Platyhelminth phylogenetics - a key to understanding parasitism?

The comparative method, the inference of biological processes from phylogenetic patterns, is founded on the reliability of the phylogenetic tree. In attempting to apply the comparative method to the understanding of the evolution of parasitism in the phylum Platyhelminthes, we have highlighted several points we consider to be of value along with many problems. We discuss four of these topics. Firstly, we view the group at a phylum level, in particular discussing the importance of establishing the sister taxon to the obligate parasite group, the Neodermata, for addressing such questions as the monophyly, parasitism or the endo or ectoparasitic nature of the early parasites. The variety of non-congruent phylogenetic trees presented so far, utilising either or both morphological and molecular data, gives rise to the suggestion that any evolutionary scenarios presented at this stage be treated as interesting hypotheses rather than well-supported theories. Our second point of discussion is the conflict between morphological and molecular estimates of monogenean evolution. The Monogenea presents several well-established morphological autapomorphies, such that morphology consistently estimates the group as monophyletic, whereas molecular sequence analyses indicate paraphyly, with different genes giving different topologies. We discuss the problem of reconciling gene and species trees. Thirdly, we use recent phylogenetic results on the tapeworms to interpret the evolution of strobilation, proglottization, segmentation and scolex structure. In relation to the latter, the results presented indicate that the higher cestodes are diphyletic, with one branch difossate and the other tetrafossate. Finally, we use a SSU rDNA phylogenetic tree of the Trematoda as a basis for the discussion of an aspect of the digenean life-cycle, namely the nature of the first intermediate host. Frequent episodes of host-switching, between gastropod and bivalve hosts or even into annelids, are indicated.

Recent advances in our knowledge of the Myxozoa

In the last few years two factors have helped to significantly advance our understanding of the Myxozoa. First, the phenomenal increase in fin fish aquaculture in the 1990s has lead to the increased importance of these parasites; in rum this has lead to intensified research efforts, which have increased knowledge of the development, diagnosis, and pathogenesis of myxozoans. The hallmark discovery in the 1980s that the life cycle of Myxobolus cerebralis requires development of an actinosporean stage in the Oligochaete. Tubifex tubifex, led to the elucidation of the life cycles of several other myxozoans. Also, the life cycle and taxonomy of the enigmatic PKX myxozoan has been resolved: it is the alternate stage of the unusual myxozoan. Tetracapsula bryosalmonae, from bryozoans. The 18S rDNA gene of many species has been sequenced, and here we add 22 new sequences to the data set. Phylogenetic analyses using all these sequences indicate that: 1) the Myxozoa are closely related to Cnidaria (also supported by morphological data), 2) marine taxa at the genus level branch separately from genera that usually infect freshwater fishes; 3) taxa cluster more by development and tissue location than by spore morphology; 4) the tetracapsulids branched off early in myxozoan evolution, perhaps reflected by their having bryozoan. rather than annelid hosts; 5) the morphology of actinosporeans offers little information for determining their myxosporean counterparts (assuming that they exist), and 6) the marine actinosporeans from Australia appear to form a clade within the platysporinid myxosporeans. Ribosomal DNA sequences have also enabled development of diagnostic tests for myxozoans. PCR and in situ hybridisation tests based on rDNA sequences have been developed for Myxobolus cerebralis. Ceratomyxa shasta. Kudoa spp,, and Tetracapsula bryosalmonae (PKX). Lectin-based and antibody tests have also been developed for certain myxozoans, such as PKX and C. shasta. We also review important diseases caused by myxozoans. which are emerging or re-emerging. Epizootics of whirling disease in wild rainbow trout (Oncorhynchus mykiss) have recently been reported throughout the Rocky Mountain states of the USA. With a dramatic increase in aquaculture of fishes using marine netpens, several marine myxozoans have been recognized or elevated in status as pathological agents. Kudoa thyrsites infections have caused severe post-harvest myoliquefaction in pen-reared Atlantic salmon (Salmo salar), and Ceratomyxa spp., Sphaerospora spp., and Myxidium leei cause disease in pen-reared sea bass (Dicentrarchus labrax) and sea bream species (family Sparidae) in Mediterranean countries.

