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PubMed2005

Glycosphingolipid structural analysis and glycosphingolipidomics.

Sphingosines, or sphingoids, are a family of naturally occurring long-chain hydrocarbon derivatives sharing a common 1,3-dihydroxy-2-amino-backbone motif. The majority of sphingolipids, as their derivatives are collectively known, can be found in cell membranes in the form of amphiphilic conjugates, each composed of a polar head group attached to an N-acylated sphingoid, or ceramide. Glycosphingolipids (GSLs), which are the glycosides of either ceramide or myo-inositol-(1-O)-phosphoryl-(O-1)-ceramide, are a structurally and functionally diverse sphingolipid subclass; GSLs are ubiquitously distributed among all eukaryotic species and are found in some bacteria. Since GSLs are secondary metabolites, direct and comprehensive analysis (metabolomics) must be considered an essential complement to genomic and proteomic approaches for establishing the structural repertoire within an organism and deducing its possible functional roles. The glycosphingolipidome clearly comprises an important and extensive subset of both the glycome and the lipidome, but the complexities of GSL structure, biosynthesis, and function form the outlines of a considerable analytical problem, especially since their structural diversity confers by extension an enormous variability with respect to physicochemical properties. This chapter covers selected developments and applications of techniques in mass spectrometric (MS) that have contributed to GSL structural analysis and glycosphingolipidomics since 1990. Sections are included on basic characteristics of ionization and fragmentation of permethylated GSLs and of lithium-adducted nonderivatized GSLs under positive-ion electrospray ionization mass spectrometry (ESI-MS) and collision-induced mass spectrometry (CID-MS) conditions; on the analysis of sulfatides, mainly using negative-ion techniques; and on selected applications of ESI-MS and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to emerging GSL structural, functional, and analytical issues. The latter section includes a particular focus on evolving techniques for analysis of gangliosides, GSLs containing sialic acid, as well as on characterizations of GSLs from selected nonmammalian eukaryotes, such as dipterans, nematodes, cestodes, and fungi. Additional sections focus on the issue of whether it is better to leave GSLs intact or remove the ceramide; on development and uses of thin-layer chromatography (TLC) blotting and TLC-MS techniques; and on emerging issues of high-throughput analysis, including the use of flow injection, liquid chromatography mass spectrometry (LC-MS), and capillary electrophoresis mass spectrometry (CE-MS).

Animals↗
PubMed2003

Echinococcosis.

Echinococcosis is a near-cosmopolitan zoonosis caused by adult or larval stages of cestodes belonging to the genus Echinococcus (family Taeniidae). The two major species of medical and public health importance are Echinococcus granulosus and Echinococcus multilocularis, which cause cystic echinococcosis and alveolar echinococcosis, respectively. Both are serious and severe diseases, the latter especially so, with high fatality rates and poor prognosis if managed incorrectly. Several reports have shown that both diseases are of increasing public health concern and that both can be regarded as emerging or re-emerging diseases. In this review we discuss aspects of the biology, life cycle, aetiology, distribution, and transmission of the Echinococcus organisms, and the epidemiology, clinical features, treatment, and diagnosis of the diseases they cause. We also discuss the countermeasures available for the control and prevention of these diseases. E granulosus still has a wide geographical distribution, although effective control against cystic echinococcosis has been achieved in some regions. E multilocularis and alveolar echinococcosis are more problematic, since the primary transmission cycle is almost always sylvatic so that efficient and cost-effective methods for control are unavailable.

Echinococcosis↗
PubMed2003

Human Taeniasis and cysticercosis in Asia.

