A Roepstorff

IT University of Copenhagen, København, Capital Region, Denmark

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Publications (134)276.72 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Eggs of the pig whipworm, Trichuris suis ova (TSO), are currently tested in human clinical trials for their potential immunomodulatory capacity. The biological potency of TSO (egg viability and infectivity) is traditionally assessed in Göttingen minipigs as the establishment of intestinal larvae after inoculation with a known number of eggs. To minimize testing in animal models, development of an in vitro egg hatching assay is proposed as a reliable, cost-effective, and a faster alternative to test the egg viability. The present study aimed to investigate the influence of different chemical, physical, and biological factors on egg hatching. Thus, in a series of experiments and in different combinations, the eggs were stimulated with glass beads, artificial gastric juice, bile salt and trypsin solution, fermentation gut medium, or stimulated with mucosal scrapings from the ileum and the large intestine of the infected and uninfected Göttingen minipig. Mechanical stimulation with glass beads presented a simple and reproducible method for egg hatching. However, incubation of eggs with mucosal scrapings from the ileum, caecum, and colon for 24 h at 38 °C significantly increased hatching.
    Parasitology Research 05/2015; DOI:10.1007/s00436-015-4476-1 · 2.33 Impact Factor
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    ABSTRACT: This histopathological study was carried out in order to investigate the cellular response in the jejunum to Ascaridia galli during the first 7 weeks of infection. Fourty-two ISA Brown chickens (7 weeks old) were infected orally with 500 embryonated A. galli eggs each while 28 chickens were left as uninfected controls. Six infected and four control chickens were necropsied at each time point 3, 7, 10, 14, 21, 28 and 42 days post-infection (dpi). Samples for histopathology were taken from three sites of the jejunoileum. Significantly higher eosinophil counts were seen in infected chickens compared to uninfected at 3, 7, 10, 14 and 28 dpi (P < 0.01). In both groups, the initial number of mast cells was high, but this high level of mast cells remained for a longer period in the infected group compared to the control group. Significantly higher counts were thus found in the infected group at 21 (P < 0.001), 28 (P < 0.01) and 42 dpi (P < 0.05). A. galli infection induced changes in the mucosal thickness as reduced villi length at 7, 10, 14, 21 and 28 dpi and in the degree of general cellular infiltration in the lamina propria of the mucosal layer. No adult worms were seen during the experiment; therefore, A. galli larvae have elicited a moderate cellular response in the lamina propria, mainly consisting of eosinophils in the early phase and later of mast cells.
    Parasitology Research 04/2015; DOI:10.1007/s00436-015-4450-y · 2.33 Impact Factor
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    ABSTRACT: An observational study has suggested that relapsing-remitting multiple sclerosis patients with helminth infections have lower disease activity and progression than uninfected multiple sclerosis patients. To evaluate the safety and efficacy on MRI activity of treatment with TSO in relapsing MS. The study was an open-label, magnetic resonance imaging assessor-blinded, baseline-to-treatment study including ten patients with relapsing forms of multiple sclerosis. Median (range) age was 41 (24-55) years, disease duration 9 (4-34) years, Expanded Disability Status Scale score 2.5 (1-5.0), and number of relapses within the last two years 3 (2-5). Four patients received no disease modifying therapy, while six patients received IFN-β. After an observational period of 8 weeks, patients received 2500 ova from the helminth Trichuris suis orally every second week for 12 weeks. Patients were followed with serial magnetic resonance imaging, neurological examinations, laboratory safety tests and expression of immunological biomarker genes. Treatment with Trichuris suis orally was well-tolerated apart from some gastrointestinal symptoms. Magnetic resonance imaging revealed 6 new or enlarged T2 lesions in the run-in period, 7 lesions in the early period and 21 lesions in the late treatment period. Two patients suffered a relapse before treatment and two during treatment. Eight patients developed eosinophilia. The expression of cytokines and transcription factors did not change. In a small group of relapsing multiple sclerosis patients, Trichuris suis oral therapy was well tolerated but without beneficial effect. © The Author(s), 2015.
