 16:1 ω6c and iso-C 17:0 3-OH and the predominant respiratory quinone was MK-6. The novel isolates were distinguished from related Chryseobacterium species by physiological and biochemical tests. The genotypic and phenotypic properties of the isolates from rainbow trout suggest their classification as representatives of a novel species of the genus Chryseobacterium, for which the name Chryseobacterium oncorhynchi sp. nov. is proposed. The type strain is 701B-08 T (=CECT 7794 T = CCUG 60105 T).

International Journal of Systematic and Evolutionary Microbiology, 2012

A taxonomic study was carried out on five Gram-staining-negative, catalase- and oxidase-positive,... more A taxonomic study was carried out on five Gram-staining-negative, catalase- and oxidase-positive, rod-shaped bacteria isolated from the gills and livers of five diseased rainbow trout. The five novel isolates were designated strains 687B-08T, 445-08, 452-08, 453B-08 and 967B-08. In phylogenetic analyses based on 16S rRNA gene sequences, the five novel strains appeared almost identical (99.0–100 % sequence similarity) and to belong to the genus Chryseobacterium . Strain 687B-08T (the strain selected to represent the five novel isolates) was found to be most closely related to Chryseobacterium oncorhynchi 701B-08T (98.9 % sequence similarity), Chryseobacterium ureilyticum F-Fue-04IIIaaaaT (98.6 %), Chryseobacterium indologenes ATCC 29897T (98.3 %), Chryseobacterium jejuense JS17-8T (98.1 %) and Chryseobacterium gleum ATCC 35910T (98.1 %). In DNA–DNA hybridizations, DNA–DNA relatedness values of 99–100 % were recorded between the five novel strains. Lower DNA–DNA relatedness values (21...

International Journal of Systematic and Evolutionary Microbiology, 2005

Five strains of an unusual Gram-negative, catalase-positive, oxidase-positive, coccobacillus-shap... more Five strains of an unusual Gram-negative, catalase-positive, oxidase-positive, coccobacillus-shaped bacterium isolated from the lungs and heart of pigs with pneumonia and pericarditis were characterized by phenotypic and molecular genetic methods. On the basis of cellular morphology and biochemical criteria, the isolates were tentatively assigned to the family Neisseriaceae, although they did not appear to correspond to any recognized genus or species. Comparative 16S rRNA gene sequencing showed that the five unidentified strains were phylogenetically highly related to each other and represent a hitherto unknown subline within the family Neisseriaceae. On the basis of both phenotypic and phylogenetic evidence, it is proposed that the unknown isolates from pigs be classified as a novel genus and species within the family Neisseriaceae, for which the name Uruburuella suis gen. nov., sp. nov. is proposed. The type strain of U. suis is 1258/02T (=CCUG 47806T=CECT 5685T).

Journal of Wildlife Diseases, 2003

Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Al... more Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Although P. multocida is not ordinarily associated with disease in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), numerous isolates were cultured in high numbers from free-ranging bighorn sheep in the Hells Canyon area of Idaho, Washington, and Oregon (USA) during the winter of 1995-96. Animals captured in Hells Canyon and held in captivity, and their offspring, also harbored P. multocida. Biochemical utilization tests on 90 isolates identified three subspecies: P. multocida multocida a (nϭ54); P. multocida multocida b (nϭ13); and P. multocida gallicida (nϭ15); and a non-speciated biotype, U 6 (nϭ8). Genomic DNA digestion with restriction endonuclease Hha I separated the isolates into 62 unique restriction fragment length polymorphism profiles. Capsular type A was predominant (72% of isolates). Only one isolate type, which may have been transmitted from a feral goat, was capsular type D, possessed the structural gene, toxA, for dermonecrotoxin detected by polymerase chain reaction, and produced toxin as determined by monoclonal antibody immunoblot. In conclusion, bighorn sheep in this study carried diverse types of generally non-toxigenic P. multocida associated with epizootic pneumonia.

Anaporc: revista de …, 2007

... Caracterización de Salmonella. Autores: Ana Isabel Vela, José Francisco Fernández-Garayzábal,... more ... Caracterización de Salmonella. Autores: Ana Isabel Vela, José Francisco Fernández-Garayzábal, Lucas Domínguez Rodríguez; Localización: Anaporc: revista de la Asociación de Porcinocultura Científica, ISSN 1697-2147, Vol. 4, Nº. 36, 2007 , págs. 34-39. Fundación Dialnet. ...

Microorganisms

Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically sig... more Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the s...

