Person:
Bezos Garrido, Javier

First Name

Javier

Last Name

Bezos Garrido

URI

https://hdl.handle.net/20.500.14352/80454

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Veterinaria

Department

Sanidad Animal

Area

Sanidad Animal

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Now showing 1 - 10 of 13

Tuberculosis in domestic ruminants: towards eradication of zoonotic tuberculosis
(Frontiers in Veterinary Science, 2024) Boschiroli, Laura; Salguero, Francisco Javier; Jones, Gareth; Bezos Garrido, Javier
Tuberculosis (TB) is a zoonosis with a significant impact on human and animal health and it remains endemic in many countries around the world. Most cases of human TB are caused by Mycobacterium tuberculosis, whilst TB in wildlife and domestic ruminants is mainly caused by animal adapted mycobacterial species such as M. bovis and M. caprae. However, the impact of zoonotic TB (zTB) has been known for a long time and resulted in the implementation of milk pasteurization as a preventive public health measure in many countries. According to data published by the WHO, more than 140,000 people are infected and more than 12,000 die every year due to zTB, particularly in the African and Southeast Asian regions. Zoonotic M. bovis transmission represents 0.4% of all cases in the EU, although this may be underestimated, as it is not always possible to confirm the TB species involved, or conduct appropriate epidemiological investigations. Livestock TB control programs were originally implemented to reduce zTB. Successes in controlling infections in livestock, together with measures to control animal products with suspected infection have contributed to lowering the annual incidence of cases of zTB in high-income countries. The incidence tends to be higher in countries where animal and public health resources are inadequate to manage costly bovine TB control programs. TB eradication is difficult to achieve mainly due to: (I) limited knowledge of the complex immunological response against the infection, (II) complex epidemiology, including several domestic and wildlife reservoirs, (III) the absence of an effective vaccine, (IV) a limited performance of the current diagnostic tests, and (V) societal aspects. Therefore, there is a need to continue working on these fields of TB research. In this Research Topic, different aspects of the epidemiology, diagnosis and control of TB in wildlife and domestic ruminants have been addressed to increase the knowledge about this important zoonotic disease. This will contribute to TB eradication in domestic ruminants, which also represents an important step to reduce the zTB.
Intradermal Tuberculin Test in Water Buffalo (Bubalus bubalis): Experimental use of Mycobacterial Antigens for the Diagnosis of Bovine Tuberculosis
(Journal of Buffalo Science, 2024) Martucciello, Alessandra; Mazzone, Piera; Napolitano, Francesco; Bezos Garrido, Javier; Grandoni, Francesco; Boniotti, Maria Beatrice; Cagiola, Monica; Cappelli, Giovanna; Di Vuolo, Gabriele; Galiero, Giorgio; Signorelli, Federica; De Carlo, Esterina
The study aims to evaluate the potential use of mycobacterial ESAT6 and CFP10 antigens, Early Secretory Proteins (ESP) in the Skin Test used for bovine tuberculosis (TB) diagnosis in Water Buffalo. A pilot study was performed on 21 buffaloes from a TB outbreak and 11 buffaloes from a TB-free herd. Three concentrations of ESAT6-CFP10 (10, 20, and 30 mg) and two of ESP (50 and 100 µg) were inoculated in the Skin Test, along with PPDB, PPDA, and PBS as a negative control. Skin thickness was measured with calipers before the test and every 24 hours for 4 days. Then, to evaluate the specificity of the antigens, a field study was conducted, and 100 buffaloes from a TB-free herd were inoculated using the best antigens concentration derived from the pilot study. In the positive buffaloes, the strongest skin response was to PPDB at 24h, with some subjects becoming inconclusive at 72 and 96 h. A peak response to PPDA at 48 hours was detected, followed by a slight decrease. The response to ESP-100 µg remained high at 24 and 48 h, then decreased, remaining positive at 72 h. In the 100 TB-free buffaloes, the best specificity was observed using ESAT6-CFP10 and ESP. ESP yielded the best results, showing higher reactivity in infected animals and no reactivity in the healthy ones at 72 h. Therefore, ESP could be an excellent candidate for further extensive studies in the buffalo species to improve Skin Test performance.
