Domínguez Bernal, Gustavo Ramón

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First Name
Gustavo Ramón
Last Name
Domínguez Bernal
Universidad Complutense de Madrid
Faculty / Institute
Sanidad Animal
Sanidad Animal
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Now showing 1 - 10 of 21
  • Publication
    Strength and medium-term impact of HisAK70 immunization in dogs: Vaccine safety and biomarkers of effectiveness for ex vivo Leishmania infantum infection
    (Elsevier, 2019-08) Fernández-Cotrina; Javier; Belinchón-Lorenzo, Silvia; Arias, Pablo; Martínez Rodrigo, Abel; Mas Zubiri, Alicia; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón
    HisAK70candidateshave successfullybeentested incutaneous (CL) andvisceral leishmaniosis (VL)mouse models.Here,weanalysedifferentbiomarkersindogtrialsafteraheterologousimmunizationstrategywitha HisAK70candidate(plasmidDNAplusadoptivetransferofperipheralblood-deriveddendriticcells(DCs)pulsed withthesamepathoantigenandCpGODNasanadjuvant)toexploretheantileishmanialactivityinanexvivo canineco-culturesysteminthepresenceofLeishmaniainfantumparasites.Inthecaninemodel,theheterologous HisAK70vaccinecoulddecreasetheinfectionindexintheDC-Tcellco-culturesystembyupto54%after30days andreachalmost67%after100dayspost-immunization,respectively,comparedtothoseobtainedinthecontrol groupofdogs.Theobservedsecurityandpotential tofight exvivoL. infantuminfectionhighlightaHisAK70 heterologousimmunizationstrategyasapromisingalternativetoevaluateitseffectivenessagainstcanineVL.
  • Publication
    Detection and antimicrobial resistance of Enterobacteriaceae other than Escherichia coli in raccoons from the Madrid region of Spain
    (SCIENDO, 2022-11-04) Orden Gutiérrez, José Antonio; Martínez Rodrigo, Abel; Vela Alonso, Ana Isabel; Fernández-Garayzábal Fernández, José Francisco; Hurtado Morillas, Clara; Mas Zubiri, Alicia; Domínguez Bernal, Gustavo Ramón
    Raccoons are an invasive alien species widely distributed in the Madrid region of Spain. These animals can carry a variety of enteric bacteria with associated antimicrobial resistance, which can infect humans and livestock. However, to our knowledge, the presence of non-E. coli Enterobacteriaceae in raccoons has not been previously studied. We conducted a study to examine the species distribution of Enterobacteriaceae isolates other than E. coli, as well as their antimicrobial resistance, in the faeces of 83 raccoons in the Madrid region. We detected 12 Enterobacteriaceae isolates other than E. coli belonging to seven different species: Citrobacter freundii (1 isolate), Citrobacter gillenii (3 isolates), Citrobacter murliniae (1 isolate), Citrobacter portucalensis (2 isolates), Enterobacter hormaechei subsp. hoffmannii (1 isolate), Hafnia paralvei (2 isolates) and Raoultella ornithinolytica (2 isolates). These isolates were found in 7 of the 83 (8.4%) animals studied. To our knowledge, this study is the first report of the presence of non-E. coli Enterobacteriaceae in raccoon faeces. All isolates but one were resistant to at least one of the 14 antimicrobials tested. Resistance to ampicillin (83.3%), amoxicillinclavulanic acid (50%) and cefoxitin (33.3%) was the most frequent. Our study indicates that raccoons are a potential source of infection with Enterobacteriaceae other than E. coli for humans and livestock in the Madrid region.
