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2009-01-01
Studies of marine mammal parasites in the Caribbean are scarce. An assessment for marine mammal endo- and ectoparasites from Puerto Rico and the Virgin Islands, but extending to other areas of the Caribbean, was conducted between 1989 and 1994. The present study complements the latter and enhances identification of anisakid nematodes using molecular markers. Parasites were collected from 59 carcasses of stranded cetaceans and manatees from 1994 to 2006, including Globicephala macrorhynchus, Kogia breviceps, Kogia sima, Lagenodelphis hosei, Mesoplodon densirostris, Peponocephala electra, Stenella longirostris, Steno bredanensis, Trichechus manatus. Tursiops truncatus, and Ziphius cavirostris. Sixteen species of endoparasitic helminthes were morphologically identified, including two species ...
Sensitive Detection and Quantification of Anisakid Parasite Residues in Food Products
2010-01-01
Abstract Anisakids are nematodes whose larval stages are often present in fish, molluscs, and crustaceans. Members of the family Anisakidae belonging to the genera Anisakis and Pseudoterranova are implicated in human infections caused by the consumption of raw or undercooked fish. Adequate cooking will kill anisakid larvae, however, killed or inactivated larvae can still cause sensitization and immunoglobulin E-dependent hypersensitivity in human. This work describes the development of DNA-based tests to detect and quantify the presence of Anisakis spp. and Pseudoterranova spp. larvae in fish and fish-derived products, including fish fillets, surimi, fish sticks, canned fish, and baby food. Primers and TaqMan MGB probes recognizing only Anisakis spp. and Pseudoterranova spp. were designed ...
Sensitive detection and quantification of anisakid parasite residues in food products.
Anisakids are nematodes whose larval stages are often present in fish, molluscs, and crustaceans. Members of the family Anisakidae belonging to the genera Anisakis and Pseudoterranova are implicated in human infections caused by the consumption of raw or undercooked fish. Adequate cooking will kill anisakid larvae, however, killed or inactivated larvae can still cause sensitization and immunoglobulin E-dependent hypersensitivity in human. This work describes the development of DNA-based tests to detect and quantify the presence of Anisakis spp. and Pseudoterranova spp. larvae in fish and fish-derived products, including fish fillets, surimi, fish sticks, canned fish, and baby food. Primers and TaqMan MGB probes recognizing only Anisakis spp. and Pseudoterranova spp. were designed on the first internal transcribed spacer 1 regions of rDNA for a real-time polymerase chain reaction assay. A commercial probe for 18S rDNA was used to detect and quantify the total eukaryotic DNA of the samples. The specificity and sensitivity of the assays were tested using reference samples prepared from mixtures made of Anisakis larvae in different quantity of codfish, and subsequent dilutions. Studies were performed to assess the ability of the test to detect and quantify anisakids in various products. Results showed that this test is able to detect anisakid DNA contained in a proportion of 1:10(5) in 1 ng of total DNA. The high prevalence of anisakids reported in main fishery species was confirmed by frequently detecting anisakids DNA in fish muscle and fish-derived products. A partial correlation was found between the number of larvae present in the viscera and the level of contamination of fish fillets. In conclusion, this molecular test is useful to detect the presence of Anisakis spp. and Pseudoterranova spp. in fish and fish-derived products and to quantify the level of contamination along the food chain, with potential applications for fish farms, fish markets, and food producers.
2008-01-01
The taxonomic composition of the metazoan parasites found in the stomach of 629 specimens of the "Patagonian toothfish" Dissostichus eleginoides Smitt, 1898, from two localities in southern Chile (Lebu 36degree00primeS; Quellon 48degree49primeS), is recorded. More than 58,000 parasite specimens belonging to 9 taxa were registered. Adult worms were represented by the hemiurids Gonocerca phycidis Manter 1925, Lecithochirium genypteri Luhe 1905, Brachyphallus crenatus (Rudolphi 1802), and Derogenes varicus (Muller 1784) and the nematode Hysterothylacium sp. Ward & Magath 1917. Larval forms of anisakid nematodes, an acanthocephalan (Corynosoma sp.), a trypanorhynch cestode (Hepatoxylon trichiuri (Holten 1802)) and an unidentified pseudophyllidean cestode were also obtained. Parasitological evi...
2009-01-01
Until now, Anisakis typica has been the sole anisakid identified by means of genetic markers from the Atlantic Ocean off Brazil. In this study we developed, and applied to larvae and adults, an A. typica-specific molecular identification protocol based on the complete intergenic transcribed spacer (ITS) region. Anisakids from the dolphin Sotalia guianensis and from the frigate tuna, Auxis thazard, caught off the coast of Brazil, were processed using two DNA extraction methods. Molecular diagnosis identified A. typica in 13 of 15 samples. Complete ITS analysis showed that the remaining two isolates were in fact A. physeteris. Maximum parsimony analysis of complete ITS region (915bp) confirmed these results. Our study verified the dominance of A. typica infecting hosts from the Brazilian coa...
2010-01-01
Between February 1996 and January 2000, parasitological examination of Scomber scombrus (n=447), Trachurus trachurus (n=175), Sardina pilchardus (n=160) and Engraulis encrasicolus (n=153) from the fishing-ground of Tarragona (NE Spain) were made for anisakids. Three anisakid species (Anisakis larva type I sensu Berland, 1961, Contracaecum sp. and Hysterothylacium aduncum) were morphologically identified. The prevalence of Anisakis larva type I sensu Berland (1961) ranged from 11.0% (S. scombrus) and 2.6% (T. trachurus) to 0.0% (S. pilchardus and E. encrasicolus). The prevalence of H. aduncum in S. scombrus was 1.1%. The prevalence of Contracaecum sp. was 2.0% in S. scombrus, and 0.7% in T. trachurus respectively. There was a positive and significant correlation between the host length and ...
2009-01-01
A new anisakid nematode, Anisakis nascettii n. sp., is described from beaked whales Mesoplodon spp. off the coast of New Zealand and South Africa. Morphological and molecular (allozymes and mtDNA cox2 sequence) data were used for diagnostic and identification purposes. Among the 19 allozymes studied, 10 were found to be unique and characteristic for A. nascettii n. sp. Analysis of allozymes demonstrated reproductive isolation from A. ziphidarum Paggi, Nascetti, Webb, Mattiucci, Cianchi & Bullini, 1998 and mtDNA cox2 sequences depict this Anisakis species as a distinct and unique entity. Key morphological diagnostic traits for A. nascettii with respect to the genetically closely related species A. ziphidarum include: spicule length, the spicule/body length ratio, the arrangement of the caud...
2008-10-01
The use of sewage or wastewater in agriculture is becoming increasingly common as a result of global water scarcity. Intestinal nematode infections have been identified as the main health risk...Full Text Available
Brassicaceous and rye cover crops altered free-living soil nematode community composition
2010-01-01
Nematode community analysis was utilized to evaluate the biofumigant or allelopathic effects of brassicaceous and rye winter cover crops on non-target nematodes in three experiments (two sites) in Maryland. The cover crop treatments included mustard blend (Sinapis alba and Brassica juncea) 'Caliente', rapeseed (B. napus) 'Essex'/'Humus', forage radish (Raphanus sativus) 'Dichon', oilseed radish (R. sativus) 'Adagio'/'Colonel', rye (Secale cereale) 'Wheeler' and a no cover crop (winter weeds) control. Soil samples (0-15cm) were collected two or three times per year and extracted nematodes were identified to genus or family. Nematode response parameters were genus, family, trophic group population density, and percent distribution of trophic groups in the entire nematode community divided in...
EVALUATION OF VITIS AESTIVALIS AND RELATED TAXA AS SOURCES OF RESISTANCE TO ROOT-KNOT NEMATODES
Development of rootstocks with resistance to root-knot nematodes (Meloidogyne spp.) is a top priority in grape breeding. Accessions of Vitis aestivalis resistant to root-knot nematodes have been identified. This species is very difficult to propagate from cuttings, essentially precluding its direct...
Phytoecdysteroids: a novel defense against plant-parasitic nematodes
2004-01-01
The original publication can be found at www.springerlink.comThe phytoecdysteroid, 20-hydroxyecdysone (20E), is a major molting hormone of invertebrates possibly including nematodes. As 20E is inducible in spinach, the defensive role against plant-parasitic nematodes was investigated. The effects of direct application on nematodes was assessed by treating cereal cyst nematode, Heterodera avenae, juveniles with concentrations of 20E from 8.2 × 10⁻⁸ to 5.2 × 10⁻⁵ M before applying to Triticum aestivum growing in sand. H. avenae, Heterodera schachtii (sugarbeet cyst nematode), Meloidogyne javanica (root-knot nematode) and Pratylenchus neglectus (root lesion nematode) were treated with 5.2 × 10⁻⁵ 20E and incubated in moist sand. To test the protective effects of 20E in plants, the latter three nematodes were applied to Spinacia oleracea in which elevated concentrations of 20E had been induced by methyl jasmonate. Abnormal molting, immobility, reduced invasion, impaired development, and death occurred in nematodes exposed to 20E either directly at concentration above 4.2 × 10⁻⁷ M or in plants. Phytoecdysteroid was found to protect spinach from plant-parasitic nematodes and may confer a mechanism for nematode resistance. Publisher: Springer Contributor: School of Agriculture, Food and Wine : Plant and Food Science Other identifier: Journal of Chemical Ecology, 2004; 30 (10):1885-1899; 0098-0331; 0020077536; 20080604175347; 000224670400001
2010-01-01
Crop growth and damage parameters (plant growth and yield, root damage and nematode population densities), believed to be associated with resistance of Musa genotypes to nematodes under field conditions, were evaluated in a field trial of 24 Musa genotypes inoculated at planting with a combination of Radopholus similis and Helicotylenchus multicinctus with the objective to identify parameters with strong association with nematode resistance and high heritability. Correlation and path analysis of the association between plant growth, yield, root damage and nematode population densities showed a strong negative association between percentage dead roots, percentage root necrosis, R. similis and H. multicinctus population densities and yield. The strongest negative association was observed bet...
An endosymbiotic bacterium in a plant-parasitic nematode: Member of a new Wolbachia supergroup
2009-01-01
Wolbachia is an endosymbiotic bacterium widely present in arthropods and animal-parasitic nematodes. Despite previous efforts, it has never been identified in plant-parasitic nematodes. Random sequencing of genes expressed by the burrowing nematode Radopholus similis resulted in several sequences with similarity to Wolbachia genes. The presence of a Wolbachia-like endosymbiont in this plant-parasitic nematode was investigated using both morphological and molecular approaches. Transmission electron microscopy, fluorescent immunolocalisation and staining with DAPI confirmed the presence of the endosymbiont within the reproductive tract of female adults. 16S rDNA, ftsZ and groEL gene sequences showed that the endosymbiont of R. similis is distantly related to the known Wolbachia supergroups. ...
We reported recently a unique fungal structure, called the spiny ball, on the vegetative hyphae of Coprinus comatus (O. F. Mll.:Fr.) Pers. Although some observations regarding the role of this structure were presented, its function remained largely unknown. In this study, we showed that purified (isolated and washed) spiny balls could immobilize and kill the free-living nematode Panagrellus redivivus Goodey highly efficiently. Scanning electron microscopy studies illustrated that the spiny structure damaged the nematode cuticle, suggesting the presence of a mechanical force during the process of nematode immobilization. Severe injuries on nematode cuticles caused the leakage of inner materials of the nematodes. When these structures were ground in liquid nitrogen, their killing efficacy against nematodes was lost, indicating that the shape and the complete structure of the spiny balls are indispensable for their function. However, extraction with organic solvents never lowered their activity against P. redivivus, and the extracts showed no obvious effect on the nematode. We also investigated whether C. comatus was able to produce toxins which would aid in the immobilization of nematodes. In total, we identified seven toxins from C. comatus that showed activity to immobilize the nematodes P. redivivus and Meloidogyne incognita (Kofoid et White) Chitwood. The chemical structures of these toxins were identified with nuclear magnetic resonance, mass spectrometry, infrared, and UV spectrum analysis. Two compounds were found to be novel. The toxins found in C. comatus are O-containing heterocyclic compounds.
2007-06-01
Full Text Available.We reported recently a unique fungal structure, called the spiny ball, on the vegetative hyphae of Coprinus comatus (O. F. Müll.:Fr.) Pers. Although some observations regarding the role of this structure were presented, its function remained largely unknown. In this study, we showed that purified (isolated and washed) spiny balls could immobilize and kill the free-living nematode Panagrellus redivivus Goodey highly efficiently. Scanning electron microscopy studies illustrated that the spiny structure damaged the nematode cuticle, suggesting the presence of a mechanical force during the process of nematode immobilization. Severe injuries on nematode cuticles caused the leakage of inner materials of the nematodes. When these structures were ground in liquid nitrogen, their killing efficacy against nematodes was lost, indicating that the shape and the complete structure of the spiny balls are indispensable for their function. However, extraction with organic solvents never lowered their activity against P. redivivus, and the extracts showed no obvious effect on the nematode. We also investigated whether C. comatus was able to produce toxins which would aid in the immobilization of nematodes. In total, we identified seven toxins from C. comatus that showed activity to immobilize the nematodes P. redivivus and Meloidogyne incognita (Kofoid et White) Chitwood. The chemical structures of these toxins were identified with nuclear magnetic resonance, mass spectrometry, infrared, and UV spectrum analysis. Two compounds were found to be novel. The toxins found in C. comatus are O-containing heterocyclic compounds.
New insights into the evolution of subtilisin-like serine protease genes in Pezizomycotina
Full Text Available.BackgroundSubtilisin-like serine proteases play an important role in pathogenic fungi during the penetration and colonization of their hosts. In this study, we perform an evolutionary analysis of the subtilisin-like serine protease genes of subphylum Pezizomycotina to find if there are similar pathogenic mechanisms among the pathogenic fungi with different life styles, which utilize subtilisin-like serine proteases as virulence factors. Within Pezizomycotina, nematode-trapping fungi are unique because they capture soil nematodes using specialized trapping devices. Increasing evidence suggests subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles. Here we also conduct positive selection analysis on the subtilisin-like serine protease genes from nematode-trapping fungi.ResultsPhylogenetic analysis of 189 subtilisin-like serine protease genes from Pezizomycotina suggests five strongly-supported monophyletic clades. The subtilisin-like serine protease genes previously identified or presumed as endocellular proteases were clustered into one clade and diverged the earliest in the phylogeny. In addition, the cuticle-degrading protease genes from entomopathogenic and nematode-parasitic fungi were clustered together, indicating that they might have overlapping pathogenic mechanisms against insects and nematodes. Our experimental bioassays supported this conclusion. Interestingly, although they both function as cuticle-degrading proteases, the subtilisin-like serine protease genes from nematode-trapping fungi and nematode-parasitic fungi were not grouped together in the phylogenetic tree. Our evolutionary analysis revealed evidence for positive selection on the subtilisin-like serine protease genes of the nematode-trapping fungi.ConclusionsOur study provides new insights into the evolution of subtilisin-like serine protease genes in Pezizomycotina. Pezizomycotina subtilisins most likely evolved from endocellular to extracellular proteases. The entomopathogenic and nematode-parasitic fungi likely share similar properties in parasitism. In addition, our data provided better understanding about the duplications and subsequent functional divergence of subtilisin-like serine protease genes in Pezizomycotina. The evidence of positive selection detected in the subtilisin-like serine protease genes of nematode-trapping fungi in the present study suggests that the subtilisin-like serine proteases may have played important roles during the evolution of pathogenicity of nematode-trapping fungi against nematodes.
2009-09-01
Full Text Available.In this study, 4847 expressed sequenced tags (ESTs) from mixed stages of the migratory plant-parasitic nematode Ditylenchus africanus (peanut pod nematode) were investigated. It is the first molecular survey of a nematode which belongs to the family of the Anguinidae (order Rhabditida, superfamily Sphaerularioidea). The sequences were clustered into 2596 unigenes, of which 43% did not show any homology to known protein, nucleotide, nematode EST or plant-parasitic nematode genome sequences. Gene ontology mapping revealed that most putative proteins are involved in developmental and reproductive processes. In addition unigenes involved in oxidative stress as well as in anhydrobiosis, such as LEA (late embryogenesis abundant protein) and trehalose-6-phosphate synthase were identified. Other tags showed homology to genes previously described as being involved in parasitism (expansin, SEC-2, calreticulin, 14-3-3b and various allergen proteins). In situ hybridization revealed that the expression of a putative expansin and a venom allergen protein was restricted to the gland cell area of the nematode, being in agreement with their presumed role in parasitism. Furthermore, 7 putative novel candidate parasitism genes were identified based on the prediction of a signal peptide in the corresponding protein sequence and homologous ESTs exclusively in parasitic nematodes. These genes are interesting for further research and functional characterization. Finally, 34 unigenes were retained as good target candidates for future RNAi experiments, because of their nematode specific nature and observed lethal phenotypes of Caenorhabditis elegans homologs.
With the availability of microarray technology, the expression profiles of thousands of genes can be monitored simultaneously to help determine the mechanisms of these biological processes. We conducted Affymetrix GeneChip microarray analyses of the Arabidopsis-cyst nematode interaction and employed a statistical procedure to analyze the resultant data, which allowed us to identify significant gene expression changes. Quantitative real-time RT-PCR assays were used to confirm the microarray analyses. The results of the expression profiling revealed 128 genes with altered steady-state mRNA levels following infection by the sugar beet cyst nematode (Heterodera schachtii; BCN), in contrast to only 12 genes that had altered expression following infection by the soybean cyst nematode (H. glycines; SCN). The expression of these 12 genes also changed following infection by BCN, i.e. we did not identify any genes regulated exclusively by SCN. The identification of 116 genes whose expression changes during successful cyst nematode parasitism by BCN suggests a potential involvement of these genes in the infection events starting with successful syncytium induction. Further characterization of these genes will permit the formulation of testable hypotheses to explain successful cyst nematode parasitism.
A new molecular technique detects Anisakis in all fish
2010-07-15
A new method enables anisakids to be detected in any fish product, from a whole fish, fresh or frozen, to tinned fish and surimi. Developed by Spanish scientists, the system is based on molecular techniques and ...
2009-01-01
Adult and fourth-stage larval nematodes found in the stomachs of farmed cod in North Norway in October 2006 were identified as Hysterothylacium cornutum (Stossich, 1904), a nematode considered to be specific at the adult stage to tunas of the genus Thunnus. As far as we are aware, this is the first report of an adult form of this nematode from any host other than members of the genus Thunnus, and is also the first report from a polar region. The two infected cod, one with one large adult worm and another with six larvae, were in a sample of 17 that had been captured from the wild about 1year before sampling and held in floating sea cages in ksfjord, Finnmark County, North Norway, where the examinations took place. No further infections were found in any other samples of wild and farmed...
2008-01-01
Rhynchophorus palmarum, the palm weevil, has been reported as a pest of palms and sugarcane plants. The Red Ring Disease is an infectious plant disease caused by nematodes. The etiological agent, Bursaphelencus (Rhadinaphelencus) cocophilus (Nematoda: Aphelenchoididae), completes its life cycle within 9 to 10days inside the palm tree. The main symptom is a permanent wilting of the plant aerial parts. Previous studies stated that B. cocophilus cohabits with other nematodes the gut of R. palmarum. The aim of this study is to identify nematodes collected from palm weevil found in coconut plantations from the north of the Rio de Janeiro State. Light (LM) and scanning electron microscopy (SEM) analyses were carried on samples of infected R. palmarum and fragments and fresh juice of infected p...
Identification and localization of food-source microbial nucleic acids inside soil nematodes
2010-01-01
Microorganisms (e.g., prokaryotes, fungi) are food sources for soil nematodes, but they can also be potential mutualists or pathogens. Understanding the linkages between microorganism and invertebrate diversity in soils requires the ability to distinguish between these microbial roles. We tested the potential of a taxon-specific fluorescent in situ hybridization (FISH) procedure for identifying and localizing microbial rRNA within the bodies of soil nematodes. Our objective was to determine whether the rate of digestion permitted detection and identification of food-source nucleic acids within the nematode digestive system (i.e., pharynges, intestines) before their breakdown. First, using laboratory cultures of Caenorhabditis elegans maintained on Escherichia coli, we were able to localize...
Epidemiology of nematode parasites of sheep around Jimma, southwestern Ethiopia
2010-01-01
An investigation was made into the epidemiology of nematode infections of sheep in two districts of Jimma zone, southwestern Ethiopia. We used two approacheslong-term monitoring of identified sheep for nematode infection and abattoir or market survey for analysis. In the first monitoring regime, we used 80 lambs [40 sheep (20 per sex) from each district (Dedo and Yebu)] averaging 45months of age. Faecal egg counts (FEC), packed cell volume (PCV) and body weight changes were monitored over a period of 1year. Additionally, faecal samples were collected (on a weekly basis) from sheep brought to abattoir/market for 1year to monitor faecal egg counts. The nematode parasite burden, as judged by FEC and PCV, was generally low indicating that the climatic conditions are not conducive t...
Comparisons of Female and Egg Assays to Identify Rotylenchulus reniformis Resistance in Cotton
2006-09-01
More plants can be screened for reniform nematode resistance each year if the time involved can be shortened. In this study, the hypothesis that female counts are as efficient as egg counts in identifying...Full Text Available
Infection of plant-parasitic nematodes by Paecilomyces lilacinus and Monacrosporium lysipagum
2006-01-01
Studying the mode of infection of a biocontrol agent is important in order to assess its efficiency. The mode and severity of infection of nematodes by a soil saprophyte Paecilomyces lilacinus (Thom) Samson and a knob-producing nematode trapping fungus Monacrosporium lysipagum (Drechsler) Subram were studied under laboratory conditions using microscopy. Infection of stationary stages of nematodes by P. lilacinus was studied with three plant-parasitic nematodes Meloidogyne javanica (Treub) Chitwood, Heterodera avenae Wollenweber and Radopholus similis (Cobb) Thorne. Paecilomyces lilacinus infected eggs, juveniles and females of M. javanica by direct hyphal penetration. The early developed eggs were more susceptible than the eggs containing fully developed juveniles. As observed by transmission electron microscopy, fungal hypha penetrated the M. javanica female cuticle directly. Paecilomyces lilacinus also infected immature cysts of H. avenae including eggs in the cysts and the eggs of R. similis. Trapping and subsequent killing of mobile stages of nematodes by M. lysipagum were studied with the above three nematodes. In addition, plant-parasitic nematodes Pratylenchus neglectus (Rensch) Chitwood and Oteifa and Ditylenchus dipsaci (Kuhn) Filipjev were tested with M. lysipagum. This fungus was shown to infect mobile stages of all the plant-parasitic nematodes. In general, juveniles except those of P. neglectus, were more susceptible to the attack than adults.20 page(s) Publisher: Springer Contributor: Macquarie University. Dept. of Chemistry and Biomolecular Sciences Relation: BioControl, Vol. 51, Issue 5, p.659-678; 10.1007/s10526-005-4242-x Other identifier: ISSN:1573-8248; mq-rm-2006000368 Language: eng
Exploring the transcriptome of the burrowing nematode Radopholus similis.