Journal of Antimicrobial Chemotherapy, 2017

already evolved to form Tn6172 (pHypo in Figure 1c). Using the tni genes and the IRl of Tn6022 to... more already evolved to form Tn6172 (pHypo in Figure 1c). Using the tni genes and the IRl of Tn6022 together with the IRr of Tn6172, this segment has transposed as a single unit into the comM gene of the ancestor of the GC2 clone. Subsequently, AbGRI1 has evolved in situ into the large variety of forms seen in current GC2 isolates, some of which have acquired additional antibiotic resistance genes, such as tet(B), bla PER or oxa23 in Tn2006 or AbaR4.

International journal of systematic and evolutionary microbiology, Jan 16, 2016

Biochemical and molecular genetic studies were performed on two novel Gram-positive catalase-nega... more Biochemical and molecular genetic studies were performed on two novel Gram-positive catalase-negative coccus-shaped organisms isolated from liquid joint samples of two pigs. The microorganisms were not identified as a recognized species based on their cellular, morphological and biochemical tests. 16S rRNA gene sequence comparison studies allowed their identifications as members of the genus Jeotgalibaca, but the organisms were different to Jeotgalibaca dankookensis, the single species of the genus. The two microorganisms shared 96.3% and 96.9% 16S rRNA gene sequence similarity values with their nearest phylogenetic relative, J. dankookensis. The novel bacterial isolates were distinguished from J. dankookensis using biochemical tests. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacteria be classified as two new species of the genus Jeotgalibaca, Jeotgalibaca porci and Jeotgalibaca arthritidis sp. nov. The type strain of Jeotgalibaca porci is 1...

PloS one, 2017

The application of MALDI-TOF MS for identifying streptococcal isolates recovered from clinical sp... more The application of MALDI-TOF MS for identifying streptococcal isolates recovered from clinical specimens of diseased pigs was evaluated. For this proposal, the MALDI BDAL Database (Bruker Daltoniks, Germany) was supplemented with the main spectrum profiles (MSP) of the reference strains of S. porci, S. porcorum and S. plurextorum associated with pneumonia and septicemia. Although these three species showed similar MALDI profiles, several peaks were recognized that can be useful for their differentiation: S. porci (4113, 6133, 7975 and 8228 m/z Da), S. plurextorum (3979, 4078, 4665, 6164, 6491, 6812, 7959 and 9330 m/z Da) and S. porcorum (3385, 3954, 4190, 6772, 7908, and 8381 m/z Da). After adding these MSPs, an evaluation was conducted to determine the accuracy of MALDI-TOF MS for the identification of streptococci from diseased pigs using 74 field isolates. Isolates were identified as S. suis, S. porcinus, S. dysgalactiae, S. hyovaginalis, S. porcorum, S. alactolyticus, S. hyointe...

PLOS ONE, 2015

Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycob... more Tuberculosis-like lesions (TBL) in pigs have been associated with microorganisms other than mycobacteria. In this work a histopathological and microbiological evaluation of TBL in pigs is shown. A total of 352 samples belonging to 171 pigs totally condemned at slaughterhouse due to generalized TBL were sampled and selected for analysis. Pyogranulomatous (56.2%) and granulomatous lesions (20.2%) were observed in all analysed organs. Most of the granulomas observed in both lymph nodes and lungs belonged to more advanced stages of development (stages III and IV) whereas in the liver and the spleen most of lesions belonged to intermediate stages (stages II and III). Different microorganisms were simultaneously detected from TBL in the 42.7% of the animals. Mycobacterium tuberculosis complex (MTC) (38%), coryneform bacteria (40.3%) and streptococci (28.1%) were the main groups of microorganisms detected after bacteriological analysis, with Trueperella pyogenes and Streptococcus suis as the most frequently isolated species. Mycobacteria belonging to MTC were the most frequently detected pathogens in granulomatous and pyogranulomatous lesions in submandibular lymph nodes (32.7%) and coryneform bacteria were the microorganisms more frequently isolated from lungs (25.9%) and spleen samples (37.2%). These results may provide new insights into the pathogenesis and diagnosis of this pathology. The importance of coryneform bacteria and streptococci in such processes must be evaluated in future studies.

