Person: Aranaz Martín, Alicia
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First Name
Alicia
Last Name
Aranaz Martín
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Sanidad Animal
Area
Sanidad Animal
Identifiers
14 results
Search Results
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Item Comparison of four different culture media for isolation and growth of type II and type I/III Mycobacterium avium subsp. paratuberculosis strains isolated from cattle and goats(Applied and Environmental Microbiology, 2006) Juan Ferré, Lucía De; Álvarez Sánchez, Julio; Romero Martínez, Beatriz; Bezos Garrido, Javier; Castellanos, Elena; Aranaz Martín, Alicia; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas JoséCulture is considered the definitive technique for Johne's disease diagnosis, and it is essential for later applications of certain molecular typing techniques. In this study, we have tested four solid media (Herrold's egg yolk medium [HEYM] with sodium pyruvate and mycobactin [HEYMm-SP], HEYM with mycobactin and without sodium pyruvate [HEYMm], Middlebrook 7H11 with mycobactin [Mm], and Löwenstein-Jensen with mycobactin [LJm]) for isolation of Mycobacterium avium subsp. paratuberculosis strains in 319 tissue samples from cattle herds and goat flocks. We have shown that each of the two main groups of M. avium subsp. paratuberculosis (type II and type I/III) has different requirements for growth in the culture media studied. The recommended solid media for isolation of type I/III strains are LJm and Mm, since the combination of both media allowed the recovery of all these strains. The most widespread culture medium, HEYM, is not suitable for the isolation of this group of M. avium subsp. paratuberculosis strains. Regarding the type II strains, HEYMm-SP was the medium where more strains were isolated, but the other three media are also needed in order to recover all type II strains. The incubation period is also related to the strain type. In conclusion, because the type of strain cannot be known in advance of culture, coupled with the fact that cattle and goats can be infected with both groups of strains, we recommend the use of the four solid media and the prolongation of the incubation period to more than 6 months to detect paratuberculous herds/flocks and to determine the true prevalence of the infection.Item Effect of paratuberculosis on the diagnosis of bovine tuberculosis in a cattle herd with a mixed infection using interferon-gamma detection assay(Veterinary Microbiology, 2009) Álvarez Sánchez, Julio; Juan Ferré, Lucía De; Bezos Garrido, Javier; Romero Martínez, Beatriz; Sáez, Jose Luis; Marqués, Sergio; Domínguez, Concepción; Mínguez, Olga; Fernández-Mardomingo, Baudilio; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas José; Aranaz Martín, AliciaInterferon-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 paratuberculosis as the gold standard to determine the infection status of every animal. A total of 218 animals were slaughtered and sampled for bacteriology after blood sampling. IFN-gamma assay showed a lower apparent sensitivity in animals with a mixed infection (50%) compared to all animals suffering tuberculosis (78.3%). This finding indicates that the presence of paratuberculosis in tuberculosis-infected herds could imply a serious impairment in the sensitivity of IFN-gamma detection test.Item Genetic diversity of Mycobacterium avium isolates recovered from clinical samples and from the environment: molecular characterization for diagnostic purposes(Journal of clinical microbiology, 2008) Álvarez Sánchez, Julio; Gómez García, Ignacio; Aranaz Martín, Alicia; Bezos Garrido, Javier; Romero Martínez, Beatriz; Juan Ferré, Lucía De; Mateos García, Ana Isabel; Gómez Mampaso, Enrique; Domínguez Rodríguez, Lucas JoséIsolation 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 out of 19 (26.3%) isolates from respiratory specimens. This strain did not cause disease in the patients. Testing best characterized the strain as M. avium subsp. hominissuis, with the unusual feature that 81.4% of these isolates lacked the IS1245 element. Contamination of certain clinical samples with an environmental strain was the most likely event; therefore, characterization of the environmental mycobacteria present in health care