Four new species of Macropodinium (Ciliophora : Litostomatea) from Australian wallabies and pademelons

Samples of Macropodinium spp. were collected from 3 new macropodid species: from 21 of 28 (75%) black-striped wallabies (Macropus dorsalis); 10 of 11 (91%) swamp wallabies (Wallabia bicolor); and 22 of 43 (51%) Tasmanian pademelons (Thylogale billardierii). The examination of ciliate morphology by silver impregnation and scanning electron microscopy led to the redescription of the genus Macropodinium and the description of 4 new species: Ma. tricresta sp. nov. and Ma. spinosus sp. nov. from M. dorsalis; Ma. maira sp. nov. from T. billardierii; and M. bicolor sp. nov. from W. bicolor; each species was strictly host specific. Cellular orientation was reinterpreted on the basis of vestibular morphology and it is concluded that Macropodinium spp. are laterally rather than dorso-ventrally compressed. The striated groove is thus dorso-ventral rather than lateral. Oral ciliation consisted of up to three bands: an adoral band composed of oblique kineties; a vestibular band of longitudinal kineties; and a preoral band of longitudinal kineties. Somatic ciliation occurred in two longitudinal bands: a dense band composed of several parallel kineties on the left side of the dorso-ventral groove; and a sparse band composed of a single kinety on the right internal side of the dorso-ventral groove. Few structures were homologous to those of other litostome ciliates, and thus the relationship of Macropodinium to other litostomes cannot yet be clearly defined.

A novel variant genotype C of hepatitis B virus identified in isolates from Australian Aborigines: complete genome sequence and phylogenetic relatedness

There have been no reports of DNA sequences of hepatitis B virus (HBV) strains from Australian Aborigines, although the hepatitis B surface antigen (HBsAg) was discovered among them. To investigate the characteristics of DNA sequences of HBV strains from Australian Aborigines, the complete nucleotide sequences of HBV strains were determined and subjected to molecular evolutionary analysis. Serum samples positive for HBsAg were collected from five Australian Aborigines. Phylogenetic analysis of the five complete nucleotide sequences compared with DNA sequences of 54 global HBV isolates from international databases revealed that three of the five were classified into genotype D and were most closely related in terms of evolutionary distance to a strain isolated from a healthy blood donor in Papua New Guinea. Two of the five were classified into a novel variant genotype C, which has not been reported previously, and were closely related to a strain isolated from Polynesians, particularly in the X and Core genes. These two strains of variant genotype C differed from known genotype C strains by 5.9-7.4% over the complete nucleotide sequence and 4.0-5.6 % in the small-S gene, and had residues Arg(122), Thr(127) and Lys(160) characteristic of serotype ayw3, which have not been reported previously in genotype C. In conclusion, this is the first report of the characteristics of complete nucleotide sequences of HBV from Australian Aborigines. These results contribute to the investigation of the worldwide spread of HBV, the relationship between serotype and genotype and the ancient common origin of Australian Aborigines.

The upper Paleozoic miospore genus Spelaeotriletes Neves and Owens, 1966, and constituent Gondwanan species