Analysis of the cattle movement network and its association with the risk of bovine tuberculosis at the farm level in Castilla y Leon, Spain
(Transboundary and Emerging Diseases, 2019) Pozo Piñol, Pilar; Bezos Garrido, Javier; Álvarez Sánchez, Julio; VanderWaal, Kimberly; Grau, Anna; de la Cruz Conty, María Luisa; Nácar, Jesus; Pérez, Andrés; Mínguez, Olga
Between-farm transmission of bovine tuberculosis (bTB) occurs mostly through fence-to-fence contact between neighbouring farms, endemic infected wildlife or movement of infected animals. Unfortunately, bTB detection is frequently delayed and identification of the source of introduction is often difficult, particularly in endemic regions. Here, we characterized the cattle movement network of Castilla y Leon, a high bTB-prevalence (1.9% at the farm level in 2015) region in Spain, over six years and analysed the distribution of bTB to ultimately assess the likelihood of spatial and movement-mediated transmission. We analysed movement and bTB data from 27,633 units located in the region, of which 87% were involved in ~1.4 million movements of ~8.8 million animals. Network-level connectivity was low, although a few highly connected units were identified. Up to 15% of the herds became bTB-positive at some point during the study, with the highest percentage found in bullfighting and beef herds. Although bTB-positive herds had a significantly higher degree and moved more cattle than negative herds. Results of the k-test, a permutation-based procedure, suggested that positive farms were not significantly clustered in the movement network. Location was a likely risk factor as bTB-positive farms tended to be located within 5 km from each other. Results suggested that movements may be a source of bTB in cattle in Castilla y Leon, although local factors may be more influential in determining risk of disease at the farm level. The description of the movement network in Castilla y Leon may be valuable for bTB surveillance in Spain. Moreover, results are useful to assess the movement-associated risk for multiple diseases.
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, Alicia
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 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.
Evaluation of the Effect of a Recent Comparative Intradermal Tuberculin Test on the Humoral Diagnosis of Paratuberculosis Using Serum and Milk Samples from Goats
(Veterinary Science, 2024) Velasco, Carlos; Ortega, Javier; Gómez-Buendía, Alberto; Grau, Anna; López, Marisol; Álvarez Sánchez, Julio; Romero Martínez, Beatriz; Juan Ferré, Lucía De; Bezos Garrido, Javier
Paratuberculosis (PTB) and tuberculosis (TB) are two mycobacterial diseases with a severe economic and health impact on domestic ruminants. The ante mortem diagnosis of PTB is hampered, among other factors, by the limited sensitivity of all the available diagnostic techniques. Since TB-infected goats subjected to the comparative intradermal tuberculin test (CITT) may experience a booster effect on their antibody titer and a potential enhancement to the sensitivity of humoral techniques for tuberculosis, in the present study we aimed to evaluate this diagnostic strategy on the humoral diagnosis of PTB in serum and milk samples collected from a caprine herd that was TB free and PTB infected. The results from 120 goats indicated a significant increase (p < 0.001) in the quantitative response detected using an ELISA technique, conducted using serum and milk samples taken 15 and 30 days after performing a CITT (day 0 of the study); although, it did not translate into a significant increase in the number of reactors during any of the testing events (0, 3,15, 30 and 60 days post-CITT). Additionally, the number of ELISA-positive animals was higher for the serum versus the milk samples at both 15 and 30 days post-CITT. The increase in the quantitative ELISA result suggested a diagnostic strategy that maximizes ELISA sensitivity, mainly using serum samples, in PTB-infected herds; although, it may depend on individual differences and the interpretation criteria.
Effect of a recent intradermal test on the specificity of P22 ELISA for the diagnosis of caprine tuberculosis
(Frontiers in Veterinary Science, 2024) Romero Martínez, Beatriz; Juan Ferré, Lucía De; Domínguez Rodríguez, Lucas José; Álvarez Sánchez, Julio; Bezos Garrido, Javier; Velasco, Carlos; Ortega, Javier; Domínguez, Mercedes; Ricón, Jaime; Moreno, Inmaculada