International Journal of Systematic and Evolutionary Microbiology, 2015

Four isolates of an unknown Gram-stain-positive, catalase-negative coccus-shaped organism, isolat... more Four isolates of an unknown Gram-stain-positive, catalase-negative coccus-shaped organism, isolated from the pharynx of four wild rabbits, were characterized by phenotypic and molecular genetic methods. The micro-organisms were tentatively assigned to the genus Streptococcus based on cellular morphological and biochemical criteria, although the organisms did not appear to correspond to any species with a validly published name. Comparative 16S rRNA gene sequencing confirmed their identification as members of the genus Streptococcus, being most closely related phylogenetically to Streptococcus porcorum 682-03T (96.9 % 16S rRNA gene sequence similarity). Analysis of rpoB and sodA gene sequences showed divergence values between the novel species and S. porcorum 682-03T (the closest phylogenetic relative determined from 16S rRNA gene sequences) of 18.1 and 23.9 %, respectively. The novel bacterial isolate could be distinguished from the type strain of S. porcorum by several biochemica...

Archivos de medicina veterinaria, 2010

Determinación de los tejidos y medios de elección para la confirmación microbiológica de los resu... more Determinación de los tejidos y medios de elección para la confirmación microbiológica de los resultados serológicos de las campañas de control de Brucella melitensis en el ganado ovino Determination of best tissues and culture media for microbiological confirmation of the serological results of the control campaigns of Brucella melitensis in sheep

Microbial Ecology, 2014

In this work, we describe the biodiversity of cloacal and pharynx culture-based bacteria (commens... more In this work, we describe the biodiversity of cloacal and pharynx culture-based bacteria (commensal and pathogenic), in 75 Eurasian griffon vultures (Gyps fulvus) from two geographic areas. We address the question of whether the cultivable microbiota of vultures is organised into assemblages occurring by chance. In addition, we assess bacterial diversity in both anatomic regions and geographic areas. Bacterial diversity was represented by 26 Gram-negative and 20 Gram-positive genera. The most common genera were Escherichia, Enterococcus, Staphylococcus, Clostridium and Lactococcus. Escherichia coli and Enterococcus faecalis were the most common species in cloacal and pharyngeal samples. Staphylococcus and Erysipelothrix were isolated from the pharynx and Salmonella and Corynebacterium from the cloacae, and no Campylobacter was isolated from the cloacal swabs. Ten cloacal swabs were positive for Salmonella, of which five isolates were Salmonella enterica serotype 4,(5),12:i:-, one isolate was S. enterica serotype Derby, three isolates were S. enterica serotype 61:k:1,5,7 and one isolate was S. enterica serotype Infantis. The null modelling approach revealed that the commensal bacteria of vultures are not structured in assemblages. On the other hand, differences in bacterial genus and species richness between cloacal and pharyngeal samples or between geographic areas were clear, with the pharynx in vultures from both geographic areas being richer. The results of this study indicate also that vultures can serve as a reservoir of certain pathogenic zoonotic bacteria. The dissemination of these zoonotic pathogens in wildlife could be prevented by periodic sanitary surveys.

Veterinary Microbiology, 2005

The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from disease... more The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from diseased pigs in Spain was determined by a microdilution method. Isolates were mostly susceptible to beta-lactam antimicrobials, aminoglycosides, enrofloxacin, novobiocin and spectinomycin. More than 87% of the S. suis isolates were resistant to tetracyclines, sulphonamides, macrolides and clindamycin. Strains of serotype 9 were significantly more resistant than strains of serotype 2 (P<0.05) to tylosin (94% versus 77%) and clindamycin (94% versus 64%). Eighty-seven percent of the S. suis isolates were resistant to at least four antimicrobials and nine isolates (6%) were resistant to at least six antimicrobials. The most frequently identified multidrug pattern involved resistance against tetracyclines, sulphonamides, macrolides and lincosamides, with 69% of the isolates exhibiting this resistotype. Fifteen out of the 22 strains of serotype 2 (68.2%), and 84 out of the 98 of the strains of serotype 9 (85.7%) exhibited this resistotype, indicating its widespread distribution among the strains of the two most frequently isolated serotypes.

Veterinary Microbiology, 2013

Veterinary Microbiology, 2012

Staphylococcus aureus is an important pathogen in humans and many animal species. The prevalence ... more Staphylococcus aureus is an important pathogen in humans and many animal species. The prevalence of different clonal types in animal species remains largely unknown. We analyzed 267 S. aureus from intramammary infections in goats (47) and sheep (220) by spa typing, multi-locus sequence typing (MLST) and antimicrobial susceptibility. The most frequent spa types in goats were t337 (N=9), t759 (N=6) and t1534 (N=5). Sheep isolates mainly belonged to spa types t1534 (N=72), t2678 (N=29) and t3576 (N=20). Eighteen novel spa-types were observed; two from goat strains, 13 from sheep and three in both species. The majority of the goat strains grouped in MLST CC133 (N=10) and ST522 (N=10), followed by CC9 (N=9), while the majority of the sheep strains were of ST522 (N=108) followed by CC133 (N=86) and CC130 (N=11). Nine new MLST types were detected; three in goat and sheep isolates (ST1739, ST1758 and ST1780), two identified in goats only (ST1740 and ST2061) and four in sheep only (ST1742, ST1743, ST1781 and ST2011). Strains showed resistance below 20% against penicillin and tetracycline; a strong association between CC-types and penicillin resistance was observed. No resistance was detected to cefoxitin, quinupristin-dalfopristin, rifampicin and vancomycin. This study suggests that ST522 is the most common S. aureus clone associated with small ruminants followed by CC133.

