The development of coalescent-based and other multilocus methods for species delimitation has facilitated the identification of cryptic species complexes across the tree of life. A recent taxonomic revision of the ecologically important soft coral genus Ovabunda validated 11morphospecies, all with type localities and overlapping geographic ranges in the Red Sea. A subsequent molecular phylogenetic analysis using mitochondrial and 28S nrDNA genes divided the genus into just two clades, with no apparent genetic distinctions among morphospecies. To further explore species boundaries among morphospecies of Ovabunda we sequenced three additional nuclear genes (ITS, ATPSα, ATPSβ), and obtained data for 1332 unlinked SNPs from restriction-site associated DNA sequencing. Both coalescent-based and allele-sharing species delimitation analyses supported four species of Ovabunda, each of which included multiple morphotypes encompassing the full range of morphological variation observed within the genus. All four species occurred over the same depth range of 5-41m, and were sympatric at sites separated by 1100km in the Red Sea. The only characters that have been found to distinguish three of the four species are diagnostic substitutions in the nuclear genome; the fourth differs by exhibiting polyp pulsation, a behavioral trait that can be assessed only in live colonies. The lack of any obvious morphological, life history, ecological or geographical differences among these four species begs the question of what drove the evolution and maintenance of reproductive isolating mechanisms in this cryptic species complex.

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In Australia, disease caused by betanodavirus has been reported in an increasing number of cultured finfish since the first report of mortalities in 1990. Partial coat protein gene sequences from the T2 or T4 regions of 8 betanodaviruses from barramundi Lates calcarifer, sleepy cod Oxyeleotris lineolata, striped trumpeter Latris lineata, barramundi cod Cromileptes altivelis, Australian bass Macquaria novemaculata and gold-spotted rockcod Epinephelus coioides from several Australian states were determined. Analysis of the 606 bp nucleotide sequences of the T2 region of 4 isolates demonstrated the close relationship with isolates from the red-spotted grouper nervous necrosis virus (RGNNV) genotype and the Cluster Ia subtype. Comparison of a smaller 289 bp sequence from the T4 region identified 2 distinct groupings of the Australian isolates within the RGNNV genotype. Isolates from barramundi from the Northern Territory, barramundi, sleepy cod, barramundi cod and gold-spotted rockcod from Queensland, and striped trumpeter from Tasmania shared a 96.2 to 99.7% nucleotide identity with each other. These isolates were most similar to the RGNNV genotype Cluster Ia. Isolates from Australian bass from New South Wales and from barramundi from South Australia shared a 98.6% sequence identity with each other. However, these isolates only shared an 85.8 to 87.9% identity with the other Australian isolates and representative RGNNV isolates. The closest nucleotide identity to sequences reported in the literature for the New South Wales and South Australian isolates was to an Australian barramundi isolate (Ba94Aus) from 1994. These 2 Australian isolates formed a new subtype within the RGNNV genotype, which is designated as Cluster Ic. KEY WORDS: Betanodavirus · Phylogeny · Nervous necrosis virus · Coat protein · Nodavirus · Molecular detection Resale or republication not permitted without written consent of the publisherDis Aquat Org 87: [151][152][153][154][155][156][157][158][159][160] 2009 like viral particles were first reported to be associated with degenerative areas of the brain and retina of larvae which had exhibited abnormal swimming behaviour (Glazebrook et al. 1990, Munday et al. 1992. The close antigenic relationship of the picorna-like virus in diseased barramundi to the betanodavirus of striped jack nervous necrosis virus (SJNNV) was reported after specific fluorescence was observed in barramundi tissue using anti-SJNNV rabbit serum in an indirect fluorescence antibody test (Munday et al. 1994). Amplification and sequencing of the T4 region of the coat protein gene of an isolate from barramundi from Australia (Ba94Aus) confirmed betanodavirus as the causative agent (Nishizawa et al. 1997). Virions are small (25 to 30 nm), non-enveloped, icosahedral in shape, and contain single-stranded, bipartite positive sense RNA. The larger RNA segment (RNA1, 3.1 kb) encodes a nonstructural protein of ~110 kDa, while the smaller RNA segment (RNA2, 1.4 kb) encodes the 42 kDa coat protein (Mori et al. 1992, Murphy et...

The evaluation of measurement uncertainties has been widely applied to the calibration of measurement instruments, whereas its application to tests, despite increasing requirements, is a more recent phenomenon. The generalization of the evaluation of measurement uncertainties to tests has been a gradual process, in line with changes in the requirements of the normative framework that regulates the accreditation of tests laboratories and also as the perceived good practices have evolved. The sole identification of the relevant sources of uncertainty was followed by the requirement to provide a simplified estimate of the measurement uncertainty, and it is now an accepted requirement to properly evaluate the expanded measurement uncertainty associated with any tests. In this study, the evaluation of measurement uncertainty associated with the determination of sulfate in water will be attempted using a procedure that includes linear regression, with the regression parameters provided with associated uncertainties, and a Monte Carlo method applied as a validation tool of the conventional mainstream evaluation method, concerning the approximations in terms of linearization of the model and the assumed shape of the output distribution introduced by this approach.