    Multiple Sclerosis 02/2015; DOI:10.1177/1352458514568173 · 4.86 Impact Factor
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    ABSTRACT: Embryonated eggs of the pig whipworm Trichuris suis (TSOee) constitute the active pharmaceutical ingredient (API) in a medicinal product explored in human clinical trials against several immune-mediated diseases. The measurement of TSO biological potency (hatchability and infectivity) is a requirement for the assessment of TSO's pharmacological potency in human clinical trials. The present study aims to validate the dose-dependent establishment of T. suis larvae in Göttingen minipigs and eventual clinical implication of a dose range (1000-10,000 TSO). Four groups of 5 minipigs were inoculated with doses of 1000, 2500, 7500, and 10,000 TSOee, respectively, to evaluate a range of concentrations of TSOee in a minipig infectivity model. Unembryonated eggs (TSOue) were added to keep the total egg number in the inoculum constant at 10,000 eggs. Two groups received 2500 and 7500 TSOee per pig without the addition of TSOue as controls. The intestinal larval establishment at 21 days post inoculation (dpi) demonstrated a clear positive linear dose-response relationship between numbers of inoculated TSOee and recovered larvae. There was a low level of variation in larval counts in all study groups. Thus, the infectivity model in minipigs within the tested dose range offers a reliable, sensitive and accurate assay for testing biological potency of TSO. Copyright © 2015 Elsevier B.V. All rights reserved.
    Veterinary Parasitology 01/2015; 208(3-4). DOI:10.1016/j.vetpar.2015.01.018 · 2.55 Impact Factor
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    ABSTRACT: Background OvaSpec is a new, fully automated, vision-based instrument for assessing the quantity (concentration) and quality (embryonation percentage) of Trichuris suis parasite eggs in liquid suspension. The eggs constitute the active pharmaceutical ingredient in a medicinal drug for the treatment of immune-mediated diseases such as Crohn's disease, ulcerative colitis, and multiple sclerosis. Methods This paper describes the development of an automated microscopy technology, including methodological challenges and design decisions of relevance for the future development of comparable vision-based instruments. Morphological properties are used to distinguish eggs from impurities and two features of the egg contents under brightfield and darkfield illumination are used in a statistical classification to distinguish eggs with undifferentiated contents (non-embryonated eggs) from eggs with fully developed larvae inside (embryonated eggs). Results For assessment of the instrument's performance, six egg suspensions of varying quality were used to generate a dataset of unseen images. Subsequently, annotation of the detected eggs and impurities revealed a high agreement with the manual, image-based assessments for both concentration and embryonation percentage (both error rates <1.0%). Similarly, a strong correlation was demonstrated in a final, blinded comparison with traditional microscopic assessments performed by an experienced laboratory technician. Conclusions The present study demonstrates the applicability of computer vision in the production, analysis, and quality control of T. suis eggs used as an active pharmaceutical ingredient for the treatment of autoimmune diseases.
    Computers in Biology and Medicine 10/2014; 53. DOI:10.1016/j.compbiomed.2014.07.009 · 1.46 Impact Factor
  • 06/2014; 4(2):93-105. DOI:10.1007/s13165-014-0061-7
  • 06/2014; 4(2):135-147. DOI:10.1007/s13165-014-0069-z
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    ABSTRACT: The aim of the present study was to investigate the population dynamics and potential interactions between Trichuris suis and Oesophagostomum dentatum in experimentally co-infected pigs, by quantification of parasite parameters such as egg excretion, worm recovery and worm location. Forty-eight helminth naïve pigs were allocated into four groups. Group O was inoculated with 20 O. dentatum L3/kg/day and Group T with 10 T. suis eggs/kg/day. Group OT was inoculated with both 20 O. dentatum L3/kg/day and 10 T. suis eggs/kg/day, while Group C was kept as an uninfected control group. All inoculations were trickle infections administered twice weekly and were continued until slaughter. Faecal samples were collected from the rectum of all pigs at day 0, and twice weekly from 2 to 9 weeks post first infection (wpi). Six pigs from each group were necropsied 5 wpi and the remaining 6 pigs from each group were necropsied 10 wpi. The faecal egg counts (FEC) and total worm burdens of O. dentatum were dramatically influenced by the presence of T. suis, with significantly lower mean FECs and worm burdens at 5 and 10 wpi compared to single infected pigs. Furthermore, in the presence of T. suis we found that O. dentatum was located more posteriorly in the gut. The changes in the Trichuris population were less prominent, but faecal egg counts, worm counts 5 wpi (57% recovered vs. 39%) and the proportion of infected animals at 10 wpi were higher in Group OT compared to Group T. The location of T. suis was unaffected by the presence of O. dentatum. These results indicate an antagonistic interaction between T. suis and O. dentatum which is dominated by T. suis.