Vaccine, 2006

Three outbreaks of clostridial disease in ruminants were reported in Spain. Out of 202,525 animal... more Three outbreaks of clostridial disease in ruminants were reported in Spain. Out of 202,525 animals in affected herds, 41,767 were infected and 22,189 died. Epidemiological investigation linked these outbreaks with vaccination with three different commercial anti-clostridial vaccines contaminated with Clostridium sordellii. Vaccines were produced by the same manufacturer. Microbiological and molecular genetic analyses confirmed this association, and isolates of C. sordellii with identical biochemical and pulsed-field gel electrophoresis (PFGE) macrorestriction patterns were isolated from both diseased animals and the epidemiologically related vaccines. Contamination of the commercial vaccines was not detected by the sterility test proposed by the European (EU) Pharmacopoeia. However, growth was obtained when the commercial vaccines were inoculated into specific culture media for Clostridium and incubated for up to 60 days.

Veterinary Record, 2002

nosa are also responsible for a significant proportion of cases of mastitis in dairy sheep (Lafi ... more nosa are also responsible for a significant proportion of cases of mastitis in dairy sheep (Lafi and others 1998, Scott and Jones 1998, Rapoport and others 1999). P aeruginosa represents less than 10 per cent of the isolates from subclinical mastitis (Lafi and others 1998), but it is responsible for outbreaks of clinical mastitis with significant sanitary and economic repercussions (Las Heras and others 1999b, Rapoport and others 1999). Molecular characterisation studies provide relevant information towards greater knowledge of the epidemiology of the different bacterial pathogens which cause mastitis, as well as for the establishment of rational control measures. These studies have generally been carried out in dairy cows with bacteria which are of higher clinical significance, mainly with S aureus (Fitzgerald and others 1997, Lange and others 1999, Buzzola and others 2001, Joo and oth-

Veterinary Record, 2011

Two hundred and five isolates of Pasteurella multocida from pigs were phenotypically and genetica... more Two hundred and five isolates of Pasteurella multocida from pigs were phenotypically and genetically characterised by determining their biovar, capsular type, virulence-associated genes and pulsed-field gel electrophoresis (PFGE) profiles. All isolates were identified as P multocida subspecies multocida and most were assigned to biovar 3 (58 per cent) and biovar 2 (39.5 per cent). Biovar 1 represented 2.4 per cent of the isolates. According to the capsular type, the great majority of the isolates (79.0 per cent) belonged to capsular type A, 18.5 per cent belonged to capsular type D and 2.4 per cent were of capsular type F. All isolates harboured ompH, psl, oma87, ptfA, nanB, nanH, tonB, hgbA, sodA and sodC genes, while none of them possessed the transferrin-binding protein gene tbpA. The prevalence of toxA, pfhaA and hgbB genes was variable (7.8, 40.5 and 60.5 per cent of the isolates, respectively). After PFGE typing, isolates of biovar 2 and 3 were grouped in two different clusters (A and B) at a level of 45 per cent similarity. In addition, isolates of biovar 2 and 3 exhibited statistically significant differences (P<0.05) in the virulence-associated hgbB and pfhA genes (biovar 3 was hgbB(+) pfhA(-), while biovar 2 was hgbB- pfhA(+)).

