Vaccination with BM86, subolesin and akirin protective antigens for the control of tick infestations in white tailed deer and red deer

Red deer (Cervus elaphus) and white-tailed deer (Odocoileus virginianus) are hosts for different tick species and tick-borne pathogens and play a role in tick dispersal and maintenance in some regions. These factors stress the importance of controlling tick infestations in deer and several methods such as culling and acaricide treatment have been used. Tick vaccines are a cost-effective alternative for tick control that reduced cattle tick infestations and tick-borne pathogens prevalence while reducing the use of acaricides. Our hypothesis is that vaccination with vector protective antigens can be used for the control of tick infestations in deer. Herein, three experiments were conducted to characterize (1) the antibody response in red deer immunized with recombinant BM86, the antigen included in commercial tick vaccines, (2) the antibody response and control of cattle tick infestations in white-tailed deer immunized with recombinant BM86 or tick subolesin (SUB) and experimentally infested with Rhipicephalus (Boophilus) microplus, and (3) the antibody response and control of Hyalomma spp. and Rhipicephalus spp. field tick infestations in red deer immunized with mosquito akirin (AKR), the SUB ortholog and candidate protective antigen against different tick species and other ectoparasites. The results showed that deer produced an antibody response that correlated with the reduction in tick infestations and was similar to other hosts vaccinated previously with these antigens. The overall vaccine efficacy was similar between BM86 (E=76%) and SUB (E=83%) for the control of R. microplus infestations in white-tailed deer. The field trial in red deer showed a 25-33% (18-40% when only infested deer were considered) reduction in tick infestations, 14-20 weeks after the first immunization. These results demonstrated that vaccination with vector protective antigens could be used as an alternative method for the control of tick infestations in deer to reduce tick populations and dispersal in regions where deer are relevant hosts for these ectoparasites.This work was supported by the Ministerio de Ciencia e Innovación, Spain (project BFU2008-01244/BMC) and the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Spain (project FAU2008-00014-00-00). D. Carreón was partially supported by a grant-in-aid from Conacyt, Mexico and the EU grant TB-STEP 212414. M. Villar was funded by the JAE-DOC program (CSIC-FSE), Spain. J.A. Moreno-Cid is a recipient of a JCCM fellowship, Spain.Peer Reviewe

Messaging performance of FIPA interaction protocols in networked embedded controllers

Agent-based technologies in production control systems could facilitate seamless reconfiguration and integration of mechatronic devices/modules into systems. Advances in embedded controllers which are continuously improving computational capabilities allow for software modularization and distribution of decisions. Agent platforms running on embedded controllers could hide the complexity of bootstrap and communication. Therefore, it is important to investigate the messaging performance of the agents whosemainmotivation is the resource allocation in manufacturing systems (i.e., conveyor system). The tests were implemented using the FIPA-compliant JADE-LEAP agent platform. Agent containers were distributed through networked embedded controllers, and agents were communicating using request and contract-net FIPA interaction protocols. The test scenarios are organized in intercontainer and intracontainer communications. The work shows the messaging performance for the different test scenarios using both interaction protocols

Embedded XML DOM parser: an approach for XML data processing on networked embedded systems with real-time requirements

Trends in control and automation show an increase in data processing and communication in embedded automation controllers. The eXtensible Markup Language (XML) is emerging as a dominant data syntax, fostering interoperability, yet little is still known about how to provide predictable real-time performance in XML processing, as required in the domain of industrial automation. This paper presents an XML processor that is designed with such real-time performance in mind. The publication attempts to disclose insight gained in applying techniques such as object pooling and reuse, and other methods targeted at avoiding dynamic memory allocation and its consequent memory fragmentation. Benchmarking tests are reported in order to illustrate the benefits of the approach

New phylogenetic insights into Saprolegniales (Oomycota, Straminipila) based upon studies of specimens isolated from Brazil and Argentina

Saprolegniales is a complex and monophyletic order of oomycetes. Their members inhabit terrestrial, freshwater, and marine ecosystems and have a worldwide distribution. In these ecosystems, they are found as saprobes, parasites, or even pathogens of animals and plants of economic importance. In this study, a concatenate phylogeny of the partial LSU and complete ITS rDNA regions is presented, including isolates from Brazil and Argentina, which were sequenced after a detailed morphological analysis. Among the sequenced species, Achlya orion, Leptolegnia eccentrica, Phragmosporangium uniseriatum, and Pythiopsis irregularis are included for the first time in a phylogeny. Our results are in agreement with the recent informal proposals outlined in taxonomic overviews of the Oomycota of G.W. Beakes and collaborators, who placed the family Verrucalvaceae into the Saprolegniales and introduced the family Achlyaceae to group Achlya s.s., Brevilegnia, Dictyuchus, and Thraustotheca. These results also support the transference of Achlya androgyna to Newbya. Leptolegnia appears as paraphyletic, with the separation of L. eccentrica from the other species of this genus. In addition, Phragmosporangium, which is herein sequenced for the first time, clustered as sister to some species of Aphanomyces, including the type species, A. stellatus.Fil: Rocha, Sarah C. O.. Instituto de Botânica. Núcleo de Pesquisa em Micologia; BrasilFil: Lopez Lastra, Claudia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Marano, Agostina Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Instituto de Botânica. Núcleo de Pesquisa em Micologia; BrasilFil: de Souza, José Ivanildo. Instituto de Botânica. Núcleo de Pesquisa em Micologia; BrasilFil: Rueda Páramo, Manuel Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Pires Zottarelli, Carmen L. A.. Instituto de Botânica. Núcleo de Pesquisa em Micologia; Brasi

