Pyruvated carrageenans from Solieria robusta and its adelphoparasite Tikvahiella candida

Solieria, the type genus of the commercially important red algal family Solieriaceae (Gigartinales), contains seven or eight species, three of which are represented in Australia. The cell-wall galactans of the most common Australian Solieria species, S. robusta (Greville) Kylin, were analysed by a combination of compositional assays, linkage analysis, and Fourier transform infrared (FTIR) and C-13 nuclear magnetic resonance (NMR) spectroscopy. They are shown to be composed predominantly of carrabiose 2,4'-disulphate units (the repeating unit of iota-carrageenan) and a significant proportion of 4',6'-pyruvated carrabiose 2-sulphate units. The constituent sugars, pyruvate content, FTIR spectrum, and linkage and substitution patterns of the galactans from Tikvahiella candida Kraft et Gabrielson, an adelphoparasite of Solieria robusta, closely resemble those of its host and furnish evidence in support of a close phylogenetic relationship between the two species.

Description of Gluconacetobacter sacchari sp. nov., a new species of acetic acid bacterium isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug

A new species of the genus Gluconacetobacter, for which the name Gluconacetobacter sacchari sp. nov. is proposed, was isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug, Saccharicoccus sacchari, found on sugar cane growing in Queensland and northern New South Wales, Australia, The nearest phylogenetic relatives in the alpha-subclass of the Proteobacteria are Gluconacetobacter liquefaciens and Gluconacetobacter diazotrophicus, which have 98.8-99.3% and 97.9-98.5% 16S rDNA sequence similarity, respectively, to members of Gluconacetobacter sacchari. On the basis of the phylogenetic positioning of the strains, DNA reassociation studies, phenotypic tests and the presence of the Q10 ubiquinone, this new species was assigned to the genus Gluconacetobacter. No single phenotypic characteristic is unique to the species, but the species can be differentiated phenotypically from closely related members of the acetic acid bacteria by growth in the presence of 0.01% malachite green, growth on 30% glucose, an inability to fix nitrogen and an inability to grow with the L-amino acids asparagine, glycine, glutamine, threonine and tryptophan when D-mannitol was supplied as the sole carbon and energy source. The type strain of this species is strain SRI 1794(T) (= DSM 12717(T)).

Comparison of acetylcholinesterase genes from cattle ticks

We describe the isolation and characterisation of two putatively new acetylcholinesterase genes from the African cattle ticks Boophilus decoloratus and Rhipicephalus appendiculatus. The nucleotide sequences of these genes had 93% homology to each other and 95% and 91% identity, respectively, to the acetylcholinesterase gene from an Australian strain of another cattle tick, Boophilus microplus. Translation of the nucleotide sequences revealed putative amino acids that are essential for acetylcholinesterase activity: the active site serine, and the histidine and glutamate residues that associate with this serine to form the catalytic triad. All known acetylcholinesterases have three sets of cysteines that form disulfide bonds; however, the acetylcholinesterase genes of these three species of ticks encode only two sets of cysteines. Acetylcholinesterases of B. microplus from South Africa, Zimbabwe, Kenya and Mexico had 98-99% identity with acetylcholinesterase from B. microplus from Australia, whereas acetylcholinesterase from B. microplus from Indonesia was identical to that from Australia. Preliminary phylogenetic analyses surprisingly indicate that the acetylcholinesterases of ticks are closer phylogenetically to acetylcholinesterases of vertebrates than they are to those of other arthropods. (C) 1999 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.

What the evolution of the amyloid protein precursor supergene family tells us about its function

The Alzheimer's disease amyloid protein precursor (APP) gene is part of a multi-gene super-family from which sixteen homologous amyloid precursor-like proteins (APLP) and APP species homologues have been isolated and characterised. Comparison of exon structure (including the uncharacterised APL-1 gene), construction of phylogenetic trees, and analysis of the protein sequence alignment of known homologues of the APP super-family were performed to reconstruct the evolution of the family and to assess the functional significance of conserved protein sequences between homologues. This analysis supports an adhesion function for all members of the APP super family, with specificity determined by those sequences which are not conserved between APLP lineages, and provides evidence for an increasingly complex APP superfamily during evolution. The analysis also suggests that Drosophila APPL and Caenorhabdotids elegans APL-1 may be a fourth APLP lineage indicating that these proteins, while not functional homologues of human APP, are similarly likely to regulate cell adhesion. Furthermore, the beta A4 sequence is highly conserved only in APP orthologues, strongly suggesting this sequence is of significant functional importance in this lineage. (C) 2000 Elsevier Science Ltd. All rights reserved.