CONTEXT: Human Taeniasis caused by the pork, Taenia solium, or beef, T saginata, tapeworm arises after eating pork or beef contaminated with metacestodes, the larval stage of these parasites. Taeniasis with T solium can lead to neurocysticercosis and threaten others by accidental ingestion of eggs released from asymptomatic Taeniasis patients. The 2003 World Health Assembly declared that T solium is of worldwide public-health importance, and that it is an eradicable parasitic disease worldwide. Adult taeniid tapeworms expelled from people in almost all Asian countries appeared to be T saginata (the so-called Asian Taenia), even though they ate pork. The organism is now named T asiatica, and has been found in Taiwan, Korea, China, Vietnam, and Indonesia. But it has been difficult to differentiate T saginata from beef and Asian Taenia from pork. STARTING POINT: Marshall Lightowlers and colleagues (Int J Parasitol 2003; 33: 1207-17) recently demonstrated that recombinant oncosphere vaccines against several taeniid cestodes, including T ovis, T saginata, T solium, and Echinococcus granulosus, are highly effective. Protection was almost 100%, in the laboratory and in the field. These researchers found several common features, including a predicted secretory signal sequence, and one or two copies of a fibronectin type III domain, each encoded by separate exons within the associated gene. WHERE NEXT? Molecular and immunological techniques, including vaccine research and development of animal models for differentiation of taeniid species in humans, have greatly advanced over the past decade. The clinical importance of infections by these taeniids, including T asiatica, in humans, and the potential for cysticercosis attributable to T asiatica in humans, needs further study.

Animals↗
PubMed1998

A mucosal IgA response, but no systemic antibody response, is evoked by intranasal immunisation of dogs with Echinococcus granulosus surface antigens iscoms.

The search for protective antigens of intestinal parasites is conditioned by the methodology used to induce a relevant local immune response against them. The present work describes the use of immuno stimulating complexes (iscoms) from tegumental antigens from protoscoleces (PSC) of the cestode Echinococcus granulosus as immunogens in dogs by the intranasal route. It also describes the evaluation of the immune response evoked at the antibody level (systemically and at a distant mucosal location) as well as at the level of antibody secreting cells in peripheral blood. Iscoms from both E. granulosus tegumental antigens and hen ovalbumin (OVA), given at 50 microg doses by intranasal route, evoked significant secretory IgA antibody responses detected in saliva. Specific IgA secreting cells in peripheral blood also increased 10-20-fold, although transiently, after primary and secondary stimulation, whereas specific IgG secreting cells in peripheral blood were only detected in some individuals after the second antigenic exposure. Generation of immune responses at a related mucosal site provides evidence of localised immunity. No significant increase in systemic antibody titers of either IgM, IgG or IgA isotype was detected in plasma as a result of the immunisation. This fact could reflect that the nasopharyngeal mucosal associated lymphoid tissue of dogs is more strictly compartmentalised than that of other mammals.

Adjuvants, Immunologic↗
PubMed2001

Alternative splicing and sequence diversity of transcripts from the oncosphere stage of Taenia solium with homology to the 45W antigen of Taenia ovis.

Genes and transcripts which show homology to the host-protective 45W antigen of Taenia ovis have been cloned from the human parasite Taenia solium. The T. solium genes cloned in this study (TSO45) show conserved genomic structural features which are also features of the T. ovis 45W gene family. The TSO45 genes consist of a four exon and three intron structure. Eight TSO45 transcripts, encoded by at least five genes, were cloned from T. solium oncospheres and comparison of their DNA sequence indicates that some transcripts have arisen by alternative splicing, the first demonstration of exon inclusion/exon skipping in cestodes. Alternative splicing occurred with respect to both exons II and III with three splice variants identified from the TSO45-1 gene and two splice variants from TSO45-5. The proteins encoded by this family of genes contain putative N-linked glycosylation sites, an amino terminal secretory signal, a hydrophobic carboxy terminal sequence characteristic of GPI-anchored proteins and fibronectin type III motifs. These features are common to their T. ovis and Taenia saginata homologues. The similarities of the TSO45 genes cloned in this study with genes encoding host-protective antigens of T. ovis and T. saginata indicates that the encoded T. solium proteins are quite possibly antigenic and have potential use as a vaccine to prevent T. solium infection in the parasite's intermediate host. In this respect, the generation of sequence diversity and hence potential antigenic diversity through alternative splicing of TSO45 genes may have implications for the use of these proteins in vaccines against T. solium cysticercosis.

Alternative Splicing↗
PubMed1997

Structural characterization of the N-glycans from Echinococcus granulosus hydatid cyst membrane and protoscoleces.

