Diversity of anaerobic fungi population (phylum Neocallimastigomycota) in the horse hindgut

This thesis aims to study the diversity of symbiotic gut anaerobic fungi (phylum Neocallimastigomycota) in the different segments of the equine hindgut (cecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and rectum). A clone library of fungal ITS1 fragments of rDNA was prepared for each hindgut segment. These fragments were sequenced and phylogenetically analysed. The major finding of this study is the prevalence of uncultured Neocallimastigales in the horse lower digestive tract, representing 81% of the hindgut fungi and the completely different diversity of gut fungi of feces compared to cecum and colon content. The fecal sample did not reflect the microbial population composition of hindgut. The fungal population similarity between LVC and feces was evidenced in this study. RVC, LDC and RDC are inhabited by unknown gut fungi. A significant proportion of uncultured sequences is highly similar to the sequences of a new group of anaerobic fungi, which is specific for the Equidae family. Moreover a new cluster of anaerobic fungi was found in the digestive tract of horse (Anglo-Arabian reared in Sardinia). The horse digestive tract is populated by unknown, not yet cultivated species of anaerobic fungi and only a limited percentage of identified sequences can be classified into known species of gut fungi. The conventional microbiological cultivation techniques underestimate the diversity of the anaerobic fungal population in the horse hindgut.</br

Strategie di sorveglianza attiva e networking per il controllo delle infezioni da Listeria monocytogenes

Aim: the aim of this project was to activate an hospital and community surveillance system of listeriosis through epidemiological prospective and retrospective longitudinal study to describe prevalence of invasive infection and phenotypically and genotipically characteristics of the strains circulating in Northern Sardinia isolated from humans, foods and environment samples. Methods: L.monocytogenes strains isolated from 2013 to 2016 in food and environment samples and patients were analyzed for presence of virulence genes. The strains were characterized genotypically (PCR, PFGE, MLST) to define the spread of the pathogen and the relationships clonality between the strains of human and food origin. Furthermore, it was characterized the immune profile of patients with invasive disease vs health controls, through antigen specific stimulation of PBMC and cytofluorimetric analysis in blood samples. Results: Samples were positive for all 7 virulence-associated genes analyzed. 51,3%, 35,1% and 13,5% were identified serotype 4b, 1/2a and 1/2b, respectively. The PFGE showed the presence of 5 pulsetype, gruped in two major clusters (A,B,C-D,E) with 31,91% similarity index (Dice). This genetic variability was confirmed by MLST which identified 5 different STs (ST 1, ST 5, ST 8, ST 101, ST 121). Cytofluorimetric analysis showed the percentage of CD8 and CD4 T-cells significantly less in patient compared to a health control (19 ± 4 vs 4 ± 0.02, p> 0.0001); although, the CD8 cells of patient produced levels of cytokines (TNFα, IL-2) higher than health control. Conclusion: the results show that the active surveillance programs could be a crucial tool for assessing the incidence of listeriosis and describe the molecular characteristics of L.monocytogenes isolated. Also, the data obtained are an important tool for study how the immune profile can promote the development of invasive disease. The results obtained and the research protocols used are disclosed through a website

Beyond the third moment? Mapping the state of qualitative tourism research

© 2019, © 2019 Informa UK Limited, trading as Taylor & Francis Group. This paper poses a central question: what is the present moment in qualitative tourism research? To answer this, the authors review the state of contemporary qualitative tourism research over the past decade (2007–2017), against current progress in the social sciences. Specifically, a systematic bibliometric analysis of tourism journals was undertaken to determine how tourism scholarship maps against Denzin and Lincoln’s social science ‘moments’. These moments prove useful in heuristically framing the development of qualitative social research and thinking; from early moments characterized by positivist ways of knowing, through to later moments where scholars struggle through and beyond the crisis of representation. Using an adapted moments-based framework, our analysis of 1541 qualitative papers across 51 tourism journals indicates that the majority of papers are emblematic of ‘early moment’ thinking. Despite clear evidence of a crisis of representation in some areas, and an increase in the number of qualitative papers published, there is a lack of ‘later moment’ thinking, methodology and/or approach. The political, social and philosophical constraints and opportunities presented by these findings are discussed