The upper Paleozoic miospore genus Spelaeotriletes Neves and Owens, 1966 is reviewed as a morpho-taxonomic entity and vis-a-vis other similarly constructed (pseudosaccate) genera - Geminospora Balme, 1962, Grandispora Hoffmeister, Staplin, and Malloy, 1955, Rhabdosporites Richardson, 1960, and Retispora Staplin, 1960. Detailed studies of numerous, mainly topotype specimens of Spelaeotriletes ybertii (Marques-Toigo, 1970) Playford and Powis, 1979 from the Lower Permian of Uruguay result in its re-diagnosis, in conjunction with a survey of its exclusively Gondwanan occurrences, particularly in South American strata extending from the Upper Carboniferous (Westphalian) into the Lower Permian, and also in Australian strata of approximately equivalent age. The characteristics of other species of Spelaeotriletes reported from upper Paleozoic deposits of Gondwana are discussed, as are their temporal representations in various broad regions of the supercontinent (South America, North Africa, Australia). These species include two, perhaps three, that, like Spelaeotriletes triangulus/arenaceus, are known also from Euramerica - S. balteatus (Playford, 1963) Higgs, 1996, S. pretiosus (Playford, 1964) Utting, 1987, and possibly S. owensii Loboziak and Alpern, 1978. Other species, such as S. benghaziensis Loboziak and Clayton, 1988, S. giganteus Loboziak and Clayton, 1988, and S. vibrissus Playford and Satterthwait, 1988, have, on present knowledge, exclusively Gondwanan occurrences. S. queenslandensis Jones and Truswell. 1992, known only from Upper Carboniferous strata of northeastern Australia, is formally reassigned on sculptural grounds to Grandispora. Not unexpectedly in a paleogeographic perspective, North Africa and South America are more closely allied with each other than with Australia in terms of shared species of Spelaeotriletes. (C) 2001 Elsevier Science Ltd. All rights reserved.

Experimental susceptibility of some reef fish species to Benedenia lutjani (Monogenea : Capsalidae), a parasite of Lutjanus carponotatus (Pisces : Lutjanidae)

The susceptibility of species of lutjanid, lethrinid and serranid fish to infection by either larval or post-larval (juvenile and adult) specimens of the capsalid monogenean Benedenia lutjani Whittington and Kearn (1993) was examined experimentally. Four species of lutjanids became infected when exposed to larvae of B. lutjani, but three species of lethrinids and four species of serranids were not susceptible to larvae under the same conditions. Variability in the intensity of infection by larvae occurred within and between lutjanid species. Few post-larval specimens of B. lutjani transferred between individuals of the specific host Lutjanus carponotatus (Richardson 1842) in 60-l aquaria and none transferred between specimens of L. carponotatus in a 7,500-l concrete tank. These results indicate that transfer of post-larval B. lutjani between individuals of the specific host is unlikely to occur in the wild. Other lutjanid species did not become infected when exposed to specimens of L. carponotatus infected heavily by post-larval B. lutjani, but two lethrinid species were susceptible to infection under the same conditions. These data indicate that different factors may mediate host-specificity for larval and post-larval B. lutjani.

A new genus of viviparous gyrodactylid (Monogenea) from the Great Barrier Reef, Australia with descriptions of seven new species

Acanthoplacatus gen. nov., a new genus of viviparous gyrodactylid, is described from the rns and skin of siganid fishes from the Great Barrier Reef, Australia. The genus is characterized by a muscular, tube-like haptor with 16 marginal hooks on the posterior margin. The ventral lobe of the haptor is located anteriorly relative to the dorsal lobe and contains a pair of hamuli and a ventral bar with posteriorly-projecting ventral bar membrane. A dorsal bar is absent. Five pairs of posterior gland cells surround the posterior terminations of the gut. The male copulatory organ is a muscular, non-eversible bulb with several spines around the distal opening. Species of Acanthoplacatus have a bilateral excretory system consisting of six pairs of flame cells and a pair of excretory bladders. Seven new species are described: Acanthoplacatus adlardi sp. nov. and A. amplihamus sp. nov. from Siganus punctatus (Forster, 1801), A. brauni sp. nov. from S. corallinus (Valenciennes, 1835), A. parvihamus sp. nov. from S. vulpinus (Schlegel and Mueller, 1845), A. puelli sp. nov. from S. puellus Schlegel, 1852, A. shieldsi sp. nov. from S. lineatus (Valenciennes, 1835) and A. sigani sp. nov. from S. fuscescens (Houttuyn, 1782). Species can be discriminated by shape and size of the hamuli, marginal hooks and ventral bar and by male copulatory organ sclerite morphology. Three species (A. brauni sp. nov., A. shieldsi sp. nov. and A. sigani sp. nov.) were assessed for seasonal variation of sclerite size. Ten of thirteen morphological characters showed seasonal variation in size for at least one of the species. The characters were longer in winter except dorsal root tissue cap width. Only one character, marginal hook length, showed significant seasonal variation for all three species. Species of Acanthoplacatus were observed to attach using only the marginal hooks and the role of hamuli in attachment is unclear. The dorsal rn of the host is the preferred site for most species but the anal fin, caudal fin and body surfaces are preferred by some species. Prevalences for species range from 57 to 100%.

