Prevalence of the scuticociliate Orchitophrya stellarum in seastars from the Pacific and Atlantic oceans.

As part of a study to investigate the use of the scuticociliate Orchitophrya stellarum as a biological control for the invasive seastar Asterias amurensis in Australia, we collected prevalence data for O. stellarum from 3 seastar species (A. amurensis, A. rubens, Pisaster ochraceus) between 1996 and 1999 from the Pacific (Australia, Japan, Korea, Canada) and Atlantic (France, Netherlands, Canada) oceans. In the Pacific Ocean, for the first time, we found O. stellarum in male A. amurensis in Korea and female A. amurensis in Japan. The parasite was not detected in the invasive A. amurensis from Australia. There was no significant difference between size of infected and uninfected male seastars, nor a correlation between biased sex ratio and parasite prevalence in populations in the Pacific or Atlantic oceans. Therefore, unlike other studies, we found size and sex ratio in seastar populations in the field are unreliable indicators of parasite impacts. Regular monitoring of infected seastar populations in the field would be useful to better understand how sex ratio varies with parasite prevalence. We recommend laboratory studies under controlled conditions to determine the effect of O. stellarum on seastar populations.

Incorporating social dimensions in planning, managing and evaluating environmental projects.

Most conservation research aims to inform management of environmental challenges, but scientific evidence is used inconsistently in environmental programmes and practice. We used semi-structured retrospective interviews to ask 12 environmental scientists and 14 practitioners (land managers, park rangers, project managers and planners from natural resource management agencies) about factors that facilitated and hindered the use of scientific input during 15 environmental projects. We used the common factors from interviews to develop a process model describing how scientific input informs programmes and practice. The model emphasised the social dimensions of environmental projects which are often overlooked when these projects are planned, managed and evaluated. It highlighted the pivotal role of relationships in achieving outcomes which include creating practical, useful products and tools, and robust, credible and trusted evidence. By clarifying the process of how scientific knowledge informs environmental programmes and practice, the model enabled us to provide guidance about how to undertake transdisciplinary work and suggest indicators to track progress. Although derived from environmental projects, the guidance is likely to apply to other fields, particularly where different disciplines work together.

Conservation of sequence in the internal transcribed spacers and 5.8S ribosomal RNA among geographically separated isolates of parasitic scuticociliates (Ciliophora, Orchitophryidae).

Nucleotide sequence from the internal transcribed spacers (ITS1 and ITS2) and the 5.8S gene from the ribosomal RNA gene cluster of isolates of the scuticociliate Orchitophrya stellarum from 4 asteroid hosts were compared. Surprisingly, these data (495 bp) were identical for O. stellarum isolated from the testes of Asterias amurensis from Japan; Pisaster ochraceus from British Columbia, Canada; Asterias rubens from The Netherlands; and Asterias vulgaris from Prince Edward Island, Canada. These sequence data were compared to those from 3 scuticociliates which parasitise crustaceans: Mesanophrys pugettensis, M. chesapeakensis and Anophryoides haemophila. No difference was found in this region between the nucleotide sequence of M. pugettensis and M. chesapeakensis. The sequence of Mesanophrys spp. differed by 9.2% in the ITS1 and 4.7% in the ITS2 from that of O. stellarum. The sequence from the ITS1 (135 bp) and ITS2 (233 bp) of A. haemophila differed by 42.6 and 20.5% respectively from those of O. stellarum. Therefore, nucleotide sequence of the ITS regions in these scuticociliates is highly conserved.

Identification of high-copy disruptors of telomeric silencing in Saccharomyces cerevisiae.

The ends of chromosomes in Saccharomyces cerevisiae initiate a repressive chromatin structure that spreads internally and inhibits the transcription of nearby genes, a phenomenon termed telomeric silencing. To investigate the molecular basis of this process, we carried out a genetic screen to identify genes whose overexpression disrupts telomeric silencing. We thus isolated 10 DOT genes (disruptor of telomeric silencing). Among these were genes encoding chromatin component Sir4p, DNA helicase Dna2p, ribosomal protein L32, and two proteins of unknown function, Asf1p and Ifh1p. The collection also included genes that had not previously been identified: DOT1, DOT4, DOT5, DOT6, and TLC1, which encodes the RNA template component of telomerase. With the exception of TLC1, all these genes, particularly DOT1 and DOT4, also reduced silencing at other repressed loci (HM loci and rDNA) when overexpressed. Moreover, deletion of the latter two genes weakened silencing as well, suggesting that DOT1 and DOT4 normally play important roles in gene repression. DOT1 deletion also affected telomere tract length. The function of Dot1p is not known. The sequence of Dot4p suggests that it is a ubiquitin-processing protease. Taken together, the DOT genes include both components and regulators of silent chromatin.

Chytridiomycosis causes amphibian mortality associated with population declines in the rain forests of Australia and Central America.

Epidermal changes caused by a chytridiomycete fungus (Chytridiomycota; Chytridiales) were found in sick and dead adult anurans collected from montane rain forests in Queensland (Australia) and Panama during mass mortality events associated with significant population declines. We also have found this new disease associated with morbidity and mortality in wild and captive anurans from additional locations in Australia and Central America. This is the first report of parasitism of a vertebrate by a member of the phylum Chytridiomycota. Experimental data support the conclusion that cutaneous chytridiomycosis is a fatal disease of anurans, and we hypothesize that it is the proximate cause of these recent amphibian declines.

The ciliate Orchitophrya cf. stellarum and other parasites and commensals of the northern Pacific seastar Asterias amurensis from Japan.

