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DNA barcoding based on a fragment of the cytochrome c oxidase subunit I (COI) gene from the mitochondrial genome is widely applied in species identification, species discovery and biodiversity stud...

The development of new genetic resources is key for biological conservation in an era when the preservation of biodiversity is crucial. Here, we report the development of microsatellites and mitochondrial markers for the redbait species Pyura herdmani and P. stolonifera, both important bioengineers along African coastlines. Specifically, we identified microsatellites by means of pyrosequencing and variable regions in the mitochondrial genome using RAD-seq libraries that were generated with Illumina sequencing. A total of 4 339 putative microsatellites were found, of which 32 were tested on individuals collected along the southeast coast of South Africa. Of those, five microsatellites cross-amplified consistently in both species, and these were tested on a total of 260 samples. Additionally, we mapped the partial mitochondrial genome of a small number of individuals from both species and identified the most-variable regions by comparing a small number of individuals from all regions where these species have been recorded (including western South Africa and northwest Africa). The genetic markers developed here can be used to conduct population genetic studies, to monitor range shifts under contemporary climate change, and to help in preserving ecosystem functioning along African coastlines.

Several studies have attempted to understand the origin and evolution of single‐exon genes (SEGs) in eukaryotic organisms, including fishes, but few have examined the functional and evolutionary relationships between SEGs and multiple‐exon gene (MEG) paralogs, in particular the conservation of promoter regions. Given that SEGs originate via the reverse transcription of mRNA from a “parental” MEGs, such comparisons may enable identifying evolutionarily‐related SEG/MEG paralogs, which might fulfill equivalent physiological functions. Here, the relationship of SEG proportion with MEG count, gene density, intron count, and chromosome size was assessed for the genome of the European sea bass, Dicentrarchus labrax. Then, SEGs with an MEG parent were identified, and promoter sequences of SEG/MEG paralogs were compared, to identify highly conserved functional motifs. The results revealed a total count of 1,585 (8.3% of total genes) SEGs in the European sea bass genome, which was correlated with MEG count but not with gene density. The significant correlation of SEG content with the number of MEGs suggests that SEGs were continuously and independently generated over evolutionary time following species divergence through retrotranscription events, followed by tandem duplications. Functional annotation showed that the majority of SEGs are functional, as is evident from their expression in RNA‐seq data used to support homology‐based genome annotation. Differences in 5′UTR and 3′UTR lengths between SEG/MEG paralogs observed in this study may contribute to gene expression divergence between them and therefore lead to the emergence of new SEG functions. The comparison of nonsynonymous to synonymous changes (Ka/Ks) between SEG/MEG parents showed that 74 of them are under positive selection (Ka/Ks > 1; p = .0447). An additional fifteen SEGs with an MEG parent have a common promoter, which implies that they are under the influence of common regulatory networks., This study investigated the relationship of SEG proportion with MEG count, gene density, intron count, and chromosome size for the genome of sea bass, Dicentrarchus labrax. Then, SEGs with an MEG parent were identified, and promoter sequences of SEG/MEG orthologs were compared, to identify highly conserved functional motifs. The results revealed a significant correlation between SEG and MEG counts over the genome and allowed identifying SEG/MEG orthologs that share the same promoter sequence, suggesting that they are under the influence of common regulatory networks.

Knowledge about the demographic histories of natural populations helps to evaluate their conservation status, and potential impacts of natural and anthropogenic pressures. In particular, estimates of effective population size obtained through molecular data can provide useful information to guide management decisions for vulnerable populations. The spotted ragged-tooth shark Carcharias taurus (also known as the sandtiger or grey nurse shark) is widely distributed in warm-temperate and subtropical waters, but has suffered severe population declines across much of its range as a result of overexploitation. Here, we used multilocus genotype data to investigate the demographic history of the South African C. taurus population. Using approximate Bayesian computation and likelihood-based importance sampling, it was found that the population underwent a historical range expansion that may have been linked to climatic changes during the late Pleistocene. There was no evidence for a recent anthropogenic decline. Together with census data suggesting a stable population, these results support the idea that fishing pressure and other threats have so far not been detrimental to the local C. Taurus population. The results reported here indicate that South Africa could possibly harbour the last remaining, relatively pristine population of this widespread but vulnerable top predator.

