14 research outputs found

    NMR metabolomics reveals effects of Cryptosporidium infections on host cell metabolome

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    Background: Cryptosporidium is an important gut microbe whose contributions towards infant and immunocompromise patient mortality rates are steadily increasing. Over the last decade, we have seen the development of various tools and methods for studying Cryptosporidium infection and its interactions with their hosts. One area that is sorely overlooked is the effect infection has on host metabolic processes. Results: Using a 1H nuclear magnetic resonance approach to metabolomics, we have explored the nature of the mouse gut metabolome as well as providing the first insight into the metabolome of an infected cell line. Statistical analysis and predictive modelling demonstrated new understandings of the effects of a Cryptosporidium infection, while verifying the presence of known metabolic changes. Of note is the potential contribution of host derived taurine to the diarrhoeal aspects of the disease previously attributed to a solely parasite-based alteration of the gut environment, in addition to other metabolites involved with host cell catabolism. Conclusion: This approach will spearhead our understanding of the Cryptosporidium-host metabolic exchange and provide novel targets for tackling this deadly parasite

    Review of Cryptosporidium and Giardia in the eastern part of Europe, 2016

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    Introduction: This paper reviews the current knowledge and understanding of Cryptosporidium spp. an d Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results: Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices and detection methodologies used. Conclusion: Upgrading and making the diagnosis/detection procedures more uniform is recommended throughout the region. Public health authorities should actively work towards increasing reporting and standardising reporting practices as these prerequisites for the reported data to be valid and therefore necessary for appropriate control plans.Peer reviewe

    Latent microsporidiosis caused by Encephalitozoon cuniculi in immunocompetent hosts: a murine model demonstrating the ineffectiveness of the immune system and treatment with albendazole.

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    BACKGROUND: Microsporidia are obligate intracellular parasites causing severe infections with lethal outcome in immunocompromised hosts. However, these pathogens are more frequently reported as latent infections in immunocompetent individuals and raises questions about the potential risk of reactivation following induced immunosuppression. AIMS: To evaluate the possibility latent microsporidiosis, efficacy or albendazole, and reactivation, the authors monitored the course of E. cuniculi infection in immunocompetent BALB/c mice and immunodeficient SCID mice using molecular methods. METHODS: Mice were per orally infected with 10(7) spores of E. cuniculi. Selected groups were treated with albendazole, re-infected or chemically immunosuppressed by dexamethasone. The presence of microsporidia in the host's organs and feces were determined using PCR methods. Changes in numbers of lymphocytes in blood and in spleen after induction of immunosuppression were confirmed using flow cytometry analysis. RESULTS: Whereas E. cuniculi caused lethal microsporidiosis in SCID mice, the infection in BABL/c mice remained asymptomatic despite parasite dissemination into many organs during the acute infection phase. Albendazole treatment led to microsporidia elimination from organs in BALB/c mice. In SCID mice, however, only a temporary reduction in number of affected organs was observed and infection re-established post-treatment. Dexamethasone treatment resulted in a chronic microsporidia infection disseminating into most organs in BALB/c mice. Although the presence of E. cuniculi in organs of albendazole- treated mice was undetectable by PCR, it was striking that infection was reactivated by immunosuppression treatment. CONCLUSION: Our results demonstrated that microsporidia can successfully survive in organs of immunocompetent hosts and are able to reactivate from undetectable levels and spread within these hosts after induction of immunosuppression. These findings stress the danger of latent microsporidiosis as a life-threatening risk factor especially for individuals undergoing chemotherapy and in transplant recipients of organs originating from infected donors

    Cryptosporidium

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    According to the World Health Organisation, cryptosporidiosis is a global diarrhoeal disease affecting millions of individuals; it is the second most common cause of infantile death in developing countries and is increasingly identified as an emerging cause of morbidity and mortality worldwide. The disease is also extremely severe in livestock, causing profuse diarrhea and considerable economic losses in farmed young animals. Given the lack of effective treatment (absence of vaccines and effective drugs) and the limited understanding of the causative parasite, cryptosporidiosis represents a major challenge in the battle against global diarrheal diseases. Currently, there are 45 described Cryptosporidium species infecting a whole spectrum of animals. In this book chapter we will present an overview of the parasite, focusing on its taxonomic status, its morphology, its prevalence and transmission. We will review both cell biological and molecular techniques currently used to investigate the biology of this parasite and we will introduce the new state-of-the-art techniques that have been established by several laboratories in the field. With the development of these new technologies, we will be able to further understand the unique biology of Cryptosporidium and its role in health and disease of its host

    Course of <i>Encepahlitozoon cuniculi</i> genotype II infection, including pattern of spore shedding and dissemination of infection to selected organs and tissues.

