Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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    Nosema apis en Nosema ceranae. Achtergrondinformatie Nosema
    Steen, J.J.M. van der - \ 2010
    Wageningen : Plant Research International
    honingbijen - apis mellifera - nosema apis - bijenziekten - infecties - insectenpathogene protozoën - nosemaziekte - protozoëninfecties - honey bees - apis mellifera - nosema apis - bee diseases - infections - entomopathogenic protozoa - nosema disease - protozoal infections
    Nosema apis en Nosema ceranae komen beide voor in de volwassen honingbij Apis mellifera. Beide nosema’s worden zowel apart als samen in de honingbij gevonden. Tegenwoordig komt N. ceranae meer voor dan N. apis. Omdat in Noord Europa, de schadelijke gevolgen van de N. ceranae besmettingen min of meer overeen komen met die van N. apis worden bij de preventie en bestrijding de twee soorten vooralsnog gelijk behandeld. Wanneer gesproken wordt over nosema worden hiermee beide nosemasoorten samen bedoeld Achtergrondinformatie: Nosema apis en Nosema ceranae komen beide voor bij A. mellifera. De eerste beschrijvingen van de parasiet N. apis bij Apis mellifera en de gevolgen van deze parasitering zijn 100 jaar geleden beschreven door Zander [18]. Besmettingen van A. mellifera met N. ceranae zijn van een meer recente datum. Fries et al. [6] hebben N. ceranae beschreven. Dat N. ceranae voorkomt bij de Europese honingbij en N. apis ook de Aziatische honingbij kan infecteren weten we sinds een paar jaar. Higes et al. [9] ontdekten in 2006 als eerste het voorkomen van N. cerana in de Europese honingbij in Spanje. N. ceranae komt al langer, zeker sinds 1998, bij de Europese honingbij voor zonder dat dit opgemerkt was [13]. In Nederland heeft R. van der Zee in 2007 de eerste N. ceranae infectie op Terschelling aangetoond. Beide parasieten kunnen op een bijenstand tegelijk voorkomen zoals in Nederland aangetoond is bij de landelijke monitoring van PRI bijen in 2008. Hieruit bleek dat N. apis op 10% van de standen voorkomt, N. ceranae op 87% en een combinatie van beide nosemasoorten is vastgesteld op 6% van de 170 bemonsterde bijenstanden. Op een aantal standen werd geen nosema gevonden
    Infections with helminths and/or protozoa in cats in animal shelters in the Netherlands
    Robben, S.R. ; Nobel, W.E. le; Dopfer, D.D.V. ; Hendrikx, W.M. ; Boersema, J.H. ; Fransen, F. ; Eysker, M. - \ 2004
    Tijdschrift voor Diergeneeskunde 129 (2004)1. - ISSN 0040-7453 - p. 2 - 6.
    katten - gezelschapsdieren - protozoëninfecties - protozoa - helminthosen - kattenziekten - infectieziekten - cats - pets - protozoal infections - protozoa - helminthoses - cat diseases - infectious diseases - dogs
    To determine the prevalence of infections with helminths and protozoa in cats in animal shelters, faecal samples from 305 cats from 22 animal shelters in the Netherlands were examined, using a centrifugation-sedimentation-flotation-technique. The association between potential risk factors and the occurrence of an infection was also tested. Infections with helminths and/or protozoa were found in 160 samples (52.5%). Toxocara cati was found in 86 cats (28.2%), Cystoisospora felis in 59 cats (19.3%), Cystoisospora rivolta in 43 cats (14.1%), Capillaria spp. in 34 cats (11.2%), Ancylostoma tubaeforma in 9 cats (3.0%), Taenia taeniaeformis in 9 cats (3.0%), Aelurostrongylus abstrusus in 8 cats (2.6%), Giardia intestinalis in 3 cats (1.0%), Dipylidium caninum in 2 cats (0.7%) and Toxoplasma gondii in 1 cat (0.3%). The highest prevalence was seen in kittens and stray cats. The main preventive factor against infection was a short stay in a shelter.
    Development of a novel subunit vaccine against East Coast fever based on the Theileria parva sporozoite surface protein p67
    Kaba, S.A. - \ 2003
    Wageningen University. Promotor(en): Just Vlak; R.W. Goldbach, co-promotor(en): Monique van Oers. - [S.l.] : S.n. - ISBN 9789058088895 - 120
    theileria parva - rundvee - protozoëninfecties - sporozoïten - oppervlakteantigenen - vaccinontwikkeling - infectieziekten - theileria parva - cattle - protozoal infections - sporozoites - surface antigens - vaccine development - infectious diseases
    Theileriaparva is an intracellular protozoan parasite and the causative agent of a lethal cattle disease, called East Coast fever (ECF). This disease poses a major constraint on improvement of cattle production in Eastern, Central andSouthern Africa, especially for smallholder farmers. The protozoa are transmitted to cattle in the form of sporozoites by the brown-ear tick, Riphicephalus appendiculatus . The sporozoites invade lymphocytes, where they develop into schizonts. In addition, they induce a large-scale uncontrolled proliferation of the lymphocytes, leading to severe clinical symptoms, like weight loss, pyrexia, anaemia, terminal respiratory distress and finally death ensues within two to three weeks, if the animal is not treated. The disease can be cured with antibiotics, and this is the basis for the current method of vaccination, called "Infection and Treatment", where animals are injected with T. parva sporozoites and are, simultaneously, treated with antibiotics over a longer period of time. This vaccination method, however, is far from convenient, since the production of large amounts of sporozoites is very time consuming and a cold environment is needed to keep the sporozoites alive. In addition, the use of a live vaccine has pertinent risks, especially when the instructions for antibiotic treatment are not strictly followed and the immunity engendered is strain-specific. 