The status of the genera Hysterolecithoides Yamaguti, 1934, Neotheletrum Gibson & Bray, 1979 and Machidatrema Leon-Regagnon, 1998 (Digenea : Hemiuroidea), including a description of M. leonae n. sp from Australian waters

A diagnosis is given for the lecithasterid genus Hysterolecithoides Yamaguti, 1934, which is now found to have two to six (possibly seven) vitelline masses. The species H. frontilatus (Manter, 1969) is returned to the genus, having been considered a member of the bunocotylid genus Neotheletrum by recent authors. It is redescribed from Siganus nebulosus, Moreton Bay, and S. doliatus, Lizard Island, Great Barrier Reef and New Caledonia, with emphasis on the presence of Juel's organ, a uterine seminal receptacle and the blind sac associated with the genital atrium. It differs from its congeners in the trajectory of the pars prostatica which recurves dorsally to the sinus-sac. Oligolecithoides Shen, 1982 is synonymised with Hysterolecithoides and O. trilobatus Shen, 1982 is synomised with H. epinepheli Yamaguti, 1934. Machidatrema Leon-Regagnon, 1998 is diagnosed, and found to be close to Hysterolecithoides, but differs in the lack of a blind-sac projecting from the dorsal genital atrium, by its tandem testes, the coiling of the uterus between the testes and the ovary, and the ventral excretory pore. M. leonae n. sp. is described from Siganus fuscescens, S. lineatus, S. doliatus, S. corallinus, S. vulpinus and Scarus globiceps at Heron Island, Queensland. It differs from its closest congener, M. akeh, in the muscular and tegumental flap over the genital pore and details of the terminal genitalia. M. chilostoma (Machida, 1980) and M. kyphosi (Yamaguti, 1970) are redescribed from Kyphosus vaigiensis from Heron Island. Neotheletrum Gibson & Bray, 1979 is diagnosed: it differs from Hysterolecithoides in its confluent excretory arms, blind seminal receptacle (no Juel's organ) and uniformly tripartite vitellarium. A cladistic analysis suggests that M. chilostoma and M. kyphosi are not best accommodated in Machidatrema, that Machidatrema (sensu stricto) is monophyletic and that Hysterolecithoides is paraphyletic. Hysterolecithoides and Machidatrema are considered hysterolecithine lecithasterids, whilst Neotheletrum is retained as an opisthadenine bunocotylid.

The status of Petalocotyle Ozaki, 1934 (Digenea : Gyliauchenidae), including the description of two new species from acanthurid fishes in Queensland, Australia

The status of Petalocotyle Ozaki, 1934 within the Gyliauchenidae Goto & Matsudaira, 1918 is reviewed. Two new species, P. adenometra from Prionurus microlepidotus (Amity Point, Queensland, Australia) and P. diverticulata from Acanthurus nigrofuscus and A. lineatus (Heron Island, Queensland, Australia), are described. The body plan of Petalocotyle conforms to that of members of the Gyliauchenidae (oral sucker absent, well-developed pharynx, complex oesophagus and characteristic male terminal genitalia), indicating justifiable inclusion in this family. A new diagnosis is given for the genus, such that Petalocotyle is now identified by the presence of an anterior, protuberant ventral sucker, long caeca, a large, sigmoid cirrus-sac containing a coiled ejaculatory duct, and an extensive vitellarium. We suggest that, of all the known genera of gyliauchenids, Petalocotyle may most closely resemble the 'archaetypal gyliauchenid', that is, it may be placed basally within the radiation of the Gyliauchenidae. However, derived characters, like diverticula in the reproductive system, indicate that some characters of individual members of Petalocotyle may be considered advanced and do not reflect an archaetypal condition. Parallels in the structure of the male and female genitalia of Robphildollfusium Paggi & Orecchia, 1963 and Petalocotyle, along with the shared morphology of the digestive tract, indicate possible phylogenetic links between the two genera. This affinity is difficult to infer using morphology alone and recommend that Robphildollfusium remain detached from the Gyliauchenidae.

