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At least 19 recordsLinked to original sources
PubMed2004

Immunomagnetic separation of pathogenic organisms from environmental matrices.

One of the most difficult challenges in the analysis of environmental samples is to separate the organism of interest from a sample that is high in background debris. Immunomagnetic separation (IMS) is one technique that has been developed to accomplish this in a rapid and reliable assay. Immunomagnetic separation (or biomagnetic separation) involves a superparamag-netic, monodispersed, polystyrene microsphere that is coated with a specific ligand. When added to a heterogeneous target suspension, the microspheres bind to the desired target. Using a powerful magnet, the microsphere-target complex is then removed from the suspension. Many different targets of interest can be isolated with this technique, including fungal/bacterial cells or spores, protozoan parasites, cellular and subcellular material, proteins, and nucleic acid products. This wide range of application makes IMS one of the most versatile techniques available for the purification of target products from heterogeneous sample matrices.

Animals↗
PubMed2002

The sequence of regulatory events in the sporulation control network of Physarum polycephalum analysed by time-resolved somatic complementation of mutants.

The developmental decision for sporulation of Physarum polycephalum plasmodia is under sensory control by environmental factors like visible light or heat shock and endogenous signals like glucose starvation. Several hours after perceiving an inductive stimulus, plasmodia become committed to sporulation; thereby, they lose their unlimited replicative potential and execute a developmental program that involves differentiation into various cell types required to form a mature fruiting body. Plasmodia are multinuclear single cells which spontaneously fuse upon physical contact. Fusion of mutant plasmodia and cytoplasmic mixing allows complementation studies to be performed at the functional level. Mutant cells altered in their ability to sporulate in response to phytochrome activation by far-red light were cured by fusion with wild-type or other mutant plasmodia. Phytochrome activation in one plasmodium and subsequent fusion with a non-induced plasmodium revealed that complementation of the two mutations depended on (i) which of two genetically distinct plasmodial cells was stimulated; and (ii) on the delay time elapsed between stimulation and cytoplasmic mixing. Such experiments allow us to determine the kinetics and the causal sequence of the regulatory events tagged by mutation.

Animals↗
PubMed2006

Infectivity of Theileria parva sporozoites following cryopreservation in four suspension media and multiple refreezing: evaluation by in vitro titration.

Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59% and 67%, respectively. In a second experiment, a stabiiate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35% per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35% loss of infectivity.

Animals↗
PubMed2005

The Bacillus subtilis spore coat provides "eat resistance" during phagocytic predation by the protozoan Tetrahymena thermophila.

Bacillus spores are highly resistant to many environmental stresses, owing in part to the presence of multiple "extracellular" layers. Although the role of some of these extracellular layers in resistance to particular stresses is known, the function of one of the outermost layers, the spore coat, is not completely understood. This study sought to determine whether the spore coat plays a role in resistance to predation by the ciliated protozoan Tetrahymena, which uses phagocytosis to ingest and degrade other microorganisms. Wild-type dormant spores of Bacillus subtilis were efficiently ingested by the protozoan Tetrahymena thermophila but were neither digested nor killed. However, spores with various coat defects were killed and digested, leaving only an outer shell termed a rind, and supporting the growth of Tetrahymena. A similar rind was generated when coat-defective spores were treated with lysozyme alone. The sensitivity of spores with different coat defects to predation by T. thermophila paralleled the spores' sensitivities to lysozyme. Spore killing by T. thermophila was by means of lytic enzymes within the protozoal phagosome, not by initial spore germination followed by killing. These findings suggest that a major function of the coat of spores of Bacillus species is to protect spores against predation. We also found that indigestible rinds were generated even from spores in which cross-linking of coat proteins was greatly reduced, implying the existence of a coat structure that is highly resistant to degradative enzymes.

Animals↗
PubMed2003

Ultrastructural studies on the development of Enteromyxum scophthalmi (Myxozoa), an enteric parasite of turbot (Scophthalmus maximus L.).