Bioinformatics Discovery of Vertebrate Cathelicidins from the Mining of Available Genomes

Due to the worrying increase in antimicrobial resistance to conventional antibiotics, the search for alternatives is becoming increasingly important. Antimicrobial peptides (AMPs), originating from natural resources, have been recognised as a novel class of antibiotics. An advantage of peptides over antibiotics is that the resistance is more difficult to attain than for conventional antibiotics. With the increasing number of genomes sequenced and available in the public domain, one alternative methodology to obtain novel AMPs is to analyse genes and proteins from genomic databases to predict and identify amino acid sequences that share similarities and molecular features with natural bioactive antimicrobial peptides. In this chapter, we summarise some of our recent results on the production of antimicrobial peptides, particularly, how we managed to identify a family of antimicrobial peptides: cathelicidins, through bioinformatics tools, from the genomes of two lower vertebrates (a reptile and a bird) available in public databases. We hope that our preliminary investigation with these novel peptides could be useful for the design of future strategies that pursue the production of antimicrobial peptides through biotechnology

Tonsils of the soft palate do not mediate the response of pigs to oral vaccination with heat-inactivated Mycobacterium bovis

Mycobacterium bovis causes animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivated M. bovis and challenge with a virulent M. bovis field strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests for in vivo TB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivated M. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. Massive M. bovis growth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased after M. bovis challenge. This information is relevant for pig production in regions that are endemic for M. bovis and for TB vaccine research.This study is a contribution to EU FP7 grant 613779 WildTBVac and to MINECO Plan Nacional grant AGL2011-30041 and FEDER.Peer Reviewe

Oral vaccination with heat inactivated Mycobacterium bovis activates the complement system to protect against tuberculosis

Tuberculosis (TB) remains a pandemic affecting billions of people worldwide, thus stressing the need for new vaccines. Defining the correlates of vaccine protection is essential to achieve this goal. In this study, we used the wild boar model for mycobacterial infection and TB to characterize the protective mechanisms elicited by a new heat inactivated Mycobacterium bovis vaccine (IV). Oral vaccination with the IV resulted in significantly lower culture and lesion scores, particularly in the thorax, suggesting that the IV might provide a novel vaccine for TB control with special impact on the prevention of pulmonary disease, which is one of the limitations of current vaccines. Oral vaccination with the IV induced an adaptive antibody response and activation of the innate immune response including the complement component C3 and inflammasome. Mycobacterial DNA/RNA was not involved in inflammasome activation but increased C3 production by a still unknown mechanism. The results also suggested a protective mechanism mediated by the activation of IFN-γ producing CD8+ T cells by MHC I antigen presenting dendritic cells (DCs) in response to vaccination with the IV, without a clear role for Th1 CD4+ T cells. These results support a role for DCs in triggering the immune response to the IV through a mechanism similar to the phagocyte response to PAMPs with a central role for C3 in protection against mycobacterial infection. Higher C3 levels may allow increased opsonophagocytosis and effective bacterial clearance, while interfering with CR3-mediated opsonic and nonopsonic phagocytosis of mycobacteria, a process that could be enhanced by specific antibodies against mycobacterial proteins induced by vaccination with the IV. These results suggest that the IV acts through novel mechanisms to protect against TB in wild boar.This research was supported by Plan Nacional I+D+I AGL2011-30041 from Ministerio de Economía y Competitividad (MINECO), Spain and FEDER. This is also a contribution to EU FP7 grant WildTBvac and the EU FP7 ANTIGONE project number 278976. R.C. Galindo was funded by MEC, Spain. B. Beltrán-Beck was supported by MINECO grant BES-2009-017401.Peer Reviewe