Infection by the tapeworm Echinococcus granulosus in the intermediate host results in the development of a hydatid cyst which contains the protoscoleces within a fluid-filled cavity enclosed by the bilayered cyst membrane. N-glycans were enzymatically released from crude extracts of homogenates of hydatid cyst membranes and protoscoleces and their structures were defined by high sensitivity fast atom bombardment mass spectrometry in conjunction with sequential exoglycosidase digestions. The major N-glycans from the cyst membrane were found to be non-charged structures having complex-type antennae and core fucosylation. The antennae are either truncated at the first N-acetylglucosamine or are extended with beta-galactose to form N-acetyllactosamine (lacNAc). A significant proportion of the lacNAc backbones are capped by alpha-galactose. The resulting Gal alpha-Gal beta-terminal structures may account for the earlier observation that antibodies against the blood group P1 epitope recognise components of hydatid cyst extracts. The complex-type N-glycans identified in the protoscoleces extracts were the same as the neutral structures found in the cyst membrane but a small proportion of high mannose structures and truncated di- and trimannosyl core structures were also identified. Sialylated N-glycans were identified as minor constituents of the cyst membrane preparation but were not observed in protoscoleces extracts. Whether the sialylated glycans are host derived or endogenously synthesized by the parasite remains to be established. This is the first reported structural analysis of N-glycans from cestodes and provides new insights into protein glycosylation in helminths.

Animals↗
PubMed1997

Phylogenetic analysis of the Monogenea and their relationships with Digenea and Eucestoda inferred from 28S rDNA sequences.

Platyhelminth phylogeny is controversial. Phylogenetic analyses of the partial domain C1 and the full domains D1 and C2 (358 nucleotides) from the 28S ribosomal RNA gene for 21 species from the Monogenea, Digenea, Cestoda, and, as the outgroup, Tricladida reveal major departures from prevailing theory. The Digenea and not the Monogenea (Monopisthocotylea and Polyopisthocotylea) form the sister group of the cestodes; the Monopisthocotylea and Polyopisthocotylea are each monophyletic, but the Monogenea do not form a monophylum; the sister group of the Digenea + Cestoda is the Polyopisthocotylea; and Monopisthocotylea are the sister group of all other parasitic flatworms.

Animals↗
PubMed1997

Sequence analysis of a gene family encoding Taenia ovis vaccine antigens expressed during embryogenesis of eggs.

The 45W vaccine antigen of the cestode parasite Taenia ovis is a member of a small gene family estimated to comprise six members. All six genes were cloned and characterised. Nucleotide sequence analysis of the different family members identified a set of closely related genes exhibiting between 75.2 and 98.6% DNA sequence homology. Intron/exon structure for the various genes constituting the family was found to be conserved. Open reading frame analysis and pairwise alignments of predicted polypeptides from the various members of the family revealed that the encoded proteins share between 51.8 and 96.5% amino acid identity with the host-protective 45W antigen. Polymerase chain reaction (PCR) primers were designed which allowed amplification of individual family members in a transcript-specific manner. These were used to investigate gene expression in different developmental stages of the parasite by reverse-transcription polymerase chain reaction (RT-PCR). All members of the gene family were expressed during the T. ovis life-cycle. 45W gene family transcription correlates with embryogenesis of eggs and expression is increased in activated eggs (oncospheres). One family member, ToW3S, was expressed in the early larvae (cysticercus) stages. Expression of genes closely related to the 45W vaccine antigen has important implications for immune evasion under selection from the application of a 45W-based vaccine.

Amino Acid Sequence↗
PubMed2002

Ecto-, endo- and haemoparasites in free-range chickens in the Goromonzi District in Zimbabwe.