Functional significance of the beta-hairpin in the insecticidal neurotoxin omega-atracotoxin-Hv1a

omega -Atracotoxin-Hv1a is an insect-specific neurotoxin whose phylogenetic specificity derives from its ability to antagonize insect, but not vertebrate, voltage-gated calcium channels. In order to help understand its mechanism of action and to enhance its utility as a lead compound for insecticide development, we used a combination of protein engineering and site-directed mutagenesis to probe the toxin for key functional regions. First, we constructed a Hairpinless mutant in which the C-terminal beta -hairpin, which is highly conserved in this family of neurotoxins, was excised without affecting the fold of the residual disulfide-rich core of the toxin. The Hairpinless mutant was devoid of insecticidal activity, indicating the functional importance of the hairpin. We subsequently developed a highly efficient system for production of recombinant toxin and then probed the hairpin for key functional residues using alanine-scanning mutagenesis followed by a second round of mutagenesis based on initial hits from the alanine scan. This revealed that two spatially proximal residues, Asn(27) and Arg(35), form a contiguous molecular surface that is essential for toxin activity. We propose that this surface of the beta -hairpin is a key site for interaction of the toxin with insect calcium channels.

Cryptococcus nyarrowii sp nov., a basidiomycetous yeast from Antarctica

in December 1997,196 soil and snow samples were collected from Vestvold Hills, Davis Base, Antarctica. Two isolates, CBS 8804 T (pink colonies) and CBS 8805 (yellow colonies), were shown by proteome analysis and DNA sequencing to represent the same species. Results from the sequencing of the D1/D2 region of the large rDNA subunit placed this species in the hymenomycetous tree in a unique sister clade to the Trichosporonalles and the Tremellalles. The clade consists of Holtermannia corniformis CBS 6979 and CBS strains 8804(T) 8805, 8016, 7712, 7713 and 7743. Morphological and physiological characteristics placed this species in the genus Cryptococcus, with characteristics including the assimilation Of D-glucuronate and myo-inositol, no fermentation, positive Diazonium blue B and urease reactions, absence of sexual reproduction and production of starch-like compounds. Fatty acid analysis identified large proportions of polyunsaturated lipids, mainly linolleic (C-18.2) and, to a lesser extent, linolenic (C-18.3) acids. On the basis of the physiological and phylogenetic data, isolates CBS 8804(T) and CBS 8805 are described as Cryptococcus nyarrowii sp. nov.

Isolation and characterization of a neprilysin-like protein from Venturia canescens virus-like particles

Maternal protein secretions from endoparasitoid wasps are evolutionary adaptations to regulate host physiology as part of an extended wasp phenotype. Virus-like particles (VLPs) produced in the calyx region of Venturia canescens wasps are involved in immune evasion of the developing parasitoid inside the host. In contrast to polydnaviruses (PDVs), VcVLPs are devoid of any nucleic acids. To understand the role of these particles in the regulation of host physiology and phylogenetic relationship between VLPs and PDVs, it is essential to identify particle proteins. In this paper, we describe the isolation and molecular cloning of a neprilysin-like gene (VcNEP) coding for a 94 kDa VcVLP protein and discuss its possible role in host regulation.

Intragenomic variation in ITS2 rDNA in the louse of humans, Pediculus humanus: ITS2 is not a suitable marker for population studies in this species

The two internal transcribed spacers (ITS) of ribosomal DNA are often used as markers of populations of insects. We studied the ITS2 of the head lice and body lice of humans, to determine whether this gene is a suitable marker of populations of these insects. ITS2 sequences were amplified by PCR from lice from four different countries: Australia, China, Japan and the USA. Direct cycle-sequencing of some of these PCR products gave equivocal nucleotide chromatograms. This indicated that some lice had more than one ITS2 sequence, so we cloned PCR products from these lice. Temperature gradient gel electrophoresis (TGGE) revealed that 50 of the 67 clones we screened had different nucleotide sequences. All lice had several ITS2 types, including those with unequivocal chromatograms. A phylogenetic tree of 15 different ITS2 sequences showed that the sequences from individual lice were not monophyletic. We conclude that the ITS2 is not a useful marker of populations for Pediculus humanus.

