Description of Eimeria vejdovskyi sp.n. and redescription of Eimeria media Kessel, 1929 from the rabbit

On the basis of oocyst morphology, endogenous cycles, prepatent period, and enzyme analysis, Eimeria media was found to be a composite species, representing two independent species differing from all other species of rabbit coccidia. One of them was designated Eimeria media Kessel, 1929 and the other was described as a new species, Eimeria vejdovskyi. E. media has relatively larger oocysts (mean size 32.88 X 19.19 micron), the endogenous development takes place in the epithelium of the ileum or of posterior jejunum, and the developmental stages gradually migrate from the crypts towards the tips of villi. E. media forms three asexual generations, the prepatent period is 9 days. Eimeria vejdovskyi sp.n. has relatively smaller oocysts (29.05 X 18.18 micron) and is localized throughout the small intestine. E. vejdovskyi sp.n. forms two asexual generations, which are localized mostly in the upper part of villi, like the gametogony. At the beginning of the endogenous development the parasites were localized mostly in the epithelium, while the second asexual generation and gametogony took place also in the lamina propria. The prepatent period was 6 days. It was found that the two species differ from one another in the electrophoretic mobility of lactate dehydrogenase and glucosophosphate isomerase.     

The life cycle of Eimeria danailovi from ducks.

The life cycle of Eimeria danailovi Gr?fner, Graubmann et Betke in experimentally infected ducks was studied by optical microscopy. The asexual generation developed in the posterior part of jejunum and in the whole ileum. The sexual stages occurred in jejunum and ileum, and, in addition, in cecum and colon. All endogenous stages were localized in the cytoplasm of epithelial cells in the apical and basal parts of villi. Two generations of meronts were found to develop, differing from one another in the number of merozoites. The meronts of the first generation were observed 2 days post infection (DPI) and contained 10-14 (on the average 11) merozoites. The second generation of meronts, containing 12-22 (on the average 16) merozoites, developed on 3 DPI. The sexual stages were found in histological preparations on 5 and 6 DPI. They appeared in the faces of experimentally infected ducks first on 5 DPI and they were shed for three days. Oocyst sporulation at room temperature lasted 2-3 days.     

Evaluations of antibody levels in Toxoplasma infection by the immunofluorescent antibody test and ELISA test

In this study 292 sera were screened by the IFAT. 46 sera with variable IFAT titres were tested with ELISA Reagent Set and microtitration tests. A comparative evaluation of the specificity and reliability of ELISA method with that of IFAT for the detection of Toxoplasma infection was done.     

Differentiation of species of Eimeria from the fowl using a computerized image-analysis system

Oocysts of Eimeria species from the fowl have been identified using a computerized image-analysis system (Leitz T.A.S. Plus Image-Analyser). The system enabled semiautomatic measurement of oocyst dimensions with subsequent species of diagnosis based on graphic statistical evaluation of size and shape of measured parasites. E. mitis, E. acervulina, E. brunetti, E. maxima, E. tenella and complex of E. necatrix and E. praecox were distinguishable both in pure cultures and in mixtures. It was not possible to distinguish E. praecox from E. necatrix using this system.     

An evaluation of a virobacterial agglutination test for the detection of plant viruses

Agglutination of Staphylococcus aureus bacteria particles, conjugated with specific virus antibodies, has been used to identify a wide range of plant viruses in crude sap extracts. The test distinguishes between seven different potyviruses in homologous and heterologous reactions, but does not distinguish different strains of bean yellow mosaic virus. The sensitivity of the virobacterial agglutination (VBA) test compares favourably with virus detection in ISEM particle trapping and local lesion assay and is only slightly less sensitive than direct ELISA tests. The test is more sensitive and simpler to user than latex particle agglutination tests.     

Development of a recombinant antibody ELISA test for the detection of Polymyxa betae and its use in resistance screening

An ELISA test was developed for the quantitative detection of the obligate parasite Polymyxa betae , the vector of Beet necrotic yellow vein virus (BNYVV), in infected sugarbeet roots. The test used monoclonal and polyclonal antibodies raised to a recombinantly expressed glutathione-S-transferase (GST) from P. betae . A close correlation was found between the number of P. betae zoospores in serially diluted suspensions and absorbance values in the ELISA test. Time-course studies of plants grown in naturally infested soils in controlled environment tests demonstrated the value of the ELISA test in screening for P. betae resistance. In preliminary tests, P. betae -resistant accessions of the wild sea beet ( Beta vulgaris ssp. maritima ), which might be used to restrict the transmission of BNYVV, were identified.     