A cross-sectional study determined the prevalence of ecto-, endo- and haemoparasites in free-range chickens from the Goromonzi District, Zimbabwe. Fifty young and 50 adult birds were selected randomly. All chickens harboured ecto- and endoparasites, and 32% were infected with haemoparasites. Eight different ectoparasites were identified; the more prevalent ones had the following prevalences (young, %; adult, %): Argas persicus (6; 14), Cnemidocoptes mutans (6; 32), Echidnophaga gallinacea (72; 74), Goniocotes gallinae (0; 22), Menacanthus stramenius (90; 88) and Menopon gallinea (24; 66). The prevalences of C. mutans, G. gallinae and M. gallinae were higher in adults compared to young chickens. The mean (+/-S.D.) number of helminth species per chicken was 6.7+/-2.0 for young chickens and 6.4+/-2.0 for adult chickens with a range of 1-10 for young chickens and a range of 1-11 for adult chickens. The most prevalent nematodes identified were (with prevalence in % for young/adult birds): Allodapa suctoria (76; 72), Ascaridia galli (48; 24), Gongylonema ingluvicola (28; 56), Heterakis gallinarum (64; 62) and Tetrameres americana (70; 62). For cestodes the prevalences were: Amoebotaenia cuneata (60; 68), Hymenolepis spp. (62; 80), Raillietina echinobothrida (66; 34), Raillietina tetragona (94; 100) and Skrjabinia cesticillus (50; 76). The young chickens had higher prevalences of A. galli and R. echinobothrida compared to adults, but lower prevalence of G. ingluvicola and S. cesticillus. Eimeria spp. oocysts were isolated in 36% of 47 investigated samples. The prevalence was 47% for young chickens and 18% for adult chickens. Prevalences (in %) of haemoparasites in young and adult chickens were: Aegyptinella pullorum (7; 6), Leucocytozoon sabrazesi (3; 1), Plasmodium gallinaceum (8; 6) and Trypanosoma avium (2; 3).

Animals↗
PubMed2001

Oral inoculation with Gymnorhynchus gigas induces anti-parasite anapyhylactic antibody production in both mice and rats and adverse reactions in challenge mice.

This study was performed to mimic human consumption of fish flesh infected with larvae of the fish cestode Gymnorhynchis gigas and examine possible side effects thereof. Both a rat and a mouse G. gigas oral inoculation model were used. The rat model was evaluated according to propensity to induce stress responses in three tissues and anaphylactic antibody production. The mouse model measured anti-G. gigas IgG, M and A (H + L) levels in intestinal fluids, fecal suspensions and serum and specific serum IgE levels by enzyme-linked immunosorbent assay. Additionally, biological activity of anaphylactic antibodies in test mice and rats were evaluated utilizing challenge reinoculation(s) and intradermal skin testing, respectively. With the rat inoculation model, we noted both occurrence of a shock response, viz. increased expression of heat shock proteins in intestine and spleen, and of immediate-type skin reactions. No positive wheals were seen on skin sites treated with PBS or soluble Trichinella spiralis extract. With the mouse model, our results showed that all body fluids tested had significantly more anti-G. gigas IgG, M and A (H + L) than their counterparts from either PBS-treated or T. spiralis-infected controls. In addition, the mouse G. gigas model had significantly higher specific serum IgE. When challenged by oral route all test mice (n = 5) manifested immediate-type signs of distress. Repeated exposure to the "allergen", produced clinical signs appearing more rapidly and persisting longer. These findings suggest that feeding on fish infected with G. gigas plerocercoids triggers the production of anaphylactic-type antibodies in both rats and mice and, by implication, possibly also in humans.

Anaphylaxis↗
PubMed1996

Species of Echinococcus: pattern and process.

Although classification and nomenclature within the cestode genus Echinococcus has, historically, been controversial, the past 20-30 years have provided a period of relative stability. Recent calls for taxonomic revision in the genus have therefore created something of a stir. In this article, Alan Lymbery and Andrew Thompson describe the reasons for the new controversy, and suggest that the problem can only be resolved by agreement on an appropriate species concept and on operational procedures for implementing that concept.

Journal Article↗
PubMed1998

Muscling in on parasitic flatworms.

The musculature of parasitic platyhelminths (monogeneans, digeneans and cestodes) presents an attractive target for chemotherapeutic intervention in that it is central to locomotory movement and attachment and serves an essential role in alimentation (feeding, translocation of food and digestive waste) and reproduction (insemination, egg formation and oviposition). Here, Gunnar Mair, Aaron Maule, Chris Shaw and David Halton review muscle organization in flatworm parasites, and outline some of the recent physiological and molecular advances that have been made in flatworm muscle research.

Journal Article↗
PubMed1999

Parasites as accumulation indicators of heavy metal pollution.