Species and regional variations in the effectiveness of antivenom against the in Vitro neurotoxicity of death adder (Acanthophis) venoms

Although viperlike in appearance and habit, death adders belong to the Elapidae family of snakes. Systemic envenomation represents a serious medical problem with antivenom, which is raised against Acanthophis antarcticus venom, representing the primary treatment. This study focused on the major Acanthophis variants from Australia and islands in the Indo-Pacific region. Venoms were profiled using liquid chromatography-mass spectrometry, and analyzed for in vitro neurotoxicity (0.3-10 mug/ml), as well as the effectiveness of antivenom. (1-5 units/ml; 10 min prior to the addition of 10 mug/ml venom). The following death adder venoms were examined: A. antarcticus (from separate populations in New South Wales, Queensland, South Australia, and Western Australia), A. hawkei, A. praelongus, A. pyrrhus, A. rugosus, A. wellsi, and venom from an unnamed species from the Indonesian island of Seram. All venoms abolished indirect twitches of the chick isolated biventer cervicis nerve-muscle preparation in a dose-dependent manner. In addition, all venoms blocked responses to exogenous acetylcholine (1 m-M) and carbachol (20 muM), but not KCl (40 mM), suggesting postsynaptic neurotoxicity. Death adder antivenom (1 unit/ml) prevented the neurotoxic effects of A. pyrrhus, A. praelongus, and A. hawkei venoms, although it was markedly less effective against venoms from A. antarcticus (NSW, SA, WA), A. rugosus, A. wellsi, and A. sp. Scram. However, at 5 units/ml, antivenom was effective against all venoms tested. Death adder venoms, including those from A. antarcticus geographic variants, differed not only in their venom composition but also in their neurotoxic activity and susceptibility to antivenom. For the first time toxicological aspects of A. hawkei, A. wellsi, A. rugosus, and A. sp. Seram venoms were studied. (C) 2001 Academic Press.

Incubation temperature, energy expenditure and hatchling size in the green turtle (Chelonia mydas), a species with temperature-sensitive sex determination

Eggs from the Heron Island, Great Barrier Reef, nesting population of green turtles (Chelonia mydas) were incubated at all-male-determining (26 degreesC) and all-female-determining (30 degreesC) temperatures. Oxygen consumption and embryonic growth were monitored throughout incubation, and hatchling masses and body dimensions were measured from both temperatures. Eggs hatched after 79 and 53 days incubation at 26 degreesC and 30 degreesC respectively. Oxygen consumption at both temperatures increased to a peak several days before hatching, a pattern typical of turtle embryos, and the rate of oxygen was higher at 30 degreesC than 26 degreesC. The total amount of energy consumed during incubation, and hatchling dimensions, were similar at both temperatures, but hatchlings from 26 degreesC had larger mass, larger yolk-free mass and smaller residual yolks than hatchlings from 30 degreesC. Because of the difference in mass of hatchlings, hatchlings from 30 degreesC had a higher production cost.

A critical comparison of the external and internal boron requirements for contrasting species in boron-buffered solution culture

Despite reports that boron (B) requirements differ among plant species there is a shortage of critical evidence to demonstrate unequivocally whether species differ in internal or external B requirements or both. The present research was conducted to establish the external and internal B requirements of three contrasting species, a woody dicot (marri), an herbaceous dicot (sunflower) and a monocot (wheat) using B-buffered solution culture. Boron-buffered solution culture provided satisfactory control of external B concentrations ranging from 0.04 to 30 muM throughout the 20- (sunflower and wheat) or 40-day (marri) growth period. At low external B concentrations (less than or equal to 0.13 muM), the growth of marri and sunflower was severely depressed but by contrast the vegetative growth of wheat plants was satisfactory and free of B deficiency symptoms. Marri and sunflower plants achieved total maximum shoot growth at greater than or equal to1.2 muM B in solutions while wheat plants did so at greater than or equal to 0.6 muM B. The critical B concentrations (mg kg(-1) dry matter) in the youngest open leaf blades of marri, sunflower and wheat plants were 17.9, 19.7 and 1.2 on 20, 10 and 10 days after transplanting (DAT), respectively. Lower internal and external B requirements of wheat were matched by a lower uptake rate of B compared to marri and sunflower.