Seahorses and pipefishes are heavily exploited for use in Traditional Chinese Medicine (TCM), and less frequently for curio markets or as aquarium fish. A number of recent studies have used DNA barcoding to identify species sold at TCM markets in East Asia, but the usefulness of this approach in determining the region of origin remains poorly explored. Here, we generated DNA barcodes of dried seahorses and pipefishes destined for TCM that were confiscated at South Africa’s largest airport because they lacked the export permits required for the CITES-listed seahorses. These were compared with published sequences and new sequences generated for Mozambican seahorses, with the aim of determining whether it is possible to identify their country of origin. All pipefishes were identified as Syngnathoides biaculeatus, a widespread Indo-Pacific species, but the published sequence data did not provide sufficient resolution to identify the region of origin. The same was true of the majority of seahorses, which could not even be identified to species level because they clustered among an unresolved species complex whose sequences were published under the names Hippocampus kuda, H. fuscus and H. capensis. The presence of a few specimens of a second seahorse, H. camelopardalis, suggests that the shipment originated from East Africa because the range of this seahorse is centred around this region, but again, it was not possible to determine their country of origin. Even though seahorses and pipefishes have high levels of genetic population structure because of their low dispersal potential, DNA barcoding was only suitable to tentatively identify species, but not their region of origin. DNA barcoding is increasingly used to identify illegally traded wildlife, but our results show that more sophisticated methods are needed to monitor and police the trade in seahorses and pipefishes.

Spinicaudatan clam shrimp are a widespread and diverse group of branchiopod crustaceans, yet few mitochondrial genomes have been published for this taxonomic group. Here, we present the mitogenome ...

Large-scale monitoring of wild populations in remote areas using traditional live-capturing methods is logistically and financially challenging. Devices that can be used to obtain biological material remotely and store it for an extended period have considerable potential to monitor population densities and health status, but their applicability remains largely unexplored. The present study describes a device that collects trace amounts of DNA from the saliva of small mammals that is deposited on the surface of a collection medium (WaxTags®). The device’s performance was evaluated on Australian brushtail possums (Trichosurus vulpecula), an invasive pest species and the most significant vector of bovine tuberculosis infective agent (Mycobacterium bovis), under field conditions in Canterbury, New Zealand. The retrieved DNA was used to amplify eight possum-specific microsatellite markers and bacterial 16S rRNA. The design is mechanically robust, and the quality of the recovered DNA was adequate for microsatellite-based identification of individual possums, estimation of population density, and partial reconstruction of their oral microbiomes as a potential indicator of health. Several medically important bacteria, including strains of environmental Mycobacterium sp., were detected. The design can be refined to monitor other animals’ populations proactively and provide different levels of information necessary to manage wild populations.

Clam shrimps (Spinicaudata) are a widespread and diverse crustacean group that frequent temporary aquatic habitats, but few complete mitochondrial genomes have been published for this group. Here, ...

Fairy shrimps (Anostraca) constitute an important component of seasonally aquatic habitats, but few complete mitochondrial genomes have been published for this group. Here, we report the mitogenome of a common southern African species, Streptocephalus cafer, from Botswana (accession number: MN720104). Low-coverage shotgun sequencing recovered two contigs 15653 bp and 1347 bp in length that are separated by a repetitive region of unknown length within the non-coding control region. The mitogenome’s GC content is 31.80%. Phylogenetic analysis using protein-coding genes confirms the sister taxon relationship of S. cafer with the only other congener whose mitogenome has been reconstructed to date, the Asian S. sirindhornae.

During austral winter, the southern and eastern coastlines of South Africa witness one of the largest animal migrations on the planet, the KwaZulu-Natal sardine run. Hundreds of millions of temperate sardines, Sardinops sagax, form large shoals that migrate north-east towards the subtropical Indian Ocean. Recent studies have highlighted the role that genetic and environmental factors play in sardine run formation. In the present study, we used massively parallel sequencing to assemble and annotate the first reference transcriptome from the liver cells of South African sardines, and to investigate the functional content and transcriptomic diversity. A total of 1,310,530 transcripts with an N50 of 1578 bp were assembled de novo. Several genes and core biochemical pathways that modulate energy production, energy storage, digestion, secretory processes, immune responses, signaling, regulatory processes, and detoxification were identified. The functional content of the liver transcriptome from six individuals that participated in the 2019 sardine run demonstrated heterogeneous levels of variation. Data presented in the current study provide new insights into the complex function of the liver transcriptome in South African sardines.

