Molecular identification of Wolbachia from the filarial nematode Mansonella perstans

Abstract

Wolbachiae are bacterial endosymbionts of insects and many filarial nematodes whose products trigger inflammation in filarial infections. The dependence of the parasites on their endosymbionts has also led to the use of antibiotics directed against the Wolbachiae, therapy that has been demonstrated to have a profound salutary effect on filarial infections. The identification of Wolbachiae in Mansonella species has been conclusively shown for Mansonella ozzardi (Mo), but not for Mansonella perstans (Mp). Using primers known to amplify the 16S ribosomal DNA of other filarial Wolbachiae, an identical 1393bp band was found in all samples tested. Sequence analysis of these samples demonstrated a single consensus sequence for Mp Wolbachia 16S rDNA that was most similar to Wolbachia sequences from other filarial nematodes. When aligned with the only other Mansonella Wolbachia sequence (Mo) there were only 8 nucleotide differences in the 1369bp overlapping sequence. Phylogenetic dendrograms, examining the relationship of the Mp Wolbachia to other Wolbachia 16S rDNA, showed that the Wolbachia tracked almost identically to the 5S rRNA of their parasite host. Wolbachia surface protein (WSP) was also demonstrated in protein extracted from Mp-containing whole blood. In advance of a treatment trial of Mp, a method for the quantitation of Mp Wolbachia was developed and used to demonstrate not only a relationship between microfilarial numbers and Wolbachia copy numbers, but also to demonstrate the effect of antibiotic on ridding Mp of Wolbachia.

Some Like It Hot: Evolution and Ecology of Novel Endosymbionts in Bat Flies of Cave-Roosting Bats (Hippoboscoidea, Nycterophiliinae)

“…At this point, no obvious congruence was observed between Wolbachia and Nycterophilia fly phylogeny (trees not shown). Supergroup F Wolbachia organisms have been detected in a range of invertebrates, including filarial nematodes (27) and blood-feeding insects such as bedbugs (45), lice (9), hippoboscid flies (10), and fleas (17), and have recently been identified as a mutualist in some bedbug populations (20).…”

Wolbachia as a bacteriocyte-associated nutritional mutualist

“…S1). For the termite strains, their phenotypic aspects are unknown (31), whereas the nematode strains are suspected to be mutualistic (32). No reproductive parasite strains have been known from the F supergroup.…”

High Prevalence of Mansonella perstans Filariasis in Rural Senegal

“…Previously, only standard PCR developed by Keiser and others in 2008 and based on the internal transcribed spacer was proposed for identification of M. perstans. 33 The first set was designed to be specific for the genus Mansonella and consists of a forward primer, 5′-CCTGCGGAAGGA TCATTAAC-3′, a reverse primer, 5′-ATCGACGGTTTAG GCGATAA-3′, and a probe, 6-FAM-CGGTGATATTCGTT GGTGTCT-TAMRA. The second in silico set is specific for M. perstans, with a forward primer, 5′-AGGATCATTAACG AGCTTCC-3′, a reverse primer, 5′-CGAATATCACCGTTAA TTCAGT-3′, and a probe, 6-FAM-TTCACTTTTATTTAGCAA CATGCA-TAMRA.…”