Introduced and cryptogenic species in Port Phillip Bay, Victoria, Australia

Port Phillip Bay (PPB) is a large (1,930 km²), temperate embayment in southern Victoria, Australia. Extensive bay-wide surveys of PPB have occurred since 1840. In 1995/1996 the Commonwealth Scientific and Industrial Research Organization (CSIRO) Centre for Research on Introduced Marine Pests (CRIMP) undertook an intensive evaluation of the region with the aims of developing a comprehensive species list of native and introduced biota and contrasting previous bay-wide assessments with a current field survey in order to detect new incursions and discern alterations to native communities. Two methods were used to meet these aims: a re-evaluation of regional museum collections and published research in PPB to identify and determine the timing of introductions; and field surveys for benthic (infauna, epifauna and encrusting) organisms between September 1995 to March 1996. One hundred and sixty introduced (99) and cryptogenic (61) species were identified representing over 13% of the recorded species of PPB. As expected, the majority of these are concentrated around the shipping ports of Geelong and Melbourne. Invasions within PPB appear to be increasing, possibly due to an increase in modern shipping traffic and an increase in aquaculture (historically associated with incidental introductions); however the records of extensive biological surveys suggest that this may, in part, be an artefact of sampling effort. In contrast to Northern Hemisphere studies, PPB (and Southern Hemisphere introductions in general) have significantly different suites of successfully invading taxa. PPB is presented as one of the most invaded marine ecosystems in the Southern Hemisphere.Communicated by M.S. Johnson, Crawley     

Using fuzzy set theory to address the uncertainty of susceptibility to drought

This paper presents the technical aspects of a new methodology for assessing the susceptibility of society to drought. The methodology consists of a combination of inference modelling and fuzzy logic applications. Four steps are followed: (1) model input variables are selected—these variables reflect the main factors influencing susceptibility in a social group, population or region, (2) fuzzification—the uncertainties of the input variables are made explicit by representing them as ‘fuzzy membership functions’, (3) inference modelling—the input variables are used to construct a model made up of linguistic rules, and (4) defuzzification—results from the model in linguistic form are translated into numerical form, also through the use of fuzzy membership functions. The disadvantages and advantages of this methodology became apparent when it was applied to the assessment of susceptibility from three disciplinary perspectives: Disadvantages include the difficulty in validating results and the subjectivity involved with specifying fuzzy membership functions and the rules of the inference model. Advantages of the methodology are its transparency, because all model assumptions have to be made explicit in the form of inference rules; its flexibility, in that informal and expert knowledge can be incorporated through ‘fuzzy membership functions’ and through the rules in the inference model; and its versatility, since numerical data can be converted to linguistic statements and vice versa through the procedures of ‘fuzzification’ and ‘defuzzification’.     

Molecular phylogeny and population structure of tideland snails in the genus Cerithidea around Japan

Phylogenetic relationships and genetic population structures were analyzed for tideland gastropods in the genus Cerithidea around Japan on the basis of partial sequence of the mitochondrial COI gene. Large genetic divergence was shown between individuals of Cerithidea cingulata in the southern Ryukyus and those in the central Ryukyus and the Japanese Islands. Haplotypes of C. cingulata from the Japanese Islands were paraphyletic with the exclusion of a monophyletic group from the central Ryukyus. Genetic differentiation of C. cingulata was also detected between Amami-Oshima Island and Okinawajima Island. No genetic divergence was found between Cerithidea rhizophorarum in the Japanese Islands and its subspecies C. rhizophorarum morchii in the Ryukyu Islands. The lack of genetic divergence of Cerithidea largillierti between continental China and Japan suggests relatively recent migration between the Japanese Islands and the Asian continent. For all three Cerithidea species distributed in both the Japanese Islands and the Ryukyu Islands, the Tokara Gap and the Kerama Gap were shown to have acted as barriers to the dispersal.     

Molecular taxonomy of vestimentiferans of the western Pacific and their phylogenetic relationship to species of the eastern Pacific

The nucleotide sequence of part (624 bp) of a mitochondrial gene for cytochrome oxidase I was determined for 46 escarpiid vestimentiferans collected from seven sites in the western Pacific and 49 individual specimens of Arcovestia ivanovi from two sites in the Manus Basin. Phylogenetic analysis, based on the newly obtained and previously reported sequences, indicated that escarpiids in the western Pacific can be divided into two tentative species, as we proposed in a previous report. While members of the first tentative species have been collected exclusively from a seep area at a depth of 300 m off the coast of central Japan, the members of the second species inhabit some sites at depths greater than 1,100 m, namely, seep areas in Japanese and Papua-New Guinean waters as well as hydrothermal vent fields in the Okinawa Trough and the Manus Basin. We detected no genetic structure among populations of the second tentative species. The first tentative species was more closely related to a species in the eastern Pacific, Escarpia spicata, and to a species in the Gulf of Mexico, Escarpia laminata, than to the second tentative species in the western Pacific. Sequences obtained from all arcovestiids were identical with the exception of those from three individuals, each of which included a single synonymous nucleotide substitution relative to the dominant haplotype, and no genetic differences were detected between specimens from the two sites in the Manus Basin.     

A comparison of different lysis buffers to assess allele dropout from single cells for preimplantation genetic diagnosis

Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999 ). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (ΔF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. Copyright © 2001 John Wiley & Sons, Ltd.