IDENTIFICACIÓN DE HELICOBACTER PYLORI A PARTIR DE LESIÓN ATEROSCLERÓTICA CORONARIA HUMANA

Introduction: The main pathogenic mechanism underlying the genesis and development of diseases of the circulatory system is atherosclerosis. It is a complex and progressive pathological process of the arterial wall that affects especially the coronary, cerebral and peripheral arteries. Currently, there is talk about "The infectious hypothesis of atherosclerosis". Helicobacter pylori infection has been one of the most researched worldwide. Objective: To identify H. pylori from atherosclerotic lesions of patients who attend the cardiovascular and angiology services of the city of Barranquilla. Methods: A descriptive cross-sectional study was carried out. In a sample of 102 participants. The atheromas were taken by expert personnel in endarterectomy and dissection of blood vessels. The specimens were embedded in 4% formaldehyde solution. DNA extraction was performed from atheromas. The molecular diagnosis of H. pylori was performed by ANIDADA PCR, evidencing a 120 bp fragment after electrophoresis in a 3.5% agarose gel in 0.5X TBE. Results: 102 samples of atheromas were obtained. One sample was positive for H. pylori (Sample # 14, 1/102). The atherosclerotic tissue was obtained from the right coronary artery. Conclusion: The infectious hypothesis of atherosclerosis has been one of the main research topics worldwide in recent decades. H. pylori infection is a risk factor, however, several studies are necessary to be able to conclude more accurately.Introducción: El principal mecanismo patogénico que subyace la génesis y desarrollo de enfermedades del sistema circulatorio es la aterosclerosis. Es un proceso patológico complejo y progresivo de la pared arterial que afecta especialmente a las arterias coronarias, cerebrales y periféricas. Actualmente se habla sobre “La hipótesis infecciosa de la aterosclerosis”. La infección por Helicobacter pylori ha sido una de las más investigadas a nivel global. Objetivo: Identificar H. pylori a partir de lesiones ateroscleróticas de pacientes que acuden a los servicios de cirugía cardiovascular y angiología de la ciudad de Barranquilla. Métodos: Se realizó un estudio descriptivo de corte transversal. En una muestra de 102 participantes. Los ateromas fueron tomados por personal experto en endarterectomía y disección de vasos sanguíneos. Los especímenes fueron embebidos en solución formaldehído al 4%. Se realizó extracción de ADN a partir de ateromas. El diagnóstico molecular de H. Pylori se realizó por PCR ANIDADA, evidenciando un fragmento de 120 pb posterior a la electroforesis en gel de agarosa al 3,5% en TBE 0,5X. Resultados: Se obtuvieron 102 muestras de ateromas. Una muestra resultó positiva para H. pylori (Muestra #14; 1/102). El tejido aterosclerótico fue obtenido a partir de arteria coronaria derecha. Conclusión: La hipótesis infecciosa de la aterosclerosis ha sido uno de los principales temas de investigación a nivel mundial en las últimas décadas. La infección por H. pylori es un factor de riesgo, sin embargo, varios estudios son necesarios para poder concluir de manera más precisa

No evidence that wild red deer (Cervus elaphus) on the Iberian Peninsula are a reservoir of Mycobacterium avium subspecies paratuberculosis infection

The potential role of red deer (Cervus elaphus) as a reservoir of Mycobacterium avium subspecies paratuberculosis (MAP) infection is largely unknown. A total of 332 wild red deer were investigated using post-mortem examination, bacteriology and serology. Only three animals (1.12%) were found to have lesions on histopathological examination and no MAP bacteria were recovered on culture. The results suggest it is unlikely that wild red deer make a significant contribution to the maintenance of MAP infection in the region. The cross-reactivity of the ELISAs used indicates this diagnostic modality is ineffective in the detection of MAP infection in this species. The implications of these results for the control of this important pathogen in both livestock and wildlife are discussed.This research was supported by MCINN Grants AGL2008-03875/FEDER and FP7EU/TB-STEP-212414.Peer Reviewe

Tick capillary feeding for the study of proteins involved in tick-pathogen interactions as potential antigens for the control of tick infestation and pathogen infection

BACKGROUND: Ticks represent a significant health risk to animals and humans due to the variety of pathogens they can transmit during feeding. The traditional use of chemicals to control ticks has serious drawbacks, including the selection of acaricide-resistant ticks and environmental contamination with chemical residues. Vaccination with the tick midgut antigen BM86 was shown to be a good alternative for cattle tick control. However, results vary considerably between tick species and geographic location. Therefore, new antigens are required for the development of vaccines controlling both tick infestations and pathogen infection/transmission. Tick proteins involved in tick-pathogen interactions may provide good candidate protective antigens for these vaccines, but appropriate screening procedures are needed to select the best candidates. METHODS: In this study, we selected proteins involved in tick-Anaplasma (Subolesin and SILK) and tick-Babesia (TROSPA) interactions and used in vitro capillary feeding to characterize their potential as antigens for the control of cattle tick infestations and infection with Anaplasma marginale and Babesia bigemina. Purified rabbit polyclonal antibodies were generated against recombinant SUB, SILK and TROSPA and added to uninfected or infected bovine blood to capillary-feed female Rhipicephalus (Boophilus) microplus ticks. Tick weight, oviposition and pathogen DNA levels were determined in treated and control ticks. RESULTS: The specificity of purified rabbit polyclonal antibodies against tick recombinant proteins was confirmed by Western blot and against native proteins in tick cell lines and tick tissues using immunofluorescence. Capillary-fed ticks ingested antibodies added to the blood meal and the effect of these antibodies on tick weight and oviposition was shown. However, no effect was observed on pathogen DNA levels. CONCLUSIONS: These results highlighted the advantages and some of the disadvantages of in vitro tick capillary feeding for the characterization of candidate tick protective antigens. While an effect on tick weight and oviposition was observed, the effect on pathogen levels was not evident probably due to high tick-to-tick variations among other factors. Nevertheless, these results together with previous results of RNA interference functional studies suggest that these proteins are good candidate vaccine antigens for the control of R. microplus infestations and infection with A. marginale and B. bigemina