    Veterinary Parasitology 10/2013; 199(1-2). DOI:10.1016/j.vetpar.2013.09.030 · 2.55 Impact Factor
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    ABSTRACT: This paper reviews the available information on the different health and animal welfare issues in organic pigs in relation to weaning. It addresses the most relevant health and welfare problems and reviews their potential hazards and associated risk factors. Regarding health, problems related to post weaning diarrhoea, cold stress, skin lesions, endoparasites and post weaning multisystemic wasting syndrome are described. Reasons for distress and frustration in weaned piglets are identified as mainly separation from the mother, a new environment, mixing and fear of humans. Finally, hazards and risk factors for health and welfare in organic weaners are related to animal characteristics, housing systems, feed/nutrition and management. Generally, it is concluded that diseases around weaning are multifactorial in nature, with several factors contributing simultaneously as stressors at the time of weaning. In order to solve problems around weaning, the complexity and the individuality of farm systems need to be taken into account.
    06/2013; DOI:10.1007/s13165-013-0054-y
  • 06/2013; 4(2):107-121. DOI:10.1007/s13165-013-0052-0
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    ABSTRACT: SUMMARY The population dynamics of Ascaridia galli was studied in 70 ISA Brown layer pullets, 42 of them were each experimentally infected with 500 embryonated A. galli eggs and 28 chickens were kept as uninfected controls. Six chickens from the infected group and 4 from the control group were necropsied at 3, 7, 10, 14, 21, 28 and 42 days post-infection (d.p.i.). The mean worm recovery varied from 11-20% of the infection dose with the highest recovery at 3 d.p.i. and the lowest at 21 and 42 d.p.i. (P < 0·05). More larvae were recovered from the intestinal wall than from the content (P < 0·0001) and intestinal content larvae were longer than those from the wall (mean length 1·6 and 1 mm, respectively, P < 0·0001). Although larvae were growing over time, a population of small-sized larvae (length < 1 mm) was recovered at all d.p.i. During the first week of infection most of the larvae were located in the anterior half of the jejunoileum but they moved posteriorly with the age of infection. Thus, a subpopulation of larvae mainly in the lumen grew with time while another subpopulation remained small and associated with the mucosa. During the infection both subpopulations moved to a more posterior localization in the gastrointestinal (GI) tract.
    Parasitology 05/2013; DOI:10.1017/S0031182013000401 · 2.35 Impact Factor
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    ABSTRACT: Anaerobic digestion of residual materials from animals and crops offers an opportunity to simultaneously produce bioenergy and plant fertilizers at single farms and in farm communities where input substrate materials and resulting digested residues are shared among member farms. A surplus benefit from this practice may be the suppressing of propagules from harmful biological pests like weeds and animal pathogens (e.g. parasites). In the present work, batch experiments were performed, where survival of seeds of seven species of weeds and non-embryonated eggs of the large roundworm of pigs, Ascaris suum, was assessed under conditions similar to biogas plants managed at meso- (37°C) and thermophilic (55°C) conditions. Cattle manure was used as digestion substrate and experimental units were sampled destructively over time. Regarding weed seeds, the effect of thermophilic conditions (55°C) was very clear as complete mortality, irrespective of weed species, was reached after less than 2 days. At mesophilic conditions, seeds of Avena fatua, Sinapsis arvensis, Solidago canadensis had completely lost germination ability, while Brassica napus, Fallopia convolvulus and Amzinckia micrantha still maintained low levels (∼1%) of germination ability after 1 week. Chenopodium album was the only weed species which survived 1 week at substantial levels (7%) although after 11d germination ability was totally lost. Similarly, at 55°C, no Ascaris eggs survived more than 3h of incubation. Incubation at 37°C did not affect egg survival during the first 48h and it took up to 10days before total elimination was reached. In general, anaerobic digestion in biogas plants seems an efficient way (thermophilic more efficient than mesophilic) to treat organic farm wastes in a way that suppresses animal parasites and weeds so that the digestates can be applied without risking spread of these pests.
