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      Immunogenicity of an Electron Beam Inactivated Rhodococcus equi Vaccine in Neonatal Foals

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          Abstract

          Rhodococcus equi is an important pathogen of foals that causes severe pneumonia. To date, there is no licensed vaccine effective against R. equi pneumonia of foals. The objectives of our study were to develop an electron beam (eBeam) inactivated vaccine against R. equi and evaluate its immunogenicity. A dose of eBeam irradiation that inactivated replication of R. equi while maintaining outer cell wall integrity was identified. Enteral administration of eBeam inactivated R. equi increased interferon-γ production by peripheral blood mononuclear cells in response to stimulation with virulent R. equi and generated naso-pharyngeal R. equi-specific IgA in newborn foals. Our results indicate that eBeam irradiated R. equi administered enterally produce cell-mediated and upper respiratory mucosal immune responses, in the face of passively transferred maternal antibodies, similar to those produced in response to enteral administration of live organisms (a strategy which previously has been documented to protect foals against intrabronchial infection with virulent R. equi). No evidence of adverse effects was noted among vaccinated foals.

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          Most cited references56

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          Effect of formalin tissue fixation and processing on immunohistochemistry.

          Although immunohistochemistry is routinely performed by many pathology laboratories, its standardization still lags behind. A major cause of variation in the reproducibility of immunohistochemical staining is induced by tissue fixation and, to a lesser degree, tissue processing. This report, stemming from the first meeting of the International Consensus Group on Standardization and Quality Control (ICGSQC) in Nice, France, summarizes the problem and suggests solutions to begin to achieve standardization of fixation and processing. Most laboratories use neutral-buffered formalin (10%) for tissue fixation which introduces cross-links, whereas coagulative fixatives are less popular. Problems with formalin fixation comprise delay of fixation and variations in the duration of the fixation mainly. Solutions to these problems could be to start fixation soon ( 24-48 hrs). For tissue processing, the most important problem is inadequate tissue dehydration prior to paraffin embedding. This can be prevented by preparing all solutions freshly every week, depending on the volume of tissue processed. If consistently applied, these procedures could eliminate some of the sources of variation in immunohistochemical stains.
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            Rhodococcus equi: an animal and human pathogen.

            Recent isolations of Rhodococcus equi from cavitatory pulmonary disease in patients with AIDS have aroused interest among medical microbiologists in this unusual organism. Earlier isolations from humans had also been in immunosuppressed patients following hemolymphatic tumors or renal transplantation. This organism has been recognized for many years as a cause of a serious pyogranulomatous pneumonia of young foals and is occasionally isolated from granulomatous lesions in several other species, in some cases following immunosuppression. The last decade has seen many advances in understanding of the epidemiology, pathogenesis, diagnosis, treatment, and immunity to infection in foals. The particular susceptibility of the foal is not understood but can be explained in part by a combination of heavy challenge through the respiratory route coinciding with declining maternally derived antibody in the absence of fully competent foal cellular immune mechanisms. R. equi is largely a soil organism but is widespread in the feces of herbivores. Its growth in soil is considerably improved by simple nutrients it obtains from herbivore manure. About one-third of human patients who have developed R. equi infections had contact in some way with herbivores or their manure. Others may have acquired infection from contact with soil or wild bird manure. R. equi is an intracellular parasite, which explains the typical pyogranulomatous nature of R. equi infections, the predisposition to infection in human patients with defective cell-mediated immune mechanisms, and the efficacy of antimicrobial drugs that penetrate phagocytic cells.
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              Protection against Tuberculosis in Eurasian Wild Boar Vaccinated with Heat-Inactivated Mycobacterium bovis