Parasites are attracting increasing interest from parasite ecologists as potential indicators of environmental quality because of the variety of ways in which they respond to anthropogenic pollution. However, until recently, little was known about the accumulation of toxins within parasites. Certain parasites, particularly intestinal acanthocephalans and cestodes of fish, can accumulate heavy metals at concentrations that are orders of magnitude higher than those in the host tissues or the environment. In this review, Bernd Sures, Roy Siddall and Horst Taraschewski discuss the recently described phenomenon of conspicuous metal accumulation by parasites and how this might be applied to environmental monitoring. They also suggest how environmental science and parasitology might profit from each other in the near future.

Animals↗
PubMed2000

Vaccination against cysticercosis and hydatid disease.

Infections with the larval stages of taeniid cestode parasites cause substantial human morbidity as well as economic losses in domestic livestock species. Despite ongoing efforts around the world, few countries have been able substantially to reduce or eradicate these infections through the use of anthelmintics and lifestyle changes. Vaccines offer an additional potential tool to assist with the control of parasite transmission. Here, Marshall Lightowlers and colleagues review the substantial progress that has been made towards developing practical vaccines against hydatid disease in sheep and cysticercosis in sheep and cattle. Recombinant antigens have been used to induce more than 90% protection against challenge infections. Such success in animals encourages investigation of the potential use of vaccines in humans to prevent hydatid disease arising from infection with Echinococcus granulosus and cysticercosis from infection with Taenia solium.

Amino Acid Sequence↗
PubMed1997

Structure-activity relationships of FMRFamide-related peptides contracting Schistosoma mansoni muscle.

This study reports the potent myoactivity of flatworm FMRFamide-related peptides (FaRPs) on isolated muscle fibers of the human blood fluke, Schistosoma mansoni. The turbellarian peptides YIRFamide (EC50 4 eta M), GYIRFamide (EC50 1 eta M), and RYIRFamide (EC50 7 eta M), all induced muscle contraction more potently than the cestode FaRP GNFFRFamide (EC50 500 eta M). Using a series of synthetic analogs of the flatworm peptides YIRFamide, GYIRFamide and RYIRFamide, the structure-activity relationships of the muscle FaRP receptor were examined. With a few exceptions, each residue in YIRFamide is important in the maintenance of its myoactivity. Alanine scans resulted in peptides that were inactive (Ala1, Ala2, Ala3 and Ala4 YIRFamide; Ala4 and Ala5 RYIRFamide) or had much reduced potencies (Ala1, Ala2 and Ala3 RYIRFamide). Substitution of the N-terminal (Tyr1) residue of YIRFamide with the non-aromatic residues Thr or Arg produced analogs with greatly reduced potency. Replacement of the N-terminal Tyr with aromatic amino acids resulted in myoactive peptides (FIRFamide, EC50 100 eta M; WIRFamide, EC50 0.5 eta M). The activity of YIRFamide analogs which possessed a Leu2, Phe2 or Met2 residue (EC50's 10, 1 and 3 eta M, respectively) instead of Ile2 was not significantly altered, whereas, YVRFamide had a greatly reduced (EC50 200 eta M) activity. Replacement of the Phe4 with a Tyr4 (YIRYamide) also greatly lowered potency. Truncated analogs were either inactive (FRFamide, YRFamide, HRFamide, RFamide, Famide) or had very low potency (IRFamide and MRFamide), with the exception of nLRFamide (EC50 20 eta M). YIRF free acid was inactive. In summary, these data show the general structural requirements of this schistosome muscle FaRP receptor to be similar, but not identical, to those of previously characterized molluscan FaRP receptors.

Amino Acid Sequence↗
PubMed1997

Urea/DTT solubilization of a recombinant Taenia ovis antigen, 45W, expressed as a GST fusion protein results in enhanced protective immune response to the 45W moiety.