  • Publication
    A further investigation of the leishmaniosis outbreak in Madrid (Spain): low-infectivity phenotype of the Leishmania infantum BOS1FL1 isolate to establish infection in canine cells
    (Elsevier, 2020-12) Viñals, Luis Miguel; Mas Zubiri, Alicia; Martínez Rodrigo, Abel; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón; Carrión Herrero, Francisco Javier
    Human leishmaniosis caused by Leishmania infantum is a zoonotic disease, with dogs as the main reservoir in Mediterranean Basin countries. The largest European outbreak of human leishmaniosis declared in the southwestern Madrid region (Spain) is characterized by unusual epidemiological and clinical features, such as the emergence of new wild reservoirs (hares and rabbits), whereas the seroprevalence, infection, and severity of canine leishmaniosis have not substantially changed since the first studies conducted in Madrid before the outbreak. Previous studies reported that L. infantum isolates from the Madrid leishmaniosis focus displayed elevated virulence in in vivo models of infection and increased infectivity in murine target cells. With the aim of studying whether changes in the host-parasite interaction and virulence profile have developed, we first assessed the behaviour of one circulating isolate of the outbreak, IPER/ES/2012/BOS1FL1 (BOS1FL1), compared to that of a well-characterized strain from canine leishmaniosis, MCAN/ES/1996/BCN150 (BCN150), in terms of infection capacity (percentage of infected cells, representing infectivity, and number of amastigotes per infected cell, representing the intensity of infection) in canine monocytes and macrophages. BCN150 displayed significantly higher infectivity (76.82 ±4.40 vs 38.58 ±2.19; P <0.0001) and intensity of infection (3.64 ±0.13 vs 1.83 ±0.12; P <0.0001) than BOS1FL1 when interacting with canine cells. Our ROS induction results did not differ significantly between the two isolates or with the responses previously described for other L. infantum isolates. Paradoxically, increased resilience to hydrogen peroxide exposure was observed for BOS1FL1 (% viability 40.62 ±5.54 vs 26.37 ±2.93; P =0.039). Finally, we demonstrated that a decreased intracellular load of BOS1FL1 was associated with increased IFN-γ (261.21 ±26.29 vs 69.80 ±9.02; P =0.0151) and decreased IL- 10 production (165.06 ±23.87 vs 264.41 ±30.58; P =0.0002). In this study, we provide the first detailed insight into the differences between the isolate BOS1FL1 from the outbreak in Madrid and the well-characterized strain BCN150 MON-1 obtained from a dog in their response to interacting with canine cells. However, further studies are necessary to shed light on the immune mechanisms resulting in BOS1FL1 exhibiting less virulent behaviour in canine cells than in cells derived from other host species.
  • Publication
    Epitope Selection for Fighting Visceral Leishmaniosis: Not All Peptides Function the Same Way
    (MDPI, 2020-07-01) Álvarez-Campos, Daniel; Martínez Rodrigo, Abel; Mas Zubiri, Alicia; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón; Carrión Herrero, Francisco Javier
    Visceral leishmaniosis (VL) caused by Leishmania infantum is a disease with an increasing prevalence worldwide. Treatments are expensive, toxic, and ineffective. Therefore, vaccination seems to be a promising approach to control VL. Peptide-based vaccination is a useful method due to its stability, absence of local side effects, and ease of scaling up. In this context, bioinformatics seems to facilitate the use of peptides, as this analysis can predict high binding affinity epitopes to MHC class I and II molecules of different species. We have recently reported the use of HisAK70 DNA immunization in mice to induce a resistant phenotype against L. major, L. infantum, and L. amazonensis infections. In the present study, we used bioinformatics tools to select promising multiepitope peptides (HisDTC and AK) from the polyprotein encoded in the HisAK70 DNA to evaluate their immunogenicity in the murine model of VL by L. infantum. Our results revealed that both multiepitope peptides were able to induce the control of VL in mice. Furthermore, HisDTC was able to induce a better cell-mediated immune response in terms of reduced parasite burden, protective cytokine profile, leishmanicidal enzyme modulation, and specific IgG2a isotype production in immunized mice, before and after infectious challenge. Overall, this study indicates that the HisDTC chimera may be considered a satisfactory tool to control VL because it is able to activate a potent CD4+ and CD8+ T-cell protective immune responses.
  • Publication
    Mitigating an undesirable immune response of inherent susceptibility to cutaneous leishmaniosis in a mouse model: the role of the pathoantigenic HISA70 DNA vaccine
    (2012-08-09) Herrero-Gil, Aldara; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón; Horcajo Iglesias, María Del Pilar; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Ordóñez Gutiérrez, Lara
    Leishmania major is the major cause of cutaneous leishmaniosis (CL) outside of the Americas. In the present study we have cloned six Leishmania genes (H2A, H2B, H3, H4, A2 and HSP70) into the eukaryotic expression vector pCMVβ-m2a, resulting in pCMV-HISA70m2A, which encodes all six pathoantigenic proteins as a single polyprotein. This expression plasmid has been evaluated as a novel vaccine candidate in the BALB/c mouse model of CL. The DNA vaccine shifted the immune response normally induced by L. major infection away from a Th2-specific pathway to one of basal susceptibility. Immunization with pCMV-HISA70m2A dramatically reduced footpad lesions and lymph node parasite burdens relative to infected control mice. Complete absence of visceral parasite burden was observed in all 12 immunized animals but not in any of the 24 control mice. Moreover, vaccinated mice produced large amounts of IFN-γ, IL-17 and NO at 7 weeks post-infection (pi), and they showed lower arginase activity at the site of infection, lower IL-4 production and a weaker humoral immune response than infected control mice. Taken together, these results demonstrate the ability of the HISA70 vaccine to shift the murine immune response to L. major infection away from an undesirable, Th2-specific pathway to a less susceptible-like pathway involving Th1 and Th17 cytokine profiles.