              Tuberculosis (TB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex continues to affect humans and animals worldwide and its control requires vaccination of wildlife reservoir species such as Eurasian wild boar (Sus scrofa). Vaccination efforts for TB control in wildlife have been based primarily on oral live BCG formulations. However, this is the first report of the use of oral inactivated vaccines for controlling TB in wildlife. In this study, four groups of 5 wild boar each were vaccinated with inactivated M. bovis by the oral and intramuscular routes, vaccinated with oral BCG or left unvaccinated as controls. All groups were later challenged with a field strain of M. bovis. The results of the IFN-gamma response, serum antibody levels, M. bovis culture, TB lesion scores, and the expression of C3 and MUT genes were compared between these four groups. The results suggested that vaccination with heat-inactivated M. bovis or BCG protect wild boar from TB. These results also encouraged testing combinations of BCG and inactivated M. bovis to vaccinate wild boar against TB. Vaccine formulations using heat-inactivated M. bovis for TB control in wildlife would have the advantage of being environmentally safe and more stable under field conditions when compared to live BCG vaccines. The antibody response and MUT expression levels can help differentiating between vaccinated and infected wild boar and as correlates of protective response in vaccinated animals. These results suggest that vaccine studies in free-living wild boar are now possible to reveal the full potential of protecting against TB using oral M. bovis inactivated and BCG vaccines.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                25 August 2014
                : 9
                : 8
                : e105367
                Affiliations
                [1 ]Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, United States of America
                [2 ]National Center for Electron Beam Research and Departments of Poultry Science and Nutrition and Food Science, Texas A&M University, College Station, Texas, United States of America
                [3 ]Antech Diagnostics, College Station, Texas, United States of America
                [4 ]Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, United States of America
                [5 ]Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, Texas A&M University, College Station, Texas, United States of America
                [6 ]Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, United States of America
                [7 ]Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America
                East Carolina University School of Medicine, United States of America
                Author notes

                Competing Interests: AIB's salary was supported by a fellowship from Fort Dodge Animal Health/Pfizer Animal Health (now Zoetis). The Morris Animal Foundation and Link Equine Research Endowment sponsored this study, and additional support provided by an unrestricted gift from Boerhinger-Ingelheim. None of the agencies or companies listed above had any role or influence in the data collection, data analysis, interpretation of results, or conclusions reported in this manuscript; none of these companies have reviewed the manuscript. Moreover, none of the authors were employed in any way (including consultancy) by any of the companies listed above. The authors declare no affiliation to products, patents, etc. related to this work with any company, including those listed above. This support does not alter our adherence to PLOS ONE policies on sharing data and materials. FNO is employed by Antech Diagnostics; however, FNO was not paid for his services and provided assistance outside of his work at Antech Diagnostics. Relevant to this work, FNO has no competing interests relating to employment, consultancy, patents, products in development, etc. The authors declare no other competing financial interests.

                Conceived and designed the experiments: AIB WM DNM CCL SDP MJBF NDC. Performed the experiments: AIB CB KBB JRB MC NDC. Analyzed the data: AIB FNO MJBF NDC. Contributed reagents/materials/analysis tools: AIB CCL MJBF NDC. Contributed to the writing of the manuscript: AIB SDP CB MC FNO WM DNM CCL MJBF NDC.

                Article
                PONE-D-14-24064
                10.1371/journal.pone.0105367
                4143214
                25153708
                db54e57a-776b-4a83-bd68-d5c98cdffa96
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 29 May 2014
                : 18 July 2014
                Page count
                Pages: 11
                Funding
                This work was supported by the Morris Animal Foundation (D13EQ-0001). Additional funding was provided by the Link Equine Research Endowment, Texas A&M University. AIB was supported by a fellowship from Fort Dodge Animal Health/Pfizer Animal Health (now Zoetis). An unrestricted gift from Boehringer-Ingelheim to the Equine Infectious Disease Laboratory at Texas A&M University provided partial support to this project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Biophysics
                Radiation Biophysics
                Biotechnology
                Applied Microbiology
                Immunology
                Clinical Immunology
                Infectious Disease Immunology
                Pulmonary Immunology
                Vaccination and Immunization
                Vaccine Development
                Vaccines
                Microbiology
                Veterinary Science
                Veterinary Diseases
                Veterinary Bacteriology
                Veterinary Medicine
                Veterinary Immunology
                Veterinary Microbiology
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper and its Supporting Information files.

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