Radopholus similis is an important nematode pest on fruit crops in the tropics. Unraveling the transcriptome of this migratory plant-parasitic nematode can provide insight in the parasitism process and lead to more efficient control measures. For the first high throughput molecular characterization of this devastating nematode, 5,853 expressed sequence tags from a mixed stage population were generated. Adding 1,154 tags from the EST division of GenBank for subsequent analysis, resulted in a total of 7,007 ESTs, which represent approximately 3,200 genes. The mean G + C content of the nucleotides at the third codon position (GC3%) was calculated to be as high as 64.8%, the highest for nematodes reported to date. BLAST-searches resulted in about 70% of the clustered ESTs having homology to (DNA and protein) sequences from the GenBank database, whereas one-third of them did not match to any known sequence. Roughly 40% of these latter sequences are predicted to be coding, representing putative novel protein coding genes. Functional annotation of the sequences by GO annotation revealed the abundance of genes involved in reproduction and development, which reflects the nematode population biology. Genes with a role in the parasitism process are identified, as well as genes essential for nematode survival, providing information useful for parasite control. No evidence was found for the presence of trans-spliced leader sequences commonly occurring in nematodes, despite the use of various approaches. In conclusion, we found three different sources for the EST sequences: the majority has a nuclear origin, approximately 1% of the EST sequences are derived from the mitochondrial transcriptome, and interestingly, 1% of the tags are with high probability derived from Wolbachia, providing the first molecular indication for the presence of this endosymbiont in a plant-parasitic nematode.
2008-01-01
Plant-parasitic nematodes are important pests of horticultural crops grown in tropical and subtropical regions of Australia. Burrowing nematode (Radopholus similis) is a major impediment to banana production and root-knot nematodes (predominantly Meloidogyne javanica and M. incognita) cause problems on pineapple and a range of annual vegetables, including tomato, capsicum, zucchini, watermelon, rockmelon, potato and sweet potato. In the early 1990s, nematode control in these industries was largely achieved with chemicals, with methyl bromide widely used on some subtropical vegetable crops, ethylene dibromide applied routinely to pineapples and non-volatile nematicides such as fenamiphos applied up to four times a year in banana plantations. This paper discusses the research and extension work done over the last 15 years to introduce an integrated pest management approach to nematode control in tropical and subtropical horticulture. It then discusses various components of current integrated pest management programs, including crop rotation, nematode monitoring, clean planting material, organic amendments, farming systems to enhance biological suppression of nematodes and judicious use of nematicides. Finally, options for improving current management practices are considered. Publisher: CSIRO Publishing Other identifier: Stirling, G.R. and Pattison, A.B. (2008) Beyond chemical dependency for managing plant-parasitic nematodes: Examples from the banana, pineapple and vegetable industries of tropical and subtropical Australia. Australasian Plant Pathology, 37 (3). pp. 254-267.
Soybean FGAM synthase promoters direct ectopic nematode feeding site activity.
Soybean cyst nematode (SCN) resistance in soybean is a complex oligogenic trait. One of the most important nematode resistance genes, rhg1, has been mapped to a distal region of molecular linkage group G in soybean. A simplified genetic system to identify soybean genes with modified expression in response to SCN led to the identification of several genes within the nematode feeding sites. The genes were mapped to reveal their linkage relationship to known QTLs associated with soybean cyst nematode (SCN) resistance. One candidate, a phosphoribosyl formyl glycinamidine (FGAM) synthase (EC 6.3.5.3) gene, mapped to the same genomic interval as the major SCN resistance gene rhg1 within linkage group G. Isolation of FGAM synthase from a soybean bacterial artificial chromosome (BAC) library revealed two highly homologous paralogs. The genes appeared to be well conserved between bacteria and humans. Promoter analysis of the two soybean homologs was carried out with the Arabidopsis thaliana - Heterodera schachtii system to investigate gene response to nematode feeding. The two promoters and their derived deletion constructions effected green fluorescent protein (GFP) expression within nematode feeding sites. The 1.0-kb promoter sequence immediately adjacent to the translation start site was sufficient to direct expression of GFP within syncytia. A wound-inducible element and a floral organ expression sequence were also identified within these promoters. Although a nematode-responsive element could not be identified, the observed expression of GFP within feeding sites supports the hypothesis that plant gene expression is redirected within feeding sites to benefit the parasite.
2007-01-01
SUMMARY Emerging evidence suggests that plant cell-wall-modifying enzymes induced by root-parasitic nematodes play important roles in feeding cell formation. We previously identified a tobacco endo-b-1,4-glucanase (cellulase) gene, NtCel7, that was strongly induced in both root-knot and cyst nematode feeding cells. To characterize further the developmental and nematode-responsive regulation of NtCel7, we isolated the NtCel7 promoter and analysed its expression over a time course of nematode infection and in response to auxin, gibberellin, ethylene and sucrose in soybean and tomato hairy roots and in Arabidopsis containing the NtCel7 promoter fused to the b-glucuronidase (GUS) reporter gene. Histochemical analyses of transgenic plant materials revealed that the NtCel7 promoter exhibited a u...
Metabolic profiling reveals local and systemic responses of host plants to nematode parasitism.
The plant parasitic beet cyst nematode Heterodera schachtii induces syncytial feeding structures in Arabidopsis roots. The feeding structures form strong sink tissues that have been suggested to be metabolically highly active. In the present study, metabolic profiling and gene targeted expression analyses were performed in order to study the local and systemic effects of nematode infection on the plant host. The results showed increased levels of many amino acids and phosphorylated metabolites in syncytia, as well as high accumulation of specific sugars such as 1-kestose that do not accumulate naturally in Arabidopsis roots. A correlation-based network analysis revealed highly activated and coordinated metabolism in syncytia compared to non-infected control roots. An integrated analysis of the central primary metabolism showed a clear coherence of metabolite and transcript levels, indicating transcriptional regulation of specific pathways. Furthermore, systemic effects of nematode infection were demonstrated by correlation-based network analysis as well as independent component analysis. 1-kestose, raffinose, alpha,alpha-trehalose and three non-identified analytes showed clear systemic accumulation, indicating future potential for diagnostic and detailed metabolic analyses. Our studies open the door towards understanding the complex remodelling of plant metabolism in favour of the parasitizing nematode.
The Equidae (the horse, Equus caballus, the ass, Equus asinus, zebras and their hybrids) are hosts to a great variety of nematode parasites, some of which can cause significant morbidity or mortality if individual hosts are untreated. Worldwide the nematode parasites of horses belong to 7 suborders, 12 families, 29 genera and 83 species. The great majority (19 of 29 genera and 64 of 83 species) are members of the family Strongylidae, which includes the most common and pathogenic nematode parasites of horses. Only the Strongylidae are included in this treatise. The Strongylidae (common name strongylids) of horses--nematodes with a well-developed buccal capsule, a mouth collar with two leaf-crowns, and a strongyloid (common name of superfamily Strongyloidea) copulatory bursa--can be separated into two subfamilies: Strongylinae (common name strongylins), usually large or medium-sized with a globular or funnel-shaped buccal capsule; and Cyathostominae (common name cyathostomins), usually small to medium-sized with a cylindrical buccal capsule. The increased attention to strongylid nematode parasites of horses has resulted in the need for updated diagnostic keys to these parasites using readily recognizable characters and the most recent literature on their systematics. Because the cyathostomins have been historically difficult to identify, and because they have emerged as the most significant nematode pathogens of horses, we provide a brief nomenclatural and taxonomic history and an introduction to the morphology of this group. This treatise is intended to serve as a basic working tool--providing easy identifications to genus and species of adult strongylid nematodes of equids. All strongylid nematodes normally parasitic in horses, the ass (and their hybrids), and zebras are included. The keys are illustrated with line drawings and halftone photomicrographs of each species. A short discussion of the systematics of the genus and species is provided for each genus following the species descriptions. Species diagnoses and a synonymy of each species is provided. Geographic distribution, prevalence, and location in host are also given for each species.
Screening 1154 ESTs from the plant-parasitic nematode Radopholus similis resulted in seven tags coding for proteins holding a transthyretin-like domain (PF01060). The seven ESTs corresponded to four different genes which were cloned from a cDNA library (accession numbers AM691117, AM691118, AM691119, AM691120). Transthyretin-like genes belong to a large family, different from the transthyretin and the transthyretin-related genes with whom they share some sequence similarity at the protein level. This similarity has caused an inconsistent use of different names and abbreviations in the past. To avoid further confusion, we introduce a standardized nomenclature for this gene family, and chose to name this barely characterized gene family ttl (as for transthyretin-like). Further examination of the identified genes, named Rs-ttl-1 to -4, showed that they are expressed in both juveniles and adults, but not in young embryos. Whole mount in situ hybridization revealed a distinct spatial expression pattern for two of the genes: Rs-ttl-1 is expressed in the tissues surrounding the vulva, whereas Rs-ttl-2 is expressed in the ventral nerve cord. The deduced protein sequences contain a putative signal peptide for secretion, pointing to an extracellular function of the mature proteins. Database screens showed that the ttl family is restricted to nematodes. Moreover, a HMMER search revealed that ESTs derived from ttl genes are more abundant in parasitic nematode libraries, with a bias towards the parasitic stages. Despite their abundance in nematodes, including the extensively studied model organism Caenorhabditis elegans, the function of TTL proteins remains obscure. Our data suggest a role in the nervous system. Even without insight into their biological function, the nematode-specific nature of this gene family makes it a promising target for nematicides or RNAi mediated control strategies against parasitic nematodes.
Cyst nematodes are sedentary plant parasites that cause dramatic cellular changes in the plant root to form feeding cells, so-called syncytia. 10A06 is a cyst nematode secretory protein that is most likely secreted as an effector into the developing syncytia during early plant parasitism. A homolog of the uncharacterized soybean cyst nematode (Heterodera glycines), 10A06 gene was cloned from the sugar beet cyst nematode (Heterodera schachtii), which is able to infect Arabidopsis (Arabidopsis thaliana). Constitutive expression of 10A06 in Arabidopsis affected plant morphology and increased susceptibility to H. schachtii as well as to other plant pathogens. Using yeast two-hybrid assays, we identified Spermidine Synthase2 (SPDS2), a key enzyme involved in polyamine biosynthesis, as a specific 10A06 interactor. In support of this protein-protein interaction, transgenic plants expressing 10A06 exhibited elevated SPDS2 mRNA abundance, significantly higher spermidine content, and increased polyamine oxidase (PAO) activity. Furthermore, the SPDS2 promoter was strongly activated in the nematode-induced syncytia, and transgenic plants overexpressing SPDS2 showed enhanced plant susceptibility to H. schachtii. In addition, in planta expression of 10A06 or SPDS2 increased mRNA abundance of a set of antioxidant genes upon nematode infection. These data lend strong support to a model in which the cyst nematode effector 10A06 exerts its function through the interaction with SPDS2, thereby increasing spermidine content and subsequently PAO activity. Increasing PAO activity results in stimulating the induction of the cellular antioxidant machinery in syncytia. Furthermore, we observed an apparent disruption of salicylic acid defense signaling as a function of 10A06. Most likely, increased antioxidant protection and interruption of salicylic acid signaling are key aspects of 10A06 function in addition to other physiological and morphological changes caused by altered polyamines, which are potent plant signaling molecules.
An endosymbiotic bacterium in a plant-parasitic nematode: member of a new Wolbachia supergroup.
Wolbachia is an endosymbiotic bacterium widely present in arthropods and animal-parasitic nematodes. Despite previous efforts, it has never been identified in plant-parasitic nematodes. Random sequencing of genes expressed by the burrowing nematode Radopholus similis resulted in several sequences with similarity to Wolbachia genes. The presence of a Wolbachia-like endosymbiont in this plant-parasitic nematode was investigated using both morphological and molecular approaches. Transmission electronmicroscopy, fluorescent immunolocalisation and staining with DAPI confirmed the presence of the endosymbiont within the reproductive tract of female adults. 16S rDNA, ftsZ and groEL gene sequences showed that the endosymbiont of R. similis is distantly related to the known Wolbachia supergroups. Finally, based on our initial success in finding sequences of this endosymbiont by screening an expressed sequence tag (EST) dataset, all nematode ESTs were mined for Wolbachia-like sequences. Although the retained sequences belonged to six different nematode species, R. similis was the only plant-parasitic nematode with traces of Wolbachia. Based on our phylogenetic study and the current literature we designate the endosymbiont of R. similis to a new supergroup (supergroup I) rather than considering it as a new species. Although its role remains unknown, the endosymbiont was found in all individuals tested, pointing towards an essential function of the bacteria.
An endosymbiotic bacterium in a plant-parasitic nematode: Member of a new Wolbachia supergroup.
Wolbachia is an endosymbiotic bacterium widely present in arthropods and animal-parasitic nematodes. Despite previous efforts, it has never been identified in plant-parasitic nematodes. Random sequencing of genes expressed by the burrowing nematode Radopholus similis resulted in several sequences with similarity to Wolbachia genes. The presence of a Wolbachia-like endosymbiont in this plant-parasitic nematode was investigated using both morphological and molecular approaches. Transmission electron microscopy, fluorescent immunolocalisation and staining with DAPI confirmed the presence of the endosymbiont within the reproductive tract of female adults. 16S rDNA, ftsZ and groEL gene sequences showed that the endosymbiont of R. similis is distantly related to the known Wolbachia supergroups. Finally, based on our initial success in finding sequences of this endosymbiont by screening an expressed sequence tag (EST) dataset, all nematode ESTs were mined for Wolbachia-like sequences. Although the retained sequences belonged to six different nematode species, R. similis was the only plant-parasitic nematode with traces of Wolbachia. Based on our phylogenetic study and the current literature we designate the endosymbiont of R. similis to a new supergroup (supergroup I) rather than considering it as a new species. Although its role remains unknown, the endosymbiont was found in all individuals tested, pointing towards an essential function of the bacteria.
2010-01-01
The filarial nematodes Brugia malayi, Wuchereria bancrofti and Onchocerca volvulus cause elephantiasis or dermatitis and blindness resulting in severe morbidity. Annually, 1.3 billion people are at risk of infection. Targeting the essential Wolbachia endobacteria of filarial nematodes with doxycycline has proven to be an effective therapy resulting in a block in embryogenesis, worm development and macrofilaricidal effects. However, doxycycline is contraindicated for a large portion of the at risk population. To identify new targets for anti-wolbachial therapy, understanding the molecular basis of the Wolbachia-filaria symbiosis is required. Using the B. malayi microarray we identified differentially expressed genes in the rodent filaria Litomosoides sigmodontis after depletion of Wolbachia...
Angiostrongylus cantonensis is a common cause of human eosinophilic meningitis. Recent outbreaks of this infection in endemic regions have prompted a need to determine the distribution of this nematode in the environment to control transmission. A. cantonensis is generally identified morphologically...
Molecular determinants archetypical to the phylum Nematoda
Full Text Available.BackgroundNematoda diverged from other animals between 600–1,200 million years ago and has become one of the most diverse animal phyla on earth. Most nematodes are free-living animals, but many are parasites of plants and animals including humans, posing major ecological and economical challenges around the world.ResultsWe investigated phylum-specific molecular characteristics in Nematoda by exploring over 214,000 polypeptides from 32 nematode species including 27 parasites. Over 50,000 nematode protein families were identified based on primary sequence, including ~10% with members from at least three different species. Nearly 1,600 of the multi-species families did not share homology to Pfam domains, including a total of 758 restricted to Nematoda. Majority of the 462 families that were conserved among both free-living and parasitic species contained members from multiple nematode clades, yet ~90% of the 296 parasite-specific families originated only from a single clade. Features of these protein families were revealed through extrapolation of essential functions from observed RNAi phenotypes in C. elegans, bioinformatics-based functional annotations, identification of distant homology based on protein folds, and prediction of expression at accessible nematode surfaces. In addition, we identified a group of nematode-restricted sequence features in energy-generating electron transfer complexes as potential targets for new chemicals with minimal or no toxicity to the host.ConclusionThis study identified and characterized the molecular determinants that help in defining the phylum Nematoda, and therefore improved our understanding of nematode protein evolution and provided novel insights for the development of next generation parasite control strategies.
Hsp-90 and the biology of nematodes
Full Text Available.BackgroundHsp-90 from the free-living nematode Caenorhabditis elegans is unique in that it fails to bind to the specific Hsp-90 inhibitor, geldanamycin (GA). Here we surveyed 24 different free-living or parasitic nematodes with the aim of determining whether C. elegans Hsp-90 was the exception or the norm amongst the nematodes. We combined these data with codon evolution models in an attempt to identify whether hsp-90 from GA-binding and non-binding species has evolved under different evolutionary constraints.ResultsWe show that GA-binding is associated with life history: free-living nematodes and those parasitic species with free-living larval stages failed to bind GA. In contrast, obligate parasites and those worms in which the free-living stage in the environment is enclosed within a resistant egg, possess a GA-binding Hsp-90. We analysed Hsp-90 sequences from fifteen nematode species to determine whether nematode hsp-90s have undergone adaptive evolution that influences GA-binding. Our data provide evidence of rapid diversifying selection in the evolution of the hsp-90 gene along three separate lineages, and identified a number of residues showing significant evidence of adaptive evolution. However, we were unable to prove that the selection observed is correlated with the ability to bind geldanamycin or not.ConclusionHsp-90 is a multi-functional protein and the rapid evolution of the hsp-90 gene presumably correlates with other key cellular functions. Factors other than primary amino acid sequence may influence the ability of Hsp-90 to bind to geldanamycin.
microRNAs: a role in drug resistance in parasitic nematodes?
2010-09-01
Full Text Available.Drug resistance in parasitic nematodes is an increasing problem worldwide, with resistance reported to all three commonly used classes of anthelmintics. Most studies to date have sought to correlate the resistant phenotype with genotypic changes in putative target molecules. Although this approach has identified mutations in several relevant genes, resistance might result from a complex interaction of different factors. Here we propose an alternative mechanism underlying the development of drug resistance based on functional differences in microRNA activity in resistant parasites. microRNAs play an important role in resistance to chemotherapeutic agents in many tumour cells and here we discuss whether they might also be involved in anthelmintic resistance in parasitic nematodes.
Host age, sex, and reproductive seasonality affect nematode parasitism in wild Japanese macaques
2010-01-01
Parasites are characteristically aggregated within hosts, but identifying the mechanisms underlying such aggregation can be difficult in wildlife populations. We examined the influence of host age and sex over an annual cycle on the eggs per gram of feces (EPG) of nematode parasites infecting wild Japanese macaques (Macaca fuscata yakui) on Yakushima Island. Five species of nematode were recorded from 434 fecal samples collected from an age-structured group of 50 individually recognizable macaques. All parasites exhibited aggregated EPG distributions. The ageinfection profiles of all three directly transmitted species (Oesophagostomum aculeatum, Strongyloides fuelleborni, and Trichuris trichiura) exhibited convex curves, but concavity better characterized the ageinfection curves of t...
2010-01-01
Entomopathogenic nematodes (EPNs) are lethal insect parasites that develop inside the body of their insect host. The EPN Steinernema riobrave infects many different types of soil insects, including larvae of the invasive citrus root weevil, Diaprepes abbreviatus, which is a key pest in citrus production. Soil characteristics have a great impact on nematode foraging efficacy and therefore on the ability of EPNs to be used successfully in biological control programs. Our specific objectives were to: (1) survey for the presence of native EPNs in California citrus groves; and (2) identify soil parameters that would allow us to predict EPN efficacy. The long-term objective of this study is to develop biological control methods to address the invasion of the citrus root weevil, D. abbreviatus, i...
2008-01-01
Endo-1,4-beta-glucanases, which can degrade cellulose, have been identified in a number of plant-parasitic nematodes, mainly sedentary endoparasites. This study reports the discovery of four different endoglucanases of glycosyl hydrolase family 5 (GHF5) in the migratory endoparasitic nematode Radopholus similis. Spatial expression of the corresponding genes was analysed by in situ hybridization, which showed the presence of transcripts in the pharyngeal gland cells. A semi-quantitative RT-PCR on different developmental stages was performed to study the temporal expression pattern. Three of the endoglucanase genes showed reduced expression in adult males compared with that in females. This could be explained by the fact that males do not feed and are considered non-parasitic. Only one of th...
Trichostrongylus colubriformis (Strongylida), a small intestinal nematode of small ruminants, is a major cause of production and economic losses in many countries. The aims of the present study were to define the transcriptome of the adult stage of T. colubriformis, using 454 sequencing technology and bioinformatic analyses, and to predict the main pathways that key groups of molecules are linked to in this nematode. A total of 21,259 contigs were assembled from the sequence data produced from a normalized cDNA library; 7876 of these contigs had known orthologues in the free-living nematode Caenorhabditis elegans, and encoded, amongst others, proteins with 'transthyretin-like' (8.8%), 'RNA recognition' (8.4%) and 'metridin-like ShK toxin' (7.6%) motifs. Bioinformatic analyses inferred that relatively high proportions of the C. elegans homologues are involved in biological pathways linked to 'peptidases' (4%), 'ribosome' (3.6%) and 'oxidative phosphorylation' (3%). Highly represented were peptides predicted to be associated with the nervous system, digestion of host proteins or inhibition of host proteases. Probabilistic functional gene networking of the complement of C. elegans orthologues (n=2126) assigned significance to particular subsets of molecules, such as protein kinases and serine/threonine phosphatases. The present study represents the first, comprehensive insight into the transcriptome of adult T. colubriformis, which provides a foundation for fundamental studies of the molecular biology and biochemistry of this parasitic nematode as well as prospects for identifying targets for novel nematocides. Future investigations should focus on comparing the transcriptomes of different developmental stages, both genders and various tissues of this parasitic nematode for the prediction of essential genes/gene products that are specific to nematodes.