    The research described in this thesis was aimed at the possibilities of developing a subunit vaccine against East Coast fever, based on the production of T. parva sporozoite surface major protein p67. This protein is present on the outside of sporozoites and plays a crucial role in the entry of sporozoites into lymphocytes and is the major antigen producing neutralising antibodies. The first objective was to produce large amounts of p67 in a near-authentic conformation. Production of recombinant p67 in bacterial expression systems had failed to produce correctly processed protein and large amounts were needed to achieve a reasonable (70 %) level of protection. The baculovirus-insect cell expression system forms a valuable alternative for the expression of large amounts of near-authentic and immunologically active proteins. Previous attempts, however, to produce p67 in insect cells resulted in low levels of recombinant protein, which had a conformation different from the native p67 protein. Again large quantities were needed to protect cattle against ECF.

    In the research described in thesis several types of novel baculovirus vectors were constructed to produce different regions of p67 in insect cells. In the first set of vectors, various domains of p67 were expressed as separate entities, but this resulted in low levels of expression. For the second set, domains of p67 were fused to the carboxy-terminus of the "green fluorescent protein" (GFP), a visible marker, leading to a considerable increase in yield of recombinant p67. In addition, GFP:p67 fusion polypeptides were recognised by a monoclonal antibody (TpM12), which was raised against native p67 and capable of neutralising sporozoites. On the contrary, only a small portion of full length, non-fused p67 expressed in insect cells was recognised by this antibody. Fusion to GFP, thus, appeared to increase the stability of p67 and to result in a more native configuration of the recombinant protein. In a third set of baculovirus vectors, N and C terminal domains of p67 were fused to the baculovirus envelope protein GP64. This resulted in the display of recombinant p67 on the outside of insect cells as well as on the surface of budded baculovirus particles. The TpM12 epitope was also conserved when p67 was fused to GP64.

    P67 could also be expressed as a secreted soluble protein. The rationale behind this experiment was to ultimately facilitate the purification of the recombinant protein. This was achieved by removal of a putative transmembrane domain and fusion of p67 to a specific signal peptide derived from honeybee melittin. Deletion of the viral genes, chitinase and v-cathepsine from the baculovirus genome enhanced the integrity and increased the stability of this secreted p67 protein. Unfortunately, the secreted form was no longer recognised by TpM12, and hence, had a conformation different from p67 in sporozoites. Therefore, the secreted p67 was not tested in further immunological studies.

    In order to select the best recombinant p67 products for extensive vaccine trials, the various fusion proteins combining domains of p67 with GFP or GP64 were tested in mice for their immunogenicity and, especially, the ability to induce neutralising antibodies. In mice, the p67 molecule, lacking both its signal peptide and transmembrane region, and fused to GFP (GFP:p67ΔSS) gave the best humoral immune response, followed by the p67 C-terminal domain coupled to GP64 (GP64:p67C). These two immunogens were tested in cattle, in combination with a water-in-oil or a saponin-based adjuvant. Also in cattle, a high level of sero-conversion was obtained using a total of 100 µg recombinant p67 for immunisation divided over two needle injections. Moreover, the antisera raised in mice and cattle neutralised the infectivity of T. parva sporozoites in an in-vitro assay. Subsequently, in Kenya Boran cattle were vaccinated with GFP:p67DSS or with GP64-p67C. After a primary immunisation followed by a single booster, T. parva stabilated sporozoites were injected to test whether the vaccines protected the animals from ECF. Eighty five percent of the animals was protected from the lethal disease (ECF) using a much lower dose of recombinant protein than was used in the earlier studies.

    The research described in this thesis exploited the versatility of the baculovirus-insect cell expression system and showed that an ECF subunit vaccine based on recombinant p67, in a better conformation and formulated in an optimal adjuvant, can be used effectively in a vaccination program. Both of the proteins tested are good candidates for the development of a commercial ECF subunit vaccine and may contribute substantially to improvement in cattle productivity and poverty alleviation in sub-SaharanAfrica

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