Caprine tuberculosis (TB) is a zoonotic disease caused by members of the Mycobacterium tuberculosis complex. TB eradication programs in goats are based on the single and comparative intradermal tuberculin tests (SITT and CITT, respectively). Antibody-based diagnostic techniques have emerged as potential diagnostic tools for TB. P22 ELISA has been previously evaluated using samples collected after the intradermal tuberculin tests to maximize the sensitivity, a phenomenon known as booster effect. However, there is no information available on whether the use of this diagnostic strategy could lead to a decrease of its specificity (Sp). The aim of the present study was to elucidate the interference effect of a recent CITT on the Sp of the P22 ELISA in serum and milk samples collected at different times after the CITT from a TB-free herd (n = 113). The number of reactors to P22 ELISA was significantly higher (p < 0.01) on serum samples collected 15 days post-CITT compared to day 0, showing a decrease in Sp from 99.1% (95% CI; 95.2–99.8%) to 88.5% (95% CI; 81.3–93.2%). The number of reactors and the quantitative values of P22 ELISA were significantly higher (p < 0.01) in serum samples compared to milk. No significant (p > 0.05) changes in the Sp of the P22 ELISA were observed throughout the different time samplings using milk No significant (p > 0.05) changes were observed on days 30 and 60 post-CITT. In conclusion, the booster effect strategy may significantly decrease the Sp of P22 ELISA in TB-free herds when serum samples are used but not when milk is tested.
Bovine tuberculosis in Spain, is it really the final countdown?
(Irish Veterinary Journal, 2023) Bezos Garrido, Javier; Sáez Llorente, José Luis; Álvarez Sánchez, Julio; Romero Martínez, Beatriz; Díez Guerrier, Alberto Antoine; Domínguez Rodríguez, Lucas José; Juan Ferré, Lucía De
Bovine tuberculosis (bTB) is a severe zoonotic disease that has major impacts on both health and the economy, and which has been subjected to specific eradication programmes in many countries for decades. This manuscript highlights the relevance of this disease in the context of the European Union (EU) and summarizes the epidemiological situation and the main tools (e.g. antemortem diagnostic tests, slaughterhouse surveillance, laboratories, comprehensive databases, etc.) used to control and eradicate bTB in the various EU countries with a focus on the situation in Spain. A comprehensive description of the specific bTB epidemiological situation in Spain is provided, together with an assessment of the evolution of different epidemiological indicators throughout the last decades. Moreover, the main features of the Spanish bTB eradication programme and its control tools are described, along with the studies carried out in Spain that have allowed the updating of and improvement to the programme over the years with the aim of eradication, which has been established for 2030.
Non-tuberculous mycobacteria: occurrence in skin test cattle reactors from official tuberculosis-free herds
(Frontiers in Veterinary Science, 2024) Gómez Buendía, Alberto; Álvarez Sánchez, Julio; Bezos Garrido, Javier; Mourelo, Jorge; Amado, Javier; Saez, José Luis; Juan Ferré, Lucía De; Romero Martínez, Beatriz
Non-tuberculous mycobacteria (NTM) are considered a relevant cause of non-specific reactions to the most widely applied bovine tuberculosis (bTB) test, the intradermal tuberculin test. In order to establish which NTM species might act as a potential source of such diagnostic interference, a collection of 373 isolates obtained from skin test positive cows from 359 officially tuberculosis-free (OTF) herds, culled in the framework of the bTB eradication campaign in Spain, were identified at the species level through PCR and Sanger sequencing of the 16S rDNA, hsp65 and rpoB genes.. Of the 308 isolates for which a reliable identification was achieved, 32 different mycobacterial species were identified, with certain species being most represented: among M. avium complex members (n = 142, 46.1%), M. avium subsp. hominissuis (98; 69.0%) was the most abundant followed by M. avium subsp. avium (33, 23.2%), and M. intracellulare(7, 4.9%). Among non-MAC members (n = 166, 53.9%), M. nonchromogenicum (85; 27.6%) and M. bourgelatii (11; 5.6%) were the predominant species. In addition, mixed results were obtained in 53 isolates presenting up to 30 different genotypes, which could be indicative of new mycobacterial species. Our results represent a first step toward characterizing the diversity of NTM species that could interfere with official diagnostic tests for bTB eradication in Spain.
Differences in skin test reactions to official and defined antigens in guinea pigs exposed to non-tuberculous and tuberculous bacteria