    Waste Management 12/2012; 33(4). DOI:10.1016/j.wasman.2012.11.001 · 3.16 Impact Factor
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    ABSTRACT: SUMMARY Separation of pig slurry into solid and liquid fractions is gaining importance as a way to manage increasing volumes of slurry. In contrast to solid manure and slurry, little is known about pathogen survival in separated liquid slurry. The viability of Ascaris suum eggs, a conservative indicator of fecal pollution, and its association with ammonia was investigated in separated liquid slurry in comparison with raw slurry. For this purpose nylon bags with 6000 eggs each were placed in 1 litre bottles containing one of the two fractions for 308 days at 5 °C or 25 °C. Initial analysis of helminth eggs in the separated liquid slurry revealed 47 Ascaris eggs per gramme. At 25 °C, egg viability declined to zero with a similar trend in both raw slurry and the separated liquid slurry by day 308, a time when at 5 °C 88% and 42% of the eggs were still viable in separated liquid slurry and raw slurry, respectively. The poorer survival at 25 °C was correlated with high ammonia contents in the range of 7·9-22·4 mm in raw slurry and 7·3-23·2 mm in liquid slurry compared to 3·2-9·5 mm in raw slurry and 2·6-9·5 mm in liquid slurry stored at 5 °C. The study demonstrates that at 5 °C, A. suum eggs have a higher viability in separated liquid slurry as compared to raw slurry. The hygiene aspect of this needs to be further investigated when separated liquid slurry is used to fertilize pastures or crops.
    Parasitology 11/2012; 140(3):1-7. DOI:10.1017/S0031182012001722 · 2.35 Impact Factor
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    ABSTRACT: Parasitic helminths have been shown to reduce inflammation in most experimental models of allergic disease, and this effect is mediated via cytokine responses. However, in humans, the effects of controlled helminth infection on cytokine responses during allergy have not been studied. The aim was to investigate whether infection with the nematode parasite Trichuris suis alters systemic cytokine levels, cellular cytokine responses to parasite antigens and pollen allergens and/or the cytokine profile of allergic individuals. In a randomized double-blinded placebo-controlled clinical trial (UMIN trial registry, Registration no. R000001298, Trial ID UMIN000001070, URL: http://www.umin.ac.jp/map/english), adults with grass pollen-induced allergic rhinitis received three weekly doses of 2500 Trichuris suis ova (n = 45) or placebo (n = 44) over 6 months. IFN-γ, TNF-α, IL-4, IL-5, IL-10 and IL-13 were quantified via cytometric bead array in plasma. Cytokines, including active TGF-β, were also quantified in supernatants from peripheral blood mononuclear cells cultured with parasite antigens or pollen allergens before, during and after the grass pollen season for a sub-cohort of randomized participants (T. suis ova-treated, n = 12, Placebo-treated, n = 10). Helminth infection induced a Th2-polarized cytokine response comprising elevated plasma IL-5 and parasite-specific IL-4, IL-5 and IL-13, and a global shift in the profile of systemic cytokine responses. Infection also elicited high levels of the regulatory cytokine IL-10 in response to T. suis antigens. Despite increased production of T. suis-specific cytokines in T. suis ova-treated participants, allergen-specific cytokine responses during the grass pollen season and the global profile of PBMC cytokine responses were not affected by T. suis ova treatment. This study suggests that cytokines induced by Trichuris suis ova treatment do not alter allergic reactivity to pollen during the peak of allergic rhinitis symptoms.
    Clinical & Experimental Allergy 11/2012; 42(11):1582-95. DOI:10.1111/j.1365-2222.2012.04063.x · 4.32 Impact Factor
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    ABSTRACT: This study was conducted to observe the localization and to compare methods for isolation of minute Ascaridia galli larvae in chicken intestine. Firstly, six 7-week-old layer pullets were orally infected with 2,000 embryonated A. galli eggs and necropsied either at 3, 5 or 7 days post infection (dpi). More than 95 % of the recovered larvae were obtained from the anterior half of the jejunoileum, suggesting this part as the initial predilection site for A. galli larvae. Secondly, the intestinal wall of one layer pullet infected with 20,000 A. galli eggs 3 days earlier was digested in pepsin-HCl for 90 min. The initial 10 min of digestion released 51 % of the totally recovered larvae and the last 30 min of continuous digestion yielded only 5 %. This indicates that the majority of larvae were located superficially in the intestinal mucosa. Thirdly, 48 7-week-old layer pullets were infected with 500 A. galli eggs and necropsied at 3 dpi to compare three different larval isolation methods from the intestinal wall, viz., EDTA incubation, agar-gel incubation and pepsin-HCl digestion, resulting in mean percentages of the recovered larvae: 14.4, 18.2 and 20.0 %, respectively (P = 0.15). As conclusion, we recommended Pepsin-HCl digestion as the method of choice for larval recovery from the intestinal wall in future population dynamics study due to high efficiency and quick and simple detection. The agar-gel method was considered to be a prerequisite for molecular and immunological investigations as the larvae were more active and fully intact.