Historical demographic events shape genetic diversity that remains evident in the genomes of contemporary populations. In the case of species that are of conservation concern, this information helps to unravel evolutionary histories that can be critical in guiding conservation efforts. The Knysna seahorse, Hippocampus capensis, is the world’s most endangered seahorse species, and it presently survives in only three estuaries on the South African south coast. Factors that contributed to the species becoming endangered are unclear; additionally, the lack of information on whether the three populations should be managed separately because of potential long-term isolation hampers effective management efforts. In the present study, we reconstructed the seahorses’ demographic history using a suite of microsatellite loci. We found that the largest population (Knysna Estuary) has colonised the other estuaries relatively recently (H. capensis depends primarily on the successful management of the Knysna population, although the other estuaries may serve as reservoirs of genetic diversity.

The estuarine pipefish, Syngnathus watermeyeri, is one of the rarest animals in Africa and occurs in only two South African estuaries. The species was declared provisionally extinct in 1994, but was later rediscovered and is currently listed by the IUCN as Critically Endangered. A conservation programme was launched in 2017, with the re-introduction of captive-bred individuals into estuaries where this species was recorded historically was the main aims. Successful captive breeding requires knowledge of the species9 dietary requirements. In the present study, we used metabarcoding of faecal DNA to identify prey species consumed by wild-captured S. watermeyeri from one of the two surviving populations. We compared the diet of the estuarine pipefish with that of the longsnout pipefish, S. temminckii, in the same estuary, to determine whether these two species compete for the same prey items. Both species occupy similar estuarine habitats, but S. temminckii has a much wider distribution and also occurs in the marine environment. Our results show that even though both pipefish species prey on three major invertebrate classes (Gastropoda, Malacostraca and Maxillopoda), the relative proportions differ. Syngnathus watermeyeri primarily targets Maxillopoda, with a single species of calanoid copepod constituting >95% of the Amplicon Sequence Variants (ASVs) identified from its faecal DNA, whereas the diet of S. temminckii mostly comprises snail and decapod crustacean larvae. Our finding supports the hypothesis that population declines and localised extirpations of S. watermeyeri during previous decades may have been the result of reductions in the abundance of calanoid copepods. Calanoids rely on freshwater pulses to thrive, but such events have become rare in the two estuaries inhabited by S. watermeyeri due to excessive freshwater abstraction for urban and agricultural use.

Este artículo contiene 7 páginas, 3 figuras., Explaining why some species are widespread, while others are not, is fundamental to biogeography, ecology, and evolutionary biology. A unique way to study evolutionary and ecological mechanisms that either limit species’ spread or facilitate range expansions is to conduct research on species that have restricted distributions. Nonindigenous species, particularly those that are highly invasive but have not yet spread beyond the introduced site, represent ideal systems to study range size changes. Here, we used species distribution modeling and genomic data to study the restricted range of a highly invasive Australian marine species, the ascidian Pyura praeputialis. This species is an aggressive space occupier in its introduced range (Chile), where it has fundamentally altered the coastal community. We found high genomic diversity in Chile, indicating high adaptive potential. In addition, genomic data clearly showed that a single region from Australia was the only donor of genotypes to the introduced range. We identified over 3,500 km of suitable habitat adjacent to its current introduced range that has so far not been occupied, and importantly species distribution models were only accurate when genomic data were considered. Our results suggest that a slight change in currents, or a change in shipping routes, may lead to an expansion of the species’ introduced range that will encompass a vast portion of the South American coast. Our study shows how the use of population genomics and species distribution modeling in combination can unravel mechanisms shaping range sizes and forecast future range shifts of invasive species., M.R. received funds from ASSEMBLE (a European Union Seventh Framework Programme research infrastructure initiative comprising a network of marine research stations) to visit the Estación Costera de Investigaciones Marinas de Las Cruces and the Pontificia Universidad Católica de Chile. P.R.T. was supported by the University of Johannesburg (Faculty Research Committee/ University Research Committee grant) and L.B.B. by the Australian Research Council. J.C.C. acknowledges financial support from the Nucleo Milenio en Conservación Marina. J.C.C. sincerely acknowledges long-term financial support (Pontificia Universidad Católica de Chile–Escondida Project) fromMinera Escondida Ltda, the last 10-y support from the Academic Chair “Arauco-UC” and the important and constant assistance and advice from Dr. PatricioManríquez. C.D.M. received funding fromthe South African Research Chairs Initiative of the Department of Science and Technology and the National Research Foundation (Grant 64801).

