87 research outputs found
Wolbachia genomes and the many faces of symbiosis
No full textHow could scientists working on the pathogenesis of filarial diseases, speciation and parthenogenesis in insects, sex-ratio deviations in crustaceans, pest control, and the evolution of bacterial genomes be united? How could a common research project attract the interest of these scientists? How could parasitology be made even more multi-disciplinary? Two workshops organized by New England Biolabs Inc. (Beverly, MA, USA) provide a simple answer to these questions: studying the genomes of Wolbachia endosymbionts
Draft genome of the filarial nematode parasite Brugia malayi
No full textParasitic nematodes that cause elephantiasis and river blindness threaten hundreds of millions of people in the developing world. We have sequenced the approximately 90 megabase (Mb) genome of the human filarial parasite Brugia malayi and predict approximately 11,500 protein coding genes in 71 Mb of robustly assembled sequence. Comparative analysis with the free-living, model nematode Caenorhabditis elegans revealed that, despite these genes having maintained little conservation of local synteny during approximately 350 million years of evolution, they largely remain in linkage on chromosomal units. More than 100 conserved operons were identified. Analysis of the predicted proteome provides evidence for adaptations of B. malayi to niches in its human and vector hosts and insights into the molecular basis of a mutualistic relationship with its Wolbachia endosymbiont. These findings offer a foundation for rational drug design
The bacterial catalase from filarial DNA preparations derives from common pseudomonad contaminants and not from Wolbachia endosymbionts
No full textWolbachia are obligatory endosymbionts in many species of filarial nematodes. Certain bacterial molecules induce antibody responses in mammalian hosts infected with filariae, while others activate inflammatory responses that contribute to pathology. These findings, coupled with antibiotic studies demonstrating the dependence of filarial embryogenesis on the presence of Wolbachia, have intensified research on Wolbachia-nematode interactions, and the effects of Wolbachia molecules on the mammalian immune system. By amplification and sequencing of 16S rDNA and catalase sequences, we show that filarial DNA samples prepared from nematodes collected under typical conditions are frequently contaminated with Pseudomonas DNA. Analysis of a published DNA fragment containing a catalase attributed to the Wolbachia of Onchocerca volvulus showed it to be most like Pseudomonas, both in terms of sequence similarity and genomic organization. Additionally, there was no obvious catalase in either of two available Wolbachia genome sequences. Contamination of filarial DNA with bacterial sequences other than Wolbachia can complicate studies of the role of these symbionts in filarial biology
Determination of Wolbachia genome size by pulsed-field gel electrophoresis
No full textGenome sizes of six different Wolbachia strains from insect and nematode hosts have been determined by pulsed-field gel electrophoresis of purified DNA both before and after digestion with rare-cutting restriction endonucleases. Enzymes SmaI, ApaI, AscI, and FseI cleaved the studied Wolbachia strains at a small number of sites and were used for the determination of the genome sizes of wMelPop, wMel, and wMelCS (each 1.36 Mb), wRi (1.66 Mb), wBma (1.1 Mb), and wDim (0.95 Mb). The Wolbachia genomes studied were all much smaller than the genomes of free-living bacteria such as Escherichia coli (4.7 Mb), as is typical for obligate intracellular bacteria. There was considerable genome size variability among Wolbachia strains, especially between the more parasitic A group Wolbachia infections of insects and the mutualistic C and D group infections of nematodes. The studies described here found no evidence for extrachromosomal plasmid DNA in any of the strains examined. They also indicated that the Wolbachia genome is circular
Total number of reads sequenced and mapped to <i>B</i>. <i>malayi</i>.
No full textTotal number of reads sequenced and mapped to B. malayi.</p
Total number of reads sequenced and mapped to <i>Wolbachia</i> from <i>B</i>. <i>malayi</i> (<i>w</i>Bm).
No full textTotal number of reads sequenced and mapped to Wolbachia from B. malayi (wBm).</p
Unfertilized, mature <i>B. malayi</i> oocytes contain a polar MTOC.
No full text<p>Mature oocytes (A,C) in meiosis I with bivalents associated with the anterior cortex, and embryos in first zygotic metaphase (B,D) are stained for DNA (blue), α-tubulin (red), and either the PCM marker Zyg-9 (A, B) or γ-tubulin (C,D) (green). In the oocyte, <i>Wolbachia</i> are associated with both poles and distributed in the cytoplasm. By the first zygotic division, <i>Wolbachia</i> are associated with the posterior pole. Scale bar = 5 µm.</p
Lack of an Interchromosomal Effect Associated with Spontaneous Recombination in Males of Drosophila Melanogaster
No full textAuthor Institution: Department of Biological Sciences, Bowling Green State UniversityIt is shown that the frequency of spontaneous male recombination in two different lines of Drosophila melanogaster (OKI and T-007) are not subject to an interchromosomal effect. Second-chromosome male recombination in these lines was not affected by heterozygosity for the multiple third-chromosome inversions In(3LR)TM3 or In(3LR)Ubxm, which do affect recombination in females. It seems, therefore, that a large fraction of spontaneous recombination in males of D. melanogaster occurs by some mechanism other than that in females. We discuss the possibility that the mechanism is chromosome breakage and reunion, and that these breakage events may be caused by a microorganism
Serodiagnosis of Echinococcus spp. infection: explorative selection of diagnostic antigens by peptide microarray
No full textBACKGROUND: Production of native antigens for serodiagnosis of helminthic infections is laborious and hampered by batch-to-batch variation. For serodiagnosis of echinococcosis, especially cystic disease, most screening tests rely on crude or purified Echinococcus granulosus hydatid cyst fluid. To resolve limitations associated with native antigens in serological tests, the use of standardized and highly pure antigens produced by chemical synthesis offers considerable advantages, provided appropriate diagnostic sensitivity and specificity is achieved.
METHODOLOGY/PRINCIPAL FINDINGS: Making use of the growing collection of genomic and proteomic data, we applied a set of bioinformatic selection criteria to a collection of protein sequences including conceptually translated nucleotide sequence data of two related tapeworms, Echinococcus multilocularis and Echinococcus granulosus. Our approach targeted alpha-helical coiled-coils and intrinsically unstructured regions of parasite proteins potentially exposed to the host immune system. From 6 proteins of E. multilocularis and 5 proteins of E. granulosus, 45 peptides between 24 and 30 amino acids in length were designed. These peptides were chemically synthesized, spotted on microarrays and screened for reactivity with sera from infected humans. Peptides reacting above the cut-off were validated in enzyme-linked immunosorbent assays (ELISA). Peptides identified failed to differentiate between E. multilocularis and E. granulosus infection. The peptide performing best reached 57% sensitivity and 94% specificity. This candidate derived from Echinococcus multilocularis antigen B8/1 and showed strong reactivity to sera from patients infected either with E. multilocularis or E. granulosus.
CONCLUSIONS/SIGNIFICANCE: This study provides proof of principle for the discovery of diagnostically relevant peptides by bioinformatic selection complemented with screening on a high-throughput microarray platform. Our data showed that a single peptide cannot provide sufficient diagnostic sensitivity whereas pooling several peptide antigens improved sensitivity; thus combinations of several peptides may lead the way to new diagnostic tests that replace, or at least complement conventional immunodiagnosis of echinococcosis. Our strategy could prove useful for diagnostic developments in other pathogens
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