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    <p><b>a)</b> SCID mice, <b>b)</b> SCID mice treated with albendazole, <b>c)</b> BALB/c mice and BALB/c mice re-infected in chronic stage of infection, <b>d)</b> BALB/c mice treated with albendazole and BALB/c mice treated with albendazole with following re-infection, <b>e)</b> BALB/c mice immunosuppressed in chronic stage of infection, <b>f)</b> BALB/c mice immunosuppressed after albendazole treatment. <b>Light-gray field</b> – albendazole treatment; <b>dark-gray field</b> – dexamethasone immunosuppression; <b>black line</b> – course of <i>E. cuniculi</i> infection; <b>black dash line</b> - course of <i>E. cuniculi</i> re-infection; <b>cross</b> – <i>E. cuniculi</i> positive organ during primarily infection; <b>ring</b> – <i>E. cuniculi</i> positive organ during re-infection; <b>black square</b> – spores shedding during primarily infection; <b>black circle</b> – spores shedding during re-infection.</p

    Design of experiments.

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    <p><b><sup>a</sup></b>inoculation with 200 ul sterilized deionised water; <b><sup>b</sup></b>infection with 10<sup>7 </sup><i>E. cuniculi</i> spores in 0.2 ml of sterilized deionised water; INF – infection; REINF – reinfection (black column); IMSUP – dexamethasone immunosuppression (highlighted in dark grey); TREAT – albendazole treatment (highlighted in light grey); <b>n1–</b> number of used animals; <b>n2</b>– number of dissected animals; <b>NS</b> – not shown; <b>x</b> – not observed due to mouse death; <b>DPI</b> – day post infection;</p

    Metabolic changes of the host-pathogen environment in a Cryptosporidium infection

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    Cryptosporidium is an important gut microbe whose contributions towards infant and immunocompromise patient mortality rates are steadily increasing. Current techniques for diagnosing, curing or simply understanding the biology of the parasite are few and far between, relying on a combination of in-silico predictions modelled on a varied and unique group of organisms and medical reports. The development of an in-vitro culture system, using COLO-680N cells, has provided the Cryptosporidium community with the opportunity to expand its toolkit for investigating this disease. One area in particular that is sorely overlooked is the metabolic alterations upon infection. Existing research is extremely limited and has already shown that significant variation can be found between the metabolome of different infected host species. Using a 1H Nuclear Magnetic Resonance approach to metabolomics, we have explored the nature of the mouse gut metabolome as well as providing the first insight into the metabolome of an infected cell line. Through a combination of Partial Least Squares Discriminant Analysis and predictive modelling, we exhibit new and potentially game changing insights into the effects of a Cryptosporidium parvum infection, while verifying the presence of known metabolic changes. Of particular note is the potential contribution of host derived taurine to the diuretic aspects of the disease previously attributed to a solely parasite based alteration of the gut environment. This practical and informative approach can spearhead our understanding of the Cryptosporidium-host metabolic exchange and thus provide novel targets for tackling this deadly parasite

    Long-Term Monitoring of Microsporidia, Cryptosporidium and Giardia Infections in Western Lowland Gorillas (Gorilla gorilla gorilla) at Different Stages of Habituation in Dzanga Sangha Protected Areas, Central African Republic