Sixteen species of Latrunculiidae Topsent, 1922, belonging to the genera Latrunculia du Bocage, 1869, Strongylodesma Lévi, 1969, Cyclacanthia Samaai & Kelly, 2004, Samaai & Kelly, 2002, are currently known from the temperate waters of South Africa. Extensive new sponge collections from the Amathole region of South Africa revealed the existence of three new species of Tsitsikamma, T. amatholensis sp. nov., T. madiba sp. nov., and T. beukesi sp. nov., and a new species of the endemic South African genus Cyclacanthia, C. rethahofmeyri sp. nov. With the recent addition of two new species of Tsitsikamma from Algoa Bay and Tsitsikamma National Park (T. michaeli Parker-Nance, 2019; T. nguni Parker-Nance, 2019) the total number of known South African Latrunculiidae is now 20 species in four genera. Here we propose two new subgenera of Tsitsikamma, Tsitsikamma Samaai & Kelly, 2002 and Clavicaulis subgen. nov., based on the morphological groups “favus” and “pedunculata” hypothesized by Parker-Nance et al. (2019). Species in the nominotypical subgenus Tsitsikamma, containing the type species, are thick encrusting to hemispherical with a rigid honeycombed choanosome, while species in the new subgenus Clavicaulis subgen. nov. have a purse or sac-like morphology with little choanosomal structure. Despite the obvious species-level differences in morphology, multivariate analysis based on spicule measurements (anisostyle length, discorhabd length, shaft and whorl length) was not able to distinguish between the proposed Tsitsikamma species, but separated known species T. favus Samaai & Kelly, 2002, T. pedunculata Samaai & Kelly, 2003, and T. scurra Samaai & Kelly, 2003, from each other. Similarly, DNA barcoding of the mitochondrial COI and the nuclear ITS of Tsitsikamma specimens failed to clearly differentiate between species, but was able to differentiate sister taxon relationships within the Latrunculiidae.

Environmental gradients between marine biogeographical provinces separate distinct faunal communities. However, the absence of absolute dispersal barriers allows numerous species to occur on both sides of such boundaries. While the regional populations of such widespread species are often morphologically indistinguishable from each other, genetic evidence suggests that they represent unique ecotypes, and likely even cryptic species, that may be uniquely adapted to their local environment. Here, we explored genomic divergence in four sympatric southern African decapod crustaceans whose ranges span the boundary between the cool-temperate west coast (south-eastern Atlantic) and the warm-temperate south coast (south-western Indian Ocean) near the southern tip of the African continent. Using genome-wide data, we found that all four species comprise distinct west coast and south coast ecotypes, with molecular dating suggesting divergence during the Pleistocene. Transcriptomic data from the hepatopancreas of twelve specimens of one of these species, the mudprawn Upogebia africana, which were exposed to either 10 °C or 20 °C, showed a clear difference in gene expression profiles between the west- and south coast ecotypes. This difference was particularly clear at 10 °C, where individuals from the south coast experienced a ‘transcriptomic shock’. This low temperature is more typical of the west coast during upwelling events, and the physiological stress experienced by the south coast ecotype under such conditions may explain its absence from that coastline. Our results shed new light on the processes involved in driving genomic divergence and incipient speciation along coastlines with porous dispersal barriers.

The recently published complete mitochondrial genome of the endangered Knysna seahorse, Hippocampus capensis Boulenger, 1900, was based on a specimen obtained from a Traditional Chinese Medicine market. As H. capensis is endemic to temperate South Africa and exceptionally rare, illegal trade to supply Asian markets would constitute a considerable extinction risk. I investigated the phylogenetic placement of the Chinese specimen using mitochondrial DNA control region and cytochrome b sequences from the H. capensis mitogenome among corresponding published sequences of H. capensis and a number of closely related seahorse species. The Chinese specimen was distinct from H. capensis and instead clustered with H. casscsio, a seahorse that was recently described from the South China Sea. The sequences of H. casscsio clustered randomly among those of specimens identified as H. fuscus, a species whose taxonomic validity is disputed, and which is considered to be a synonym of the widespread Indo-Pacific seahorse H. kuda. Given that the morphological identification of seahorses is difficult, it is recommended that the publication of new species descriptions and genomic resources be preceded by a comprehensive comparison with the available molecular data. The taxonomy of seahorses is far from resolved, and cutting-edge molecular studies will not improve this situation if they do not take existing information into consideration.