Full Text Available.BackgroundThe fungivorus nematode, Aphelenchus avenae is widespread in soil and is found in association with decaying plant material. This nematode is also found in association with plants but its ability to cause plant disease remains largely undetermined. The taxonomic position and intermediate lifestyle of A. avenae make it an important model for studying the evolution of plant parasitism within the Nematoda. In addition, the exceptional capacity of this nematode to survive desiccation makes it an important system for study of anhydrobiosis. Expressed sequence tag (EST) analysis may therefore be useful in providing an initial insight into the poorly understood genetic background of A. avenae.ResultsWe present the generation, analysis and annotation of over 5,000 ESTs from a mixed-stage A. avenae cDNA library. Clustering of 5,076 high-quality ESTs resulted in a set of 2,700 non-redundant sequences comprising 695 contigs and 2,005 singletons. Comparative analyses indicated that 1,567 (58.0%) of the cluster sequences had homologues in Caenorhabditis elegans, 1,750 (64.8%) in other nematodes, 1,321(48.9%) in organisms other than nematodes, and 862 (31.9%) had no significant match to any sequence in current protein or nucleotide databases. In addition, 1,100 (40.7%) of the sequences were functionally classified using Gene Ontology (GO) hierarchy. Similarity searches of the cluster sequences identified a set of genes with significant homology to genes encoding enzymes that degrade plant or fungal cell walls. The full length sequences of two genes encoding glycosyl hydrolase family 5 (GHF5) cellulases and two pectate lyase genes encoding polysaccharide lyase family 3 (PL3) proteins were identified and characterized.ConclusionWe have described at least 2,214 putative genes from A. avenae and identified a set of genes encoding a range of cell-wall-degrading enzymes. This EST dataset represents a starting point for studies in a number of different fundamental and applied areas. The presence of genes encoding a battery of cell-wall-degrading enzymes in A. avenae and their similarities with genes from other plant parasitic nematodes suggest that this nematode can act not only as a fungal feeder but also a plant parasite. Further studies on genes encoding cell-wall-degrading enzymes in A. avenae will accelerate our understanding of the complex evolutionary histories of plant parasitism and the use of genes obtained by horizontal gene transfer from prokaryotes.
Effects of root- and stolen-infecting fungi on root-colonizing nematodes of white clover
2002-01-01
The effects on white clover (Trifolium repens ) of different combinations of the nematodes Meloidogyne trifoliophila , Helicotylenchus dihystera and Heterodera trifolii and nine stolon-infecting and three root-infecting fungi were studied in a glasshouse experiment. The presence of the fungus Phytophthora megasperma alone increased (P < 0·001) root-rot severity and reduced (P < 0·001) plant growth. Other species combinations, such as Phoma nebulosa and Alternaria alternata , interacted and increased root-rot severity. Combinations of P. megasperma with Pythium irregulare , and P. nebulosa with Phoma medicaginis or A. alternata , increased M. trifoliophila populations. Several other fungi (P. irregulare , P. nebulosa , Colletotrichum coccodes , Macrophomina phaseolina , P. medicaginis and Phoma sp.) interacted with the nematode M. trifoliophila causing severe root-knot symptoms. The results indicated that fungi and nematodes interacted to cause root and stolon rot and reduced yields, and that poor persistence of white clover in pastures is likely to be a problem with a complex etiology. Publisher: Oxford: Blackwell Publishing Relation: Plant Pathology Other identifier: URN:ISSN:0032-0862
Rhynchophorus palmarum, the palm weevil, has been reported as a pest of palms and sugarcane plants. The Red Ring Disease is an infectious plant disease caused by nematodes. The etiological agent, Bursaphelencus (Rhadinaphelencus) cocophilus (Nematoda: Aphelenchoididae), completes its life cycle within 9 to 10 days inside the palm tree. The main symptom is a permanent wilting of the plant aerial parts. Previous studies stated that B. cocophilus cohabits with other nematodes the gut of R. palmarum. The aim of this study is to identify nematodes collected from palm weevil found in coconut plantations from the north of the Rio de Janeiro State. Light (LM) and scanning electron microscopy (SEM) analyses were carried on samples of infected R. palmarum and fragments and fresh juice of infected plants with B. cocophilus. Observations of R. palmarum fecal material made by LM and SEM showed three species cohabiting these samples, being also present in fresh juice and fragments of infected coconut tree: B. cocophilus, Teratorhabditis palmarum (Nematoda: Rhabditidae) and Diplogasteritus sp (Nematoda: Diplogasteridae). These findings confirm previous studies, which related that R. palmarum own a varied nematode fauna. Nematodes associated to B. cocophilus probably could be co-participates of the etiology of the Red Ring disease.
2010-01-01
The main aim of this work was to assign the cuticular lipids identified in a parasitic nematode and to distinguish those originating from its host. The hypothesis that long-chained fatty acids and sterols are imported by the parasite in the absence of certain enzymes was also tested. The organisms (Anisakis simplex and Gadus morhua) were extracted in petroleum ether and dichloromethane. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF) was used to identify unknown components, and electrospray ionization mass spectrometry (ESI/MS) to verify recognized groups of lipids. The lipid classes identified in the surface layer were free saturated and unsaturated fatty acids, triacylglycerols, sterols and non-polar sphingolipids (ceramides, sphingoid bases). The most abund...
Sequence mining and transcript profiling to explore cyst nematode parasitism
Full Text Available.BackgroundCyst nematodes are devastating plant parasites that become sedentary within plant roots and induce the transformation of normal plant cells into elaborate feeding cells with the help of secreted effectors, the parasitism proteins. These proteins are the translation products of parasitism genes and are secreted molecular tools that allow cyst nematodes to infect plants.ResultsWe present here the expression patterns of all previously described parasitism genes of the soybean cyst nematode, Heterodera glycines, in all major life stages except the adult male. These insights were gained by analyzing our gene expression dataset from experiments using the Affymetrix Soybean Genome Array GeneChip, which contains probeset sequences for 6,860 genes derived from preparasitic and parasitic H. glycines life stages. Targeting the identification of additional H. glycines parasitism-associated genes, we isolated 633 genes encoding secretory proteins using algorithms to predict secretory signal peptides. Furthermore, because some of the known H. glycines parasitism proteins have strongest similarity to proteins of plants and microbes, we searched for predicted protein sequences that showed their highest similarities to plant or microbial proteins and identified 156 H. glycines genes, some of which also contained a signal peptide. Analyses of the expression profiles of these genes allowed the formulation of hypotheses about potential roles in parasitism. This is the first study combining sequence analyses of a substantial EST dataset with microarray expression data of all major life stages (except adult males) for the identification and characterization of putative parasitism-associated proteins in any parasitic nematode.ConclusionWe have established an expression atlas for all known H. glycines parasitism genes. Furthermore, in an effort to identify additional H. glycines genes with putative functions in parasitism, we have reduced the currently known 6,860 H. glycines genes to a pool of 788 most promising candidate genes (including known parasitism genes) and documented their expression profiles. Using our approach to pre-select genes likely involved in parasitism now allows detailed functional analyses in a manner not feasible for larger numbers of genes. The generation of the candidate pool described here is an important enabling advance because it will significantly facilitate the unraveling of fascinating plant-animal interactions and deliver knowledge that can be transferred to other pathogen-host systems. Ultimately, the exploration of true parasitism genes verified from the gene pool delineated here will identify weaknesses in the nematode life cycle that can be exploited by novel anti-nematode efforts.
2010-01-01
Filarial parasites are responsible for several serious human diseases with symptoms such as lymphoedema, elephantiasis, and blindness. An understanding of how these parasites pass through developmental checkpoints may suggest potential targets for intervention. A useful model system for the study of the human parasites is the closely related nematode, D. immitis, the causative agent of dog heartworm disease. Ecdysteroids have been identified in filarial nematodes and have been shown to have a biological affect both on molting and microfilarial production. The ecdysteroid, 20-hydroxyecdysone and its receptor, EcR, have a well-characterized developmental role in insects, where it is involved in the control of molting and metamorphosis. We have identified a D. immitis nuclear receptor, DiEcR ...
In rape ( Brassica napus), no resistance to the beet cyst nematode (BCN) Heterodera schachtii is available. This study was carried out to determine the specific chromosome(s) of resistant radish ( Raphanus sativus) carrying the gene(s) for nematode resistance as a prequisite to convert rape from a host into a trap crop for this pest. A Raphanobrassica progeny of 25 plants was analyzed which segregated for all nine chromosomes of the Raphanus genome in a genetic background of synthetic rape. The number of radish chromosomes was determined by fluorescence in situ hybridization, using the Raphanus-specific DNA probe pURsN; and their type was identified by chromosome-specific randomly amplified polymorphic DNA markers. Five different multiple rape-radish chromosome additions (comprising the whole set of nine radish chromosomes, a-i) were selected and crossed to rape. For each cross-progeny, the number of cysts on plant roots was counted 42 days after inoculation with a L2 larvae suspension. Simultaneously, the plants were characterized for the presence or absence of individual radish chromosomes, using sets of chromosome-specific markers. Thus, the effect of each radish chromosome on cyst number was tested. Chromosome d had a major resistance effect, whereas the presence/absence of the other radish chromosomes had nearly no influence on cyst number. Plants with added chromosome d showed a resistance level comparable with that of the radish donor parent. The analysis in the cross to rape of a plant monosomic only for chromosome d confirmed the strong effect of this chromosome on nematode resistance. A further experiment comprising seven crosses using winter rape breeding lines and monosomic addition line d as pollen parent provided the same results on a broader genetic basis. In each case, the added chromosome d in a single dosage caused nearly the full resistance of the radish donor. Resistance was independent of the glucosinolate content in the roots. The possibilities for stabilizing BCN resistance in rape and its use for other crops and nematodes are discussed.
Full Text Available.BackgroundThe success of a biological control agent depends on key traits, particularly reproductive potential, environmental tolerance, and ability to be cultured. These traits can deteriorate rapidly when the biological control agent is reared in culture. Trait deterioration under laboratory conditions has been widely documented in the entomopathogenic nematode (EPN) Heterorhabditis bacteriophora (Hb) but the specific mechanisms behind these genetic processes remain unclear. This research investigates the molecular mechanisms of trait deterioration of two experimental lines of Hb, an inbred line (L5M) and its original parental line (OHB). We generated transcriptional profiles of two experimental lines of Hb, identified the differentially expressed genes (DEGs) and validated their differential expression in the deteriorated line.ResultsAn expression profiling study was performed between experimental lines L5M and OHB of Hb with probes for 15,220 ESTs from the Hb transcriptome. Microarray analysis showed 1,185 DEGs comprising of 469 down- and 716 up-regulated genes in trait deteriorated nematodes. Analysis of the DEGs showed that trait deterioration involves massive changes of the transcripts encoding enzymes involved in metabolism, signal transduction, virulence and longevity. We observed a pattern of reduced expression of enzymes related to primary metabolic processes and induced secondary metabolism. Expression of sixteen DEGs in trait deteriorated nematodes was validated by quantitative reverse transcription-PCR (qRT-PCR) which revealed similar expression kinetics for all the genes tested as shown by microarray.ConclusionAs the most closely related major entomopathogen to C. elegans, Hb provides an attractive near-term application for using a model organism to better understand interspecies interactions and to enhance our understanding of the mechanisms underlying trait deterioration in biological control agents. This information could also be used to improve the beneficial traits of biological control agents and better understand fundamental aspects of nematode parasitism and mutualism.
BackgroundThe success of a biological control agent depends on key traits, particularly reproductive potential, environmental tolerance, and ability to be cultured. These traits can deteriorate rapidly when the biological control agent is reared in culture. Trait deterioration under laboratory conditions has been widely documented in the entomopathogenic nematode (EPN) Heterorhabditis bacteriophora (Hb) but the specific mechanisms behind these genetic processes remain unclear. This research investigates the molecular mechanisms of trait deterioration of two experimental lines of Hb, an inbred line (L5M) and its original parental line (OHB). We generated transcriptional profiles of two experimental lines of Hb, identified the differentially expressed genes (DEGs) and validated their differential expression in the deteriorated line.ResultsAn expression profiling study was performed between experimental lines L5M and OHB of Hb with probes for 15,220 ESTs from the Hb transcriptome. Microarray analysis showed 1,185 DEGs comprising of 469 down- and 716 up-regulated genes in trait deteriorated nematodes. Analysis of the DEGs showed that trait deterioration involves massive changes of the transcripts encoding enzymes involved in metabolism, signal transduction, virulence and longevity. We observed a pattern of reduced expression of enzymes related to primary metabolic processes and induced secondary metabolism. Expression of sixteen DEGs in trait deteriorated nematodes was validated by quantitative reverse transcription-PCR (qRT-PCR) which revealed similar expression kinetics for all the genes tested as shown by microarray.ConclusionAs the most closely related major entomopathogen to C. elegans, Hb provides an attractive near-term application for using a model organism to better understand interspecies interactions and to enhance our understanding of the mechanisms underlying trait deterioration in biological control agents. This information could also be used to improve the beneficial traits of biological control agents and better understand fundamental aspects of nematode parasitism and mutualism.
The role of pseudo-endoglucanases in the evolution of nematode cell wall-modifying proteins.
2010-04-24
In this article, the characterization and evolution of pseudo-endoglucanases and a putative expansin-like gene in the migratory nematode Ditylenchus africanus are described. Four genes were cloned with a very high similarity to the endoglucanase Da-eng1, which, however, lack a part of the catalytic domain most probably due to homologous recombination. Owing to this deletion, at least one of the catalytic residues of the corresponding protein is missing, and hence these genes are possibly pseudogenes. In two of the pseudo-endoglucanase genes, the deletions cause a frameshift (Da-engdel2, Da-engdel4), while two others (Da-engdel1, Da-engdel3) code for protein sequences with an intact carbohydrate-binding module (CBM). Recombinant proteins for Da-ENG1, Da-ENGDEL1, and Da-ENGDEL3 were demonstrated to bind to cellulose, while only Da-ENG1 showed cellulose-degrading activity. This indicates that Da-ENGDEL1 and Da-ENGDEL3 which lack cellulase activity, could still exert a function similar to cellulose-binding proteins (CBPs). Next to the pseudo-endoglucanases, a putative expansin-like gene (Da-exp1) was identified, consisting of a signal peptide, an expansin-like domain, and a CBM. This domain structure was never found before in nematode expansin-like proteins. Interestingly, the CBM of the expansin-like gene is very similar to the endoglucanase CBMs, and a conserved intron position in the CBM of nematode endoglucanases, expansin-like genes, and CBPs indicates a common origin for these domains. This suggests that domain shuffling is an important mechanism in the evolution of cell wall-modifying enzymes in nematodes.
The Wolbachia endosymbiont as an anti-filarial nematode target
2010-07-01
Full Text Available.Human disease caused by parasitic filarial nematodes is a major cause of global morbidity. The parasites are transmitted by arthropod intermediate hosts and are responsible for lymphatic filariasis (elephantiasis) or onchocerciasis (river blindness). Within these filarial parasites are intracellular alpha-proteobacteria, Wolbachia, that were first observed almost 30 years ago. The obligate endosymbiont has been recognized as a target for anti-filarial nematode chemotherapy as evidenced by the loss of worm fertility and viability upon antibiotic treatment in an extensive series of human trials. While current treatments with doxycycline and rifampicin are not practical for widespread use due to the length of required treatments and contraindications, anti-Wolbachia targeting nevertheless appears a promising alternative for filariasis control in situations where current programmatic strategies fail or are unable to be delivered and it provides a superior efficacy for individual therapy. The mechanisms that underlie the symbiotic relationship between Wolbachia and its nematode hosts remain elusive. Comparative genomics, bioinfomatic and experimental analyses have identified a number of potential interactions, which may be drug targets. One candidate is de novo heme biosynthesis, due to its absence in the genome sequence of the host nematode, Brugia malayi, but presence in Wolbachia and its potential roles in worm biology. We describe this and several additional candidate targets, as well as our approaches for understanding the nature of the host-symbiont relationship.
Metabolic profiling reveals local and systemic responses of host plants to nematode parasitism
2010-06-01
Full Text Available.The plant parasitic beet cyst nematode Heterodera schachtii induces syncytial feeding structures in Arabidopsis roots. The feeding structures form strong sink tissues that have been suggested to be metabolically highly active. In the present study, metabolic profiling and gene targeted expression analyses were performed in order to study the local and systemic effects of nematode infection on the plant host. The results showed increased levels of many amino acids and phosphorylated metabolites in syncytia, as well as high accumulation of specific sugars such as 1-kestose that do not accumulate naturally in Arabidopsis roots. A correlation-based network analysis revealed highly activated and coordinated metabolism in syncytia compared to non-infected control roots. An integrated analysis of the central primary metabolism showed a clear coherence of metabolite and transcript levels, indicating transcriptional regulation of specific pathways. Furthermore, systemic effects of nematode infection were demonstrated by correlation-based network analysis as well as independent component analysis. 1-kestose, raffinose, α,α-trehalose and three non-identified analytes showed clear systemic accumulation, indicating future potential for diagnostic and detailed metabolic analyses. Our studies open the door towards understanding the complex remodelling of plant metabolism in favour of the parasitizing nematode.
Codiostomum struthionis is a nematode parasite of the ostrich caecum. Little is known about its pathology, being considered by many authors as a non-pathogenic parasite. Infections by C. struthionis are sometimes overlooked because its eggs are indistinguishable from another ostrich nematode, Libyostrongylus spp. Fecal cultures and infective larvae identification are necessary for proper identification. The aim of this study is to provide improved morphological characterization of adults and infective larvae of C. struthionis. Ten caeca of adult ostriches were collected and washed in 0.09% saline solution. Male and female nematodes were collected and quantified separately. Nematodes were fixed in A.F.A. for optical microscopy or fixed in Karnovsky solution for scanning electron microscopy. To obtain infective larvae, fecal samples were collected at sites of high concentration of parasites in the caeca and fecal cultured. The resultant larvae were identified and measured with light microscope at 400x. Nine of the 10 slaughtered ostriches were parasitized by C. struthionis. All nematodes were found in the distal third of the caeca. A total of 566 parasites were recovered (234 males and 332 females). All the cultured larvae had characteristics of C. struthionis (rounded cephalic region with a flat extremity, an acute larvae tail termination and a long and filamentous sheath tail). All the adult parasites were characterized as C. struthionis. Through the analysis of the infective larvae it was determined that the morphology of the larvae tail was the best trait to use in the distinction of this species (live bird diagnosis).
We previously isolated a partial soybean cDNA clone whose transcript abundance is increased upon infection by the sedentary, endoparasitic soybean cyst nematode Heterodera glycines. We now isolated the corresponding full-length cDNA and determined that the predicted gene product was similar to the group of cofactor-dependent phosphoglycerate mutase/bisphosphoglycerate mutase enzymes (PGM/bPGM; EC 5.4.2.1/5.4.2.4). We designated the corresponding soybean gene GmPGM. PGM and bPGM are key catalysts of glycolysis that have been well characterized in animals but not plants. Using the GmPGM cDNA sequence, we identified a homologous Arabidopsis thaliana gene, which we designated AtPGM. Histochemical GUS analyses of transgenic Arabidopsis plants containing the AtPGM promoter ::GUS construct revealed that the AtPGM promoter directs GUS expression in uninfected plants only to the shoot and root apical meristems. In infected plants, GUS staining also is evident in the nematode feeding structures induced by the cyst nematode Heterodera schachtii and by the root-knot nematode Meloidogyne incognita. Furthermore, we discovered that the AtPGM promoter was down-regulated by abscisic acid and hydroxyurea, whereas it was induced by sucrose, oryzalin, and auxin, thereby revealing expression characteristics typical of genes with roles in meristematic cells. Assessment of the auxin-inducible AUX1 gene promoter (a gene coding for a polar auxin transport protein) similarly revealed feeding cell and meristem expression, suggesting that auxin may be responsible for the observed tissue specificity of the AtPGM promoter. These results provide first insight into the possible roles of PGM/bPGM in plant physiology and in plant-pathogen interactions.
Plant-parasitic cyst nematodes secrete a complex of cell wall-digesting enzymes, which aid in root penetration and migration. The soybean cyst nematode Heterodera glycines also produces a cellulose binding protein (Hg CBP) secretory protein. To determine the function of CBP, an orthologous cDNA clone (Hs CBP) was isolated from the sugar beet cyst nematode Heterodera schachtii, which is able to infect Arabidopsis thaliana. CBP is expressed only in the early phases of feeding cell formation and not during the migratory phase. Transgenic Arabidopsis expressing Hs CBP developed longer roots and exhibited enhanced susceptibility to H. schachtii. A yeast two-hybrid screen identified Arabidopsis pectin methylesterase protein 3 (PME3) as strongly and specifically interacting with Hs CBP. Transgenic plants overexpressing PME3 also produced longer roots and exhibited increased susceptibility to H. schachtii, while a pme3 knockout mutant showed opposite phenotypes. Moreover, CBP overexpression increases PME3 activity in planta. Localization studies support the mode of action of PME3 as a cell wall-modifying enzyme. Expression of CBP in the pme3 knockout mutant revealed that PME3 is required but not the sole mechanism for CBP overexpression phenotype. These data indicate that CBP directly interacts with PME3 thereby activating and potentially targeting this enzyme to aid cyst nematode parasitism.
[Gastro-intestinal helminths detected by fecal examination in stray dogs in the Aydin province]
Fecal specimens of a total of 200 dogs were examined by native, Fulleborn's floatation and Benedek's sedimentation methods to determine the prevalence of gastro-intestinal helminth infections in stray dogs in the Aydin Municipality Animal Shelter and the Ku?adasi Municipality Animal Shelter. Helminth infections were encountered in 82 (41%) of the fecal samples examined. One cestode egg and four nematode eggs were found in the infected fecal samples. The helminth eggs found were identified as follows: Taenia spp. (7.5%), Toxacara spp. (20%), Toxascaris leonina (1%), Uncinaria stenocephala (21%) and Trichuris vulpis (1.5%). No trematode eggs and nematode larvae were found in this study.
Zoonotic intravitreal Onchocerca in Hungary
2005-01-01
Objective To report the case of a 65-year-old male patient from western Hungary who presented with rapidly progressive peripheral visual field (VF) loss and the sensation of an actively moving object in his central VF. Design Interventional case report. Method/intervention A live nematode was removed from the anterior vitreous cavity by pars plana vitrectomy. Results The worm was successfully removed surgically, and the patient had an uneventful recovery. The nematode was identified as an immature filaria, most likely a member of the genus Onchocerca. Conclusions Only 3 previous reports exist of human infection of the eye caused by zoonotic Onchocerca, 2 involving the subconjunctiva and 1 the cornea. Although rare, zoonotic onchocercal infection of the eye must be considered a differential...
2009-01-01
Gastrointestinal nematodes were recovered from thirty four Gracilinanus agilis from forty four collected in Pantanal, Mato Grosso do Sul State, Brazil. Two hundred seventy four spirurids were recovered from the esophagus, stomach and intestines, comprising three species from three different genera. These were identified as Pterygodermatites (Paucipectines) jagerskioldi, Spirura guianensis and Physaloptera herthameyerae n. sp. is first described. This is the first record of nematodes of the genera Physaloptera and Spirura in hosts of the genus Gracilinanus. The high prevalence of spirurids in 72.3% of the G. agilis collected probably is influenced by the arboreal and diet behaviors.
