Wolbachia infections of tephritid fruit flies: molecular evidence for five distinct strains in a single host species

Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and can induce cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing or feminization in their hosts. Here, we report phylogenetic relationships of Wolbachia in tephritid fruit flies based on wsp gene sequences. We also report, for the first time, five distinct strains of Wolbachia in Bactrocera ascita sp. B. Four of the five Wolbachia strains found in this species were in the same groups as those found in other tephritid fruit flies, suggesting possible horizontal transmission of Wolbachia from other fruit flies into B. ascita sp. B. The unreliability of wsp-specific group primers demonstrated in this study suggests that these primers might be useful only for preliminary identification of Wolbachia. Final determination of group affiliation needs to be verified with wsp sequence data.

wsp Gene sequences from the Wolbachia of filarial nematodes

Wolbachia endosymbiotic bacteria are widespread in arthropods and are also present in filarial nematodes. Almost all filarial species so far examined have been found to harbor these endosymbionts. The sequences of only three genes have been published for nematode Wolbachia (i.e., the genes coding for the proteins FtsZ and catalase and for 16S rRNA). Here we present the sequences of the genes coding for the Wolbachia surface protein (WSP) from the endosymbionts of eight species of filaria. Complete gene sequences were obtained from the endosymbionts of two different species, Dirofilaria immitis and Brugia malayi. These sequences allowed us to design general primers for amplification of the wsp gene from the Wolbachia of all filarial species examined. For these species, partial WSP sequences (about 600 base pairs) were obtained with these primers. Phylogenetic analysis groups these nematode wsp sequences into a coherent cluster. Within the nematode cluster, wsp-based Wolbachia phylogeny matches a previous phylogeny obtained with ftsZ gene sequences, with a good consistency of the phylogeny of hosts (nematodes) and symbionts (Wolbachia). In addition, different individuals of the same host species (Dirofilaria immitis and Wuchereria bancrofti) show identical wsp gene sequences.

Phylogenetic position of Chitinophaga pinensis in the Flexibacter-Bacteroides-Cytophaga phylum

Comparison of the 16S rRNA gene sequence determined for Chitinophaga pinensis showed that this species is most closely related to Flexibacter filiformis in the Flexibacter-Bacteroides-Cytophaga phylum, These two chitinolytic bacteria, which are characterized by transformation into spherical bodies on ageing, belong to a strongly supported lineage that also includes Cytophaga arvensicola, Flavobacterium ferrugineum and Flexibacter sancti, The lineage is distinct from the microcyst-forming species Sporocytophaga myxococcoides.

Description of Skermanella parooensis gen. nov., sp. nov. to accommodate Conglomeromonas largomobilis subsp. parooensis following the transfer of Conglomeromonas largomobilis subsp. largomobilis to the genus Azospirillum

As a consequence of the transfer of the type species Conglomeromonas largomobilis subsp. largomobilis to the genus Azospirillum, the name of the genus Conglomeromonas must be changed in accordance with Rule 37a(1) of the International Code of Nomenclature of Bacteria. Consequently, it is proposed that the subspecies Conglomeromonas largomobilis subsp, parooensis be transferred to the genus Skermanella gen, nov. as the type species Skermanella parooensis gen, nov., sp, nov. This taxon belongs to an isolated subline of descent in the Azospirillum branch of the alpha-Proteobacteria. The spelling of the specific epithet of Azospirillum largomobile is corrected to Azospirillum largimobile.

Genus I. Chrysiogenes

This volume includes numerous introductory chapters on the basic concepts and practice of prokaryotic taxonomy in addition to detailed descriptions of the Archaea, phototrophic bacteria and some very deep bacterial phyla. Introductory chapters include the topics of classification of prokaryotes, the concept of bacterial speciation, numerical and polyphasic taxonomy, bacterial nomenclature and the etymology of prokaryotic names, nucleic acid probes and their application in environmental microbiology, culture collections, and the intellectual property of prokaryotes. The first Road Map to the prokaryotes is included as well as an overview of the phylogenetic backbone and taxonomic framework for prokaryotic systematics.