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    Background Infectious diseases pose one of the greatest threats to endangered species, and a risk of gastrointestinal parasite transmission from humans to wildlife has always been considered as a major concern of tourism. Increased anthropogenic impact on primate populations may result in general changes in communities of their parasites, and also in a direct exchange of parasites between humans and primates. Aims To evaluate the impact of close contact with humans on the occurrence of potentially zoonotic protists in great apes, we conducted a long-term monitoring of microsporidia, Cryptosporidium and Giardia infections in western lowland gorillas at different stages of the habituation process, humans, and other wildlife in Dzanga-Sangha Protected Areas in the Central African Republic. Results We detected Encephalitozoon cuniculi genotypes I and II (7.5%), Enterocytozoon bieneusi genotype D and three novel genotypes (gorilla 1–3) (4.0%), Giardia intestinalis subgroup A II (2.0%) and Cryptosporidium bovis (0.5%) in gorillas, whereas in humans we found only G. intestinalis subgroup A II (2.1%). In other wild and domestic animals we recorded E. cuniculi genotypes I and II (2.1%), G. intestinalis assemblage E (0.5%) and C. muris TS03 (0.5%). Conclusion Due to the non-specificity of E. cuniculi genotypes we conclude that detection of the exact source of E. cuniculi infection is problematic. As Giardia intestinalis was recorded primarily in gorilla groups with closer human contact, we suggest that human-gorilla transmission has occurred. We call attention to a potentially negative impact of habituation on selected pathogens which might occur as a result of the more frequent presence of humans in the vicinity of both gorillas under habituation and habituated gorillas, rather than as a consequence of the close contact with humans, which might be a more traditional assumption. We encourage to observe the sections concerning hygiene from the IUCN best practice guidelines for all sites where increased human-gorilla contact occurs

    Follow-up investigation into Cryptosporidium prevalence and transmission in Western European dairy farms

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    Cryptosporidium parvum is an enteric parasite and a major contributor to acute enteritis in calves worldwide, causing an important economic burden for farmers. This parasite poses a major public health threat through transmission between livestock and humans. Our previous pilot study in Western Europe revealed a high prevalence of Cryptosporidium in calves of dairy farms. In the sequel study herein, 936 faecal samples were collected from the same 51 dairy farms across Belgium, France, and the Netherlands. Following DNA extraction, Cryptosporidium screening was carried out using nested-PCR amplification targeting the SSU rRNA gene. All positive samples were sequenced, and phylogenetic analyses were used to identify the Cryptosporidium spp. present. The 60 kDa glycoprotein (gp60) gene was also sequenced to determine the C. parvum subtypes present. Prevalence of Cryptosporidium ranged from 23.3% to 25%, across the three countries surveyed. The parasite was found in most of the farms sampled, with 90.2% testing positive. Cryptosporidium parvum, C. bovis, C. ryanae and C. andersoni were all identified, with the former being the most predominant, representing 71.4% of all infections. Cryptosporidium parvum was associated with pre-weaned calves, while other species were associated with older animals. Subtyping of gp60 gene revealed nine subtypes, eight of which have previously been reported to cause clinical disease in humans. Similarly to the first study, vertical transmission was not a major contributor to Cryptosporidium spread. Our study highlights the need for further investigation into cryptosporidiosis transmission, and future studies will require a One Health approach to reduce the impact of this disease

    Cross-border investigations on the prevalence and transmission dynamics of Cryptosporidium species in dairy cattle farms in western mainland Europe

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    Cryptosporidium is comprised an apicomplexan parasitic protist, which infects a wide range of hosts, causing cryptosporidiosis. In cattle farms, the incidence of cryptosporidiosis results in high mortality in calves leading to considerable economic loss in the livestock industry. Infected animals may also act as a major reservoir of Cryptosporidium spp., in particular C. parvum, the most common cause of cryptosporidiosis in calves. This poses a significant risk to other farms via breeding centres, to trading of livestock and to human health. This study, funded by the Interreg-2-seas programme, is a part of a global project aimed at strategies to tackle cryptosporidiosis. To reach this target, it was essential to determine whether prevalence was dependent on the studied countries or if the issue was borderless. Indeed, C. parvum occurrence was assessed across dairy farms in certain regions of Belgium, France and the Netherlands. At the same time, the animal-to-animal transmission of the circulating C. parvum subtypes was studied. To accomplish this, 1084 faecal samples, corresponding to 57 dairy-farms from all three countries, were analysed. Well-established protocols amplifying the 18S rDNA and gp60 genes fragments, followed by DNA sequencing, were used for the detection and subtyping C. parvum; the DNA sequences obtained were further characterised using a combination of bioinformatics and phylogenetics methods. Our results show 25.7%, 24.9% and 20.8% prevalence of Cryptosporidium spp. in Belgium, France and the Netherlands respectively. Overall, 93% of the farms were Cryptosporidium positive. The gp60 subtyping demonstrated a significant number of the C. parvum positives belonged to the IIa allelic family, which has been also detected in humans. Consequently, this study highlights how widespread is C. parvum in dairy farms and endorses cattle as a major carrier of zoonotic C. parvum subtypes, which subsequently pose a significant threat to human health
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