The common brushtail possum (Trichosurus vulpecula), introduced from Australia in the mid-nineteenth century, is an invasive species in New Zealand where it is widespread and forms the largest self-sustained reservoir of bovine tuberculosis (Mycobacterium bovis) among wild populations. Conservation and agricultural authorities regularly apply a series of population control measures to suppress brushtail possum populations. The evolutionary consequence of more than half a century of intensive population control operations on the species’ genomic diversity and population structure is hindered by a paucity of available genomic resources. This study is the first to characterise the functional content and diversity of brushtail possum liver and brain cerebral cortex transcriptomes. Raw sequences from hepatic cells and cerebral cortex were assembled into 58,001 and 64,735 transcripts respectively. Functional annotation and polymorphism assignment of the assembled transcripts demonstrated a considerable level of variation in the core metabolic pathways that represent potential targets for selection pressure exerted by chemical toxicants. This study suggests that the brushtail possum population in New Zealand harbours considerable variation in metabolic pathways that could potentially promote the development of tolerance against chemical toxicants.

Mitochondrial DNA (mtDNA) has long been used to date the divergence between species, and to explore the time when species’ effective population sizes changed. The idea that mitochondrial DNA is useful for molecular dating rests on the premise that its evolution is neutral. This premise was questionable to begin with, and even though it has long been challenged, the evidence against clock-like evolution of mtDNA is usually ignored. Here, we present a particularly clear and simple example to illustrate the implications of violations of the assumption of selective neutrality. DNA sequences were generated for the mtDNA COI gene and the nuclear 28S rRNA of two closely related and widely distributed rocky shore snails whose geographical ranges are defined by different thermal preferences. To our knowledge, this is the first study to use nuclear rRNA sequence for studying species-level genealogies instead of phylogenetics, presumably because this marker is considered to be uninformative at this taxonomic level. Even though the COI gene evolves at least an order of magnitude faster, which was reflected in high inter-specific divergence, intraspecific genetic variation was similar for both markers. As a result, estimates of population expansion times based on mismatch distributions were completely different for the two markers. Assuming that 28S evolves effectively clock-like, these findings likely illustrate variation-reducing purifying selection in mtDNA at the species level, and an elevated divergence rate caused by divergent selection between the two species. Although these two selective forces together make mtDNA suitable as a DNA barcoding marker because they create a ‘barcoding gap’, estimates of demographic change can be expected to be highly unreliable. Our study contributes to the growing evidence that the utility of mtDNA beyond DNA barcoding is limited.

Lovenula raynerae is the largest known African freshwater copepod. To date, it has only been sampled from ephemeral freshwater ecosystems. This paper reports the complete mitochondrial genome of L....

Cryptopygus antarcticus travei (Collembola) is a springtail endemic to the sub-Antarctic Prince Edward Islands. The mitogenome of C. a. travei has a length of 15,743 bp and comprises 13 protein-cod...

Tullbergia bisetosa is a springtail (Collembola) native to the sub-Antarctic Prince Edward Islands. Unlike most other springtails, it has acquired a euedaphic (living within the soil) life form. In...

Isotomurus maculatus (Collembola, Isotomidae) is a springtail with a large distribution. This species has been introduced to the sub-Antarctic Prince Edward Islands. In this study, the mitogenome o...

The penaeid prawns Fenneropenaeus indicus and Metapenaeus monoceros support shallow-water prawn fisheries in the south-west Indian Ocean. They are sympatric and have similar life histories, includi...

Two endemic southern African pipefish species (Teleostei: Syngnathidae) co-occur in estuaries on the southeast coast of South Africa. The larger longsnout pipefish, Syngnathus temminckii , is abundant and has a wide range that comprises coastal and estuarine habitats in all three of the region's marine biogeographic provinces. In contrast, the smaller estuarine pipefish S . watermeyeri is critically endangered, and confined to a few warm-temperate estuaries. Here, we explore reasons for these considerable differences in conservation status. Fecundity is related to fish size, with large live-bearing S . temminckii males carrying up to 486 developing eggs/embryos, compared to a maximum of only 44 recorded for S . watermeyeri . Loss of submerged seagrass habitats due to episodic river flooding appears to be correlated with the temporary absence of both species from such systems. Prolonged cessation in river flow to estuaries can cause a collapse in estuarine zooplankton stocks, a food resource that is important to pipefish species. The greater success of S . temminckii when compared to S . watermeyeri can be attributed to the former species' wider geographic distribution, fecundity, habitat selection and ability to use both estuaries and the marine environment as nursery areas. Genetic data indicate that this has resulted in a much smaller long-term effective population size of S . watermeyeri , a situation that has persisted since the beginning of the present interglacial period. Syngnathus watermeyeri is thus naturally more susceptible to anthropogenic disturbances, which have resulted in an alarming reduction in its contemporary population size. Possible measures to promote the conservation of S . watermeyeri are presented.