Evidence for an independent radiation of endosymbiotic litostome ciliates within Australian marsupial herbivores

The recent discovery of isotrichid-like ciliates occurring as endosymbionts in macropodid marsupials posed interesting questions in regard to both their phyletic origin (all previous records confined to eutherian mammals) and their morphological evolution (Australian forms possibly representing missing links between previously described genera). The SSU rRNA gene was sequenced for three species (Dasytricha dehorityi, D. dogieli, and Batricha tasmaniensis) and aligned against representatives of all major ciliate classes. The Australian species did not group with the other isotrichid species but instead formed an independent radiation. Discrepancies between recent global phylogenies of the phylum Ciliophora were examined by manipulation of the aligned sequence data set. Sources of conflict between these studies did not stem from differences in outgroup choice or phylogenetic reconstruction methods. Differences in the application of confidence limits and primary sequence alignment have probably resulted in the reporting of spurious associations which are not supported by more conservative confidence or alignment methodology. At present, the ciliate subphylum Intramacro-nucleata is an unresolved polytomy which may be due to deficiencies in the SSU rRNA gene sequence dataset or indicate that the ciliates radiated into their extant classes by rapid burst-like evolution. (C) 2001 academic Press.

New arsenite-oxidizing bacteria isolated from Australian gold mining environments - Phylogenetic relationships

Nine novel arsenite-oxidizing bacteria have been isolated from two different gold mine environments in Australia. Four of these organisms grow chemolithoautotrophically with oxygen as the terminal electron acceptor, arsenite as the electron donor, and carbon dioxide-bicarbonate as the sole carbon source. Five heterotrophic arsenite-oxidizing bacteria were also isolated, one of which was found to be both phylogenetically and physiologically identical to the previously described heterotrophic arsenite oxidizer misidentified as Alcaligenes faecalis. The results showed that this strain belongs to the genus Achromobacter. Phylogenetically, the arsenite-oxidizing bacteria fall within two separate subdivisions of the Proteobacteria. Interestingly, the chemolithoautotrophic arsenite oxidizers belong to the alpha-Proteobacteria, whereas the heterotrophic arsenite oxidizers belong to the beta-Proteobacteria.

Candidatus "Scalindua brodae", spec. nov., Candidatus "Scalindua wagneri", spec. nov., two new species of anaerobic ammonium oxidizing bacteria

Anaerobic ammonium oxidation (anammox) is both a promising process in wastewater treatment and a long overlooked microbial physiology that can contribute significantly to biological nitrogen cycling in the world's oceans. Anammox is mediated by a monophyletic group of bacteria that branches deeply in the Planctomycetales. Here we describe a new genus and species of anaerobic ammonium oxidizing planctomycetes, discovered in a wastewater treatment plant (wwtp) treating landfill leachate in Pitsea, UK. The biomass from this wwtp showed high anammox activity (5.0 +/- 0.5 nmol/mg protein/min) and produced hydrazine from hydroxylamine, one of the unique features of anammox bacteria. Eight new planctomycete 16S rRNA gene sequences were present in the 16S rRNA gene clone library generated from the biomass. Four of these were affiliated to known anammox 16S rRNA gene sequences, but branched much closer to the root of the planctomycete line of descent. Fluorescence in situ hybridization (FISH) with oligonucleotide probes specific for these new sequences showed that two species (belonging to the same genus) together made up > 99% of the planctomycete population which constituted 20% of the total microbial community. The identification of these organisms as typical anammox bacteria was confirmed with electron microscopy and lipid analysis. The new species, provisionally named Candidatus Scalindua brodae and Scalindua wagneri considerably extend the biodiversity of the anammox lineage on the 16S rRNA gene level, but otherwise resemble known anammox bacteria. Simultaneously, another new species of the same genus, Candidatus Scalindua sorokinii, was detected in the water column of the Black Sea, making this genus the most widespread of all anammox bacteria described so far.