Anaerobic fungal communities differ along the horse digestive tract

Anaerobic fungi are potent fibre degrading microbes in the equine hindgut, yet our understanding of their diversity and community structure is limited to date. In this preliminary work, using a clone library approach we studied the diversity of anaerobic fungi along six segments of the horse hindgut: caecum, right ventral colon (RVC), left ventral colon (LVC), left dorsal colon (LDC), right dorsal colon (RDC) and rectum. Of the 647 ITS1 clones, 61.7 % were assigned to genus level groups that are so far without any cultured representatives, and 38.0 % were assigned to the cultivated genera Neocallimastix (35.1 %), Orpinomyces (2.3 %), and Anaeromyces (0.6 %). AL1 dominated the group of uncultured anaerobic fungi, particularly in the RVC (88 %) and LDC (97 %). Sequences from the LSU clone library analysis of the LDC, however, split into two distinct phylogenetic clusters with low sequence identity to Caecomyces sp. (94–96 %) and Liebetanzomyces sp. (92 %) respectively. Sequences belonging to cultured Neocallimastix spp. dominated in LVC (81 %) and rectum (75.5 %). Quantification of anaerobic fungi showed significantly higher concentrations in RVC and RDC compared to other segments, which influenced the interpretation of the changes in anaerobic fungal diversity along the horse hindgut. These preliminary findings require further investigation.</p

Monogenic diseases that can be cured by liver transplantation

While the prevalence of most diseases caused by single-gene mutations is low and defines them as rare conditions, all together, monogenic diseases account for approximately 10 in every 1000 births according to the World Health Organisation. Orthotopic liver transplantation (LT) could offer a therapeutic option in monogenic diseases in two ways: by substituting for an injured liver or by supplying a tissue that can replace a mutant protein. In this respect, LT may be regarded as the correction of a disease at the level of the dysfunctional protein. Monogenic diseases that involve the liver represent a heterogeneous group of disorders. In conditions associated with predominant liver parenchymal damage (i.e., genetic cholestatic disorders, Wilson's disease, hereditary hemochromatosis, tyrosinemia, α1 antitrypsin deficiency), hepatic complications are the major source of morbidity and LT not only replaces a dysfunctional liver but also corrects the genetic defect and effectively cures the disease. A second group includes liver-based genetic disorders characterised by an architecturally near-normal liver (urea cycle disorders, Crigler-Najjar syndrome, familial amyloid polyneuropathy, primary hyperoxaluria type 1, atypical haemolytic uremic syndrome-1). In these defects, extrahepatic complications are the main source of morbidity and mortality while liver function is relatively preserved. Combined transplantation of other organs may be required, and other surgical techniques, such as domino and auxiliary liver transplantation, have been attempted. In a third group of monogenic diseases, the underlying genetic defect is expressed at a systemic level and liver involvement is just one of the clinical manifestations. In these conditions, LT might only be partially curative since the abnormal phenotype is maintained by extrahepatic synthesis of the toxic metabolites (i.e., methylmalonic acidemia, propionic acidemia). This review focuses on principles of diagnosis, management and LT results in both paediatric and adult populations of selected liver-based monogenic diseases, which represent examples of different transplantation strategies, driven by the understanding of the expression of the underlying genetic defect. © 2013 European Association for the Study of the Liver

Beyond the third moment? Mapping the state of qualitative tourism research

This paper poses a central question: what is the present moment in qualitative tourism research? To answer this, the authors review the state of contemporary qualitative tourism research over the past decade (2007–2017), against current progress in the social sciences. Specifically, a systematic bibliometric analysis of tourism journals was undertaken to determine how tourism scholarship maps against Denzin and Lincoln’s social science ‘moments’. These moments prove useful in heuristically framing the development of qualitative social research and thinking; from early moments characterized by positivist ways of knowing, through to later moments where scholars struggle through and beyond the crisis of representation. Using an adapted moments-based framework, our analysis of 1541 qualitative papers across 51 tourism journals indicates that the majority of papers are emblematic of ‘early moment’ thinking. Despite clear evidence of a crisis of representation in some areas, and an increase in the number of qualitative papers published, there is a lack of ‘later moment’ thinking, methodology and/or approach. The political, social and philosophical constraints and opportunities presented by these findings are discussed

Monocyte Phenotypes and Physical Activity in Patients with Carotid Atherosclerosis