2006-01-01
SUMMARY Parasitic nematodes are constantly exposed to the immune effector mechanisms of their hosts. One strategy of the worms to cope with these defence reactions is the secretion of modulatory proteins that down-regulate cell-mediated immune responses. We analysed the proliferation of mesenteric lymph node cells of mice infected with Heligmosomoides polygyrus and showed that cellular proliferation was strongly suppressed in the chronic phase of infection. To identify proteins of H. polygyrus that are involved in parasite-induced immunomodulation, worm extract and culture supernatant of adult H. polygyrus were fractionated by gel chromatography and activity of each fraction was determined. One of the fractions (fraction 9) of worm extract as well as worm secretory products inhibited the a...
2010-01-01
The genome of Caenorhabditis elegans includes six homologs of matrix metalloproteinases (MMPs). The C. elegans MMP gene zmp-1 has recently been shown to be involved in anchor cell invasion during post-embryonic vulval development. Here, we identified H19M22.3 (zmp-2) as a pleiotropic MMP gene regulating disease resistance, molting, larval development, and fecundity. Zmp-2(RNAi) nematodes showed significant lifespan reduction during infection with pathogenic Photorhabdus luminescence. Moreover, we observed molting defects indicating a direct or regulative role in extracellular matrix degradation during ecdysis, delayed larval to adult development, and reduced offspring production in hermaphrodite adults. GFP-expressing nematodes revealed predominant expression of zmp-2 in multiple cells dur...
Photorhabdus temperata subsp. stackebrandtii subsp. nov. (Enterobacteriales: Enterobacteriaceae)
2010-01-01
The bacterial symbiont of the entomopathogenic nematode Heterorhabditis bacteriophora strain GPS11 was characterized by 16S rRNA gene sequence and physiological traits. The phylogenetic tree built upon 16S rRNA gene sequences clustered the GPS11 bacterial isolate with Photorhabdus temperata strains which have been previously isolated from Heterorhabditis species. The phylogenetic tree further identified four subgroups in P. temperata, and the relationships among these subgroups were confirmed by gyrase subunit B (gyrB) gene sequence analysis. The subgroup containing the GPS11 bacterial isolate differs from other subgroups in sequences of 16S rRNA and gyrB gene, physiological traits, nematode host species, and geographic origin. Therefore, the subgroup comprising the GPS11 bacterial isolate...
Parasite Prevalence, Abundance, and Diversity in a Semi-free-ranging Colony of Mandrillus sphinx
2007-01-01
We examined parasite prevalence, abundance of protozoan cysts and helminth eggs in the feces, and number of parasitic taxa in a population of mandrills (Mandrillus sphinx) in semi-free-ranging conditions in their habitat range, with respect to the annual cycle, sex, age, dominance rank, and female reproductive status. We identified 3 taxa of amebic protozoa (Entamoeba coli, E. histolytica/dispar complex, and Endolimax nana), 1 ciliate protozoa (Balantidium coli), and various nematodes. Prevalence ranged from 1 observation in 874 samples for Trichuris and Mammomonogamus (nematodes) to 100% for Entamoeba coli and Endolimax nana. Daily observation, consistency of fecal samples, and periodic veterinary examination indicated that the mandrills were all healthy, suggesting that the presence of i...
Neurodegenerative disorders: Insights from the nematode Caenorhabditis elegans
2010-01-01
Neurodegenerative diseases impose a burden on society, yet for the most part, the mechanisms underlying neuronal dysfunction and death in these disorders remain unclear despite the identification of relevant disease genes. Given the molecular conservation in neuronal signaling pathways across vertebrate and invertebrate species, many researchers have turned to the nematode Caenorhabditis elegans to identify the mechanisms underlying neurodegenerative disease pathology. C. elegans can be engineered to express human proteins associated with neurodegeneration; additionally, the function of C. elegans orthologs of human neurodegenerative disease genes can be dissected. Herein, we examine major C. elegans neurodegeneration models that recapitulate many aspects of human neurodegenerative disease...
2009-01-01
Intensification of animal production systems has led to an increased reliance on effective anthelmintics to control parasitic worms. However, the excessive and continued use of these ''wormers'' can lead to high selection pressures and have resulted in increased reports of emerging nematode populations exhibiting resistance to all of the main anthelmintic classes. Faecal egg count reduction tests (FECRT) were conducted on six farms in England and Wales according to standardised guidelines produced by the World Association for the Advancement of Parasitology. Selected farms were identified from a network of 40 study farms participating in a study investigating implementation of Sustained Control of Parasites in Sheep (SCOPS) principles. Resistance to the larval development test (LDT) to eit...
2010-01-01
Gastrointestinal nematodes of the family Trichostrongylidae are the most important parasites of sheep, goats and other ruminants worldwide. Of this group, Haemonchus contortus is usually the most damaging species, particularly in warmer climates or during the summer. It is therefore useful to be able to rapidly differentiate infections with this nematode from other, less pathogenic, species. However, aside from Nematodirus spp., there are only subtle differences between the egg morphology within the trichostrongyles, making it very difficult to identify eggs to the species level. It has been shown previously that peanut agglutinin specifically binds to Haemonchus eggs and not those of other trichostrongyle species. By using this lectin conjugated to fluorescein isothiocyanate, binding to H...
Immune mechanisms of resistance to gastrointestinal nematode infections in sheep
2010-01-01
Summary Infections with gastrointestinal nematode parasites are a major problem for the sheep industry in Australia and New Zealand and have been the subject of intensive research to define mechanisms of resistance. The ability to take continuous biopsy samples of infected organs and cannulate both afferent and efferent lymphatics of draining lymph nodes has been particularly useful in illuminating the kinetics of immune responses at the site of infection. Distinct localized immune responses were shown to occur within and between sheep breeds at different sensitization regimes, as well as at different developmental stages of the parasite within the host. Using localized antibodies derived from mucus and lymph nodes, two major antigens have been identified on the infective L3 stage, which m...
HAIRY-like Transcription Factors and the Evolution of the Nematode Vulva Equivalence Group
2006-01-01
SummaryBackgroundNematode vulva formation provides a paradigm to study the evolution of pattern formation and cell-fate specification. The Caenorhabditis elegans vulva is generated from three of six equipotent cells that form the so-called vulva equivalence group. During evolution, the size of the vulva equivalence group has changed: Panagrellus redivivus has eight, C. elegans six, and Pristionchus pacificus only three cells that are competent to form vulval tissue. In P. pacificus, programmed cell death of individual vulval precursor cells alters the size of the vulva equivalence group.ResultsWe have identified the genes controlling this cell-death event and the molecular mechanism of the reduction of the vulva equivalence group. Mutations in Ppa-hairy, a gene that is unknown from C. eleg...
Density-dependent immune responses against the gastrointestinal nematode Strongyloides ratti
2007-01-01
Negative density-dependent effects on the fitness of parasite populations are an important force in their population dynamics. For the parasitic nematode Strongyloides ratti, density-dependent fitness effects require the rat host immune response. By analysis of both measurements of components of parasite fitness and of the host immune response to different doses of S. ratti infection, we have identified specific parts of the host immune response underlying the negative density-dependent effects on the fitness of S. ratti. The host immune response changes both qualitatively from an inflammatory Th1- to a Th2-type immune profile and the Th2-type response increases quantitatively, as the density of S. ratti infection increases. Parasite survivorship was significantly negatively related to the...
Cutaneous periocular Habronema infection in a dromedary camel (Camelus dromedarius)
2010-01-01
Abstract A 6-year-old castrated dromedary camel (Camelus dromedarius) presented with a non-healing, severely pruritic, ulcerative fibrotic plaque located at the medial canthus. Histological examination of surgical biopsies identified degenerating nematode larvae within eosinophilic granulomas. Treatment involved repeated debridement of the lesion, injectable ivermectin and anti-inflammatory therapies, and injectable and topical antibiotics. A specially constructed mask with goggles to prevent the camel from continuing to self-traumatize the eye and lesion was also placed. Full recovery occurred approximately 1 month after diagnosis. Because of the location of the lesion, time of year, the gross and microscopic characteristics of the lesion, the presence of a likely nematode larva and the r...
Crossover suppressors and balanced recessive lethals in Caenorhabditis elegans. [Nematode]
Two dominant suppressors of crossing over have been identified following x-ray treatment of the small nematode C. elegans. They suppress crossing over in linkage group II (LGII) about 100-fold and 50-fold and are both tightly linked to LGII markers. One, called C1, segregates independently of all other linkage groups and is homozygous fertile. The other is a translocation involving LGII and X. The translocation also suppresses crossing over along the right half of X and is homozygous lethal. C1 has been used as a balancer of LGII recessive lethal and sterile mutations induced by EMS. The frequencies of occurrence of lethals and steriles were approximately equal. Fourteen mutations were assigned to complementation groups and mapped. They tended to map in the same region where LGII visibles are clustered.
Crossover suppressors and balanced recessive lethals in Caenorhabditis elegans. [Nematode]
1978-01-01
Two dominant suppressors of crossing over have been identified following x-ray treatment of the small nematode C. elegans. They suppress crossing over in linkage group II (LGII) about 100-fold and 50-fold and are both tightly linked to LGII markers. One, called C1, segregates independently of all other linkage groups and is homozygous fertile. The other is a translocation involving LGII and X. The translocation also suppresses crossing over along the right half of X and is homozygous lethal. C1 has been used as a balancer of LGII recessive lethal and sterile mutations induced by EMS. The frequencies of occurrence of lethals and steriles were approximately equal. Fourteen mutations were assigned to complementation groups and mapped. They tended to map in the same region where LGII visibles are clustered.
2009-01-01
Cooperia oncophora is an important parasitic nematode of cattle with a wide distribution in temperate areas. Twenty Holstein nematode-naive bull calves were experimentally infected with approximately 100,000 infective L3s and infection was allowed to progress for 7, 14, 28, 42 days, respectively. This experiment was conducted to identify putative recognition and inflammatory pathways in the host-parasite relationship. Gene expression profiles of the small intestine were compared using a high-density bovine 60 mer oligo microarray. A total of 310 genes were differentially expressed during the course of infection. The pathways and regulatory networks significantly impacted by the infection were analysed. A total of 22 canonical pathways and nine regulatory networks were significantly affecte...
A pathogenic parasite interferes with phagocytosis of insect immunocompetent cells
2010-01-01
Phagocytosis activity of hemocytes of the host Galleria mellonella (Lepidoptera) was modulated by the infection of the entomopathogenic nematode Steinernema feltiae (Rahbditida) and was found to be correlated with the opsonization of bacteria by hemolymph factors. The presence of nematodes resulted in a significative decrease in phagocytosis of bacteria by host hemocytes, both in in vivo and in in vitro assays. Host interacting proteins (HIPs), which appear to function as opsonic factors and are essential to perform immune responses, were removed by S. feltiae from host hemolymph, by means of its epicuticle binding properties. Host humoral factors sequestered by the parasite have been identified by monodimensional and 2D electrophoretic analysis. The data suggest that S. feltiae, living in...
A metabolic switch is involved in lifestyle decisions in Photorhabdus luminescens
2010-01-01
Summary Photorhabdus luminescens is a species of Gram-negative bacteria that is pathogenic to insects while also maintaining a mutualistic association with nematodes from the family Heterorhabditis. P. luminescens elaborates an extensive secondary metabolism during the post-exponential phase of growth that includes the production of an antibiotic called 3-5-dihydroxy-4-isopropylstilbene (ST), an anthraquinone pigment (AQ) and bioluminescence. In this study we identified a mutant that was unable to produce ST, AQ and light. This mutation was found to be in the mdh gene, encoding malate dehydrogenase, a key enzyme in the tricarboxylic acid (TCA) cycle. Interestingly the mdh mutant was unaffected in virulence but was unable to support nematode growth and development in vivo or in vitro. This ...
2009-01-01
Southern root-knot nematode (Meloidogyne incognita) is a major problem in vegetable production in China due to the expansion of plastic tunnel and solar greenhouse. Using resistant cultivars is an effective approach to control the disease. Nine genes, Mi-1 to Mi-9, have been reported and only Mi-1 has been successfully used in tomato breeding. However, Mi-1 is inactive at a temperature above 28degreeC. In order to identify sources for heat-stable resistance to southern root-knot nematode, 53 genotypes of tomato (Solanum spp.) were inoculated with an isolate of M. incognita in the growth chamber at 28 or 32degreeC for initial screening. 28 lines had less than 25 galls and were considered as resistant candidates. The top 60% (16 in total) of resistant candidates obtained from each temperatur...
The aim of this study was to evaluate the influence of different doses of Heterorhabditis amazonensis RSC-5 on the biological parameters of engorged females of Rhipicephalus (Boophilus) microplus. The female ticks, individually identified, were divided into six groups of 20 each and exposed to the following nematode concentrations: 0, 75, 150, 300, 600, and 1,200/female. The following parameters were observed: initial weight, final weight, alteration weight, egg mass weight, pre-oviposition period, oviposition period, survival period, incubation period, hatching percentage, reproductive efficiency index (REI), nutritional efficiency index (NI), and percentage of control. There were no significant differences observed for the initial weight, pre-oviposition period, incubation period, and NI (p > 0.05) between the various treated groups and the control group. However, there were significant differences (p 0.05). The highest control rate was 67.8%, obtained in the treatment with a concentration of 300 entomopathogenic nematodes/female. The results demonstrate that this nematode has a deleterious effect on the majority of the biological parameters analyzed.
Haemonchus contortus: molecular characterisation of a small heat shock protein
2003-01-01
A cDNA encoding a predicted small heat shock protein, HSP20, was isolated from the parasitic nematode Haemonchus contortus. This cDNA encoded a predicted protein of 156 amino acids, which had high sequence identity with other nematode small heat shock proteins. Southern blot analysis of genomic DNA suggested that in H. contortus HSP20 is encoded by a single copy gene. The HSP20 transcript and protein were expressed in the infective larvae (L3), early L4 and adult stages, but expression was not increased by heat shock treatment. In situ hybridisation analysis was used to localise expression of HSP20 mRNA in the adult parasite. Similar HSPs (heat shock protein) were detected by Western blotting in Ancylostoma caninum, Dictyocaulus viviparus, and Toxocara canis, but not in Trichostronglyus colubriformis. The conservation of HSP20 in several different nematode species may reflect its importance to parasites that require mammalian hosts as a part of their development. Publisher: Academic Press Other identifier: swin:15156 Language: English Source: Experimental Parasitology, Vol. 104, no. 304 (Jul-Aug 2003), pp. 96-103 Rights: Copyright © 2003 Elsevier B.V. All rights reserved.
2009-01-01
Infected calves from two different rural estates in Brazil were studied to assess the anthelmintic efficacy of oral trichlorfon against naturally occurring ivermectin resistant parasitic nematode strains. In experiment I, infected animals were from a region where ivermectin resistant populations of Haemoncus placei, Cooperia punctata, Cooperia spatulata and Trichuris discolor have recently been identified. Six calves with natural gastrointestinal nematode infections were treated with 48.5mg/kg aqueous trichlorfon administered orally and six calves acted as a non-treated control group. In experiment II 24 naturally infected calves were selected to enter one of four treatment groups, six animals each received: 48.5mg/kg oral trichlorfon; 200mg/kg subcutaneous 1% ivermectin; 630mg/kg subcutan...
2009-01-01
Anguina paludicola sp. n. is described from Polypogon monspeliensis from southeastern South Australia and Lachnagrostis filiformis from northern New South Wales, following an examination of nematode populations by allozyme electrophoresis and morphological analysis. Genetic differences were consistent with those expected from conspecific allopatric populations and were based on geography rather than host. Morphological differences between nematodes from P. monspeliensis and L. filiformis were inconsistent and varied between different gall types on the same host plant. The new species is characterised by adults with irregular and variable tail termini, which appear lobed; females may have a process of vestigial ovary extending from the post uterine sac towards the tail; males are dorsally curved with a single reflexed testis; and the haploid chromosome complement is n=18.Terry Bertozzi and Kerrie A. Davieshttp://www.mapress.com/zootaxa/2009/f/z02060p046f.pdf Publisher: Magnolia Press Contributor: School of Agriculture, Food and Wine Other identifier: Zootaxa, 2009;(2060):33-46; 1175-5326; 0020090400; 000264719700003 Language: en Rights: Copyright 2009 Magnolia Press
The experimental establishment of ruminant nematodes in European hares (Lepus europaeus)
2009-01-01
Philip Stott, Michael O’Callaghan, Peter Phillips and Ari Verbyla Publisher: Elsevier Science BV Contributor: School of Agriculture, Food and Wine Other identifier: Veterinary Parasitology, 2009; 159(1):82-85; 0304-4017; 0020083957; 10.1016/j.vetpar.2008.10.016; 000262663600014 Language: en
2006-03-31
DescriptionTo use 'omic' technologies (transcriptomics, proteonics, mechanistics and demographics) to catalogue the response cascades in model and sentinel soil organisms exposed to a range of pollutants of concern. Central to achieving this goal will be the exploitation of the vast bioinformatic resources available for a model organism (the nematode C. elegans) to identify and characterise pollutant responses shared with a sentinel invertebrate (the earthworm L. rubellus). This unique data set will identi [continued...]
2006-03-08
DescriptionTo use 'omic' technologies (transcriptomics, proteonics, mechanistics and demographics) to catalogue the response cascades in model and sentinel soil organisms exposed to a range of pollutants of concern. Central to achieving this goal will be the exploitation of the vast bioinformatic resources available for a model organism (the nematode C. elegans) to identify and characterise pollutant responses shared with a sentinel invertebrate (the earthworm L. rubellus). This unique data set will identi [continued...]
2005-06-30
DescriptionTo use 'omic' technologies (transcriptomics, proteonics, mechanistics and demographics) to catalogue the response cascades in model and sentinel soil organisms exposed to a range of pollutants of concern. Central to achieving this goal will be the exploitation of the vast bioinformatic resources available for a model organism (the nematode C. elegans) to identify and characterise pollutant responses shared with a sentinel invertebrate (the earthworm L. rubellus). This unique data set will identi [continued...]
2004-06-30
DescriptionTo use 'omic' technologies (transcriptomics, proteonics, mechanistics and demographics) to catalogue the response cascades in model and sentinel soil organisms exposed to a range of pollutants of concern. Central to achieving this goal will be the exploitation of the vast bioinformatic resources available for a model organism (the nematode C. elegans) to identify and characterise pollutant responses shared with a sentinel invertebrate (the earthworm L. rubellus). This unique data set will identi [continued...]
2005-12-31
Objectives'DescriptionTo use 'omic' technologies (transcriptomics, proteonics, mechanistics and demographics) to catalogue the response cascades in model and sentinel soil organisms exposed to a range of pollutants of concern. Central to achieving this goal will be the exploitation of the vast bioinformatic resources available for a model organism (the nematode C. elegans) to identify and characterise pollutant responses shared with a sentinel invertebrate (the earthworm L. rubellus). This unique data set will identi [continued...]
Autophagy Shows Its Animal Side
2010-01-01
Most autophagy genes have been discovered in the single-celled yeast Saccharomyces cerevisiae, and little is known about autophagy genes that are specific to multicellular animals. In this issue, Tian et al. (2010) now identify four new autophagy genes: one specific to the nematode Caenorhabditis elegans and three conserved from worms to mammals.
Arabidopsis small RNAs and their targets during cyst nematode parasitism.
Plant-parasitic cyst nematodes induce the formation of specialized feeding cells in infected roots, which involves plant developmental processes that have been shown to be influenced by microRNAs (miRNAs) and other small RNAs. This observation provided the foundation to investigate the potential involvement of small RNAs in plant-cyst nematode interactions. First, we examined the susceptibilities of Arabidopsis DICER-like (dcl) and RNA-dependent RNA polymerase (rdr) mutants to the sugar beet cyst nematode Heterodera schachtii. The examined mutants exhibited a trend of decreased susceptibility, suggesting a role of small RNAs mediating gene regulation processes during the plant-nematode interaction. Second, we generated two small RNA libraries from aseptic Arabidopsis roots harvested at 4 and 7 days after infection with surface-sterilized H. schachtii. Sequences of known miRNAs as well as novel small interfering (si)RNAs were identified. Following this discovery, we used real-time reverse-transcriptase polymerase chain reaction to quantify a total of 15 Arabidopsis transcripts that are known targets of six of the different miRNA families found in our study (miR160, miR164, miR167, miR171, miR396, and miR398) in inoculated and noninoculated Arabidopsis roots. Our analyses showed mostly negative correlations between miRNA accumulation and target gene mRNA abundance, suggesting regulatory roles of these miRNAs during parasitism. Also, we identified a total of 125 non-miRNA siRNAs. Some of these siRNAs perfectly complement protein-coding mRNAs or match transposon or retrotransposon sequences in sense or antisense orientations. We further quantified a group of siRNAs in H. schachtii-inoculated roots. The examined siRNAs exhibited distinct expression patterns in infected and noninfected roots, providing additional evidence for the implication of small RNAs in cyst nematode parasitism. These data lay the foundation for detailed analyses of the functions of small RNAs during phytonematode parasitism.
2010-01-01
Full Text Available. Background. A comparative microarray investigation was done using detection call methodology (DCM) and differential expression analyses. The goal was to identify genes found in specific cell populations that were eliminated by differential expression analysis due to the nature of differential expression methods. Laser capture microdissection (LCM) was used to isolate nearly homogeneous populations of plant root cells. Results. The analyses identified the presence of 13,291 transcripts between the 4 different sample types. The transcripts filtered down into a total of 6,267 that were detected as being present in one or more sample types. A comparative analysis of DCM and differential expression methods showed a group of genes that were not differentially expressed, but were expressed at detectable amounts within specific cell types. Conclusion. The DCM has identified patterns of gene expression not shown by differential expression analyses. DCM has identified genes that are possibly cell-type specific and/or involved in important aspects of plant nematode interactions during the resistance response, revealing the uniqueness of a particular cell population at a particular point during its differentiation process.
