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      Microbiota composition and inflammatory immune responses upon peroral application of the commercial competitive exclusion product Aviguard ® to microbiota-depleted wildtype mice

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          Abstract

          Non-antibiotic feed additives including competitive exclusion products have been shown effective in reducing pathogen loads including multi-drug resistant strains from the vertebrate gut. In the present study we surveyed the intestinal bacterial colonization properties, potential macroscopic and microscopic inflammatory sequelae and immune responses upon peroral application of the commercial competitive exclusion product Aviguard ® to wildtype mice in which the gut microbiota had been depleted by antibiotic pre-treatment. Until four weeks following Aviguard® challenge, bacterial strains abundant in the probiotic suspension stably established within the murine intestines. Aviguard® application did neither induce any clinical signs nor gross macroscopic intestinal inflammatory sequelae, which also held true when assessing apoptotic and proliferative cell responses in colonic epithelia until day 28 post-challenge. Whereas numbers of colonic innate immune cell subsets such as macrophages and monocytes remained unaffected, peroral Aviguard ® application to microbiota depleted mice was accompanied by decreases in colonic mucosal counts of adaptive immune cells such as T and B lymphocytes. In conclusion, peroral Aviguard® application results i.) in effective intestinal colonization within microbiota depleted mice, ii.) neither in macroscopic nor in microscopic inflammatory sequelae and iii.) in lower colonic mucosal T and B cell responses.

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          Most cited references22

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          World Health Organization Global Estimates and Regional Comparisons of the Burden of Foodborne Disease in 2010

          Illness and death from diseases caused by contaminated food are a constant threat to public health and a significant impediment to socio-economic development worldwide. To measure the global and regional burden of foodborne disease (FBD), the World Health Organization (WHO) established the Foodborne Disease Burden Epidemiology Reference Group (FERG), which here reports their first estimates of the incidence, mortality, and disease burden due to 31 foodborne hazards. We find that the global burden of FBD is comparable to those of the major infectious diseases, HIV/AIDS, malaria and tuberculosis. The most frequent causes of foodborne illness were diarrheal disease agents, particularly norovirus and Campylobacter spp. Diarrheal disease agents, especially non-typhoidal Salmonella enterica, were also responsible for the majority of deaths due to FBD. Other major causes of FBD deaths were Salmonella Typhi, Taenia solium and hepatitis A virus. The global burden of FBD caused by the 31 hazards in 2010 was 33 million Disability Adjusted Life Years (DALYs); children under five years old bore 40% of this burden. The 14 subregions, defined on the basis of child and adult mortality, had considerably different burdens of FBD, with the greatest falling on the subregions in Africa, followed by the subregions in South-East Asia and the Eastern Mediterranean D subregion. Some hazards, such as non-typhoidal S. enterica, were important causes of FBD in all regions of the world, whereas others, such as certain parasitic helminths, were highly localised. Thus, the burden of FBD is borne particularly by children under five years old–although they represent only 9% of the global population–and people living in low-income regions of the world. These estimates are conservative, i.e., underestimates rather than overestimates; further studies are needed to address the data gaps and limitations of the study. Nevertheless, all stakeholders can contribute to improvements in food safety throughout the food chain by incorporating these estimates into policy development at national and international levels.
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            The European Union One Health 2018 Zoonoses Report

            (2019)
            Abstract This report of the European Food Safety Authority and the European Centre for Disease Prevention and Control presents the results of zoonoses monitoring activities carried out in 2018 in 36 European countries (28 Member States (MS) and 8 non‐MS). The first and second most commonly reported zoonoses in humans were campylobacteriosis and salmonellosis, respectively. The European Union (EU) trend for confirmed human cases of these two diseases was stable during 2014–2018. The proportion of human salmonellosis cases due to Salmonella Enteritidis was at the same level in 2018 as in 2017. Of the 27 reporting MS, 16 met all Salmonella reduction targets for poultry, whereas 11 MS failed meeting at least one. The EU flock prevalence of target Salmonella serovars in breeding hens, laying hens, broilers and fattening turkeys decreased during recent years but stalled in breeding turkeys. Salmonella results from Competent Authorities for pig carcasses and for poultry tested through National Control Programmes were more frequently positive compared with food business operators. Shiga toxin‐producing Escherichia coli (STEC) infections in humans were the third most commonly reported zoonosis in the EU and increased from 2014 to 2018. Yersiniosis was the fourth most frequently reported zoonosis in humans in 2018 with a stable trend in 2014–2018. The number of reported confirmed listeriosis cases further increased in 2018, despite Listeria rarely exceeding the EU food safety limit tested in ready‐to‐eat food. In total, 5,146 food‐ and waterborne outbreaks were reported. Salmonella was the most commonly detected agent with S. Enteritidis causing one in five outbreaks. Salmonella in eggs and egg products was the highest risk agent/food pair. A large increase of human West Nile virus infections was reported in 2018. The report further updates on bovine tuberculosis, Brucella, Trichinella, Echinococcus, Toxoplasma, rabies, Coxiella burnetii (Q fever) and tularaemia.
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              Gram-negative bacteria aggravate murine small intestinal Th1-type immunopathology following oral infection with Toxoplasma gondii.