Enzyme variants of Eimeria parasitizing the domestic fowl and possibilities of species diagnostics.

Electrophoretic variation of the enzymes lactate dehydrogenase (LDH) and glucosephosphate isomerase (GPI) of Eimeria parasitizing the domestic fowl in Czechoslovakia is summarized and the differentiation of species of poultry coccidia is discussed. A new method for evaluation of zymograms of coccidial enzymes is presented. This method enables the results of different experiments to be compared by calculating standardized rates of mobility of each enzyme band relative to the positions of reference variants coded LDH-8 or GPI-9.     

Eimeria burdai sp. n. (Apicomplexa: Eimeriidae), a new parasite species from subterranean African silvery mole-rat, Heliophobius argenteocinereus

A new coccidian parasite of the genus Eimeria Schneider, 1875 is described from the subterranean African silvery mole-rat Heliophobius argenteocinereus Peters, 1846. Oocysts of Eimeria burdai sp. n. were subspherical to broadly ellipsoidal 17.8 (16-19) x 14.1 (12-15), with a shape index 1.2 (1.1-1.4). Oocyst wall was bilayered, smooth and colourless, approximately 1.0 thick. Outer layer was significantly thicker than inner one. A micropyle and oocyst residuum were absent. One or two ellipsoidal or spherical polar granules were present. Sporocysts were ellipsoidal, 10.8 (9-12) x 6.2 (5-8) with a shape index 1.7 (1.5-1.9). Sporocyst wall was single-layered, thin, smooth and colourless, with small Stieda body at the pointed end. In freshly sporulated oocysts, spherical sporocyst residuum was composed of small granules enclosed by a thin membrane. Sporozoites were elongate, lying length-wise in the long axis of the sporocyst, partially curled around each other, with single large refractile body located posteriorly. Faintly distinguishable nucleus was in the central part of the sporozoite. This eimerian represents the first coccidian species described from subterranean African silvery mole-rat (Rodentia: Bathyergidae).     

The endogenous development, described by light and electron microscopy, of Eimeria jamescooki sp. n. (Apicomplexa: Eimeriidae) from the skink Cryptoblepharus virgatus

Eimeria jamescooki sp. n. was recovered from the skink Cryptoblepharus virgatus (Garman) found on the grounds of James Cook University, Townsville (type locality), North Queensland, Australia. Oocysts were 17.5-25.0 (22.1 +/- 1.9) x 15-22.5 (17.7 +/- 1.6) microm and sporocysts 6.25-10.0 (7.9 +/- 1.15) x 3.75-6.25 (5.3 +/- 1.0) microm in size. Endogenous stages are described from histological material examined by light microscope and by transmission electron microscope. Both merogony stages and gamonts were found to develop in the cytoplasm of the anterior gut mucosal epithelium. Meront progeny were comprised of 10 to 21 merozoites. Premature macrogamonts were elongate; some host cells contained two elongate macrogamonts. Unique to the presently described species were the Golgi "plaques" and an enclosure of tubuli. Mature macrogamonts and young oocysts ranged in size from 14 x 7 to 21 x 11 microm and contained two types of wall-forming bodies, canaliculi and amylopectin granules. Differentiating microgamonts conformed in fine structure with that observed in other eimerians. Their sizes increased from 15.4 x 4.2 to 28 x 8.4 microm while dividing to over 70 nuclei, which formed a corresponding yield of microgametes.     

An in vitro feeding assay to test acaricides for control of hard ticks

Animal husbandry could not be practised over large areas of the planet without acaricides. The prevention of tick bite and the transmission of diseases requires the use of pesticides, but this contributes to the development of tick resistance against acaricides. This drives the quest for new molecules that target physiological processes crucial to tick survival. In vivo trials involve multiple repetitions because of inherent variations between host animals, requiring large amounts of test products and ticks. An in vitro alternative should permit the testing of the ability of a product to restrict attachment and feeding by ticks at precise doses. In this paper an in vitro feeding system is described where the European tick Ixodes ricinus L. feeds on blood through a cellulose rayon-reinforced silicone membrane. The membrane Shore hardness is modified to imitate the elastic retraction forces of skin that ensure the closing of tick penetration sites on the membrane to prevent bleeding. Tick attachment (75-100%) is achieved by adding chemical and mechanical stimuli to the membrane. Survival curves for different doses of fipronil and ivermectin tested with the method showed highly reproducible acaricide effects within 5-7 days. Significant effects are recorded down to ppb levels in blood. Standardised tests can be made with blood from the same donor animal or culture medium under the membrane.     