Development of key soil health indicators for the Australian banana industry
2008-01-01
To improve the sustainability and environmental accountability of the banana industry there is a need to develop a set of soil health indicators that integrate physical, chemical and biological soil properties. These indicators would allow banana growers, extension and research workers to improve soil health management practices. To determine changes in soil properties due to the cultivation of bananas, a paired site survey was conducted comparing soil properties under conventional banana systems to less intensively managed vegetation systems, such as pastures and forest. Measurements were made on physical, chemical and biological soil properties at seven locations in tropical and sub-tropical banana producing areas. Soil nematode community composition was used as a bioindicator of the biological properties of the soil. Soils under conventional banana production tended to have a greater soil bulk density, with less soil organic carbon (C) (both total C and labile C), greater exchangeable cations, higher extractable P, greater numbers of plant-parasitic nematodes and less nematode diversity, relative to less intensively managed plant systems. The organic banana production systems at two locations had greater labile C, relative to conventional banana systems, but there was no significant change in nematode community composition. There were significant interactions between physical, chemical and nematode community measurements in the soil, particularly with soil C measurements, confirming the need for a holistic set of indicators to aid soil management. There was no single indicator of soil health for the Australian banana industry, but a set of soil health indicators, which would allow the measurement of soil improvements should include: bulk density, soil C, pH, EC, total N, extractable P, ECEC and soil nematode community structure. Publisher: Elsevier B.V. Other identifier: Pattison, A.B. and Moody, P.W. and Badcock, K.A. and Smith, L.J. and Armour, J.A. and Rasiah, V. and Cobon, J.A. and Gulino, L.-M. and Mayer, R. (2008) Development of key soil health indicators for the Australian banana industry. Applied Soil Ecology, 40 (1). pp. 155-164.
Influence of metalaxyl on three nematodes of citrus.
Metalaxyl significantly reduced population of Pratylenchus coffeae, Radopholus similis, and Tylenchulus semipenetrans in roots of Citrus limon (rough lemon) under greenhouse conditions. Postinoculation treatment of rough lemon seedlings was not as effective i n reducing nematode populations as was treatment before inoculation. Fewer nematodes infected metalaxyl-treated roots than nontreated roots. However, incubation of nematodes in metalaxyl did not inhibit nematode motility or their ability to locate and infect roots. Cellular responses to nematode injection differed between treated and nontreated tissues. Metalaxyl appeared to confer nematode contraol by modifying citrus roots such that a normally susceptible rootstock became tolerant.
2006-06-30
DescriptionSoil nematode populations may be controlled by fungi; V. chlamydosporium, the causal agent of the decline of several nematode pests in a range of soils, is very variable. It is unclear if nematode suppression is due to one effective isolate or several distinct isolates acting together. Molecular, biochemical and population studies will investigate the spatio-temporal dynamics of different fungal isolates in soil, rhizosphere and nematode eggmasses. Determining isolate diversity in these nich [continued...]
2010-01-01
Rma, a dominant root-knot nematode resistance gene introduced into tetraploid peanut (Arachis hypogaea) from a synthetic allotetraploid donor (TxAG-6), has been widely deployed in modern cultivars. The genomic location and borders of the alien chromosome segment introgressed from TxAG-6 into NemaTAM (a BC7-derived introgression line) and other modern cultivars carrying Rma have not been genetically mapped, and resistance gene candidates (RGCs) have not been identified for Rma. Our study focused on densely populating the alien introgression with codominant DNA markers, identifying and mapping the borders of the alien introgression carried by NemaTAM, and identifying RGCs for Rma. Altogether, 2,847 simple sequence repeat (SSR) and 380 single strand conformational polymorphism (SSCP) markers ...
Full Text Available.BackgroundWolbachia are among the most abundant symbiotic microbes on earth; they are present in about 66% of all insect species, some spiders, mites and crustaceans, and most filarial nematode species. Infected filarial nematodes, including many pathogens of medical and veterinary importance, depend on Wolbachia for proper development and survival. The mechanisms behind this interdependence are not understood. Interestingly, a minority of filarial species examined to date are naturally Wolbachia-free.Methodology/Principal FindingsWe used 454 pyrosequencing to survey the genomes of two distantly related Wolbachia-free filarial species, Acanthocheilonema viteae and Onchocerca flexuosa. This screen identified 49 Wolbachia-like DNA sequences in A. viteae and 114 in O. flexuosa. qRT-PCR reactions detected expression of 30 Wolbachia-like sequences in A. viteae and 56 in O. flexuosa. Approximately half of these appear to be transcribed from pseudogenes. In situ hybridization showed that two of these pseudogene transcripts were specifically expressed in developing embryos and testes of both species.Conclusions/SignificanceThese results strongly suggest that the last common ancestor of extant filarial nematodes was infected with Wolbachia and that this former endosymbiont contributed to their genome evolution. Horizontally transferred Wolbachia DNA may explain the ability of some filarial species to live and reproduce without the endosymbiont while other species cannot.
Sedentary plant-parasitic nematodes are able to induce specialized feeding structures in the root system of their host plants by triggering a series of dramatic cellular responses. These changes presumably are accompanied by a reprogramming of gene expression. To monitor such changes, a variety of promoter-gusA fusion constructs were introduced into Arabidopsis and tobacco. Transgenic plants were analysed histochemically for GUS activity in the nematode feeding structures after infection with either Heterodera schachtii or Meloidogyne incognita. Promoters of the Cauliflower Mosaic Virus 35S gene, the bacterial nopaline synthase, rooting loci (rol) and T-cyt genes and the plant-derived phenylalanine ammonia-lyase I gene, which are highly active in non-infected roots, were all downregulated in the feeding structures as indicated by the strong decrease of GUS activity inside these structures. Less stringent downregulation was observed with chimeric gusA fusion constructs harbouring truncated rolB and rolC promoter sequences. Similar observations were made with transgenic Arabidopsis lines that carried randomly integrated promoterless gusA constructs to identify regulatory sequences in the plant genome. Most of the lines that were selected for expression in the root vascular cylinder demonstrated local downregulation in feeding structures after infection with H. schachtii. The reverse pattern of GUS activity, a blue feeding structure amidst unstained root cells, was also found in several lines. However, GUS activity that was entirely specific for the feeding structures was not observed. Our data show that the expression of a large number of genes is influenced during the development of the nematode feeding structures.
Diversity and activity of sugar transporters in nematode-induced root syncytia
2009-07-01
The plant-parasitic nematode Heterodera schachtii stimulates plant root cells to form syncytial feeding structures which synthesize all nutrients required for successful nematode development....Full Text Available
2008-01-01
Root-lesion nematodes (Pratylenchus thornei Sher and Allen and P. neglectus ( Rensch) Filipijev and Schuurmans Stekhoven) cause substantial yield loss to wheat crops in the northern grain region of Australia. Resistance to P. thornei for use in wheat breeding programs was sought among synthetic hexaploid wheats (2n= 6x = 42, AABBDD) produced through hybridisations of Triticum turgidum L. subsp. durum ( Desf.) Husn ( 2n= 4x = 28, AABB) with Aegilops tauschii Coss. ( 2n= 2x = 14, DD). Resistance was determined for the synthetic hexaploid wheats and their durum and Ae. tauschii parents from the numbers of nematodes in the roots of plants grown for 16 weeks in pots of pasteurised soil inoculated with P. thornei. Fifty-nine (32%) of 186 accessions of synthetic hexaploid wheats had lower numbers of nematodes than Gatcher Selection 50a (GS50a), a partially resistant bread wheat. Greater frequencies of partial resistance were present in the durum parents (72% of 39 lines having lower nematode numbers than GS50a) and in the Ae. tauschii parents (55% of 53 lines). The 59 synthetic hexaploids were re- tested in a second experiment along with their parents. In a third experiment, 11 resistant synthetic hexaploid wheats and their F-1 hybrids with Janz, a susceptible bread wheat, were tested and the F(1)s were found to give nematode counts intermediate between the respective two parents. Synthetic hexaploid wheats with higher levels of resistance resulted from hybridisations where both the durum and Ae. tauschii parents were partially resistant, rather than where only one parent was partially resistant. These results suggest that resistance to P. thornei in synthetic hexaploid wheats is polygenic, with resistances located both in the D genome from Ae. tauschii and in the A and/or B genomes from durum. Five synthetic hexaploid wheats were selected for further study on the basis of ( 1) a high level of resistance to P. thornei of the synthetic hexaploid wheats and of both their durum and Ae. tauschii parents, ( 2) being representative of both Australian and CIMMYT ( International Maize and Wheat Improvement Centre) durums, and ( 3) being representative of the morphological subspecies and varieties of Ae. tauschii. These 5 synthetic hexaploid wheats were also shown to be resistant to P. neglectus, whereas GS50a and 2 P. thornei-resistant derivatives were quite susceptible. Results of P. thornei resistance of F(1)s and F(2)s from a half diallel of these 5 synthetic hexaploid wheats, GS50a, and Janz from another study indicate polygenic additive resistance and better general combining ability for the synthetic hexaploid wheats than for GS50a. Published molecular marker studies on a doubled haploid population between the synthetic hexaploid wheat with best general combining ability ( CPI133872) and Janz have shown quantitative trait loci for resistance located in all 3 genomes. Synthetic hexaploid wheats offer a convenient way of introgressing new resistances to P. thornei and P. neglectus from both durum and Ae. tauschii into commercial bread wheats. Publisher: CSIRO Publishing Other identifier: Thompson, J.P. (2008) Resistance to root-lesion nematodes (Pratylenchus thornei and P-neglectus) in synthetic hexaploid wheats and their durum and Aegilops tauschii parents. Australian Journal of Agricultural Research, 59 (5). pp. 432-446.
Analysis of nematode-responsive promoters in sugar beet hairy roots.
One of the strategies to make crops resistant to the beet cyst nematode Heterodera schachtii is the destruction of the feeding site or syncytium. This can be achieved by local expression of the cytotoxic barnase gene under control of a nematode-inducible plant promoter that is active in the syncytium. Expression of barnase outside the feeding site has to be neutralized by its inhibitor barstar driven from a constitutive promoter that is downregulated in the syncytium. Several promoters that are upregulated in feeding structures were identified using the promoter tagging strategy in Arabidopsis thaliana (Barthels et al., 1997) or by differential cDNA screening in tomato (Van der Eycken et al., 1996). Nematode downregulated promoters in Arabidopsis were described by Goddijn et al. (1993). Five nematode-induced promoters (ARM1, 1164, 728, 25 and Lemmi9) and four downregulated promoters (CaMV35S, the nopaline synthase promoter (nos) and the rooting loci promoters RolC and RolD) fused to the beta-glucuronidase (gus) reporter gene were introduced into sugar beet hairy roots by transformation with Agrobacterium rhizogenes to evaluate their expression pattern. All upregulated promoters were found to be active at the base of lateral roots. The 728 and 25 promoter were as well active in root tips. In the 25-gus lines GUS could also be detected in the vascular tissue, while the ARM1 promoter was also active in sugar beet callus. The Lemmi9 promoter and the 4 constitutive promoters were active in the entire root. The transgenic hairy roots were inoculated with Heterodera schachtii and at different time-points (4, 8, 15, 22 days after inoculation; dpi) GUS analysis was performed on the infected roots. For the ARM1, 1164 and 728 promoter the highest gus expression level in syncytia was observed at 8 dpi. In 4 days old syncytia of the 25-gus lines the intensity of the GUS signal was of the same extent as the non-specific vascular signal. In later stages it even disappeared from the feeding sites. The gus expression level in syncytia of Lemmi9-gus hairy roots was equal to that in control roots. The RolC and 35S promoter were found to be downregulated at 8 dpi, the RolD promoter at 15 dpi and the nos promoter already at 4 dpi.
Candida albicans, the most common human pathogenic fungus, can establish a persistent lethal infection in the intestine of the microscopic nematode Caenorhabditis elegans. The C. elegans–C. albicans infection model was previously adapted to screen for antifungal compounds. Modifications to this screen have been made to facilitate a high-throughput assay including co-inoculation of nematodes with C. albicans and instrumentation allowing precise dispensing of worms into assay wells, eliminating two labor-intensive steps. This high-throughput method was utilized to screen a library of 3,228 compounds represented by 1,948 bioactive compounds and 1,280 small molecules derived via diversity-oriented synthesis. Nineteen compounds were identified that conferred an increase in C. elegans survival, including most known antifungal compounds within the chemical library. In addition to seven clinically used antifungal compounds, twelve compounds were identified which are not primarily used as antifungal agents, including three immunosuppressive drugs. This assay also allowed the assessment of the relative minimal inhibitory concentration, the effective concentration in vivo, and the toxicity of the compound in a single assay.
Genetic analysis of resistance to root-lesion nematode (Pratylenchus thornei) in wheat
2004-01-01
The inheritance of resistance to root-lesion nematode was investigated in five synthetic hexaploid wheat lines and two bread wheat lines using a half-diallel design of F-1 and F-2 crosses. The combining ability of resistance genes in the synthetic hexaploid wheat lines was compared with the performance of the bread wheat line 'GS50a', the source of resistance to Pratylenchus thornei used in Australian wheat breeding programmes. Replicated glasshouse trials identified P. thornei resistance as polygenic and additive in gene action. General combining ability (GCA) of the parents was more important than specific combining ability (SCA) effects in the inheritance of P. thornei resistance in both F-1 and F-2 populations. The synthetic hexaploid wheat line 'CPI133872' was identified as the best general combiner, however, all five synthetic hexaploid wheat lines possessed better GCA than 'GS50a'. The synthetic hexaploid wheat lines contain novel sources of P. thornei resistance that will provide alternative and more effective sources of resistance to be utilized in wheat breeding programmes. Publisher: Blackwell Verlag Contributor: C. Jung; W.E. Weber Coverage: 2004-01-01T00:00:00Z
2007-01-01
Filaria martis causes a poorly known subcutaneous filariosis in mustelids. Few information is available about lesions that F. martis causes in beech martens, on its morphology, biology and the occurrence of the infection. From 1997 to 2006, 29 beech martens from two sites of southern Italy (Sites A and B) have been necropsied. Ectoparasites and nematodes were collected and morphologically identified. A variable region of the cytochrome c oxidase subunit 1 (cox1) of F. martis has been characterised to compare females presenting caudal tips smooth without spines (i.e. Morphotype 1Mrph. 1) and with spines (i.e. Mrph. 2). All ticks collected were identified as Haemaphysalis erinacei. Eleven animals from Site A were found infected by F. martis nematodes in subcutaneous tissue in both membran...
Cryopreservation of Pratylenchys spp.
1995-12-01
Abstract: Optimal conditions for cryopreserving of populations of root lesion nematode (Pratylenchus spp.) were determined. Nematode survival was achieved using glycerol pre-treatments...Full Text Available
[Enterobiasis vaginal. Report of one case]
The nematode Enterobius vermicularis is normally found within the human gastrointestinal tract. Pregnant females migrate towards the anus to lay their eggs. During this nocturnal migration some worms find another adjacent orifices, most commonly the female genitourinary tract. The presented case is a 34-year-old woman referred to the Colposcopy Unit of the Antiguo Hospital Civil of Guadalajara Fray Antonio Alcalde by cervical erosion. With the colposcopy a live worm in the vagina was visualized; it was identified as a gravid female of Enterobius vermicularis by microscopy. The diagnosis of ectopic enterobiasis was an incidental finding during the colposcopic examination of the patient with diagnosis of ectopy.
Zinc sulfide in intestinal cell granules of Ancylostoma caninum adults
1991-04-01
A source of confusion has existed since the turn of the century about the reddish brown, weakly birefringent 'sphaerocrystals' located in the intestines of strongyle nematodes, Strongylus and Ancylostoma. X-ray diffraction and energy dispersive spectrometric analyses were used for accurate determination of the crystalline order and elemental composition of the granules in the canine hookworm Ancylostoma caninum. The composition of the intestinal pigmented granules was identified unequivocally as zinc sulfide. It seems most probable that the granules serve to detoxify high levels of metallic ions (specifically zinc) present due to the large intake of host blood.
2007-01-20
DescriptionIn the life-cycle of C. elegans there is a developmental choice (dauer or non-dauer development), modulated by the worms' environment. This is an example of a phenotypically plastic trait, i.e. an environmentally induced change in phenotype. Isolates of C. elegans vary in their phenotypic plasticity of dauer development. We propose to identify the loci controlling this phenotypic plasticity by a combined analysis of gene expression (using whole genome microarrays) and mapping the quantitative tr [continued...]
MicroRNAs of parasites: current status and future perspectives
2010-01-01
MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs regulating gene expression in eukaryotes at the post-transcriptional level. The complex life cycles of parasites may require the ability to respond to environmental and developmental signals through miRNA-mediated gene expression. Over the past 17years, thousands of miRNAs have been identified in the nematode Caenorhabditis elegans and other parasites. Here, we review the current status and potential functions of miRNAs in protozoan, helminths, and arthropods, and propose some perspectives for future studies.
Intraocular Gnathostoma spinigerum: a case report.
Live intraocular nematode is a rare occurrence that is mostly reported in South East Asian countries. Herewith we report such a case from Nayagarh district of Odisha. A 28 year old female presented with swelling, redness, lacrimation, pain & diminished vision of left eye since 2 1/2 years. Slit lamp examination revealed a worm piercing iris muscle. The worm was removed by paracentesis of anterior chamber and sent to the Department of Microbiology. It was identified to be Gnathostoma spinigerum basing on the typical morphology of its cephalic end. The patient responded completely to oral albendazole therapy.
Intraocular Gnathostoma spinigerum: a case report
Full Text Available.Live intraocular nematode is a rare occurrence that is mostly reported in South East Asian countries. Herewith we report such a case from Nayagarh district of Odisha. A 28 year old female presented with swelling, redness, lacrimation, pain & diminished vision of left eye since 2 1/2 years. Slit lamp examination revealed a worm piercing iris muscle. The worm was removed by paracentesis of anterior chamber and sent to the Department of Microbiology. It was identified to be Gnathostoma spinigerum basing on the typical morphology of its cephalic end. The patient responded completely to oral albendazole therapy.
Gnathostoma hispidum infection in a Korean man returning from China.
Human Gnathostoma hispidum infection is extremely rare in the world literature and has never been reported in the Republic of Korea. A 74-year-old Korean man who returned from China complained of an erythematous papule on his back and admitted to our hospital. Surgical extraction of the lesion and histopathological examination revealed sections of a nematode larva in the deep dermis. The sectioned larva had 1 nucleus in each intestinal cell and was identified as G. hispidum. The patient recalled having eaten freshwater fish when he lived in China. We designated our patient as an imported G. hispidum case from China.
Strongylus vulgaris is an important strongyle nematode with high pathogenic potential infecting horses world-wide. Several decades of intensive anthelmintic use has virtually eliminated clinical disease caused by S. vulgaris, but has also caused high levels of anthelmintic resistance in equine small strongyle (cyathostomin) nematodes. Recommendations aimed at limiting the development of anthelmintic resistance by reducing treatment intensity raises a simultaneous demand for reliable and accurate diagnostic tools for detecting important parasitic pathogens. Presently, the only means available to differentiate among strongyle species in a faecal sample is by identifying individual L3 larvae following a two week coproculture procedure. The aim of the present study is to overcome this diagnostic obstacle by developing a fluorescence-based quantitative PCR assay capable of identifying S. vulgaris eggs in faecal samples from horses. Species-specific primers and a TaqMan probe were designed by alignment of published ribosomal DNA sequences of the second internal transcribed spacer of cyathostomin and Strongylus spp. nematodes. The assay was tested for specificity and optimized using genomic DNA extracted from identified male worms of Strongylus and cyathostomin species. In addition, eggs were collected from adult female worms and used to evaluate the quantitative potential of the assay. Statistically significant linear relationships were found between egg numbers and cycle of threshold (Ct) values. PCR results were unaffected by the presence of cyathostomin DNA in the sample and there was no indication of PCR inhibition by faecal sources. A field evaluation on faecal samples obtained from four Danish horse farms revealed a good agreement with the traditional larval culture (kappa-value=0.78), but with a significantly higher performance of the PCR assay. An association between Ct values and S. vulgaris larval counts was statistically significant. The present assay can reliably and semi-quantitatively detect minute quantities of S. vulgaris eggs in faecal samples.
2009-06-01
Full Text Available.With the post-genomic era came a dramatic increase in high-throughput technologies, of which transcriptional profiling by microarrays was one of the most popular. One application of this technology is to identify genes that are differentially expressed in response to different environmental conditions. These experiments are constructed under the assumption that the differentially expressed genes are functionally important in the environment where they are induced. However, whether differential expression is predictive of functional importance has yet to be tested. Here we have addressed this expectation by employing Caenorhabditis elegans as a model for the interaction of native soil nematode taxa and soil bacteria. Using transcriptional profiling, we identified candidate genes regulated in response to different bacteria isolated in association with grassland nematodes or from grassland soils. Many of the regulated candidate genes are predicted to affect metabolism and innate immunity suggesting similar genes could influence nematode community dynamics in natural systems. Using mutations that inactivate 21 of the identified genes, we showed that most contribute to lifespan and/or fitness in a given bacterial environment. Although these bacteria may not be natural food sources for C. elegans, we show that changes in food source, as can occur in environmental disturbance, can have a large effect on gene expression, with important consequences for fitness. Moreover, we used regression analysis to demonstrate that for many genes the degree of differential gene expression between two bacterial environments predicted the magnitude of the effect of the loss of gene function on life history traits in those environments.
Delivery of macromolecules to plant parasitic nematodes using a tobacco rattle virus vector.
Plant parasitic nematodes cause significant damage to crops on a worldwide scale. These nematodes are often soil dwelling but rely on plants for food and to sustain them during reproduction. Complex interactions occur between plants and nematodes during the nematode life cycle with plant roots developing specialized feeding structures through which nematodes withdraw nutrients. Here we describe a novel method for delivering macromolecules to feeding nematodes using a virus-based vector [tobacco rattle virus (TRV)]. We show that the parasitic nematode Heterodera schachtii will ingest fluorescent proteins transiently expressed in plant roots infected with a TRV construct carrying the appropriate protein sequence. A prerequisite for this delivery is the presence of replicating virus in root tips prior to the formation of nematode-induced syncytia. We show also that TRV vectors expressing nematode gene sequences can be used to induce RNAi in the feeding nematodes.
2009-01-01
SUMMARY In this article, we describe the analysis of over 9000 expressed sequence tags (ESTs) from cDNA libraries obtained from various life cycle stages of Globodera pallida. We have identified over 50 G. pallida effectors from this dataset using bioinformatics analysis, by screening clones in order to identify secreted proteins up-regulated after the onset of parasitism and using in situ hybridization to confirm the expression in pharyngeal gland cells. A substantial gene family encoding G. pallida SPRYSEC proteins has been identified. The expression of these genes is restricted to the dorsal pharyngeal gland cell. Different members of the SPRYSEC family of proteins from G. pallida show different subcellular localization patterns in plants, with some localized to the cytoplasm and others...