The spatial distribution of a species’ genetic diversity can provide insights into underlying evolutionary, ecological and environmental processes, and can contribute information towards the delineation of conservation units. The Knysna seahorse, Hippocampus capensis, is endangered and occurs in only three estuaries on the warm-temperate south coast of South Africa: Knsyna, Keurbooms and Swartvlei. Population sizes in the latter two estuaries have been very small for a prolonged period of time, and the populations residing in them may thus benefit from translocations as a means of increasing population sizes and possibly also genetic diversity. However, information on whether these three estuaries constitute distinct conservation units that warrant separate management is presently lacking. Here, we used genetic information from mitochondrial (control region) and nuclear microsatellite loci to assess the genetic diversity and spatial structure across the three estuaries, and also whether translocations should be included in the management plan for the Knysna seahorse. Although each population had a unique combination of alleles, and clustering methods identified the Swartvlei Estuary as being distinct from the others, levels of genetic admixture were high, and there was no evidence for reciprocal monophyly that would indicate that each estuary has a unique demographic history. On these grounds, we suggest recognising the three populations as a single evolutionarily significant unit (ESU), and encourage translocations between them to ensure the species’ long-term survival.

The ‘Abundant-Centre Hypothesis’ is a well-established but controversial hypothesis stating that the abundance of a species is highest at the centre of its range and decreases towards the edges, where conditions are unfavourable. As genetic diversity depends on population size, edge populations are expected to show lower intra-population genetic diversity than core populations, while showing high inter-population genetic divergence. Here, the genetic implications of the Abundant-Centre Hypothesis were tested on two coastal mussels from South Africa that disperse by means of planktonic larvae, the native Perna perna and the invasive Mytilus galloprovincialis. Genetic structure was found within P. perna, which, together with evidence from Lagrangian particle simulations, points to significant reductions in gene flow between sites. Despite this, the expected diversity pattern between centre and edge populations was not found for either species. We conclude that the genetic predictions of the Abundant-Centre Hypothesis are unlikely to be met by high-dispersal species with large population sizes, and may only become evident in species with much lower levels of connectivity.

The cosmopolitan lamniform shark Carcharias taurus (commonly known as the ragged-tooth, grey nurse or sand tiger shark) is threatened by overexploitation in parts of its range. Return migrations of females to specific nursery areas suggest that females exhibit reproductive philopatry, a behaviour that over time might lead to genetically isolated subpopulations over various spatial scales. To investigate genetic evidence for reproductive philopatry, genetic data from mitochondrial and microsatellite markers were generated for 104 young-of-the-year and juvenile sharks. Comparing the smallest versus the largest young sharks revealed a pattern of size-related differentiation between nurseries that was only found in the smaller size class. This not only confirms reproductive philopatry of their mothers, but is also in line with previous observations of larger juvenile sharks increasing their migration range and moving between sites. Our results highlight the need to target young-of-the-year sharks when investigating reproductive philopatry to exclude roaming individuals that obscure size-related signals of genetic differentiation. Given the species’ high susceptibility to overexploitation, the evidence for reproductive philopatry is of direct importance to the management and conservation of C. taurus worldwide. As many nursery areas as possible should be protected to ensure that the number of locally resident juveniles and the pool of the returning females remain stable in the long term. This may warrant protected areas, or time-area closures, prohibiting exploitation in the nursery areas during pupping season.

National Research Foundation of South Africa (BvV), the University of Johannesburg and the DST-NRF SARChi Chair of Behavioural Ecology and Physiology (NCB). Electrophoresis of microsatellite markers was done at the Analytical Facility based at Stellenbosch University.