Results and experiences from the first EU proficiency test for the detection of Phytophthora ramorum1

The Crop Protection unit of the Institute for Agricultural and Fisheries Research offers plant disease diagnostic services through its ‘Diagnostic Centre for Plants’ (ILVO‐DCP). ILVO‐DCP has requested accreditation (ISO 17025) for a number of diagnostic detection methods in bacteriology, mycology, entomology and nematology. Accreditation forms an essential part of quality control programs such as ISO17025, which can in part be realized by proving the laboratory's competence in inter‐laboratory proficiency or ring tests. In 2006, ILVO‐DCP organized such a proficiency test for the detection of Phytophthora ramorum. The protocol was developed using the standard ILAC‐G13:2000, and defined rules for participation, sample preparation and transport, communication, fraud prevention and reporting. Eight European diagnostic centres participated in the proficiency test including ILVO‐DCP. Each participant received one set of 10 coded samples, each sample consisting of either leaves or stems that were artificially inoculated with either the target or an alternative organism. Participants could use one or more methods listed in the EPPO diagnostic protocol PM 7/66. They had to report within a specific timeframe and received a detailed report of their performance. The success rate of the proficiency test was 100%. This paper lists some of the experiences gained from organising this type of proficiency test.     

Description of Eimeria arabukosokokensis sp. n. (Apicomplexa: eimeriidae) from Telescopus semiannulatus (Serpentes: Colubridae) with notes on eimerian coccidia from snakes of Eastern Kenya

Parasitological examination of faeces of 26 snakes kept in Bio-Ken Snake Farm, Watamu, Kenya revealed new species of Eimeria Schneider, 1875 in Telescopus semiannulatus Smith, 1849. Oocysts of Eimeria arabukosokokensis sp. n. are cylindrical 26.8 (25-29) x 15.1 (14-16) microm with smooth, bilayered oocyst wall and a single polar granule. The broadly ellipsoidal sporocysts average 9.3 (8.5-10) x 7.1 (6.5-7.5) microm and possess single-layered wall composed of two plates joined by longitudinal suture. Caryospora cf. regentensis Daszak et Ball, 2001 is reported from Dendroaspis angusticeps (Smith, 1849) and two additional forms of Caryospora Léger, 1904 are reported and morphologically characterised from a single specimen of Psammophis orientalis Broadley, 1977. Systematic status of Caryospora spp. in sub-Saharan Psammophis Boie, 1827 is discusses and all species reported by various authors to date are suggested to be treated as species inquirendae until more detailed data on these parasites and their hosts are available.     

A description of two new species of coccidia (Apicomplexa: Eimeriidae) from African reptiles with nomenclatural corrections for two Caryospora and one Eimeria species from snakes

Two new species of coccidian parasites are described from African reptiles. Oocysts of Eimeria foulshami sp. n. from the plated lizard Gerrhosaurus major bottegoi Del Prato of Sudan are ellipsoidal, 24.1 x 14.9 (23-26.5 x 14-17.8) microm with a bilayered, colourless oocyst wall and lack polar granules. The ellipsoidal sporocysts average 8.6 x 4.6 (7-10.6 x 4.4-7) microm and possess a prominent, globular, sporocyst residuum. Oocysts of Caryospora regentensis sp. n. from the Eastern green mamba Dendroaspis augusticeps Smith, 1849 [corrected] of Kenya are spherical to subspherical, 16.8 x 16.4 (16-17.6 x 15-17.2) microm with a bilayered oocyst wall and a single polar granule. The ellipsoidal sporocysts average 13.0 x 10.3 (10.2-14 x 9.2-11) microm and possess a Stieda and substieda body and a prominent globular sporocyst residuum. Oocysts of Caryospora legeri Hoare, 1933 are reported from a hissing sand snake, Psammophis sibilans sibilans L. from Nigeria, representing a new geographical record. The oocysts are slightly larger than the type, but otherwise identical. Caryospora psammophi Bray, 1960 and C. hermae Bray, 1960 from Psammophis sibilans phillipsi, oocysts of which are morphologically similar to and overlap in dimensions with C. legeri Hoare, 1933, are synonymised with the latter species. Eimeria samiae Iskander et Tadros, 1979 is emended to E. samyadeli to reflect the gender of the person the species was named after and because E. sani is preoccupied. In addition to these findings, Eimeria bohemi Modry, Slapeta et Koudela, 2000 and oocysts of an unidentified spherical Eimieria sp. are reported from Chamaeleo dilepis dilepis Leach from Cameroon.