Adaptation trials of Atriplex and Maireana species and their response to saline waterlogged conditions in Pakistan

Utilization of salt affected wasteland by growing forage shrubs has enormous economic and environmental implication for developing countries like Pakistan, where approximately 6.3 million ha of the land is salt affected. Considering the importance of Atriplex and Maireana species, research has been conducted using their different species on the salt affected soils of Faisalabad. Most of Atriplex and Maireana species survived under the environmental conditions of Faisalabad and gave the good yield in the form of forage. Some of these species are woody and can be used for fuel purposes. Sixteen genotypes of Atriplex and Maireana were tested for their tolerance to waterlogging in order to identify halophytic fodder shrubs suitable for growth on secondary salt-affected and waterlogged farmland. The physiological and morphological responses of the species tested were typical of species with a generally poor tolerance to waterlogging. Despite this, some species (eg A. Amnicola) were surprisingly resistant, surviving up to five months of waterlogging at moderate salinity and high evapotranspirational demand. The most resistant species, A amnicola maintained higher transpiration rates, leaf water potentials and shoot extension rates than most other species during five weeks of waterlogging, and a return to control levels more quickly than other species after plots were drained. Although little morphological adaptation to waterlogged conditions was detected, a shallow and extensive lateral root system and the formation of many short aerenchymatous adventitious roots from procumbent branches appeared to advantage A. Amnicola in an environment highly heterogeneous in salinity and low in oxygen concentration. Shallow fibrous rooted species were quickly killed by waterlogging, although the procumbent branches of some individuals survived as clones if they developed adventitious roots.

Mitochondrial gene content, arrangement and composition compared in African and Asian schistosomes

Complete sequences were obtained for the coding portions of the mitochondrial (mt) genomes of Schistosoma mansoni (NMRI strain, Puerto Rico; 14415 bp), S. japonicum (Anhui strain, China; 14085 bp) and S. mekongi (Khong Island, Laos; 14072 bp). Each comprises 36 genes: 12 protein-encoding genes (cox1-3, nad1-6, nad4L, atp6 and cob); two ribosomal RNAs, rrnL (large subunit rRNA or 16S) and rrnS (small subunit rRNA or 12S); as well as 22 transfer RNA (tRNA) genes. The atp8 gene is absent. A large segment (9.6 kb) of the coding region (comprising 14 tRNAs, eight complete and two incomplete protein-encoding genes) for S. malayensis (Baling, Malaysian Peninsula) was also obtained. Each genome also possesses a long non-coding region that is divided into two parts (a small and a large non-coding region, the latter not fully sequenced in any species) by one or more tRNAs. The protein-encoding genes are similar in size, composition and codon usage in all species except for cox1 in S. mansoni (609 aa) and cox2 in S. mekongi (219 an), both of which are longer than homologues in other species. An unexpected finding in all the Schistosoma species was the presence of a leucine zipper motif in the nad4L gene. The gene order in S. mansoni is strikingly different from that seen in the S. japonicum group and other flatworms. There is a high level of identity (87-94% at both the nucleotide and amino acid levels) for all protein-encoding genes of S. mekongi and S. malayensis. The identity between genes of these two species and those of S. japonicum is less (56-83% for amino acids and 73-79 for nucleotides). The identity between the genes of S. mansoni and the Asian schistosomes is far less (33-66% for amino acids and 54-68% for nucleotides), an observation consistent with the known phylogenetic distance between S. mansoni and the other species. (C) 2001 Elsevier Science B.V. All rights reserved.