facilities should be performed to discard false-positive isolations in nonsterile samples, mainly urine samples. Molecular techniques applied in this study demonstrated their usefulness for this purpose.Item Assessment of diagnostic tools for eradication of bovine tuberculosis in cattle co-infected with Mycobacterium bovis and M. avium subsp. paratuberculosis(Veterinary Research, 2006) Aranaz Martín, Alicia; Juan Ferré, Lucía De; Bezos Garrido, Javier; Álvarez Sánchez, Julio; Romero Martínez, Beatriz; Lozano, Francisco; Paramio, José L.; López-Sánchez, Jesús; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas JoséThe intradermal tuberculin (IDTB) test and the interferon-gamma (IFN-$\gamma$) assay are used worldwide for detection of bovine tuberculosis in cattle, but little is known about the effect of co-infecting agents on the performance of these diagnostic tests. This report describes a field trial conducted in a cattle herd with dual infection (bovine tuberculosis and paratuberculosis) during 3.5 years. It has been based on a strategic approach encompassing serial parallel testing (comparative IDTB test, the IFN-$\gamma$ assay and serology of paratuberculosis) that was repeated 8 times over the period, and segregation of animals into two herds. The IDTB test detected 65.2% and the IFN-$\gamma$ test detected 69.6% of the Mycobacterium bovis culture-positive cattle. However, the IDTB test performed better during the first part of the trial, while the IFN-$\gamma$ test was the only method that detected infected animals during the following three samplings. The number of false positive reactors with the IDTB and/or the IFN-$\gamma$ tests was remarkably high compared to other reports, and could be caused by cross-reactivity with M. avium subsp. paratuberculosis. Also, the M. bovis isolates from cattle and wildlife from the same property were characterised using molecular techniques to disclose an epidemiological link. The IDTB test may not be appropriate to eradicate bovine tuberculosis in herds with dual mycobacterial infections. This report highlights the need to use several diagnostic techniques for the accurate detection of M. bovis infected animals in these herds.Item Molecular epidemiology of Types I/III strains of Mycobacterium avium subspecies paratuberculosis isolated from goats and cattle(Veterinary Microbiology, 2006) Juan Ferré, Lucía De; Álvarez Sánchez, Julio; Aranaz Martín, Alicia; Rodríguez Bertos, Antonio Manuel; Romero Martínez, Beatriz; Bezos Garrido, Javier; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas JoséMolecular characterization of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolates classifies them into three groups: cattle or Type II, sheep or Type I, and intermediate or Type III. To avoid problems associated with characterization of extremely slow growth strains, PCR-based techniques that divide the M. a. paratuberculosis strains in two main groups (cattle or Type II, and sheep or Types I/III) can be performed. The objectives of this study were to characterize the M. a. paratuberculosis isolates identified by different PCR-based tests (IS1311-PCR and restriction endonuclease analysis, PCR test based on a DNA sequence difference, and a PCR aimed at three Type I-specific loci), and to determine the clinical and epidemiological implications of Types I/III M. a. paratuberculosis strains in livestock. One hundred and fifty-eight M. a. paratuberculosis strains from domestic ruminants were analyzed. One hundred and six M. a. paratuberculosis isolates (61 from goats and 45 from cattle) were classified as Type II strains; and 52 (29 from cows, 20 from goats, and three from sheep) were included in the Types I/III. The Types I/III M. a. paratuberculosis strains were associated to Spanish native breeds. The majority of these animals had not been in direct or indirect contact with sheep flocks infected with M. a. paratuberculosis. This fact should be taken into account when implementing paratuberculosis control programs.Item Single nucleotide polymorphisms in the IS900 sequence of Mycobacterium avium subsp. paratuberculosis are strain type specific(Journal of clinical microbiology, 2009) Castellanos, Elena; Aranaz Martín, Alicia; Juan Ferré, Lucía De; Álvarez Sánchez, Julio; Rodríguez, Sabrina; Romero Martínez, Beatriz; Bezos