maT - A clade of transposons intermediate between mariner and Tc1

A group of transposons, named maT, with characteristics intermediate between mariner and Tc1 transposons, is described. Two defective genomic copies of MdmaT from the housefly Musca domestica, with 85% identity, were found flanking and imbedded in the MdalphaE7 esterase gene involved in organophosphate insecticide resistance. Two cDNA clones, with 99% identity to each other and 72%-89% identity to the genomic copies were also obtained, but both represented truncated versions of the putative open reading frame. A third incomplete genomic copy of MdmaT was also identified upstream of the putative M. domestica period gene. The MdmaT sequences showed high identity to the transposable element Bmmar1 from the silk-worm moth, Bombyx mori, and to previously unidentified sequences in the genome of Caenorhabditis elegans. A total of 16 copies of full-length maT sequences were identified in the C elegans genome, representing three variants of the transposon, with 34%-100% identity amongst them. Twelve of the copies, named CemaT1, were virtually identical, with eight of them encoding a putative full length, intact transposase. Secondary structure predictions and phylogenetic analyses confirm that maT elements belong to the mariner-Tc1 superfamily of transposons, but their intermediate sequence and predicted structural characteristics suggest that they belong to a unique clade, distinct from either mariner-like or Tc1-like elements.

Cryptococcus statzelliae sp nov and three novel strains of Cryptococcus victoriae, yeasts isolated from Antarctic soils

A morphological and physiological characterization of yeast strains CBS 8908, CBS 8915, CBS 8920, CBS 8925(T) and CBS 8926, isolated from Antarctic soils, was performed. Phylogenetic analyses of the sequences of the D1/D2 regions and the adjacent internal transcribed spacer (ITS) regions of the large-subunit rDNA of these strains placed them into the Tremellales clade of the Hymenomycetes. The sequence data identified strains CBS 8908, CBS 8915 and CBS 8920 as belonging to the species Cryptococcus victoriae. Strains CBS 8925(T) and CBS 8926 were found to represent an unique clade within the Hymenomycetes, with Dioszegia crocea CBS 6714(T) being their closest phylogenetic relative. Fatty acid composition and proteome fingerprint data for these novel strains were also obtained. No sexual state was observed. A novel basidiomycetous species, Cryptococcus statzelliae, is proposed for strains CBS 8925(T) and CBS 8926.

Chimeric 16S rDNA sequences of diverse origin are accumulating in the public databases

A significant number of chimeric 16S rDNA sequences of diverse origin were identified in the public databases by partial treeing analysis. This suggests that chimeric sequences, representing phylogenetically novel non-existent organisms, are routinely being overlooked in molecular phylogenetic surveys despite a general awareness of PCR-generated artefacts amongst researchers.

Virus or not? Phylogenetics of polydnaviruses and their wasp carriers

Our current, still limited, understanding of the comparative biology and evolution of polydnaviruses (PDVs) is reviewed, especially in the context of the possible origins of these parasitoid viruses and of their coevolution with carrier wasps. A hypothetical scenario of evolution of PDVs from ascovirus (or ascovirus-like) ancestors is presented, with examples of apparent extant transitional forms. PDVs appear, in the case of bracoviruses, to show phylogenetic relationships that mirror those of their wasp carriers: with ichno-viruses, the picture is less clear. Ongoing sequencing studies of entire PDV genomes from diverse wasp species are likely to greatly contribute to our understanding of PDV evolution. (C) 2003 Elsevier Science Ltd. All rights reserved.

Humans, dogs and parasitic zoonoses - Unravelling the relationships in a remote endemic community in Northeast India using molecular tools

Canine parasitic zoonoses pose a continuing public health problem, especially in developing countries and communities that are socioeconomically disadvantaged. Our study combined the use of conventional and molecular epidemic, logical tools to determine the role of dogs in transmission of gastrointestinal (GI) parasites such as hookworms, Giardia and Ascaris in a parasite endemic teagrowing community in northeast India. A highly sensitive and specific molecular tool was developed to detect and differentiate the zoonotic species of canine hookworm eggs directly from faeces. This allowed epidemiological screening of canine hookworm species in this community to be conducted with ease and accuracy. The zoonotic potential of canine Giardia was also investigated by characterising Giardia duodenalis recovered from humans and dogs living in the same locality and households at three different loci. Phylogenetic and epidemiological analysis provided compelling evidence to support the zoonotic transmission of canine Giardia. Molecular tools were also used to identify the species of Ascaris egg present in over 30% of dog faecal samples. The results demonstrated the role of dogs as a significant disseminator and environmental contaminator of Ascaris lumbricoides in communities where promiscuous defecation practices exist. Our study demonstrated the usefulness of combining conventional and molecular parasitological and epidemiological tools to help solve unresolved relationships with regards to parasitic zoonoses.