(Scientific reports, 2023) Fernández Veiga, Leire; Fuertes, Miguel; Geijo, María V.; Pérez de Val, Bernat; Vidal, Enric; Michelet, Lorraine; Boschiroli, María Laura; Gómez Buendía, Alberto; Bezos Garrido, Javier; Jones, Gareth J.; Vordermeier, Martin; Juste, Ramón A.; Garrido, Joseba M.; Sevilla, Iker A.
The single and comparative intradermal tuberculin tests (SITT and CITT) are official in vivo tests for bovine tuberculosis (TB) diagnosis using bovine and avian purified protein derivatives (PPD-B and PPD-A). Infection with bacteria other than Mycobacterium tuberculosis complex (MTC) can result in nonspecific reactions to these tests. We evaluated the performance of the skin test with PPDs and new defined antigens in the guinea pig model. A standard dose (SD) of Rhodococcus equi, Nocardia sp., M. nonchromogenicum, M. monacense, M. intracellulare, M. avium subsp. paratuberculosis, M. avium subsp. avium, M. avium subsp. hominissuis, M. scrofulaceum, M. persicum, M. microti, M. caprae and M. bovis, and a higher dose (HD) of M. nonchromogenicum, M. monacense, M. intracellulare, M. avium subsp. paratuberculosis were tested using PPD-B, PPD-A, P22, ESAT-6-CFP-10-Rv3615c peptide cocktail long (PCL) and fusion protein (FP). The SD of R. equi, Nocardia sp., M. nonchromogenicum, M. monacense, M. intracellulare and M. avium subsp. paratuberculosis did not cause any reactions. The HD of M. nonchromogenicum, M. monacense, M. intracellulare, and M. avium subsp. paratuberculosis and the SD of M. avium subsp. hominissuis, M. scrofulaceum and M. persicum, caused nonspecific reactions (SIT). A CITT interpretation would have considered M. avium complex and M. scrofulaceum groups negative, but not all individuals from M. nonchromogenicum HD, M. monacense HD and M. persicum SD groups. Only animals exposed to M. bovis and M. caprae reacted to PCL and FP. These results support the advantage of complementing or replacing PPD-B to improve specificity without losing sensitivity.
Detection of caprine paratuberculosis (Johne's disease) in pre- and post-vaccinated herds: morphological diagnosis, lesion grading, and bacterial identification
(Frontiers in Veterinary Science, 2024) Stefanova, Elena Plamenova; Sierra, Eva; Fernández, Antonio; Quesada Canales, Óscar; Paz Sánchez, Yania; Colom Rivero, Ana; Espinosa de los Monteros, Antonio; Herráez, Pedro; Domínguez Rodríguez, Lucas José; Bezos Garrido, Javier; Pérez Sancho, Marta; Moreno, Inmaculada; Risalde, María A.; Andrada, Marisa
Samples from the mesenteric lymph nodes (MS LNs) and ileocecal valves (ICV) of 105 goats, comprising 61 non-vaccinated and 44 vaccinated against Mycobacterium avium subspecies paratuberculosis (MAP), were collected at slaughter from a farm with a confirmed history of paratuberculosis (PTB). These goats had subclinical infections. PTB-compatible lesions in the MS LNs, ICV lamina propria (LP), and Peyer's patches (PPs) were graded separately. Furthermore, the load of acid-fast bacilli was quantified using Ziehl-Neelsen staining (ZN), MAP antigens by immunohistochemistry (IHC), and MAP DNA by PCR targeting the IS900 sequence. Gross PTB-compatible lesions were found in 39% of the goats, with 31.72% vaccinated (V) and 68.29% non-vaccinated (nV). Histopathological lesions induced MAP were observed in 58% of the animals, with 36.07% vaccinated and 63.93% non-vaccinated. The inclusion of histopathology as a diagnostic tool led to a 28% increase in diagnosed cases in MS LNs and 86.05% in ICV. Grade IV granulomas with central mineralization and necrosis were the most common lesions in MS LNs. In the ICV, mild granulomatous enteritis with multifocal foci of epithelioid macrophages was predominant, occurring more frequently in the PPs than in the LP. Furthermore, statistical differences in the presence of histopathological lesions between vaccinated and non-vaccinated goats were noted in MS LNs, ICV LPs, and ICV PPs. Non-vaccinated animals showed higher positivity rates in ZN, IHC, and PCR tests, underscoring the benefits of anti-MAP vaccination in reducing PTB lesions and bacterial load in target organs. Our findings emphasize the necessity of integrating gross and histopathological assessments with various laboratory techniques for accurate morphological and etiological diagnosis of PTB in both vaccinated and non-vaccinated goats with subclinical disease. However, further studies are required to refine sampling protocols for subclinical PTB in goats to enhance the consistency of diagnostic tools.