Atherosclerosis is associated with low-grade inflammation involving circulating monocytes. It has been shown that the levels of intermediate pro-inflammatory monocytes are associated with cardiovascular mortality and risk of ischemic stroke. It also has been shown that physical activity (PA) decreases inflammation markers, incidence of strokes, and mortality. In this cross-sectional study, we tested the effect of PA on circulating monocytes phenotype rate. A total of 29 patients with a carotid stenosis &gt; 50% were recruited. Levels of physical activity (MET.min/week) were measured by the GPAQ questionnaire, arterial samples of blood were collected to analyze monocyte phenotype (classical, intermediate and non-classical) assessed by flow cytometry, and venous blood samples were used to dose antioxidant activity and oxidative damage. Antioxidant capacity was reduced and oxidative damage increased in patients. There was a significant decrease in the percentage of classical and intermediate monocytes in moderately active patients as compared with non-active and highly active patients. Inversely, the rate of non-classical monocytes increased in moderately active patients. Intense PA appears to blunt the beneficial effects of moderate PA. Our study also suggests that PA could be beneficial in such patients by reducing the rate of intermediate monocytes known to predict the risk of ischemic stroke and by increasing the non-classical monocytes involved in lesions&rsquo; healing. Nevertheless, a longitudinal study would be necessary to confirm this hypothesis

Association between physical activity and sedentary behaviour on carotid atherosclerotic plaques: an epidemiological and histological study in 90 asymptomatic patients

International audienceObjective Carotid atherosclerotic plaques are a source of emboli for stroke. 'Unstable' carotid atherosclerotic plaques may have intraplaque haemorrhages, neovessels, prevalent macrophages, excessive calcium deposits, a large lipid core and a thin fibrous cap. Regular physical activity (PA) may lower the risk of plaques becoming unstable. We evaluated the association of both PA and sedentary behaviour (SB) with carotid plaque histopathology.Methods 90 asymptomatic patients who were undergoing carotid endarterectomy for carotid artery narrowing identified on ultrasound reported their PA and SB by questionnaires. We calculated PA intensity in MET (metabolic equivalent of task)-min/week. For analysis, the population was divided into tertiles according to PA (T1PA: the less PA patients; T2PA: the intermediate PA patients; T3PA: the most physically active patients) (T1PA900 and <900 MET-min/week, respectively). All the other features that associate with plaque instability (eg, neovessels, macrophages, etc) did not differ by level of PA or SB.Conclusion In this cross-sectional study of asymptomatic patients who underwent endarterectomy (i) higher reported PA, (ii) intensity of PA and (iii) lower reported SB were associated with lower prevalence of intraplaque haemorrhage. This could be a mechanism whereby PA protects against cerebrovascular disease (stroke) and death

A Case of <em>Listeria monocytogenes</em> ST-219 Meningo-Encephalitis

Listeriosis is a foodborne disease characterized by high hospitalization and fatality rates, especially in vulnerable groups including elderly subjects, pregnant women, etc. We report on the first case of Listeria monocytogenes ST-219 meningo-encephalitis in a woman aged 83 years. An epidemiological and molecular investigation was performed to detect the source of infection and the virulence factors associated with L. monocytogenes invasiveness in this patient. All environmental- and clinical-associated isolates were found to belong to serotype 4b and ST-219 as well as possessing actA, prfA, hlyA, and rrn virulence genes. Antibiotic susceptibility testing also detected resistance to cotrimoxazole, clindamycin, erythromycin, and oxacillin in these isolates. Conventional and molecular surveillance of listeriosis cases, based on the systematic assessment of spatio-temporal trends, virulence genes, and antimicrobial susceptibility testing patterns, are key to preventing and controlling the emergence and spread of L. monocytogenes strains, including hypervirulent clones

Listeria spp. and Listeria monocytogenes contamination in ready-to-eat sandwiches collected from vending machines

Ready-to-eat (RTE) food is characterised by a long shelf-life at refrigerated temperature and can be consumed as such, without any treatment. The aim of the work was to evaluate the presence of Listeria spp. and Listeria monocytogenes in RTEs collected from refrigerated vending machines placed in hospital environment and accessible to the hospitalised patients. In 4 different sampling, 55 RTEs were collected from vending machines of six hospitals located in different areas of Sardinia region. All the samples were characterised by similar manufacturing process, such as the use of modified atmosphere packaging and belonged to 5 different producers. Listeria spp. was not countable using the enumeration method in all of the analysed samples. Using the detection method, Listeria spp. was recovered from 9 sandwich samples. Interestingly, 3 of these samples (5.5%) made by the manufacturer, were positive for L. monocytogenes contamination. The risk related to the L. monocytogenes presence in RTEs proportionally increases when food is introduced in susceptible environments, such as hospitals and consumed by susceptible people. Although the RTEs analysed showed values that complied with the European microbiological criteria for foodstuffs, the availability of these products in a susceptible environment should be carefully checked. Therefore, in order to limit the possible exposition to L. monocytogenes, more information on the risk related to RTE consumption should be provided to the hospitalised patients