Aim We provide an overview of the location and ages of coastal phylogeographical breaks in southern Australian planktonic dispersers, and test the hypothesis that the absence of such breaks in some species is an artefact of insufficient resolution of genetic markers when such breaks evolved comparatively recently. Location Temperate coastal Australia. Methods We generated a large (> 1500 individuals) data set from rapidly evolving microsatellite markers for two codistributed Australian coastal gastropods, and compared it with mitochondrial DNA data. Both study species, the snail Nerita atramentosa and the limpet Siphonaria diemenensis, have long planktonic dispersal phases, and neither taxon exhibits substantial regional genetic structure on the basis of mitochondrial DNA. We tested for the presence of genetic structure by means of AMOVA, Bayesian clustering (structure) and iterated realloction (flock). Results There was no compelling evidence for the existence of more than one evolutionary lineage in either species. Main conclusions Discrepancies in the phylogeographical structuring of co-distributed intertidal taxa cannot be attributed to insufficient marker resolution for the two species considered here, and likely reflect a combination of abiotic and biotic factors that include porous dispersal barriers, life history and species age/history. It appears that contemporary oceanography does not explain the presence of phylogeographical breaks, but may serve to maintain breaks that evolved earlier. Deep genetic divergence in some of the previously studied coastal invertebrates suggests that these could be cryptic species, in which case competitive exclusion may play a role in constraining species biogeography.

Distinguishing the relative influence of historic (i.e. natural) versus anthropogenic factors in metapopulation structure is an important but often overlooked step in management programs of threatened species. Biotas in freshwater wetlands and floodplains, such as those in the Murray–Darling Basin (MDB)—one of Australia’s most impacted ecosystems, are particularly susceptible to anthropogenic fragmentation. Here we present a comprehensive multilocus assessment of genetic variation in the threatened southern pygmy perch Nannoperca australis (578 individuals; 45 localities; microsatellite, allozyme and mitochondrial DNA datasets), an ecological specialist with low dispersal potential. We assess patterns of spatial structure and genetic diversity in populations spanning the highly fragmented MDB and test whether recent anthropogenic modification has disrupted range-wide connectivity. We detected strong and hierarchical population structure, very low genetic diversity and lack of contemporary gene flow across the MDB. In contrast, the apparent absence of pronounced or long-term phylogeographic structure suggests that observed population divergences generally do not reflect deeply historic natural fragmentation. Coalescent-based analyses supported this inference, revealing that divergence times between populations from the upper and lower MDB fall into the period of European settlement. It appears that the observed contemporary isolation of populations is partly explained by the severe modification of the MDB post-dating the onset of European settlement. Our integrated approach substantially improves the interpretation of how fragmentation impacts present-day biodiversity. It also provides novel contributions for risk-assessing management actions in the context of captive breeding and translocations of small freshwater fishes, a group of increasing global conservation concern.

All multicellular organisms host a wide diversity of microorganisms in and on their bodies, which are collectively known as their microbiome. Characterising microbial communities that inhabit different body niches in wild animals is critical to better understand the dynamics of microbiome diversityand its functional significance. The current study is the first to apply massively parallel sequencing of 16S rRNA to characterise the microbial diversity and functional content of oral microbiota in two of New Zealand’s most important invasive mammals, the omnivorous common brushtail possum (Trichosurus vulpecula) and the carnivorous stoat (Mustela erminea). In total, strains of bacteria belonging to 19 different phyla, 27 classes, 52 orders, 103 families, 163 genera and 51 known species were identified from the oral cavities of the study species. Strains of the phyla Proteobacteria, Firmicutes, Bacteroidetes, Fusobacteria, and Actinobacteria dominated the core oral microbial diversity in both species, while other taxa were comparatively less abundant. Despite invasive populations typically demonstrating limited genetic variation, intraspecific variation of the core bacterial taxa in the oral microbiota was considerable. This suggests that a complex interaction between genetic, physiological, and environmental factors determines the diversity of the study species’oral microbiome.

Background: Mitochondrial DNA (mtDNA) has long been used to date historical demographic events. The idea that it is useful for molecular dating rests on the premise that its evolution is neutral. Even though this idea has long been challenged, the evidence against clock-like evolution of mtDNA is often ignored. Here, we present a particularly clear and simple example to illustrate the implications of violations of the assumption of selective neutrality. Methods: DNA sequences were generated for the mtDNA COI gene and the nuclear 28S rRNA of two closely related rocky shore snails, and species-level variation was compared. Nuclear rRNA is not usually used to study intraspecific variation in species that are not spatially structured, presumably because this marker is assumed to evolve so slowly that it is more suitable for phylogenetics. Results: Even though high inter-specific divergence reflected the faster evolutionary rate of COI, intraspecific genetic variation was similar for both markers. As a result, estimates of population expansion times based on mismatch distributions differed between the two markers by millions of years. Conclusions: Assuming that 28S evolution is more clock-like, these findings can be explained by variation-reducing purifying selection in mtDNA at the species level, and an elevated divergence rate caused by diversifying selection between the two species. Although these two selective forces together make mtDNA suitable as a marker for species identifications by means of DNA barcoding because they create a ‘barcoding gap’, estimates of demographic change based on this marker can be expected to be highly unreliable. Our study contributes to the growing evidence that the utility of mtDNA sequence data beyond DNA barcoding is limited.