To identify the pathogens and possible biological control agents for the introduced seastar Asterias amurensis, we examined seastars from source populations in central and northern Japan. In particular, we sought the scuticociliate Orchitophrya cf. stellarum. The ciliate was found in male A. amurensis from five sites. We also found the caprellid amphipod Caprella astericola on A. amurensis and Distolasterias nipon from Nemuro Bay. The copepod Scottomyzon gibberum was found on A. amurensis from Usujiri and polychaete scaleworms Arctonoe vittata were found on A. amurensis from Murohama and Nemuro Bay. Of these parasites and commensals, Orchitophrya cf. stellarum is the most likely agent for biological control of A. amurensis in Australian waters; however, its ability to regulate seastar populations is uncertain.

A novel clade of protistan parasites near the animal-fungal divergence.

Sequences of nuclear-encoded small-subunit rRNA genes have been determined for representatives of the enigmatic genera Dermocystidium, Ichthyophonus, and Psorospermium, protistan parasites of fish and crustaceans. The small-subunit rRNA genes from these parasites and from the "rosette agent" (also a parasite of fish) together form a novel, statistically supported clade. Phylogenetic analyses demonstrate this clade to diverge near the animal-fungal dichotomy, although more precise resolution is problematic. In the most parsimonious and maximally likely phylogenetic frameworks inferred from the most stably aligned sequence regions, the clade constitutes the most basal branch of the metazoa; but within a limited range of model parameters, and in some analyses that incorporate less well-aligned sequence regions, an alternative topology in which it diverges immediately before the animal-fungal dichotomy was recovered. Mitochondrial cristae of Dermocystidium spp. are flat, whereas those of Ichthyophonus hoferi appear tubulovesiculate. These results extend our understanding of the types of organisms from which metazoa and fungi may have evolved.

Use of molecular data to discriminate pseudocerotid turbellarians.

Nucleotide sequence data from the Internal Transcribed Spacer 1 (ITS1) in the ribosomal RNA (rRNA) gene cluster were used to determine the utility of molecular data to discriminate species and genera of pseudocerotid turbellarians. We sequenced 388, 379 and 415 bp from the ITS1 of Pseudoceros jebborum, Pseudoceros paralaticlavus and Pseudobiceros gratus respectively, to give an aligned sequence for this region of 421 positions. The nucleotide sequence of the ITS1 of Pseudoceros jebborum differed from that of Pseudoceros paralaticlavus by 6% (24/421 positions) and from that of Pseudobiceros gratus by 36% (152/421 positions). These results indicate that sequence data from the ITS1 will be a useful taxonomic tool to discriminate pseudocerotid flatworms.

Phylogenetic position of the genus Perkinsus (Protista, Apicomplexa) based on small subunit ribosomal RNA.

Parasites of the genus Perkinsus destroy marine molluscs worldwide. Their phylogenetic position within the kingdom Protista is controversial. Nucleotide sequence data (1792 bp) from the small subunit rRNA gene of Perkinsus sp. from Anadara trapezia (Mollusca: Bivalvia) from Moreton Bay, Queensland, was used to examine the phylogenetic affinities of this enigmatic genus. These data were aligned with nucleotide sequences from 6 apicomplexans, 3 ciliates, 3 flagellates, a dinoflagellate, 3 fungi, maize and human. Phylogenetic trees were constructed after analysis with maximum parsimony and distance matrix methods. Our analyses indicate that Perkinsus is phylogenetically closer to dinoflagellates and to coccidean and piroplasm apicomplexans than to fungi or flagellates.

Use of stresscath for diagnosing stress incontinence.

The modified fluid bridge (flow) test (MFBT) is a simple procedure for detecting bladder neck incompetency using a fluid-filled catheter. The Stresscath (Hollister) female diagnostic catheter is an improved version of the MFBT. We evaluated the ability of the Stresscath to detect bladder neck incompetence in the first 66 female patients referred for urodynamics. The absence or presence of stress incontinence was based on the results from history and physical examinations. The results from the Stresscath correlated with the history showed that the Stresscath had a sensitivity of 0.73 and a specificity of 0.88 in determining stress incontinence due to bladder neck incompetency. The results from the Stresscath correlated with the physical examination (Marshall test) showed that the Stresscath had a sensitivity of 0.86 and a specificity of 0.87 in determining stress incontinence due to bladder neck incompetence. The Stresscath was at least as accurate as the urethral pressure profile in the diagnosis of stress incontinence, at less capital cost.

Diazepam-stress interactions in the rat: effects on autoanalgesia and a plus-maze model of anxiety.

On six occasions spaced at least a week apart, two groups of rats were subjected to a variety of stressful conditions consisting of a restraint/bright light complex, either alone or in combination with a tail pinch, whole-body inversion, or partial immersion in cold water. One of these groups was injected with diazepam (2.0 mg/kg) 30 min prior to the stressors, while the other group experienced the drug in their home cages the following day. A third group also received the diazepam but was not exposed to the stressors. In three test sessions all animals were injected with either diazepam or saline and were then exposed to a novel stressor: a plus-maze used as a screening device for anxiolytic drugs. This was immediately followed by a tail-flick measure of analgesia. The longest tail-flick latencies, indicating stress-induced analgesia ("autoanalgesia"), were observed in the group that had not been exposed to stress prior to testing. The other two groups exhibited substantially shorter latencies but did not differ from one another, thus showing a "stress inoculation" effect that was uninfluenced by diazepam. In the plus-maze, diazepam tends to increase the amount of time rats will spend in the two exposed arms of the maze relative to the two enclosed arms. This effect was significantly attenuated in the group that had previously experienced the variety of stressors after a diazepam injection, suggesting a learned association between drug and stress that resulted in a diminution of the drug's anxiolytic property.