The ultrastructure of the developmental stages of Enteromyxum scophthalmi is described. Scarce intracellular, early uninucleated stages appeared within intestinal epithelial cells whereas proliferative stages were abundant both intraepithelially and in the intestinal lumen. In the proliferative stages, food reserves were abundant in the cytoplasm of P cells and consisted mostly of carbohydrates in the intraepithelial stages and lipid inclusions in the luminal stages. Sporogenesis could occur in enveloped cells or by direct division or clustering of generative cells. The abundance, shape and size of mitochondria as well as the number and shape of their cristae were very variable in the different developmental stages. The cristae were usually tubular and sometimes plate-like, discoidal or lamellar. True flat cristae were not observed. We found elements of closed (cryptomitosis) and open mitosis as well as structures reminiscent of microtubule organising centres, hitherto not described in myxosporeans. The significance of these findings is discussed in relation to the taxonomic and phylogenetic position of the Myxozoa.

Animals↗
PubMed2004

Ultrastructural characteristics of the sporogenesis of genus Myxobolus infecting some Nile fishes in Egypt.

This study presents the ultrastructural characteristics of sporogenesis in the genus Myxobolus which infects four economically important Egyptian fishes: Oreochromis niloticus niloticus, Barbus bynni, Labeo niloticus and Hydrocynus forskalii, specimens of which were collected at Giza city. Although the cysts were observed in different organs, the pattern of sporogenesis was the same. Moreover, the sporogenesis found in the present study followed the usual pattern valid for most of the myxosporean genera studied.

Animals↗
PubMed2006

Differential display analysis of gene expression in two immunologically distinct strains of Eimeria maxima.

Gene expression during sporulation and sporozoite excystation of two strains of Eimeria maxima was analyzed using the mRNA differential display technique. The two strains, the Guelph strain (GS) and a single sporocyst-derived strain (M6) from Florida, have been shown to be immunologically distinct. We isolated and cloned a 453-bp complimentary DNA (cDNA) fragment (GS-453) found only in GS sporozoites. In GS, this mRNA begins to be expressed during the earliest stages of oocyst sporulation and is continuously expressed up to and including in the excysted sporozoite. In all Northern blots, digoxigenin (DIG)-labeled GS-453 probe recognized an mRNA of approximately 1.6 kb from GS but not from RNA of M6. Southern blots using various endonucleases and probed with DIG-labeled GS-453 demonstrated that the genomes of both strains contained sufficiently similar sequences to permit hybridization with the probe, but the pattern of hybridization differed between the two strains. Extensive searches of the GenBank, European Molecular Biology Laboratory, and various apicomplexan expressed sequence tag databases using the DNA or inferred amino acid sequences of GS-453 cDNA clone did not identify similarity to any existing sequences.

Animals↗
PubMed2006

The effect of cohabitation of Tubifex tubifex (Oligochaeta: Tubificidae) populations on infections to Myxobolus cerebralis (Myxozoa: Myxobolidae).

The competitive interactions between susceptible and resistant Tubifex tubifex (Oligochaeta: Tubificidae) exposed to Myxobolus cerebralis (Myxozoa: Myxobolidae) infections were investigated in two laboratory trials. Competition was assessed by the total parasite production over the course of the trials in mixed and pure cultures of M. cerebralis exposed worms, and by the genetic analyses of worms from the control and experimental groups at the beginning and end of the experiments. Mixed cultures of resistant and susceptible worms showed a 70% reduction in production of parasites released when compared with pure cultures of susceptible worms. In studies with laboratory and field-collected oligochaetes the mixed cultures at the end of the cohabitation experiments were dominated by resistant Tubifex from lineage V (HB strain) this strain of Tubifex has a competitive advantage over worms from other lineages. The results of this study suggest that certain species of Tubifex may be dead-end hosts to M. cerebralis by absorbing or inactivating the parasite and may also show greater survival compared to susceptible oligochaetes in certain whirling disease enzootic habitats.