              Oral infection of susceptible mice with Toxoplasma gondii results in Th1-type immunopathology in the ileum. We investigated gut flora changes during ileitis and determined contributions of gut bacteria to intestinal inflammation. Analysis of the intestinal microflora revealed that ileitis was accompanied by increasing bacterial load, decreasing species diversity, and bacterial translocation. Gram-negative bacteria identified as Escherichia coli and Bacteroides/Prevotella spp. accumulated in inflamed ileum at high concentrations. Prophylactic or therapeutic administration of ciprofloxacin and/or metronidazole ameliorated ileal immunopathology and reduced intestinal NO and IFN-gamma levels. Most strikingly, gnotobiotic mice in which cultivable gut bacteria were removed by quintuple antibiotic treatment did not develop ileitis after Toxoplasma gondii infection. A reduction in total numbers of lymphocytes was observed in the lamina propria of specific pathogen-free (SPF), but not gnotobiotic, mice upon development of ileitis. Relative numbers of CD4(+) T cells did not differ in naive vs infected gnotobiotic or SPF mice, but infected SPF mice showed a significant increase in the frequencies of activated CD4(+) T cells compared with gnotobiotic mice. Furthermore, recolonization with total gut flora, E. coli, or Bacteroides/Prevotella spp., but not Lactobacillus johnsonii, induced immunopathology in gnotobiotic mice. Animals recolonized with E. coli and/or total gut flora, but not L. johnsonii, showed elevated ileal NO and/or IFN-gamma levels. In conclusion, Gram-negative bacteria, i.e., E. coli, aggravate pathogen-induced intestinal Th1-type immunopathology. Thus, pathogen-induced acute ileitis may prove useful to study bacteria-host interactions in small intestinal inflammation and to test novel therapies based on modulation of gut flora.

                Author and article information

                Journal
                Eur J Microbiol Immunol (Bp)
                Eur J Microbiol Immunol (Bp)
                EUJMI
                European Journal of Microbiology & Immunology
                Akadémiai Kiadó (Budapest )
                2062-509X
                2062-8633
                27 July 2020
                14 October 2020
                : 10
                : 3
                : 139-146
                Affiliations
                Institute of Microbiology, Infectious Diseases and Immunology, Gastrointestinal Microbiology Research Group, Charité – University Medicine Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health , Berlin, Germany
                Author notes
                *Corresponding author. Department of Microbiology, Infectious Diseases and Immunology, Campus Benjamin Franklin, FEM, Charité – University Medicine Berlin, Garystr. 5, D-14195, Berlin, Germany. Tel.: +49 30 450524318. E-mail: markus.heimesaat@ 123456charite.de

                †Contributed equally.

                Author information
                https://orcid.org/0000-0001-6399-651X
                Article
                10.1556/1886.2020.00012
                7592517
                32750026
                74274fed-203d-4d5d-81a4-53f161615c43
                © 2020, The Authors

                Open Access statement. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License ( https://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and reproduction in any medium for non-commercial purposes, provided the original author and source are credited, a link to the CC License is provided, and changes - if any - are indicated.

                History
                : 04 June 2020
                : 18 June 2020
                Page count
                Figures: 7, Tables: 0, Equations: 0, References: 24, Pages: 8
                Categories
                Original Research Paper

                competitive exclusion product,aviguard®,colonization resistance,murine gut microbiota,inflammatory immune responses,host-pathogen-interaction,probiotic formulations,enteropathogenic infection

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