Traversing a wormhole to combat Parkinson’s disease
2008-01-01
Full Text Available.Human movement disorders represent a significant and unresolved societal burden. Among these, the most prevalent is Parkinson’s disease (PD), a disorder afflicting millions worldwide. Despite major advances, stemming primarily from human genetics, there remains a significant gap in our understanding of what factors underlie disease susceptibility, onset, and progression. Innovative strategies to discern specific intracellular targets for subsequent drug development are needed to more rapidly translate basic findings to the clinic. Here we briefly review the recent contributions of research using the nematode roundworm Caenorhabditis elegans as a model system for identifying and characterizing gene products associated with PD. As a microscopic but multicellular and genetically tractable animal with a well-defined nervous system and an experimentally tenable lifespan, C. elegans affords significant advantages to researchers attempting to determine causative and therapeutic factors that influence neuronal dysfunction and age-associated neurodegeneration. The rapidity with which traditional genetic, large-scale genomic, and pharmacological screening can be applied to C. elegans epitomizes the utility of this animal for disease research. Moreover, with mature bioinformatic and functional genomic data readily available, the nematode is well positioned to play an increasingly important role in PD-associated discoveries.
Traversing a wormhole to combat Parkinson’s disease
Human movement disorders represent a significant and unresolved societal burden. Among these, the most prevalent is Parkinson’s disease (PD), a disorder afflicting millions worldwide. Despite major advances, stemming primarily from human genetics, there remains a significant gap in our understanding of what factors underlie disease susceptibility, onset, and progression. Innovative strategies to discern specific intracellular targets for subsequent drug development are needed to more rapidly translate basic findings to the clinic. Here we briefly review the recent contributions of research using the nematode roundworm Caenorhabditis elegans as a model system for identifying and characterizing gene products associated with PD. As a microscopic but multicellular and genetically tractable animal with a well-defined nervous system and an experimentally tenable lifespan, C. elegans affords significant advantages to researchers attempting to determine causative and therapeutic factors that influence neuronal dysfunction and age-associated neurodegeneration. The rapidity with which traditional genetic, large-scale genomic, and pharmacological screening can be applied to C. elegans epitomizes the utility of this animal for disease research. Moreover, with mature bioinformatic and functional genomic data readily available, the nematode is well positioned to play an increasingly important role in PD-associated discoveries.
Radiation-induced gene expression in the nematode caenorhabditis elegans
2002-12-01
We used the nematode C. elegans to characterize the genotoxic and cytotoxic effects of ionizing radiation in a simple animal model emphasizing the unique effects of charged particle radiation. Here we demonstrate by reverse transcription polymerase chain reaction (RT-PCR) differential display and whole genome microarray hybridization experiments that gamma rays, accelerated protons and iron ions at the same physical dose lead to unique transcription profiles. 599 of 17871 genes analyzed (3.4%) showed differential expression 3 hrs after exposure to 3 Gy of radiation. 193 were up-regulated, 406 were down-regulated and 90% were affected only by a single species of radiation. A novel statistical clustering technique identified the regulatory relationships between the radiation-modulated genes and showed that genes affected by each radiation species were associated with unique regulatory clusters. This suggests that independent homeostatic mechanisms are activated in response to radiation exposure as a function of track structure or ionization density. (author)
Isolation of a gene from Arabidopsis thaliana related to nematode feeding structures.
Using a promoter tagging approach, a gene upregulated in nematode feeding structures (NFS) of Heterodera schachtii was identified in Arabidopsis thaliana plants. Sequence analysis of the transgenic line bearing gus reporter gene and the wild-type plant revealed that the T-DNA had been inserted into the promoter of the gene, however, with transcription start points at different sites for the gus reporter gene and for the endogenous gene. This tagged gene, designated pyk20, encodes a transcript of 2.6 kb. Southern blot analysis revealed a single gene copy for pyk20 in the Arabidopsis C-24 genome. Other cruciferous plants were shown to possess pyk20 or homologous genes. The predicted amino acid sequence of the PYK20 protein contains 695 residues with a molecular mass of 78 kDa and includes a glutamine-rich domain in the C-terminal region. IAA and kinetin treatment increased the level of the pyk20 transcript in the plant, whereas ABA treatment and temperature stress reduced the pyk20 transcript level. In-situ hybridisation and Northern blot analysis revealed that the gene is expressed in NFS. Based on homologies of the glutamine-rich domain, the biological role of the pyk20 gene product as a transcription factor is assumed.
Intestinal helminths of feral cat populations from urban and suburban districts of Qatar
2010-01-01
A survey of the helminths of 658 adult cats from feral urban and suburban populations in Qatar was conducted across all months in 2006 and 2007. Six species of helminths were identified, comprising two cestodes (Taenia taeniaeformis [73.6%] and Diplopylidium acanthotetra [47.1%]) and four nematodes (Ancylostoma tubaeforme [14.7%], Physaloptera praeputialis [5.2%], Toxocara cati [0.8%] and Toxascaris leonina [0.2%]), and 83% of cats were infected with at least one of these. The average number of species harboured was 1.4 and the average worm burden was 55.8worms/cat. The vast majority of worms (97.6%) were cestodes, nematodes being relatively rare. Prevalence and abundance of infections were analyzed, taking into consideration four factors: year (2006 and 2007), site (urban and suburban), s...
2009-01-01
Summary Elucidating the molecular mechanisms that created ancient complex traits like insect wings is difficult. Fortunately, some complex traits have arisen recently. For example, hermaphroditic reproduction evolved independently many times during recent nematode evolution [1-3]. Although C. elegans hermaphrodites require fog-2 [4], which encodes an F box protein that regulates the translation of tra-2 mRNAs [5, 6], the related species C. briggsae lacks fog-2 [7]. We identified a critical regulator of hermaphrodite development in C. briggsae, named she-1. Analysis of double mutants indicates that she-1 acts upstream of tra-2 in C. briggsae, just as fog-2 does in C. elegans. Molecular cloning shows that she-1 encodes a novel F box protein that was created by a recent gene duplication. Wher...
2010-01-01
The gene transcripts encoding both the AF8 and AF2 neuropeptides of the nematode Ascaris suum have been identified, cloned, and sequenced. The AF8 transcript (afp-3) encodes five identical copies of AF8; each peptide-encoding region is flanked by the appropriate dibasic or monobasic cleavage processing sites. The AF2 transcript (afp-4) encodes three identical copies of AF2 along with the appropriate cleavage sites. In contrast, the afp-1 transcript (Edison et al. [1997] Peptides 18:929-935) encodes six different AF peptides (AF3, 4, 10, 13, 14, 20) which all share a -PGVLRFamide C-terminus but have different N-terminal sequences. By using in situ hybridization, gene transcript expression patterns of afp-1, afp-3, and afp-4 (As-flp-18, As-flp-6, and As-flp-14, respectively, in the naming co...
2007-01-01
Identification and characterization of IgE-inducing antigens are important for elucidating the mechanisms involved in IgE-mediated immune responses in allergic diseases and parasite infections. While many allergens have been characterized, little is known about parasite antigens inducing specific IgE following infection. In order to identify antigens from the nematode Nippostrongylus brasiliensis, we generated an IgE-producing B cell hybridoma from N. brasiliensis-infected C57BL/6 mice and constructed a cDNA phage display library from N. brasiliensis. We successfully cloned and expressed an N. brasiliensis antigen (Nb-Ag1) that showed specific binding to anti-N. brasiliensis IgE. Nb-Ag1 localized to the pharynx of adult N. brasiliensis, suggesting that Nb-Ag1 is a potential pharyngeal glan...
2010-05-01
Full Text Available.Identifying the spatial variability and risk areas for southern root-knot nematode [Meloidogyne incognita (Kofoid & White) Chitwood] (RKN) is key for site-specific management (SSM) of cotton (Gossypium hirsutum L.) fields. The objectives of this study were to: (i) determine the soil properties that influence RKN occurrence at different scales; and (ii) delineate risk areas of RKN by indicator kriging. The study site was a cotton field located in the southeastern coastal plain region of the USA. Nested semivariograms indicated that RKN samples, collected from a 50×50 m grid, exhibited a local and regional scale of variation describing small and large clusters of RKN population density. Factorial kriging decomposed RKN and soil properties variability into different spatial components. Scale dependent correlations between RKN data showed that the areas with high RKN population remained stable though the growing season. RKN data were strongly correlated with slope (SL) at local scale and with apparent soil electrical conductivity deep (EC
Electro-worming: The Behaviors of Caenorhabditis (C.) elegans in DC and AC Electric Fields
2010-10-18
The video showcases how C. elegans worms respond to DC and AC electrical stimulations. Gabel et al (2007) demonstrated that in the presence of DC and low frequency AC fields, worms of stage L2 and larger propel themselves towards the cathode. Rezai et al (2010) have demonstrated that this phenomenon, dubbed electrotaxis, can be used to control the motion of worms. In the video, we reproduce Rezai's experimental results. Furthermore, we show, for the first time, that worms can be trapped with high frequency, nonuniform electric fields. We studied the effect of the electric field on the nematode as a function of field intensity and frequency and identified a range of electric field intensities and frequencies that trap worms without apparent adverse effect on their viability. Worms tethered by dielectrophoresis (DEP) avoid blue light, indicating that at least some of the nervous system functions remain unimpaired in the presence of the electric field. DEP is useful to dynamically confine nematodes for observations, sort them according to size, and separate dead worms from live ones.
Cloning and characterization of XiR1, a locus responsible for dagger nematode resistance in grape
2010-01-01
The dagger nematode, Xiphinema index, feeds aggressively on grape roots and in the process, vectors grapevine fanleaf virus (GFLV) leading to the severe viral disease known as fanleaf degeneration. Resistance to X. index and GFLV has been the key objective of grape rootstock breeding programs. A previous study found that resistance to X. index derived from Vitis arizonica was largely controlled by a major quantitative trait locus, XiR1 (X. index Resistance 1), located on chromosome 19. The study presented here develops high-resolution genetic and physical maps in an effort to identify the XiR1 gene(s). The mapping was carried out with 1,375 genotypes in three populations derived from D8909-15, a resistant selection from a cross of V. rupestris A. de Serres (susceptible)V. arizonica b...
2010-01-01
In the present study, the determination and computer-assisted characterization of ribosomal DNA (rDNA) sequence data were performed to improve classification and diagnostics of nematodes of the genus Ditylenchus. Dot-plot analyses of the ITS1-58S-ITS2 segments revealed the presence of short repeated DNA motifs localized in the ITS1 region of D. destructor, whilst no similar DNA repeats were identified in other members of the genus. Additionally, three-domain secondary structure models were inferred for ITS2 transcripts, providing a valuable resource for complex phylogenetic studies. Finally, alignment of the ITS-rDNA sequence data and subsequent oligonucleotide combinations trials led to the development of taxon-specific amplified regions. Together, this is the first report in which the th...
The filarial nematodes Brugia malayi, Wuchereria bancrofti and Onchocerca volvulus cause elephantiasis or dermatitis and blindness resulting in severe morbidity. Annually, 1.3 billion people are at risk of infection. Targeting the essential Wolbachia endobacteria of filarial nematodes with doxycycline has proven to be an effective therapy resulting in a block in embryogenesis, worm development and macrofilaricidal effects. However, doxycycline is contraindicated for a large portion of the at risk population. To identify new targets for anti-wolbachial therapy, understanding the molecular basis of the Wolbachia-filaria symbiosis is required. Using the B. malayi microarray we identified differentially expressed genes in the rodent filaria Litomosoides sigmodontis after depletion of Wolbachia which might have a role in symbiosis. The microarray data were filtered for regulated genes with a false discovery rate or = 2-fold-change. Most of the genes were differentially expressed at day 36 of tetracycline treatment, when 99.8% of Wolbachia were depleted. Several classes of genes were affected, including genes for translation, transcription, folding/sorting of proteins, motility, structure and metabolic and signalling pathways. Quantitative PCR validated 60% of the genes found to be regulated in the microarray. A nuclear encoded heme-binding protein of the globin family was up-regulated upon loss of Wolbachia. Interestingly, mitochondrial encoded subunits of respiratory chain complexes containing heme and riboflavin were also up-regulated. No change in the expression of these genes was seen in tetracycline treated Wolbachia-free Acanthocheilonema viteae. As Wolbachia synthesise heme and filaria do not, we hypothesise that without the endosymbionts no functional heme-containing enzymes can be formed, leading to loss of energy metabolism which then results in up-regulation of the mitochondrial encoded subunits in an attempt to correct the deviation from homeostasis. Our results support targeting the Wolbachia heme synthesis pathway for the discovery of new anti-filarial drugs.
2007-01-01
Gastrointestinal nematode infections of livestock animals are prevalent and costly problems worldwide. Currently, infections are controlled by anthelmintic chemicals but increasing drug resistance has prompted research interest to shift towards alternative methods of control such as vaccine development and selection of worm-resistant animals. The present study analyses proteins from Trichostrongylus colubriformis infective Us that are recognised by IgG of immune sheep. Following protein separation via two-dimensional electrophoresis and Western blot probing with plasma from sheep resistant to T colubriformis, mass spectrometry-based proteomic analyses were used to identify immuno-reactive protein spots. We were able to identify 28 immune targets, including aspartyl protease inhibitor, enolase, chaperone proteins, galectin, glycolytic enzymes, kinase, phosphatase and structural muscle proteins such as myosin, paramyosin, calponin and DIM-1. The data suggest that immune responses to T colubriformis are dispersed over a relatively large number of parasite antigens, including several cytoplasmically expressed proteins. The results have new implications for understanding the molecular mechanisms that underpin host-parasite interaction during gastrointestinal nematode infections. (c) 2007 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved. Publisher: Pergamon-elsevier Science Ltd Coverage: 2007-11-01T00:00:00Z
Filaria martis causes a poorly known subcutaneous filariosis in mustelids. Few information is available about lesions that F. martis causes in beech martens, on its morphology, biology and the occurrence of the infection. From 1997 to 2006, 29 beech martens from two sites of southern Italy (Sites A and B) have been necropsied. Ectoparasites and nematodes were collected and morphologically identified. A variable region of the cytochrome c oxidase subunit 1 (cox1) of F. martis has been characterised to compare females presenting caudal tips smooth without spines (i.e. Morphotype 1-Mrph. 1) and with spines (i.e. Mrph. 2). All ticks collected were identified as Haemaphysalis erinacei. Eleven animals from Site A were found infected by F. martis nematodes in subcutaneous tissue in both membranous capsules or free under the inner skin surface. The most important morphological characters of F. martis have been reported and discussed. The molecular analysis showed 100% homology among cox1 sequences from Mrph. 1 and 2 thus indicating that the shape of female posterior edge may vary among specimens of F. martis. The results here presented provide new insights into the biology, ecology and morphological characteristics of this scantly known nematode.
Full Text Available.BackgroundSome organisms can survive extreme desiccation by entering a state of suspended animation known as anhydrobiosis. The free-living mycophagous nematode Aphelenchus avenae can be induced to enter anhydrobiosis by pre-exposure to moderate reductions in relative humidity (RH) prior to extreme desiccation. This preconditioning phase is thought to allow modification of the transcriptome by activation of genes required for desiccation tolerance.ResultsTo identify such genes, a panel of expressed sequence tags (ESTs) enriched for sequences upregulated in A. avenae during preconditioning was created. A subset of 30 genes with significant matches in databases, together with a number of apparently novel sequences, were chosen for further study. Several of the recognisable genes are associated with water stress, encoding, for example, two new hydrophilic proteins related to the late embryogenesis abundant (LEA) protein family. Expression studies confirmed EST panel members to be upregulated by evaporative water loss, and the majority of genes was also induced by osmotic stress and cold, but rather fewer by heat. We attempted to use RNA interference (RNAi) to demonstrate the importance of this gene set for anhydrobiosis, but found A. avenae to be recalcitrant with the techniques used. Instead, therefore, we developed a cross-species RNAi procedure using A. avenae sequences in another anhydrobiotic nematode, Panagrolaimus superbus, which is amenable to gene silencing. Of 20 A. avenae ESTs screened, a significant reduction in survival of desiccation in treated P. superbus populations was observed with two sequences, one of which was novel, while the other encoded a glutathione peroxidase. To confirm a role for glutathione peroxidases in anhydrobiosis, RNAi with cognate sequences from P. superbus was performed and was also shown to reduce desiccation tolerance in this species.ConclusionsThis study has identified and characterised the expression profiles of members of the anhydrobiotic gene set in A. avenae. It also demonstrates the potential of RNAi for the analysis of anhydrobiosis and provides the first genetic data to underline the importance of effective antioxidant systems in metazoan desiccation tolerance.
2002-07-01
Full Text Available.Parelaphostrongylus tenuis is a neurotropic nematode common in white-tailed deer (Odocoileus virginianus) of eastern North America. This parasite is the causative agent of a debilitating neurologic disease in atypical hosts, including domestic livestock. In order to identify proteins of potential significance in the host-parasite relationship, a cDNA library was produced from adult P. tenuis mRNA. Screening the library with antisera from infected red deer (Cervus elaphus elaphus) and immunized AO strain rats, we identified clones with sequence similarities to aspartyl protease inhibitors from several parasitic nematodes. Antibody that was generated against this recombinant protein of P. tenuis (Pt-API-1) detected the native protein in E/S products, in muscle and gonad, and on the surface of the cuticle of adult male and female P. tenuis. The native protein was detected in internal structures of first-stage (L1) and third-stage (L3) larvae. Reverse transcription-PCR confirmed expression of Pt-api-1 in L1, L3, and adult male and female worms. Expression of Pt-API-1 throughout the life cycle of P. tenuis suggests an essential function. Antibodies specific for recombinant Pt-API-1 were detected by enzyme-linked immunosorbent assay in sera from 12 red deer experimentally infected with P. tenuis. Antibodies were detected within 28 to 56 days postinfection. Responses were sustained or biphasic in animals with patent infections, consistent with expression of Pt-API-1 by L1. Our results are compatible with findings in other parasitic nematodes showing that aspartyl protease inhibitors are highly immunogenic.
Diversifying selection and host adaptation in two endosymbiont genomes
2007-01-01
Background: The endosymbiont Wolbachia pipientis infects a broad range of arthropod and filarial nematode hosts. These diverse associations form an attractive model for understanding host: symbiont coevolution. Wolbachia's ubiquity and ability to dramatically alter host reproductive biology also form the foundation of research strategies aimed at controlling insect pests and vector-borne disease. The Wolbachia strains that infect nematodes are phylogenetically distinct, strictly vertically transmitted, and required by their hosts for growth and reproduction. Insects in contrast form more fluid associations with Wolbachia. In these taxa, host populations are most often polymorphic for infection, horizontal transmission occurs between distantly related hosts, and direct fitness effects on hosts are mild. Despite extensive interest in the Wolbachia system for many years, relatively little is known about the molecular mechanisms that mediate its varied interactions with different hosts. We have compared the genomes of the Wolbachia that infect Drosophila melanogaster, wMel and the nematode Brugia malayi, wBm to that of an outgroup Anaplasma marginale to identify genes that have experienced diversifying selection in the Wolbachia lineages. The goal of the study was to identify likely molecular mechanisms of the symbiosis and to understand the nature of the diverse association across different hosts. Results: The prevalence of selection was far greater in wMel than wBm. Genes contributing to DNA metabolism, cofactor biosynthesis, and secretion were positively selected in both lineages. In wMel there was a greater emphasis on DNA repair, cell division, protein stability, and cell envelope synthesis. Conclusion: Secretion pathways and outer surface protein encoding genes are highly affected by selection in keeping with host: parasite theory. If evidence of selection on various cofactor molecules reflects possible provisioning, then both insect as well as nematode Wolbachia may be providing substances to hosts. Selection on cell envelope synthesis, DNA replication and repair machinery, heat shock, and two component switching suggest strategies insect Wolbachia may employ to cope with diverse host and intra-host environments. Publisher: BioMed Central Ltd Contributor: Norton, M. Coverage: 2007-01-01T00:00:00Z
2012-03-31
DescriptionNematode populations may be regulated in natural and disturbed ecosystems by microbial natural enemies. In some intensive production systems e.g. cereal monoculture, natural microbial enemies have increased to levels that control specific nematode pests. These suppressive soils are slow to develop and difficult to exploit but provide a valuable resource for studying nematode interactions with the rhizosphere microbial community and for isolation of potential biological control agents. This proje [continued...]
2009-01-01
Bananas cultivated for export all belong to Cavendish cultivars and are all recognized as very susceptible to nematodes, particularly to the burrowing nematode Radopholus similis and the lesion nematode Pratylenchus coffeae. Even if there have been many changes in the management of banana nematodes in large commercial banana plantations, chemical control still remains most often the last resort method to manage the nematodes, although the number of registered products is definitely declining. Therefore, nematode control though genetic improvement is gaining new interest worldwide. In this study, 55 banana accessions mostly diploids from the Musa acuminata genome group (AA) but including some triploid accessions (AAA), some diploids of the Musa balbisiana genome group (BB) and some interspe...
2008-01-01
Wolbachia are bacteria present within the tissues of most filarial nematodes. Filarial nematode survival is known to be affected by immune responses generated during filarial nematode infection and immune responses to Wolbachia can be found in different species harbouring filarial nematode infections, including humans. Using the rodent filarial model Litomosoides sigmodontis, we show that pre-exposure to wolbachia surface protein in a Th1 context (but not in a Th2-context) enhances worm survival on subsequent challenge. This study suggests that despite abundant evidence that pro-inflammatory reactions to the endosymbiont have detrimental effects on the both the nematode and mammalian host, they may under some circumstances be beneficial to the nematode.
2006-01-01
The common soil inhabiting nematophagous fungus Paecilomyces lilacinus (Thom) Samson and the nematode trapping fungus Monacrosporium lysipagum (Drechsler) Subram were assayed for their ability to reduce the populations of three economically important plant-parasitic nematodes in pot trials. The fungi were tested individually and in combination against the root-knot nematode Meloidogyne javanica (Treub) Chitwood, cereal cyst nematode Heterodera avenae Wollenweber, or burrowing nematode Radopholus similis (Cobb) Thorne on tomato, barley and tissue cultured banana plants, respectively. In all cases, nematode populations were controlled substantially by both individual and combined applications of the fungi. Combined application of P. lilacinus and M. lysipagum reduced 62% of galls and 94% of ...
Secretions of plant-parasitic nematodes which are released into plant tissue may play critical roles in plant-nematode interactions. The identification and characterization of these molecules are of fundamental importance and may help to facilitate the development of novel strategies to interfere with nematode infection of plants and thereby decrease nematode-induced damage to crops. An antibody-based approach was used to isolate molecules present on the nematode surface and in nematode secretions. Monoclonal antibodies (MAbs) were produced to secretions and to whole Heterodera avenae 2nd-stage juveniles; several of these MAbs recognized molecules present in nematode secretions produced in vitro. Three of these molecules have been partly characterized in H. avenae, Globodera rostochiensis, G. pallida and Meloidogyne incognita. A MAb reacting with the surfaces of these nematodes recognized antigens of different molecular weight in each of the species tested. This difference in antigenicity might be related to specific functions in these nematodes. Preliminary results show that this antibody also localized the antigen in root cells surrounding the feeding site induced by M. incognita in Arabidopsis thaliana.