  • Publication
    Mechanisms of resistance and susceptibility to experimental visceral leishmaniosis: BALB/c mouse versus syrian hamster model
    (2011-02-23) Nieto, Ana; Madrid-Elena, Nadia; Carrión Herrero, Francisco Javier; Domínguez Bernal, Gustavo Ramón; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La
    Several animal models have been established to study visceral leishmaniosis (VL), a worldwide vector-borne disease affecting humans and domestic animals that constitutes a serious public health problem. BALB/c mice and Syrian hamsters are the most widely used experimental models. In this paper, we summarize the advantages and disadvantages of these two experimental models and discuss the results obtained using these models in different studies of VL. Studies using the BALB/c mouse model have underscored differences between the liver and spleen in the course of VL, indicating that pathological evaluation of the visceral organs is essential for understanding the immune mechanisms induced by Leishmania infantum infection. The main goal of this review is to collate the relevant literature on Leishmania pathogenesis into a sequence of events, providing a schematic view of the main components of adaptive and innate immunity in the liver and spleen after experimental infection with L. infantum or L. donovani. This review also presents several viewpoints and reflections about some controversial aspects of Leishmania research, including the choice of experimental model, route of administration, inoculum size and the relevance of pathology (intimately linked to parasite persistence): a thorough understanding of which is essential for future VL research and the successful development of efficient control strategies for Leishmania spp.
  • Publication
    Engineering of a live Salmonella enterica serovar Choleraesuis negative-marker strain that allows serological differentiation between immunised and infected animals
    (Elsevier, 2016-07-01) Herrero-Gil, Aldara; Carrión Herrero, Francisco Javier; Orden Gutiérrez, José Antonio; Fuente López, Ricardo De La; Domínguez Bernal, Gustavo Ramón
    The usefulness of Salmonella vaccine vehicles is limited by the fact that control programmes relying on Salmonella bacteriology and serology cannot differentiate infected animals from vaccinated ones, an ability referred to as DIVA (differentiating infected from vaccinated animals). As a first step towards Salmonella-based DIVA vaccines, the ompA gene was deleted in live attenuated ΔphoP and ΔrpoS vaccine strains. The ompA gene is present in all Salmonella enterica serovars and it encodes an abundant, highly immunogenic outer membrane protein. The double mutant ΔphoP ΔompA and ΔrpoS ΔompA strains showed similar virulence attenuation, safety and immunogenicity in a mouse model of infection as the parental ΔphoP and ΔrpoS strains. Sera from mice inoculated with the double mutant strains failed to recognise OmpA in Western blots of outer membrane extracts, whereas the protein was recognised by sera from mice inoculated with wild-type Salmonella or a mixture of double mutant and parental strains. These data suggest that OmpA can be a suitable negative marker for DIVA vaccines.