Animals↗
PubMed2004

[Three new species of Myxosporidia (Myxozoa: Myxosporea) parasites of Citharinus citharus (Geoffroy Saint-Hilaire, 1809) (Citharinidae) in Chad (Central Africa)].

The study of Citharinus citharus (Geoffroy Saint-Hilaire, 1809) (Teleost, Citharinidae) captured in the Chari river in Chad revealed the presence of three new myxosporidian which are described in the present paper: Thelohanellus taguii sp. nov., a systematic parasite; Myxobolus mbailaoi sp. nov., which affects operculum, skin and intestine, Myxobolus charii sp. nov., a parasite of the skin.

Animals↗
PubMed2005

Characterization of ichthyocidal activity of Pfiesteria piscicida: dependence on the dinospore cell density.

The ichthyocidal activity of Pfiesteria piscicida dinospores was examined in an aquarium bioassay format by exposing fish to either Pfiesteria-containing environmental sediments or clonal P. piscicida. The presence of Pfiesteria spp. and the complexity of the microbial assemblage in the bioassay were assessed by molecular approaches. Cell-free water from bioassays that yielded significant fish mortality failed to show ichthyocidal activity. Histopathological examination of moribund and dead fish failed to reveal the skin lesions reported elsewhere. Fish larvae within "cages" of variable mesh sizes were killed in those where the pore size exceeded that of Pfiesteria dinospores. In vitro exposure of fish larvae to clonal P. piscicida indicated that fish mortality was directly proportional to the dinospore cell density. Dinospores clustered around the mouth, eyes, and operculi, suggesting that fish health may be affected by their direct interaction with skin, gill epithelia, or mucous surfaces. Molecular fingerprinting revealed the presence of a very diverse microbial community of bacteria, protists, and fungi within bioassay aquaria containing environmental sediments. Some components of the microbial community were identified as potential fish pathogens, preventing the rigorous identification of Pfiesteria spp. as the only cause of fish death. In summary, our results strongly suggest (i) that this aquarium bioassay format, which has been extensively reported in the literature, is unsuitable to accurately assess the ichthyocidal activity of Pfiesteria spp. and (ii) that the ichthyocidal activity of Pfiesteria spp. is mostly due to direct interactions of the zoospores with fish skin and gill epithelia rather than to soluble factors.

Animals↗
PubMed2002

Myxobolus species (Myxozoa), parasites of fishes in the Okavango River and Delta, Botswana, including descriptions of two new species.

Fieldwork was conducted in 1998 and 1999 in the Okavango River and Delta and a total of 275 fishes representing 31 species were examined for the presence of myxosporean parasites. A total of seven myxosporeans of the genus Myxobolus Bütschli, 1882 were found infecting the fishes. Two new species namely Myxobolus etsatsaensis sp. n. from Barbus thamalakanensis Fowler, 1935 and M. paludinosus sp. n. from Barbus paludinosus Peters, 1852 are described. Myxobolus africanus Fomena, Bouix et Birgi, 1985, M. camerounensis Fomena, Marqués et Bouix, 1993, M. hydrocyni Kostoïngue et Toguebaye, 1994, M. nyongana (Fomena, Bouix et Birgi, 1985) and M. tilapiae Abolarin, 1974 are recorded for the first time in Botswana and descriptions of these species are provided.

Animals↗
PubMed2006

The myxozoan fauna of Fundulus diaphanus (Cyprinodontidae) from freshwater localities in eastern North America: prevalence, community structure, and geographic distribution.