The Veterinary Journal, 2010

This work aims to investigate the presence of Streptococcus suis in wild rabbits. A total of 65 S... more This work aims to investigate the presence of Streptococcus suis in wild rabbits. A total of 65 S. suis isolates were recovered from 33.3% of the wild rabbits examined. Most isolates (86.2%) belong to genotype cps9. These isolates were further characterized by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and virulence genotyping. Overall, S. suis exhibited a low genetic diversity. Only 5 genetic profiles were obtained by PFGE and most isolates (71.4%) were included in two pulsotypes that were also widely distributed among the wild rabbit population. MLST analysis assigned all cps9 isolates into three new singlestones (ST216, ST217 and ST284), which were not genetically related to the European ST87 and Spanish ST61 widespread swine clones, indicating a different genetic background for the S. suis isolates from wild rabbits and pigs. Wild rabbit isolates exhibited the genotype mrpÀ/epfÀ/slyÀ, different from those showed by most of the swine S. suis isolates of the ST87 and ST61 clones. None of the S. suis isolated from wild rabbits exhibited the genotype cps2/mrp+/epf+/sly+ associated with human infections. These results indicate that S. suis isolates from wild rabbits are not genetically related with prevalent clones usually associated with infections in pigs or humans in Europe and do not exhibit either their virulence genotypes. Therefore, although wild rabbits could represent an unknown reservoir of this pathogen, they could not represent a potential risk for pigs or humans.

Systematic and Applied Microbiology, 2012

Genotypic and phenotypic analyses were performed on five Gram-negative, catalase and oxidaseposit... more Genotypic and phenotypic analyses were performed on five Gram-negative, catalase and oxidasepositive, rod-shaped bacteria isolated from the gill and liver of four rainbow trout. Studies based on comparative 16S rRNA gene sequence analysis showed that the five new isolates shared 99.8-100% sequence similarity and that they belong to the genus Chryseobacterium. The nearest phylogenetic neighbours of the strain 701B-08 T were Chryseobacterium ureilyticum F-Fue-04IIIaaaa T (99.1% 16S rRNA gene sequence similarity) and Chryseobacterium joosteii LMG 18212 T (98.6%). DNA-DNA hybridization values between the five isolates were 91-99% and ranged from 2 to 53% between strain 701B-08 T and the type strains of phylogenetically closely related species of Chryseobacterium. Strain 701B-08 T had a DNA G + C content of 36.3 mol%, the major fatty acids were iso-C 15:0 , iso-C 17:1 ω9c, C 16:1 ω6c and iso-C 17:0 3-OH and the predominant respiratory quinone was MK-6. The novel isolates were distinguished from related Chryseobacterium species by physiological and biochemical tests. The genotypic and phenotypic properties of the isolates from rainbow trout suggest their classification as representatives of a novel species of the genus Chryseobacterium, for which the name Chryseobacterium oncorhynchi sp. nov. is proposed. The type strain is 701B-08 T (=CECT 7794 T = CCUG 60105 T).

International Journal of Systematic and Evolutionary Microbiology, 2012

A taxonomic study was carried out on five Gram-staining-negative, catalase- and oxidase-positive,... more A taxonomic study was carried out on five Gram-staining-negative, catalase- and oxidase-positive, rod-shaped bacteria isolated from the gills and livers of five diseased rainbow trout. The five novel isolates were designated strains 687B-08T, 445-08, 452-08, 453B-08 and 967B-08. In phylogenetic analyses based on 16S rRNA gene sequences, the five novel strains appeared almost identical (99.0–100 % sequence similarity) and to belong to the genus Chryseobacterium . Strain 687B-08T (the strain selected to represent the five novel isolates) was found to be most closely related to Chryseobacterium oncorhynchi 701B-08T (98.9 % sequence similarity), Chryseobacterium ureilyticum F-Fue-04IIIaaaaT (98.6 %), Chryseobacterium indologenes ATCC 29897T (98.3 %), Chryseobacterium jejuense JS17-8T (98.1 %) and Chryseobacterium gleum ATCC 35910T (98.1 %). In DNA–DNA hybridizations, DNA–DNA relatedness values of 99–100 % were recorded between the five novel strains. Lower DNA–DNA relatedness values (21...

International Journal of Systematic and Evolutionary Microbiology, 2005

Five strains of an unusual Gram-negative, catalase-positive, oxidase-positive, coccobacillus-shap... more Five strains of an unusual Gram-negative, catalase-positive, oxidase-positive, coccobacillus-shaped bacterium isolated from the lungs and heart of pigs with pneumonia and pericarditis were characterized by phenotypic and molecular genetic methods. On the basis of cellular morphology and biochemical criteria, the isolates were tentatively assigned to the family Neisseriaceae, although they did not appear to correspond to any recognized genus or species. Comparative 16S rRNA gene sequencing showed that the five unidentified strains were phylogenetically highly related to each other and represent a hitherto unknown subline within the family Neisseriaceae. On the basis of both phenotypic and phylogenetic evidence, it is proposed that the unknown isolates from pigs be classified as a novel genus and species within the family Neisseriaceae, for which the name Uruburuella suis gen. nov., sp. nov. is proposed. The type strain of U. suis is 1258/02T (=CCUG 47806T=CECT 5685T).