1996-12-31
Juvenile and adult nematodes emerged from onion and garlic samples on the 3 rd week after irradiation with doses up to 0.5 kGy and from potato treated with doses up to 2.0 kGy. However, irradiation of onion infected with Ditylenchus dipsaci caused the inhibition of the development and growth of juvenile nematodes to mature forms. Doses of gamma radiation ranging from 0.1 to 0.5 kGy had only a slight effect, if any, on the development and growth of D. dipsaci nematodes infecting garlic, but they increased juvenile mortality. Gamma radiation at doses up to 2.0 kGy induced increased mortality of nematode juveniles of the potato tuber nematode, D. destructor but less so inhibited their development to mature forms. Nematodes were found to be resistant to irradiation treatment. Therefore the use of gamma irradiation for nematode disinfestation of agricultural products seems to be impractical, if the aim of the treatment is to kill these pests within a few weeks. The level of radiation required to kill nematodes in infected plants would damage plant tissues so that the further storage of vegetables will be impossible. (author). 22 refs, 3 figs, 2 tabs.
Advisory services for nematode pests
2002-01-01
This book, which has been compiled by members of the Australasian Association of Nematologists, provides general guidelines on how an advisory service for nematodes should operate. It is hoped that these guidelines will help improve the quality of advice on nematode pests available to farmers in Australia and New Zealand. Although the book is mainly written for those who are operating nematode diagnostic services, it will be useful to their clients. It gives some indication of the procedures involved and the issues that should be addressed by those providing professional advice on nematode problems. Publisher: RIRDC Language: en-au Rights: Copyright Rural Industries Research and Development Corporation. All rights reserved.
Cyst nematodes induce a metabolically highly active syncytial cell complex in host roots. The syncytia are symplastically isolated. Because they form a strong sink, assimilates must be imported via the apoplast, thus suggesting that specific membrane-bound sugar transport proteins are expressed and activated. To identify possible candidate genes, transgenic Arabidopsis plants expressing different reporter genes under the control of different promoters from Arabidopsis sugar transporter genes were infected with the beet cyst nematode (Heterodera schachtii). With polymerase chain reaction, 13 additional sugar transporters were tested for their presence in the syncytia through the use of a syncytium-specific cDNA library. Analysis of the infected roots showed that the promoter of the sucrose (Suc) transporter AtSUC2 gene that codes for a companion cell-specific Suc transporter in noninfected plants was found to be expressed in syncytia. Its expression patterns in beta-glucuronidase and green fluorescent protein plants were monitored. Syncytium-specific gene expression was confirmed by reverse transcriptase-polymerase chain reaction. Results support the idea that AtSUC2 mediates the transmembrane transfer of Suc. AtSUC2 is the first disaccharide carrier described to be activated by pathogens.
Males of citrus-parasitic Radopholus citrophilus (FL1) were mated with non-citrus-parasitic R. similis (FL5) females. Progeny inherited a 2.4-kb sequence tag site (DK#1) and the ability to reproduce in citrus from the paternal parent (FLl); both traits were absent in the maternal line (FL5). The hybrid progeny produced offspring in roots of citrus seedlings over an 8-month period and therefore were considered reproductively viable. Genomic DNA hybridization studies indicated that one or more copies of DK#1 were present in R. citrophilus FL1. It is not likely that DK#1 represents a citrus parasitism gene because it was amplified from some burrowing nematode isolates that did not parasitize citrus and because DK#1 contains no open reading frames. Inability to reliably test individual nematodes for their ability to parasitize citrus was a constraint to obtaining F2 data required for definitive genetic characterization of citrus parasitism in burrowing nematodes, and alternate approaches will be required. Although the physical relationship of DK#1 and the citrus parasitism locus remains undefined, results of controlled mating studies using these parameters as genetic markers enabled us to identify hybrid F progeny. Therefore, R. similis and R. citrophilus are not sibling species since gene flow between the two does not appear to be restricted via geographic isolation (sympatric in Florida) or by genetics.
2009-09-01
Full Text Available.The carbohydrate larval antigen, CarLA, is present on the exposed surface of all strongylid nematode infective L3 larvae tested, and antibodies against CarLA can promote rapid immune rejection of incoming Trichostrongylus colubriformis larvae in sheep. A library of ovine recombinant single chain Fv (scFv) antibody fragments, displayed on phage, was prepared from B cell mRNA of field-immune sheep. Phage displaying scFvs that bind to the surface of living exsheathed T. colubriformis L3 larvae were identified, and the majority of worm-binding scFvs recognized CarLA. Characterization of greater than 500 worm surface binding phage resulted in the identification of nine different anti-CarLA scFvs that recognized three distinct T. colubriformis CarLA epitopes based on blocking and additive ELISA. All anti-CarLA scFvs were specific to the T. colubriformis species of nematode. Each of the three scFv epitope classes displayed identical Western blot recognition patterns and recognized the exposed surface of living T. colubriformis exsheathed L3 larvae. Surprisingly, each of the anti-CarLA scFvs was able to bind to only a subset of worms. Double-labelling indirect immunofluorescence revealed that the three classes of anti-CarLA scFvs recognize distinct, non-overlapping, T. colubriformis sub-populations. These results demonstrate that individual T. colubriformis L3 larvae display only one of at least three distinct antigenic forms of CarLA on their surface at any given time, and suggest that antigenic variation within CarLA is likely a mechanism of immune evasion in strongylid nematodes.
2008-01-01
'Shiraz' is a popular red wine grape variety grown in NSW, Australia, and is susceptible to citrus nematode (Tylenchulus semipenetrans). The extent of damage, particularly yield loss, or the damage threshold level of T. semipenetrans in 'Shiraz' is not known. In this study we investigated the population dynamics and the effects of a range of population densities of T. semipenetrans on yield of 'Shiraz' in a naturally infested vineyard across three growing seasons. Results showed that the population density of T. semipenetrans J2 in soil did not increase or decrease consistently during the trial period. However, the population densities varied significantly (P spring > autumn > winter. Yield from vines with T. semipenetrans J2 population densities greater than 9,000·kg-1 dry soil (average population over 11,000·kg-1 dry soil) was 15 % lower compared to vines with 500-3,000 (average 932) T. semipenetrans J2·kg-1 dry soil in 2004 but not in 2001 and 2003. Regression analysis showed a linear trend (r = -0.36) on yield decrease with the increase of T. semipenetrans densities in soil in 2004, when the 'Shiraz' vines were eight years old. Pruning weight was reduced by 18.7 to 22.9 % when nematode population densities were greater than 12,000·kg-1 dry soil (average population over 19,000·kg-1 dry soil). Publisher: Siebeldingen, ALLEMAGNE (1957) (Revue): Institut fr Rebenzchtung Geilweilerhof der Bundesanstalt fr Zchtungsforschung an Kulturplanzen Relation: Vitis Other identifier: URN:ISSN:0042-7500
Parasitic nematodes of the group Cyathostominae are an important cause of disease in horses. This group consists of approximately 50 species, all of which have similar life cycles that involve encystment of larval stages in the large intestinal wall. Encysted larvae can persist for months to years and, occasionally, large numbers can accumulate and emerge synchronously to cause severe pathology, resulting in diarrhoea, weight loss, colic and/or oedema. This syndrome, known as larval cyathostominosis, can be fatal in up to 50% of cases, despite treatment. There is no diagnostic method that enables detection of the encysted larval burden. Previously, we identified two native antigen complexes that showed utility as diagnostic markers for the estimation of cyathostomin encysted larval burdens. This paper reports the identification of a likely protein component of one of these antigen complexes. The protein, designated cyathostomin gut-associated larval antigen-1 (Cy-GALA-1), was isolated by immunoscreening a cyathostomin mixed-species, larval complementary DNA library using sera from experimentally-infected horses. The resultant recombinant protein, rCy-GALA-1, was expressed in Escherichia coli and shown to be a target of serum IgG(T) responses in experimentally- and naturally-infected horse populations. Transcription of Cy-gala-1 was restricted to cyathostomin encysted larvae and the presence of native protein was limited to developing larval stages. Importantly, rCy-GALA-1 exhibited no reactivity to serum from horses mono-specifically infected with other helminth species, nor did antisera, raised to the recombinant protein, bind to adult stage extracts of heterologous species. Immunohistochemical experiments located Cy-GALA-1 to the nematode gut. A region of the gene encoding orthologous GALA sequences was isolated from 10 separate cyathostomin species, indicating the ubiquity of the protein in this nematode group.
2007-01-01
An unusual feature of nematodes is the covalent attachment of immunomodulatory phosphorylcholine (PC) moieties to N-type glycans. Our previous work on the filarial nematode glycoprotein ES-62 has enabled us to predict the identity of enzymes necessary for PC-N-glycan biosynthesis. Here, we addressed these predictions using gene knockout technology applied to C. elegans and present two pieces of confirmatory data. Employing a triple null mutant worm lacking all three genes that encode active UDP-N-acetyl-d-glucosamine: α-3-d-mannoside β1, 2-N-acetylglucosaminyltransferase I (GnT I) we have confirmed our earlier prediction that a crucial step in the generation of the substrate for PC transfer is addition of terminal GlcNAc to the α1-3-linked mannose residue of the glycan by GnT I. Second, by silencing genes responsible for expressing enzymes of the Kennedy pathway of phosphatidylcholine biosynthesis by RNA interference (RNAi), we have confirmed our belief for a role for diacylglycerol: choline phosphotransferase (CPT) in PC-N-glycan biosynthesis. Publisher: http://dx.doi.org/10.1016/j.molbiopara.2007.08.009; Elsevier BV; Netherlands Contributor: A P Waters, C E Clayton Relation: 1; Molecular and Biochemical Parasitology; 88; 91; N; 157 Other identifier: 0166-6851 Language: en_AU Rights: Y
Climate Change Promotes the Emergence of Serious Disease Outbreaks of Filarioid Nematodes
2010-08-01
Full Text Available.Filarioid parasites represent major health hazards with important medical, veterinary, and economic implications, and considerable potential to affect the everyday lives of tens of millions of people globally (World Health Organization, 2007). Scenarios for climate change vary latitudinally and regionally and involve direct and indirect linkages for increasing temperature and the dissemination, amplification, and invasiveness of vector-borne parasites. High latitude regions are especially influenced by global climate change and thus may be prone to altered associations and dynamics for complex host-pathogen assemblages and emergence of disease with cascading effects on ecosystem structure. Although the potential for substantial ecological perturbation has been identified, few empirical observations have emanated from systems across the Holarctic. Coincidental with decades of warming, and anomalies of high temperature and humidity in the sub-Arctic region of Fennoscandia, the mosquito-borne filarioid nematode Setaria tundra is now associated with emerging epidemic disease resulting in substantial morbidity and mortality for reindeer and moose. We describe a host-parasite system that involves reindeer, arthropods, and nematodes, which may contribute as a factor to ongoing declines documented for this ungulate species across northern ecosystems. We demonstrate that mean summer temperatures exceeding 14°C drive the emergence of disease due to S. tundra. An association between climate and emergence of filarioid parasites is a challenge to ecosystem services with direct effects on public health, sustainability of free-ranging and domestic ungulates, and ultimately food security for subsistence cultures at high latitudes.
2001-01-01
Possible induction of resistance to the clover cyst nematode, Heterodera trifolii, in white clover, Trifolium repens, by application of two chemical inducers and of soil-borne Pseudomonas-like spp. and Bacillus cereus was investigated. Salicylic acid and benzo(1,2,3)thiadiazole-7-carbothioic acid-S-methyl ester, applied as a root drench in growth cabinet soil bioassays, affected development of H. trifolii in cvs Haifa and Grasslands Huia. Treatments reduced fecundity of the nematodes, increasing the proportions of distorted females and of females with fewer eggs compared to water-treated controls. Application of pectinolytic fluorescent pseudomonad strains P29 and P80, and B. cereus strain B1, induced a response similar to that resulting from the chemical induction. Both live and dead cells of P29, but not cell-free culture filtrate, induced these effects on H. trifolii. A non-pectinolytic, fluorescent pseudomonad, strain P37, had no effect. From the nature of the responses, it is concluded that the effects of both the chemical and bacterial agents have similarities to resistance. From the timing of applications and known properties of both agents, these effects seem similar to induced systemic resistance.Kempster, Valerie N.; Davies, Kerrie A.; Scott, Eileen S. Publisher: Brill Academic Contributor: Applied and Molecular Ecology; School of Agriculture, Food and Wine : Plant and Food Science Other identifier: Nematology, 2001; 3 (1):35-43; 1388-5545; 0020010056 Language: en_US
Biochemistry and molecular biology of the Caenorhabditis elegans dauer larva
Biochemical and molecular techniques have been used to study the formation and recovery of the developmentally arrested, non-feeding dauer stage of the nematode Caenorhabditis elegans. While investigating developmental transitions in energy metabolism, a major metabolite isolated from perchloric acid extracts has been identified as a modified uridine nucleotide. The compound was isolated by gel filtration and ion-exchange chromatography and its structure was determined by {sup 1}H NMR and {sup 13}C NMR spectroscopy. This compound is the most abundant metabolite detected in {sup 31}PMR spectra of perchloric acid extracts from growing larvae. In the absence of phosphoarginine or phosphocreatine, this modified nucleotide may have an important function in the nematode's energy metabolism, and it may also be found in several other invertebrates. During recovery from the dauer stage, metabolic activation is accompanied by a decrease in intracellular pH (pH{sub i}). Although metabolic activation has been associated with an alkaline pH{sub i} shift in other organisms, in vivo {sup 31}P NMR analysis of recovering dauer larvae shows a pH{sub i} decrease from {approximately}7.3 to {approximately}6.3 within 3 hr after the animals encounter food. This shift occurs before feeding begins, and coincides with, or soon follows, the development commitment to recover from the dauer stage, suggesting that control of pH{sub i} may be important in the regulation of larval development in nematodes. A library enriched for sequences expressed specifically during the L2d (predauer) stage was made by selecting plaques from a genomic lambda library that hybridized to subtracted L2d cDNA probes. Ultimately, three clones that were shown to hybridize only to L2d RNA were selected.
Biochemistry and molecular biology of the Caenorhabditis elegans dauer larva
1989-01-01
Biochemical and molecular techniques have been used to study the formation and recovery of the developmentally arrested, non-feeding dauer stage of the nematode Caenorhabditis elegans. While investigating developmental transitions in energy metabolism, a major metabolite isolated from perchloric acid extracts has been identified as a modified uridine nucleotide. The compound was isolated by gel filtration and ion-exchange chromatography and its structure was determined by {sup 1}H NMR and {sup 13}C NMR spectroscopy. This compound is the most abundant metabolite detected in {sup 31}PMR spectra of perchloric acid extracts from growing larvae. In the absence of phosphoarginine or phosphocreatine, this modified nucleotide may have an important function in the nematode's energy metabolism, and it may also be found in several other invertebrates. During recovery from the dauer stage, metabolic activation is accompanied by a decrease in intracellular pH (pH{sub i}). Although metabolic activation has been associated with an alkaline pH{sub i} shift in other organisms, in vivo {sup 31}P NMR analysis of recovering dauer larvae shows a pH{sub i} decrease from {approximately}7.3 to {approximately}6.3 within 3 hr after the animals encounter food. This shift occurs before feeding begins, and coincides with, or soon follows, the development commitment to recover from the dauer stage, suggesting that control of pH{sub i} may be important in the regulation of larval development in nematodes. A library enriched for sequences expressed specifically during the L2d (predauer) stage was made by selecting plaques from a genomic lambda library that hybridized to subtracted L2d cDNA probes. Ultimately, three clones that were shown to hybridize only to L2d RNA were selected.
The anesthetic action of ethanol analyzed by genetics in Caenorhabditis elegans
Acute exposure to ethanol causes paralysis at high concentrations in the nematode Caenorhabditis elegans. We set out to elucidate the mechanism of the anesthetic action of ethanol by genetic approaches. We identified nine mutations that conferred reduced sensitivity to ethanol after chemical, irradiation, or transposon insertion mutagenesis. Of these nine, we further characterized five mutations that defined four genes, jud-1-jud-4. Analysis of the phenotypes of the animals heterozygous for two unlinked genes revealed that jud-1 and jud-3 act synergistically in a gene dose-dependent manner. We cloned jud-4 and found that it encodes a protein with limited homology to human Homer proteins. jud-4 was expressed in the hypodermis and vulva muscles, suggesting that this gene acts in tissues directly exposed to the external environment. Characterization of the other mutations identified in this study will facilitate the elucidation of the molecular mechanism for the anesthetic action of ethanol.
OutFOXing the Grim Reaper: Novel Mechanisms Regulating Longevity by Forkhead Transcription Factors
Studies in the nematode Caenhorhabditis elegans have contributed to understanding the mechanisms that control life-span and aging. Recent experiments have employed RNAi to knock down gene expression, microarray analysis to identify genes involved in the insulin/insulin-like growth factor pathway, and bioinformatics screening to identify new Forkhead-regulated genes. The results from these experiments suggest that members of the Forkhead transcription factor family along with heat shock factors may coordinately regulate production of small heat shock proteins to control how cell stress influences life-span, and provide new links between metabolism and longevity, as well as indicate that antimicrobial genes may be involved in the aging process.
Molecular analysis and comparative morphology to resolve a complex of cryptic Xiphinema species
2010-01-01
Gutierrez-Gutierrez, C., Palomares-Rius, J.E., Cantalapiedra-Navarrete, C., Landa, B.B., Esmenjaud, D. &Castillo, P. (2010). Molecular analysis and comparative morphology to resolve a complex of cryptic Xiphinema species. -Zoologica Scripta, 39, 483-498. During nematode surveys in cultivated and natural environments in southern Spain nine populations of parthenogenic Xiphinema species tentatively identified as Xiphinema cf. pyrenaicum and one population morphologically close to Xiphinema turcicum were detected. Surveys in southern France also identified one population resembling X. pyrenaicum. We developed a comparative study among these related Xiphinema species, including topotypes of two species of this group previously synonymized, viz. Xiphinema hispanum and Xiphinema sphaerocephalum,...
Previously, we identified Arabidopsis thaliana mutant rhd1-4 as hypersusceptible to the sugar beet cyst nematode Heterodera schachtii. We assessed rhd1-4 as well as two other rhd1 alleles and found that each exhibited, in addition to H. schachtii hypersusceptibility, decreased root length, increased root hair length and density, and deformation of the root epidermal cells compared with wild-type A. thaliana ecotype Columbia (Col-0). Treatment of rhd1-4 and Col-0 with the ethylene inhibitors 2-aminoethoxyvinylglycine and silver nitrate and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid suggests that the rhd1-4 hypersusceptibility and root morphology phenotypes are the result of an increased ethylene response. Assessment of known ethylene mutants further support the finding that ethylene plays a role in mediating A. thaliana susceptibility to H. schachtii because mutants that overproduce ethylene (eto1-1, eto2, and eto3) are hypersusceptible to H. schachtii and mutants that are ethylene-insensitive (etr1-1, ein2-1, ein3-1, eir1-1, and axr2) are less susceptible to H. schachtii. Because the ethylene mutants tested show altered susceptibility and altered root hair density and length, a discrimination between the effects of altered ethylene signal transduction and root hair density on susceptibility was accomplished by analyzing the ttg and gl2 mutants, which produce ectopic root hairs that result in greatly increased root hair densities while maintaining normal ethylene signal transduction. The observed normal susceptibilities to H. schachtii of ttg and g12 indicate that increased root hair density, per se, does not cause hypersusceptibility. Furthermore, the results of nematode attraction assays suggest that the hypersusceptibility of rhd1-4 and the ethylene-overproducing mutant eto3 may be the result of increased attraction of H. schachtii-infective juveniles to root exudates of these plants. Our findings indicate that rhd1 is altered in its ethylene response and that ethylene signal transduction positively influences plant susceptibility to cyst nematodes.
2010-01-01
Faecal Egg Count Reduction Tests (FECRTs) for macrocyclic lactone (ML) and levamisole (LEV) drenches were conducted on two dairy farms in the subtropical, summer rainfall region of eastern Australia to determine if anthelmintic failure contributed to severe gastrointestinal nematode infections observed in weaner calves. Subtropical Cooperia spp. were the dominant nematodes on both farms although significant numbers of Haemonchus placei were also present on Farm 2. On Farm 1, moxidectin pour-on (MXD) drenched at 0.5 mg kg-1 liveweight (LW) reduced the overall Cooperia burden by 82% (95% confidence limits, 37-95%) at day 7 post-drench. As worm burdens increased rapidly in younger animals in the control group (n = 4), levamisole was used as a salvage drench and these calves withdrawn from the trial on animal welfare grounds after sample collection at day 7. Levamisole (LEV) dosed at 6.8 mg kg-1 LW reduced the worm burden in these calves by 100%, 7 days after drenching. On Farm 2, MXD given at 0.5 mg kg-1 LW reduced the faecal worm egg count of cooperioids at day 8 by 96% (71-99%), ivermectin oral (IVM) at 0.2 mg kg-1 LW by 1.6% (-224 to 70%) and LEV oral at 7.1 mg kg-1 LW by 100%. For H. placei the reductions were 98% (85-99.7%) for MXD, 0.7% (-226 to 70%) for IVM and 100% for LEV. This is the first report in Australia of the failure of macrocyclic lactone treatments to control subtropical Cooperia spp. and suspected failure to control H. placei in cattle. Publisher: Elsevier Other identifier: Lyndal-Murphy, M. and Rogers, D.J. and Ehrlich, W.K. and James, P.J. and Pepper, P.M. (2010) Reduced efficacy of macrocyclic lactone treatments in controlling gastrointestinal nematode infections of weaner dairy calves in subtropical eastern Australia. Veterinary Parasitology, 168 (1-2). pp. 146-150.