Vaccination and challenge infection experiments were conducted in sheep using different forms of a recombinant protein (45W) from the cestode parasite Taenia ovis. 45W was expressed in Escherichia coli as a fusion protein with glutathione-S-transferase (45W-GST) and was produced as both soluble protein and insoluble inclusion bodies. Vaccination of animals with either the soluble or inclusion body derived protein resulted in the immune response being predominantly directed to the GST moiety of 45W-GST. Conversely, vaccination with 45W-GST which had been solubilized/treated with urea and dithiothreitol (DTT), elicited enhanced responses to the 45W moiety and significantly reduced responses to GST. Vaccination with all forms of 45W-GST protected sheep against experimental T. ovis infection. However, protection was highly correlated with anti-45W antibody levels and these were significantly higher in animals vaccinated with the urea/DTT treated form of 45W-GST. It is suggested that recombinant proteins expressed either with or without fusion partners may stimulate enhanced immune responses when incorporated in vaccine formulations in a denatured/reduced state.

Animals↗
PubMed2000

Relationship between helminth species diversity, intensity of infection and breeding management in dairy goats.

Helminth infection, species diversity (proportion of each species in the community), species number, intensity of infection and anthelmintic resistance were investigated in 16 dairy-goat farms of south-western France. The aim of the study was to estimate which breeding management factors may affect diversity and intensity of infection. Anthelmintic resistance was investigated because this factor may modify composition of nematode communities. Farm history and breeding management were recorded by means of a questionnaire. Natural and experimental infections were compared to determine which strategy was the best to estimate diversity index and number of species from nematode community of a whole herd. A total of 17 species of helminths, among which 14 nematodes, one cestode (Moniezia spp.) and two trematodes (Paramphistomum daubneyi and Dicrocoelium lanceolatum) were recovered in the 26 necropsied culled goats. One culled goat worm burden represented conveniently species number but not species diversity (Shannon index) of parasite community harboured in the whole herd. Experimental sheep infection with larvae from pooled faeces of 15 goats was the best strategy to estimate species diversity in farms. Diversity could be predicted from the number of farms from which goats originated at constitution of the herd, the duration of goat winter withdrawal from pastures, and intensity of infection. Anthelmintic resistance was correlated with the number of farms of origin, area of permanent pastures and intensity of infection. The strategy adopted for the herd constitution was a critical step as species diversity and risk of anthelmintic resistance increased with the number of farms of origin. Intensity of infection was negatively correlated with species diversity. This suggested that more diverse communities were better controlled by the host, in agreement with the widespread statement that diversity would beget stability.

Abomasum↗
PubMed2002

Epidemiological studies on gastrointestinal helminths of dromedary (Camelus dromedarius) in semi-arid lands of eastern Ethiopia.

A total of 752 dromedaries were examined and 75% were found to be harbouring nematode eggs. The mean EPG was 1831 and the range was from 100 to 21,200. The prevalence rates in the four-age groups examined were 59.6% (3-7 years), 72.4% (8-12 years), 76.1% (13-17 years) and 83.9% (18-22 years). The prevalence rate for females and males were 77.6 and 64.8%, respectively and for long dry, short rainy, short dry and long rainy seasons were 66, 80, 69 and 82.6%, respectively. The mean EPG of faeces was significantly (P<0.01) higher for older animals compared to other group of younger animals (3-7 years), for females compared to males, and for rainy compared to dry season. Sixteen dromedary gastrointestinal organs were used for identification and counts of helminths. Among the adult worms identified, from the abomasum, Haemonchus longistipes had a 94% prevalence rate. From the small intestine Trichostrongylus colubriformis, Trichostrongylus probolurus, Impalaia tuberculata and Strongyloides papillosus were identified with prevalence rates of 75, 25, 63 and 20%, respectively. Moreover, cestodes such as Moniezia benedeni, Moniezia expansa, Avitellina spp. and Stilesia globipunctata with prevalence rates of 31, 13, 25 and 19%, respectively, were identified. I. tuberculata was identified for the first time in this country from a dromedary.The pathological lesions were more pronounced in higher infestations. Infestation level over approximately 1000 of H. longistipes and 15,000 in mixed infection of T. colubriformis and I. tuberculata, caused gross lesions of ulcerated and hyperaemic mucosa, and the odour of the fluid were fetid. The microscopic lesions observed were sloughing of epithelium, necrosis of glands, atrophy and loss of villi, haemorrhages and cellular infiltration mainly of eosinophiles and lymphocytes.

Age Factors↗