Are Ichthyosporea animals or fungi? Bayesian phylogenetic analysis of elongation factor 1α of Ichthyophonus irregularis

Ichthyosporea is a recently recognized group of morphologically simple eukaryotes, many of which cause disease in aquatic organisms. Ribosomal RNA sequence analyses place Ichthyosporea near the divergence of the animal and fungal lineages, but do not allow resolution of its exact phylogenetic position. Some of the best evidence for a specific grouping of animals and fungi (Opisthokonta) has come from elongation factor 1alpha, not only phylogenetic analysis of sequences but also the presence or absence of short insertions and deletions. We sequenced the EF-1alpha gene from the ichthyosporean parasite Ichthyophonus irregularis and determined its phylogenetic position using neighbor-joining, parsimony and Bayesian methods. We also sequenced EF-1alpha genes from four chytrids to provide broader representation within fungi. Sequence analyses and the presence of a characteristic 12 amino acid insertion strongly indicate that I. irregularis is a member of Opisthokonta, but do not resolve whether I. irregularis is a specific relative of animals or of fungi. However, the EF-1alpha of I. irregularis exhibits a two amino acid deletion heretofore reported only among fungi. (C) 2003 Elsevier Science (USA). All rights reserved.

Ptychogyliauchen, a new genus of Gyliauchenidae (Platyhelminthes: Digenea) from siganid fishes of the Indo-West Pacific

We propose a new genus of the Gyliauchenidae Fukui, 1929 ( Digenea), Ptychogyliauchen, gen. nov., for four new species that infect Indo-West Pacific siganid fishes. Ptychogyliauchen, gen. nov. is a morphologically distinctive genus, diagnosed principally by the presence of a highly convoluted oesophagus, which generally exceeds the total body length of the worm, and by the unusual folded structure of the ejaculatory duct. Ptychogyliauchen thetidis, sp. nov. is designated as the type species, and is described from the intestine of Siganus punctatus (Siganidae) from Heron Island, Great Barrier Reef, Queensland, Australia. Ptychogyliauchen himinglaeva, sp. nov. is described from the intestine of Siganus corallinus ( Siganidae) from Lizard Island, Great Barrier Reef, Queensland, Australia. Ptychogyliauchen leucothea, sp. nov. is described from the intestine of S. argenteus, and further recorded from S. fuscescens, off Ningaloo, Western Australia, Australia. Ptychogyliauchen thistilbardi, sp. nov. is described from the intestine of S. doliatus from Noumea, New Caledonia, and is also found in S. argenteus, S. canaliculatus, S. corallinus and S. spinus from Noumea, New Caledonia, and Moorea, Tahiti, French Pacific. Ptychogyliauchen thistilbardi, sp. nov. also occurs in the intestine of Chaetodon citrinellus (Chaetodontidae) from Moorea. A key to species is provided. All species have been described following morphological examination using light microscopy, and specimens of P. thetidis, sp. nov., P. leucothea, sp. nov. and P. thistilbardi, sp. nov. have been characterised using molecular methods. Sequences were obtained for a combination of nuclear ribosomal (28S (D1-D3) and ITS2) and mitochondrial (ND1) genes. A phylogenetic analysis of sequenced specimens of Ptychogyliauchen, gen. nov. was conducted using species of Petalocotyle Ozaki, 1934 for outgroup comparison. This analysis, based on alignments of the ITS2 and 28S (D1-D3) rDNA genes, supports monophyly of the geographically widespread P. thistilbardi, sp. nov., which is known from both siganid and chaetodontid hosts. We discuss the taxonomy of the genus and the host associations of each species and the group.

The life cycle of Paracardicoloides yamagutii Martin, 1974 (Digenea: Sanguinicolidae)

The sanguinicolids Paracardicoloides yamagutii Martin, 1974 and Plethorchis acanthus Martin, 1975 were obtained from their definitive hosts, Anguilla reinhardtii Steindachner and Mugil cephalus Linnaeus (respectively) in the tributaries of the Brisbane River, Queensland, Australia. Two putative sanguinicolid cercariae were collected from a hydrobiid gastropod, Posticobia brazieri Smith, in the same waters. The two cercariae differ markedly in size and the form of their sporocysts. Both putative cercariae develop in the digestive gland of Po. brazieri. The ITS2 rDNA region from these sanguinicolids and a Clinostomum species (utilised as an outgroup due to the close morphological similarities between the cercarial stages of the Clinostomidae and the Sanguinicolidae) were sequenced and aligned. Comparison of the ITS2 sequences showed one cercaria to be that of P. yamagutii. This is the first sanguinicolid life history determined by a molecular method. P. yamagutii is the fourth sanguinicolid known to utilise a freshwater hydrobiid gastropod as its intermediate host. ITS2 rDNA is effective in distinguishing sanguinicolids at the species level.