  • Publication
    Short communication: Isolation frequency of bacteria causing lymphadenitis and abscesses in small ruminants in central Spain
    (Elsevier, 2017-07-10) Heras, Moisés de las; Torrijos, Carlos; Carrión Herrero, Francisco Javier; Fuente López, Ricardo De La; Díez De Tejada Martín, María Paloma; Pérez Sancho, Marta; Orden Gutiérrez, José Antonio; Domínguez Bernal, Gustavo Ramón
    Infectious lymphadenitis in small ruminants caused by Corynebacterium pseudotuberculosis and Staphylococcus aureus subsp. anaerobius are widely distributed throughout the world, and result in significant economic losses. Trueperella pyogenes has also been associated with lymphadenitis in sheep and goats. In order to determinate the isolation frequency of the different agents associated with lymphadenitis and abscesses, 171 pus samples (135 from sheep and 36 from goats) from 46 flocks were investigated. Isolated bacteria were identified by MALDI-TOF method. S. aureus subsp. anaerobius was the most frequently detected agent. It was identified in 76 animals (59 sheep and 17 goats) from 24 of the surveyed flocks. Of these infected animal, 25 (32,9%) were over one year old, confirming that abscess disease may occur in a significant percentage of adult animals. C. pseudotuberculosis was identified in 45 of the sampled animals (36 sheep and 9 goats) from 24 flocks. Only 5 of animals suffering caseous lymphadenitis were under one year old. T. pyogenes was isolated from 17 animals (13 sheep and 4 goats) in 11 flocks. Seven of these samples were taken from subcutaneous abscesses located in not lymph nodes regions. A notable finding of this work is the isolation of Actinomyces hyovaginalis from 5 of the samples analyzed all of them taken from subcutaneous abscesses located in superficial lymph nodes regions. Thus, T. pyogenes and A. hyovaginalis should be included in the differential diagnosis of lymphadenitis in small ruminants. In 19 of the 46 surveyed flocks at least two of the four agents were detected, which underlines the need to analyze samples from several animals to reach an accurate diagnosis in flocks affected by lymphadenitis.
  • Publication
    Subtilase Cytotoxin-Coding Genes in Verotoxin-Producing Escherichia coli Strains from Sheep and Goats Differ from Those from Cattle
    (2011-09-24) Carrión Herrero, Francisco Javier; Orden Gutiérrez, José Antonio; Horcajo Iglesias, María Del Pilar; Fuente López, Ricardo De La; Ruiz Santa Quiteria Serrano De La Cruz, José Antonio; Domínguez Bernal, Gustavo Ramón
    Subtilase cytotoxin (SubAB) from verotoxin (VT)-producing Escherichia coli (VTEC) strains was first described in the 98NK2 strain and has been associated with human disease. However, SubAB has recently been found in two VT-negative E. coli strains (ED 591 and ED 32). SubAB is encoded by two closely linked, cotranscribed genes (subA and subB). In this study, we investigated the presence of subAB genes in 52 VTEC strains isolated from cattle and 209 strains from small ruminants, using PCR. Most (91.9%) VTEC strains from sheep and goats and 25% of the strains from healthy cattle possessed subAB genes. The presence of subAB in a high percentage of the VTEC strains from small ruminants might increase the pathogenicity of these strains for human beings. Some differences in the results of PCRs and in the association with some virulence genes suggested the existence of different variants of subAB. We therefore sequenced the subA gene in 12 strains and showed that the subA gene in most of the subAB-positive VTEC strains from cattle was almost identical (about 99%) to that in the 98NK2 strain, while the subA gene in most of the subAB-positive VTEC strains from small ruminants was almost identical to that in the ED 591 strain. We propose the terms subAB1 to describe the SubAB-coding genes resembling that in the 98NK2 strain and subAB2 to describe those resembling that in the ED 591 strain.
  • Publication
    Differences in virulence gene expression between atypical enteropathogenic Escherichia coli strains isolated from diarrheic and healthy ruminants
    (2012-06-07) Carrión Herrero, Francisco Javier; Horcajo Iglesias, María Del Pilar; Domínguez Bernal, Gustavo Ramón; Fuente López, Ricardo De La; Ruiz Santa Quiteria Serrano De La Cruz, José Antonio; Orden Gutiérrez, José Antonio
    Differences in the pathogenicity of atypical enteropathogenic Escherichia coli (EPEC) strains may be due, at least partially, to different expression patterns of some virulence genes. To investigate this hypothesis, the virulence gene expression patterns of 6 atypical EPEC strains isolated from healthy and diarrheic ruminants were compared using quantitative real-time reverse transcription polymerase chain reaction after growing the bacteria in culture medium alone or after binding it to HeLa epithelial cells. Some virulence genes in strains from diarrheic animals were upregulated relative to their expression in strains from healthy animals. When bacteria were cultured in the presence of HeLa cells, the ehxA and efa1/lifA genes, previously associated with the production of diarrhea, were expressed at higher levels in strains from diarrheic animals than in strains from healthy animals. Thus, the expression levels of some virulence genes may help determine which atypical EPEC strains cause diarrhea in ruminants.