A Transcriptomic analysis of the adult stage of the bovine lungworm, Dictyocaulus viviparus
2007-01-01
Lungworms of the genus Dictyocaulus (family Dictyocaulidae) are parasitic nematodes of major economic importance. They cause pathological effects and clinical disease in various ruminant hosts, particularly in young animals. Dictyocaulus viviparus, called the bovine lungworm, is a major pathogen of cattle, with severe infections being fatal. In this study, we provide first insights into the transcriptome of the adult stage of D. viviparus through the analysis of expressed sequence tags (ESTs). Results: Using our EST analysis pipeline, we estimate that the present dataset of 4436 ESTs is derived from 2258 genes based on cluster and comparative genomic analyses of the ESTs. Of the 2258 representative ESTs, 1159 (51.3%) had homologues in the free-living nematode C. elegans, 1174 (51.9%) in parasitic nematodes, 827 (36.6%) in organisms other than nematodes, and 863 (38%) had no significant match to any sequence in the current databases. Of the C. elegans homologues, 569 had observed 'non-wildtype' RNAi phenotypes, including embryonic lethality, maternal sterility, sterility in progeny, larval arrest and slow growth. We could functionally classify 776 (35%) sequences using the Gene Ontologies (GO) and established pathway associations to 696 (31%) sequences in Kyoto Encyclopedia of Genes and Genomes (KEGG). In addition, we predicted 85 secreted proteins which could represent potential candidates for developing novel anthelmintics or vaccines. Conclusion: The bioinformatic analyses of ESTs data for D. viviparus has elucidated sets of relatively conserved and potentially novel genes. The genes discovered in this study should assist research toward a better understanding of the basic molecular biology of D. viviparus, which could lead, in the longer term, to novel intervention strategies. The characterization of the D. viviparus transcriptome also provides a foundation for whole genome sequence analysis and future comparative transcriptomic analyses.13 page(s) Publisher: Biomed Central Ltd Contributor: Macquarie University. Dept. of Chemistry and Biomolecular Sciences Relation: BMC genomics, Vol. 8, Issue 311, p.1-13; 10.1186/1471-2164-8-311 Other identifier: ISSN:1471-2164; mq-rm-2007002480 Language: eng Rights: Copyright 2007 Ranganathan et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
2009-11-01
Full Text Available.Heterodera glycines, the soybean cyst nematode (SCN), causes the most damaging chronic disease of soybean (Glycine max). Host resistance requires the resistance allele at rhg1. Resistance destroys the giant cells created in the plant's roots by the nematodes about 24 to 48 h after commencement of feeding. In addition, 4 to 8 d later, a systemic acquired resistance develops that discourages later infestations. The molecular mechanisms that control the rhg1-mediated resistance response appear to be multigenic and complex, as judged by transcript abundance changes, even in near isogenic lines (NILs). This study aimed to focus on key posttranscriptional changes by identifying proteins and metabolites that were increased in abundance in both resistant and susceptible NILs. Comparisons were made among NILs 10 d after SCN infestation and without SCN infestation. Two-dimensional gel electrophoresis resolved more than 1,000 protein spots on each gel. Only 30 protein spots with a significant (P < 0.05) difference in abundance of 1.5-fold or more were found among the four treatments. The proteins in these spots were picked, trypsin digested, and analyzed using quadrupole time-of-flight tandem mass spectrometry. Protein identifications could be made for 24 of the 30 spots. Four spots contained two proteins, so that 28 distinct proteins were identified. The proteins were grouped into six functional categories. Metabolite analysis by gas chromatography-mass spectrometry identified 131 metabolites, among which 58 were altered by one or more treatment; 28 were involved in primary metabolism. Taken together, the data showed that 17 pathways were altered by the rhg1 alleles. Pathways altered were associated with systemic acquired resistance-like responses, including xenobiotic, phytoalexin, ascorbate, and inositol metabolism, as well as primary metabolisms like amino acid synthesis and glycolysis. The pathways impacted by the rhg1 allelic state and SCN infestation agreed with transcript abundance analyses but identified a smaller set of key proteins. Six of the proteins lay within the same small region of the interactome identifying a key set of 159 interacting proteins involved in transcriptional control, nuclear localization, and protein degradation. Finally, two proteins (glucose-6-phosphate isomerase [EC 5.3.1.9] and isoflavone reductase [EC 1.3.1.45]) and two metabolites (maltose and an unknown) differed in resistant and susceptible NILs without SCN infestation and may form the basis of a new assay for the selection of resistance to SCN in soybean.
Studies were conducted to determine the curative and residual efficacy of avermectins at controlling plant-parasitic nematodes when injected into the pseudostem of banana, Musa acuminata cv. Cavendish. In addition, we determined the lowest concentration of avermectins that provided satisfactory efficacy as protectants when injected into banana pseudostems. Experiments were conducted with a root-knot nematode, Meleidogyne javanica, and the burrowing nematode, Radopholus similis. Injections (1 ml) of >/= 100 mug a.i./plant of abamectin into pseudostems were effective at controlling M. javanica and R. similis, and were comparable to control achieved with a conventional chemical nemaficide, fenamiphos, in a protectant assay. Abamecfin injections of 250 and 500 mug a.i./plant were effective at reducing nematode infections 28 to 56 days after inoculation. Abamectin was more effective than ivermectin at controlling nematodes after nematode populations were established in banana roots. Injections of between 100 and 1,000 mug a.i./plant were effective at controlling nematodes for at least 56 days after treatment. These studies confirmed earlier results and demonstrated that abamecfin has potential for controlling nematode parasites on banana when injected into the pseudostem.
Cloning and characterization of XiR1, a locus responsible for dagger nematode resistance in grape
2010-08-01
Full Text Available.The dagger nematode, Xiphinemaindex, feeds aggressively on grape roots and in the process, vectors grapevine fanleaf virus (GFLV) leading to the severe viral disease known as fanleaf degeneration. Resistance to X. index and GFLV has been the key objective of grape rootstock breeding programs. A previous study found that resistance to X. index derived from Vitis arizonica was largely controlled by a major quantitative trait locus, XiR1 (X. index Resistance 1), located on chromosome 19. The study presented here develops high-resolution genetic and physical maps in an effort to identify the XiR1 gene(s). The mapping was carried out with 1,375 genotypes in three populations derived from D8909-15, a resistant selection from a cross of V. rupestris A. de Serres (susceptible) × V. arizonica b42-26 (resistant). Resistance to X. index was evaluated on 99 informative recombinants that were identified by screening the three populations with two markers flanking the XiR1 locus. The high-resolution genetic map of XiR1 was primarily constructed with seven DNA markers developed in this study. Physical mapping of XiR1 was accomplished by screening three bacterial artificial chromosome (BAC) libraries constructed from D8909-15, V. vinifera Cabernet Sauvignon and V. arizonica b42-26. A total of 32 BAC clones were identified and the XiR1 locus was delineated within a 115 kb region. Sequence analysis of three BAC clones identified putative nucleotide binding/leucine-rich repeat (NB-LRR) genes. This is the first report of a closely linked major gene locus responsible for ectoparasitic nematode resistance. The markers developed from this study are being used to expedite the breeding of resistant grape rootstocks.
2008-01-01
The synthetic banana hybrid FB920 (Musa spp., AAA group) has been designed to be resistant to Sigatoka and Black Leaf Streak Diseases, caused respectively by Mycosphaerella musicola and Mycosphaerella fijiensis. In addition to these features, the hybrids seem less susceptible to plant-parasitic nematodes, especially the burrowing nematodes (Radopholus similis) and the lesion nematode (Pratylenchus coffeae) than classic Cavendish cultivars, such as Williams, Grande Naine, or Poyo. This genetic modification drastically reduces pesticide use. Herein, we used the SIMBA model (a model to simulate phenology, growth, and plant-parasitic nematode/banana interactions) to examine the population dynamics of plant-parasitic nematodes in cropping systems with hybrid FB920 in various initial conditions....
2010-01-01
Endoparasites can alter their host's heavy metal concentrations by sequestering metals in their own tissues. Contracaecum spp. (a nematode), but not Drepanocephalus spathans (a trematode), were bioaccumulating mercury to concentrations 1.5 times above cormorant hosts. Nematodes did not have significantly greater stable nitrogen isotope values (d^1^5N) than their hosts, which is contradictory to prey-predator trophic enrichment studies, but is in agreement with other endoparasite-host relationships. However, Contracaecum spp. d^1^3C values were significantly greater than their hosts, which suggest that nematodes were consuming host tissues. Nematodes were accumulating and thus sequestering some of their cormorant hosts' body burden of methyl mercury; however, they were not dramatically redu...
2009-01-01
SUMMARY In cool temperate areas, such as Scotland, sheep are infected by a variety of nematodes but the dominant nematode is Teladorsagia circumcincta. Resistant animals have one or more of the following features: fewer adult nematodes, more inhibited larvae, shorter adult nematodes and decreased production of nematode eggs. In lambs at the end of the first grazing season, the heritability of adult worm length is very strong, whereas the heritability of egg production is moderate. The heritability of worm number is low while there is no detectable genetic variation in the number of inhibited larvae. The major mechanisms underlying resistance to T. circumcinctaappear to be the IgA mediated suppression of worm growth and the mast cell mediated regulation of worm number. Mast cell responses a...
Entomopathogenic nematode infectivity enhancement using physical and chemical stressors
2006-01-01
Entomopathogenic nematode in vivo production depends on a high level of successful infection during the inoculation process. Some host-nematode combinations do not provide sufficient or consistent infection levels and are, therefore, not suitable for in vivo production. Our objective was to determine whether infection levels could be improved through host or nematode exposure to physical and chemical stressors. Elevated stress may compromise host insect defenses, thus increasing its vulnerability to nematode infection. Specifically, we investigated the effects of stressors on Heterorhabditis bacteriophora infection of Tenebrio molitor larva. The stressors included temperature extremes, insecticidal oils and metal cations. Most chemical stressor-nematode combinations caused high host mortal...
Cryopreservation of Radopholus similis, a tropical plant-parasitic nematode
2007-01-01
For obligate plant-parasitic nematodes, cryopreservation has advantages over the usual preservation methods on whole plants or axenic culture systems, because the latter two are labourious and time and space consuming. In addition, cross contamination among different isolates can occur easily. Moreover, specific genetic studies require maintenance of the original population. The nematode under investigation, Radopholus similis, is a plant-parasitic nematode from the humid tropics. Therefore, any treatment at low temperatures is likely to add extra stress to the nematode, making the development of a cryopreservation protocol extremely difficult. In this paper, we describe experiments to achieve a successful cryopreservation protocol for the tropical nematode R. similis using vitrification s...
Connection between the decline of spruce and occurrence of animal pests, especially nematodes
1986-12-01
In various regions of Bavaria, affected by the decline of spruce, attack by insects and especially nematodes was examined on diseased and healthy spruces. A connection between harmful forest insects and the decline of spruce did not become evident, neither over wide areas nor by examination of single trees. Attack by nematodes was examined in soil and wood samples and also in fine feeder roots of diseased and healthy trees. Plant-parasitic nematodes were not found in the wood and in feeder roots. Although root-parasitic nematodes were present in soil samples, their density was too little to account for a direct damage to spruce. They occurred likewise in samples from healthy and diseased trees. Plant-parasitic nematodes can thus be excluded as a possible causal agent for the decline of spruce.
Activation of geminivirus V-sense promoters in roots is restricted to nematode feeding sites
2010-01-01
SUMMARY Obligate sedentary endoparasitic nematodes, such as the root-knot and cyst nematodes, elicit the differentiation of specialized nematode nurse or feeding cells [nematode feeding sites (NFS), giant cells and syncytia, respectively]. During NFS differentiation, marked changes in cell cycle progression occur, partly similar to those induced by some geminiviruses. In this work, we describe the activation of V-sense promoters from the Maize streak virus (MSV) and Wheat dwarf virus (WDV) in NFS formed by root-knot and cyst nematodes. Both promoters were transiently active in microinjection experiments. In tobacco and Arabidopsis transgenic lines carrying promoter-b-glucuronidase fusions, the MSV V-sense promoter was activated in the vascular tissues of aerial plant parts, primarily leaf ...
Acanthocytes of Stropharia rugosoannulata Function as a Nematode-Attacking Device
2006-04-01
Full Text Available.Efficient killing of nematodes by Stropharia rugosoannulata Farlow ex Murrill cultures was observed. This fungus showed the ability to immobilize the free-living nematode Panagrellus redivivus Goodey within minutes and to immobilize the pine wilt nematode Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle within hours on agar plates. Moreover, P. redivivus worms were completely degraded by the fungus within 24 to 48 h. The cultures of S. rugosoannulata studied shared the characteristic of abundantly producing cells with finger-like projections called acanthocytes. We showed that the nematode-attacking activity of this fungus is carried out by these spiny acanthocytes and that mechanical force is an important factor in the process. Furthermore, the growth and nematode-attacking activity of the fungus in soil were also determined, and our results suggest that acanthocytes are functional in soil.
2010-01-01
Nematodes are the most abundant metazoans of deep-sea benthic communities, but knowledge of their distribution is limited relative to larger organisms. Whilst some aspects of nematode processing techniques, such as extraction, have been extensively studied, other key elements have attracted little attention. We compared the effect of (1) mesh size (63, 45, and 32μm) on estimates of nematode abundance, biomass, and body size, and (2) microscope magnification (50× and 100×) on estimates of nematode abundance at bathyal sites (250–3100m water depth) on the Challenger Plateau and Chatham Rise, south-west Pacific Ocean. Variation in the effectiveness of these techniques was assessed in relation to nematode body size and environmental parameters (water depth, sedi...
2010-01-01
Anthelmintic resistant cattle nematodes have been reported in different regions around the world. However, in Western Europe the assessment of the problem relies largely on case reports and no prevalence data based on wide-scale surveys are available. Therefore, we performed a survey to (1) screen for reduced anthelmintic efficacy in Belgian and German cattle farms; (2) evaluate the usefulness of a simplified faecal egg count reduction test (FECRT), where efficacies are based on the mean FECs of 10 at random collected faecal samples pre- and post-treatment per farm and (3) identify possible risk factors for reduced anthelmintic efficacy. Of 88 farms included in this study, 84 farms used macrocyclic lactones (MLs). A FECR
The anesthetic action of ethanol analyzed by genetics in Caenorhabditis elegans
2008-01-01
Acute exposure to ethanol causes paralysis at high concentrations in the nematode Caenorhabditis elegans. We set out to elucidate the mechanism of the anesthetic action of ethanol by genetic approaches. We identified nine mutations that conferred reduced sensitivity to ethanol after chemical, irradiation, or transposon insertion mutagenesis. Of these nine, we further characterized five mutations that defined four genes, jud-1-jud-4. Analysis of the phenotypes of the animals heterozygous for two unlinked genes revealed that jud-1 and jud-3 act synergistically in a gene dose-dependent manner. We cloned jud-4 and found that it encodes a protein with limited homology to human Homer proteins. jud-4 was expressed in the hypodermis and vulva muscles, suggesting that this gene acts in tissues directly exposed to the external environment. Characterization of the other ...
2009-01-01
Synergistic combinations of biological and chemical insecticides might yield promising alternatives for soil insect pest management. In turfgrass of the Northeast U.S., control of root-feeding scarab larvae is highly dependent on conventional insecticides. Studies on interactions between entomopathogenic nematodes and neonicotinoid insecticides, however, demonstrate the feasibility of synergies as an approach for reduced-risk curative control. To understand the breadth of potential synergies, we screened numerous combinations of biological control agents with sublethal doses of neonicotinoids against third instars. Interactions were characterized as synergistic, additive or antagonistic. The most promising combinations identified in laboratory bioassays were advanced to greenhouse pot stud...
2010-01-01
Radopholus similis is a serious threat to black pepper (Piper nigrum L.) cultivation being the main causal organism of slow decline disease. Because of its migratory nature most fungal and bacterial antagonists are ineffective in suppressing R. similis. The presence of a number of endophytic bacteria in black pepper tissues has been proved in earlier studies. This study was undertaken to evaluate the bacteria isolated from black pepper for suppressing R. similis. In vitro and in vivo screenings were used initially to identify the efficient strains of endophytic bacteria that suppress R. similis. Seventy four isolates of endophytic bacteria obtained from black pepper were screened against R. similis by various bioassays. Results of the in vitro experiments were inconclusive and did not matc...
Proteomic analysis uncovers a metabolic phenotype in C. elegans after nhr-40 reduction of function
2008-01-01
Caenorhabditis elegans has an unexpectedly large number (284) of genes encoding nuclear hormone receptors, most of which are nematode-specific and are of unknown function. We have exploited comparative two-dimensional chromatography of synchronized cultures of wild type C. elegans larvae and a mutant in nhr-40 to determine if proteomic approaches will provide additional insight into gene function. Chromatofocusing, followed by reversed-phase chromatography and mass spectrometry, identified altered chromatographic patterns for a set of proteins, many of which function in muscle and metabolism. Prompted by the proteomic analysis, we find that the penetrance of the developmental phenotypes in the mutant is enhanced at low temperatures and by food restriction. The combination of our phenotypic and proteomic analysis strongly suggests that NHR-40 provides a link ...
2008-01-01
The immunoscreening of Brugia malayi adult cDNA library with pooled endemic normal sera identified several seroreactive clones including, EC-SOD which contained a 612bp insert and showed significant nucleotide and deduced amino acid sequence homologies with superoxide dismutase (SOD) of other nematode parasites. The SODs are known to play an important role in the protection of parasite against reactive oxygen species of the host. The coding region of the B. malayi EC-SOD (BmEC-SOD) was cloned and expressed in Escherichia coli followed by affinity purification on nickel agarose resin. Staining of native polyacrylamide gel for SOD activity of the expressed recombinant protein revealed that SOD activity inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, indicating...
Production of eosinophil chemoattractant activity by ovine gastrointestinal nematodes
2005-01-01
Eosinophilia is a well documented feature of helminth infections but the precise nature of the interaction between parasite and eosinophil remains an enigma. This paper describes experiments demonstrating that ruminant gastrointestinal trichostrongyles produce potent chemoattractant activity for ovine bone marrow-derived eosinophils in vitro. This activity was initially identified as a constituent of whole worm extracts of third and fourth larval (L3, L4), and adult stages of Teladorsagia circumcincta, and adult Haemonchus contortus. Similar activity was detected in excretory/secretory (E/S) material derived from live T. circumcincta L3. Subsequently, by adapting the assay technique to incorporate live worms directly into the system, it was shown that L3 of both T. circumcincta and H. cont...
Phylogenomics of caspase-activated DNA fragmentation factor
2007-01-01
The degradation of nuclear DNA by DNA fragmentation factor (DFF) is a key step in apoptosis of mammalian cells. Using comparative genomics, we have here determined the evolutionary history of the genes encoding the two DFF subunits, DFFA (also known as ICAD) and DFFB (CAD). Orthologs of DFFA and DFFB were identified in Nematostella vectensis, a representative of the primitive metazoan clade cnidarians, and in various vertebrates and insects, but not in representatives of urochordates, echinoderms, and nematodes. The domains mediating the interaction of DFFA and DFFB, a caspase cleavage site in DFFA, and the amino acid residues critical for endonuclease activity of DFFB were conserved in Nematostella. These findings suggest that DFF has been a part of the primordial apoptosis system of the eumetazoan common ancestor and that the ancient cell death machinery has degenerated ...
2007-01-01
The tissue-invasive nematode Onchocerca volvulus causes skin and eye pathology in human onchocerciasis. While the adult females reside sessile in subcutaneous nodules, the microfilariae are abundantly released from the nodules, males and juvenile worms migrate through the host tissue. Matrix-degrading metallo- and serine proteinases have been detected in excretory-secretory worm products that may be essential for migration of the mobile stages. In this study, a 1713bp long cDNA encoding for a putative proteinase of O. volvulus has been isolated. The predicted protein sequence includes a signal peptide indicating secretion to the extracellular space, a propeptide, an astacin-like protease domain, an EGF-like and a CUB-domain, thereby identifying the protein as a member of the astacin family...
Gastrointestinal parasites of working donkeys of Ethiopia
2010-01-01
The general prevalence and population composition of gastrointestinal and pulmonary helminths of working donkeys were studied. For the purpose 2935 working donkeys were coprologically examined for nematode and cestode, and 215 donkeys for trematode infections. Seven donkeys that died due to various health problems or were euthanased on a welfare ground were necropsied and the parasites were recovered and identified to the species level. The study was conducted during the periods 1996-1999. Coprological examination revealed 99% strongyle, 80% Fasciola, 51% Parascaris, 30% Gastrodiscus, 11% Strongyloides westeri, 8% cestodes and 2% Oxyuris equi infection prevalence. Over 55% of donkeys had more than 1000 eggs per gram of faeces (epg). Forty two different species of parasites consisting of 33...
Gastrointestinal motor disturbance in rabbits experimentally infected with Strongyloides papillosus
2008-01-01
Strongyloides papillosus is a common nematode in ruminants, and the rabbit is the only susceptible experimental animal that has been identified to date. It is known that heavy infection with S. papillosus causes death in a number of animals. However, even though a number of fatal cases have been reported, the mechanism by which S. papillosus infection leads to death remains unknown. In this study, the pathogenic effect of S. papillosus infection on gastrointestinal motility in infected rabbits was investigated by radiographic means. Gastrointestinal motility in rabbits experimentally infected with S. papillosus was assessed by contrast radiography after oral administration of barium sulfate on days 11 (group A) and 13 (group B) of infection. Body weight, food intake, fecal weight and egg c...
Climate Change Promotes the Emergence of Serious Disease Outbreaks of Filarioid Nematodes
2010-01-01
Filarioid parasites represent major health hazards with important medical, veterinary, and economic implications, and considerable potential to affect the everyday lives of tens of millions of people globally (World Health Organization, 2007). Scenarios for climate change vary latitudinally and regionally and involve direct and indirect linkages for increasing temperature and the dissemination, amplification, and invasiveness of vector-borne parasites. High latitude regions are especially influenced by global climate change and thus may be prone to altered associations and dynamics for complex host-pathogen assemblages and emergence of disease with cascading effects on ecosystem structure. Although the potential for substantial ecological perturbation has been identified, few empirical obs...
Caenorhabditis elegans proteomics comes of age
2010-01-01
Caenorhabditis elegans, a free-living soil nematode, is an ideal model system for studying various physiological problems relevant to human diseases. Despite its short history, C. elegans proteomics is receiving great attention in multiple research areas, including the genome annotation, major signaling pathways (e.g. TGF-b and insulin/IGF-1 signaling), verification of RNA interference-mediated gene targeting, aging, disease models, as well as peptidomic analysis of neuropeptides involved in behavior and locomotion. For example, a proteome-wide profiling of developmental and aging processes not only provides basic information necessary for constructing a molecular network, but also identifies important target proteins for chemical modulation. Although C. elegans has a simple body system an...
A model to die for: signaling to apoptotic cell removal in worm, fly and mouse
2010-01-01
Programmed cell death is used during developmental morphogenesis to eliminate superfluous cells or cells with inappropriate developmental potential (e.g., self-reactive immune cells, tumorigenic cells). Recent work in genetic models has led to a number of key observations, revealing signal transduction pathways and identifying new roles for genes previously studied in corpse removal (e.g., removal of broken synapses in the nervous system). Further, studies using mouse models have suggested a role for removal of apoptotic cells in the establishment or maintenance of immune tolerance. In this review, we survey current knowledge of phagocytic pathways derived from studies in the nematode (Caenorhabditis elegans), the fly (Drosophila melanogaster), and mouse (Mus musculus) model systems.
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