    Parasitology Research 08/2012; 111(6). DOI:10.1007/s00436-012-3079-3 · 2.33 Impact Factor
  • Neurology 04/2012; 78(Meeting Abstracts 1):S30.005-S30.005. DOI:10.1212/WNL.78.1_MeetingAbstracts.S30.005 · 8.30 Impact Factor
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    ABSTRACT: The normal habitat of the parasitic stages of Ascaridia galli is in the small intestine of poultry but the exact localization is poorly understood. Therefore, a histological study was conducted in order to localize the larvae during the early phase of infection. Six layer pullets seven-week old were infected orally with 20,000 embryonated A. galli eggs each, whereas four chickens were left as un-infected controls. At necropsy 3 days after infection the first half of jejunum/ileum was divided into two equally sized sections (J1 and J2). After taking samples for histology from the middle of J1 and J2 and the junction between these determined JX, the two sections were subjected to parasitological examination. A higher number of A. galli larvae were recovered from section J2 than J1 and the majority of larvae were recovered from the most profound layers. Based on histology 144 larvae were identified and their location was noted. The highest number of larvae was observed in the JX sample as compared to J1 and J2 (P<0.001). Most of them were located in the profound crypt zone of the mucosa (51%) as compared to the other zones (P<0.05). The number of larvae was higher in the lumen (63%) compared to the epithelium (32%) and lamina propria (5%) (P<0.001). A significantly higher number of eosinophils were found in lamina propria of the infected group compared to the control group (P<0.001). This experiment clearly showed that only few larvae had penetrated the epithelium and were positioned in the lamina propria at 3 days post infection. It was far more common that the larvae were localized within the epithelium or in the lumen of the crypts. It is therefore suggested that at least in this early phase "mucosal phase" is a more appropriate term to be used for the A. galli larval localization as compared to the term "histotrophic phase" currently used in many textbooks.
    Veterinary Parasitology 10/2011; 185(2-4):186-93. DOI:10.1016/j.vetpar.2011.10.025 · 2.55 Impact Factor
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    ABSTRACT: Symptoms after human infection with the helminth Trichuris suis have not previously been described. Exposure to helminths has been suggested as immune therapy against allergy and autoimmune diseases. We randomized adults with allergic rhinitis to ingest a dose of 2500 T. suis eggs or placebo every 21 days for 168 days (total 8 doses) in a double-blind clinical trial. In a previous publication, we reported a lack of efficacy and a high prevalence of adverse gastrointestinal reactions. The aim of the present study was to present a detailed description of the adverse event data and post-hoc analyses of gastrointestinal reactions. Adverse events and severity (mild, moderate, severe) were recorded daily by subjects, classified by organ using MedDRA 10.0, and event rates compared between subjects on T. suis treatment vs. subjects on placebo. T. suis-specific serum IgG antibodies were measured by a fluoroenzymeimmunoassay (Phadia ApS). During 163 days complete follow-up, subjects ingesting T. suis eggs (N = 49) had a three to 19-fold higher rate of events (median duration, 2 days) with gastrointestinal reactions (moderate to severe flatulence, diarrhea, and upper abdominal pain) compared with placebo subjects (N = 47). The highest incidence of affected subjects was seen from the first few days and until day 42 (3rd dose): 63% vs. 29% for placebo; day 163: 76% vs. 49% for placebo. Seroprevalences increased concurrently in the T. suis group: Day 59, 50%; day 90, 91%; day 170, 93%. The combined duration of episodes with onset before day 42 was ≤14 days in 80% of affected subjects. Age, gender, total IgE, and recent intestinal symptoms at baseline did not predict gastrointestinal side effects. In conclusion, during the first 2 months, repeated ingestions of 2500 T. suis eggs caused frequent gastrointestinal reactions lasting up to 14 days, whereas 4 months further treatment mainly provoked a subclinical stimulation. Trial registration University hospital Medical Information Network trial registry Reg. no. R000001298, Trial ID UMIN000001070.
    PLoS ONE 08/2011; 6(8):e22346. DOI:10.1371/journal.pone.0022346 · 3.53 Impact Factor

Publication Stats

2k Citations
276.72 Total Impact Points

Institutions

  • 2007–2015
    • IT University of Copenhagen
      København, Capital Region, Denmark
  • 2004–2009
    • Åbo Akademi University
      • Department of Biology
      Turku, Varsinais-Suomi, Finland
  • 2003
    • Universidad de Extremadura
      • Facultad de Veterinaria
      Cáceres, Extremadura, Spain
  • 1989–1998
    • Royal Agricultural University
      Cicester, England, United Kingdom
  • 1997
    • University of Tartu
      Dorpat, Tartu, Estonia