Membership and richness of infracommunities and component communities of myxozoan fauna of the banded killifish (Fundulus diaphanus) from freshwater localities in Ontario, Quebec, New York State, New Brunswick, Nova Scotia, and Maryland were studied. Five species of parasites were found: Myxobolus diaphanus (Fantham, Porter, and Richardson, 1940) (connective tissue throughout the body and head), Myxobolus funduli (Kudo, 1918) (interlamellar), Myxobolus neurophilus (Guilford, 1963) (optic tectum of the brain), Myxobolus sp. (connective tissue, typically adjacent to vertebrae), and Sphaerospora sp. (kidney tubules). The most abundant species locally and regionally was M. diaphanus, occurring at prevalences of 14.2 to 93.3% at 6 of 9 localities. Myxobolus funduli and Myxobolus sp. were at 3 and 2 localities respectively, while M. neurophilus and Sphaerospora each occurred at single localities. Four of the 5 myxozoans appear to be specific to fundulids, the exception being M. neurophilus, which is typically a parasite of Perca flavescens. Mean infracommunity richness was 0-1.2. Component community richness was 0-3 species. The fauna is similar in composition to that described from the spottail shiner (Notropis hudsonius) in the Great Lakes in being dominated by histozoic myxobolids and in having maximum prevalence at any single locality correlate positively with geographical distribution. Moreover, mean infracommunity richness was correlated with percentage of hosts infected with any species at a locality, and maximum infracommunity richness was correlated with component community richness. Probably because fewer species of myxozoans of fundulids occur in the regional pool, myxozoan communities encountered in the present study are generally less rich than those described from N. hudsonius. It appears that dispersal of relatively resilient myxospores through such a mechanism as piscivory effectively distributes these parasites over the landscape, while the more delicate actinospores serve to ensure colonization by amplifying species' prevalence at a specific locality and thereby contributing to initial establishment. As such, these types of myxozoans, though they are autogenic, having their entire life cycle normally completed within the aquatic environment, behave more like allogenic parasites that rely on birds and mammals as definitive hosts.

Animals↗
PubMed2005

Effects of fish age versus size on the development of whirling disease in rainbow trout.

We examined the effects of both fish age and size on the development of resistance to whirling disease in Erwin strain rainbow trout. Previously, we demonstrated that juvenile rainbow trout became resistant to development of the disease when first exposed to triactinomyxons of the parasite Myxobolus cerebralis at about 9 wk post-hatch when raised at 12 degrees C, but ages and sizes of fish used in that experiment were confounded (Ryce EKN, Zale AV, MacConnell E [2004] Dis Aquat Org 59:225-233). In this study, rainbow trout of the same age and different sizes, and the same size and different ages, were exposed to the parasite to distinguish the influences of age and size. Fish were reared at 3 different water temperatures prior to exposure to produce groups with different growth rates and were exposed to the parasite at 7 or 9 wk post-hatch. Disease severity was affected by both age and size at first exposure, but the effects were not independent. An increase in fork length from 36 to 40 mm among fish exposed at 7 wk post-hatch did not confer increased resistance, but the same increase in size at 9 wk post-hatch did. Similarly, an increase in age from 7 to 9 wk post-hatch among fish exposed at 36 mm fork length did not confer increased resistance, but the same increase in age at 40 mm did. Rainbow trout must be both 9 wk post-hatch or older and at least 40 mm in fork length at time of exposure to exhibit enhanced resistance to whirling disease. Resistance to disease was not associated with the level of skeletal ossification.

Age Factors↗
PubMed2006

Myxozoan pathogens in cultured Malaysian fishes. II. Myxozoan infections of redtail catfish Hemibagrus nemurus in freshwater cage cultures.

Cage-cultured Asian redtail catfish Hemibagrus nemurus (Valenciennes, 1840), a popular food fish in Southeast Asia, proved to be infected by 3 myxozoan species. All the 3 species belonged to the genus Henneguya: 2 were identified as H. mystusia Sarkar, 1985 and H. hemibagri Tchang et Ma, 1993, while the other was described as H. basifilamentalis sp. n. All plasmodia were found in the gills and were characterised by a specific site selection. H. mystusia formed plasmodia in the multi-layered epithelium between the gill lamellae and in the non-lamellar edge of the gill filaments, while H. hemibagri developed in the capillary network of the lamellae. H. basifilamentalis sp. n. had large oval plasmodia located deep among the filaments just above the gill arch.

Animals↗