Garrido, Javier; Stevenson, Karen; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas JoséInsertion 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.Item Interference of paratuberculosis with the diagnosis of tuberculosis in a goat flock with a natural mixed infection(Veterinary Microbiology, 2008) Álvarez Sánchez, Julio; Juan Ferré, Lucía De; Bezos Garrido, Javier; Romero Martínez, Beatriz; Sáez, Jose Luis; Reviriego Gordejo, F.J.; Briones Dieste, Víctor; Moreno Romo, Miguel Ángel; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas José; Aranaz Martín, AliciaDetection 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-γ) 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-γ assay detected 71% (62.4–78.6, 95% C.I.) of the infected animals; the single intradermal cervical comparative tuberculin (SICCT) test detected only 42.7% (34.1–51.7, 95% C.I.) of infected animals. The highest level of sensitivity was obtained when SIT test and IFN-γ assay were combined in parallel (90.8%, 84.5–95.2, 95% C.I.). Sensitivities of the tests were also assessed by comparing animals suffering tuberculosis and animals with a mixed infection; tests were found to be more effective in the former group. Paratuberculosis seems to have a major effect in the sensitivity of the diagnostic tests under study, and therefore must be taken into account; in particular, the use of the SICCT test should be questioned when both tuberculosis and paratuberculosis are present.Item Limitations of spoligotyping and variable-number tandem-repeat typing for molecular tracing of Mycobacterium bovis in a high-diversity setting(Journal of clinical microbiology, 2011) Rodríguez Campos, Sabrina; Aranaz Martín, Alicia; Juan Ferré, Lucía De; Sáez Llorente, José Luis; Romero Martínez, Beatriz; Bezos Garrido, Javier; Jiménez, Antonio; Mateos García, Ana Isabel; Domínguez Rodríguez, Lucas JoséThis study describes the attempt to trace the first Mycobacterium bovis outbreak in alpacas (Lama pacos) in Spain by spoligotyping and variable-number tandem-repeat (VNTR) analysis. Due to high genotype diversity, no matching source was identified, but local expansion of a clonal group was found and its significance for molecular tracing is discussed.Item Splitting of a prevalent Mycobacterium bovis spoligotype by variable-number tandem-repeat typing reveals high heterogeneity in an evolving clonal group(Journal of clinical microbiology, 2013) Rodriguez Campos, Sabrina; Navarro, Yurena; Romero Martínez, Beatriz; Juan Ferré, Lucía De; Bezos Garrido, Javier; Mateos García, Ana Isabel; Golby, Paul; Smith, Noel H; Hewinson, Glyn R; García de Viedma, Darío; Aranaz Martín, Alicia; Domínguez Rodríguez, Lucas JoséMycobacterium bovis populations in countries with persistent bovine tuberculosis usually show a prevalent spoligotype with a wide geographical distribution. This study applied mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing to a random panel of 115 M. bovis isolates that are representative of the most frequent spoligotype in the Iberian Peninsula, SB0121. VNTR typing targeted nine loci: ETR-A (alias VNTR2165), ETR-B (VNTR2461), ETR-D (MIRU4, VNTR580), ETR-E (MIRU31, VNTR3192), MIRU26 (VNTR2996), QUB11a (VNTR2163a), QUB11b (VNTR2163b), QUB26 (VNTR4052), and QUB3232 (VNTR3232). We found a high degree of diversity among the studied isolates (discriminatory index [D] = 0.9856), which were split into 65 different MIRU-VNTR types. An alternative short-format MIRU-VNTR typing targeting only the four loci with the highest variability values was found to offer an equivalent discriminatory index. Minimum spanning trees using the MIRU-VNTR data showed the hypothetical evolution of an apparent clonal group. MIRU-VNTR analysis was also applied to the isolates of 176 animals from 15 farms infected by M. bovis SB0121; in 10 farms, the analysis revealed the coexistence of two to five different MIRU types differing in one to six loci, which highlights the frequency of undetected heterogeneity.Item Mycobacterium avium subspecies paratuberculosis in fallow deer and wild boar in Spain(Veterinary Record, 2005) Álvarez Sánchez, Julio; Juan Ferré, Lucía De; Briones Dieste, Víctor; Romero Martínez, Beatriz; Aranaz Martín, Alicia; Fernández-Garayzábal Fernández, José Francisco; Mateos García, Ana Isabel