Tests for isolation by distance (IBD) are the most commonly used method of assessing spatial genetic structure. Many studies have exclusively used mitochondrial DNA (mtDNA) sequences to test for IBD, but this marker is often in conflict with multilocus markers. Here, we report a review of the literature on IBD, with the aims of determining (a) whether significant IBD is primarily a result of lumping spatially discrete populations, and (b) whether microsatellite datasets are more likely to detect IBD when mtDNA does not. We also provide empirical data from four species in which mtDNA failed to detect IBD by comparing these with microsatellite and SNP data. Our results confirm that IBD is mostly found when distinct regional populations are pooled, and this trend disappears when each is analysed separately. Discrepancies between markers were found in almost half of the studies reviewed, and microsatellites were more likely to detect IBD when mtDNA did not. Our empirical data rejected the lack of IBD in the four species studied, and support for IBD was particularly strong for the SNP data. We conclude that mtDNA sequence data are often not suitable to test for IBD, and can be misleading about species’ true dispersal potential. The observed failure of mtDNA to reliably detect IBD, in addition to being a single-locus marker, is likely a result of a selection-driven reduction in genetic diversity obscuring spatial genetic differentiation.

The Wild Coast in south-eastern South Africa is strongly influenced by the warm, southward-flowing Agulhas Current. This current has a significant impact on dispersal in the coastal biota of the region, and facilitates high levels of connectivity among populations. However, it is not known how the region's high-velocity hydrology affects genetic population structure in endemic estuarine species, populations of which are frequently isolated from the sea. Here, we compared genetic structure in two estuarine crabs of the family Hymenosomatidae. Both are presumed to have low dispersal potential, but they differ in terms of their life histories. Hymenosoma longicrure has abbreviated larval development and can complete its entire life cycle within estuaries, whereas Neorhynchoplax bovis is a direct developer that lacks planktonic larvae. Using DNA sequence data from the mitochondrial COI gene and the intron of the nuclear ANT gene, we found that levels of genetic structure differ considerably between the specie...

Species that disperse by means of planktonic larvae are typically not genetically structured along environmentally homogeneous coastlines. In contrast, those that lack a planktonic dispersal phase, or species with a short (

Southern Africa is a marine biodiversity hotspot that not only comprises faunal elements from the Atlantic and Indian Oceans, but also large numbers of endemic species. Using mitochondrial and nuclear DNA sequence data, we explored whether genetic structure in the endemic coastal goby Psammogobius knysnaensis, a species whose range straddles both biomes, is linked to the boundary between the two oceans. Subtle genetic structure was identified between Atlantic and Indian Ocean populations, with genetic diversity being lower in the Atlantic, and particularly on the west coast. Our results point to partial isolation between the populations associated with each biome that is most likely driven by the region’s oceanography, but unlike in other species, there is no evidence for distinct regional evolutionary lineages that are likely adapted to the environmental conditions prevalent in each region. The exclusive presence of P. knysnaensis in sheltered habitats (estuaries and lagoons) may protect this species from the severe impacts of cold water upwelling on the west coast.

Five to seven subspecies of Antarctic Tern (Sterna vittata) are recognised, with at least three (S. v. vittata, S. v. tristanensis and S. v. sanctipauli) wintering in South Africa. Morphological characters used to define these subspecies are not perfectly reliable, but fidelity to nesting site suggests they could be genetically distinct. We used morphological data and DNA to investigate the validity of subspecies. We further used stable isotope analysis of feather samples collected from the non-breeding grounds in South Africa to attempt to ascertain the population of origin. Nuclear and mitochondrial DNA sequence data identified two major genetic clades: one mostly comprised individuals partially or completely matching the morphological description of S. v. tristanensis, the other included individuals from S. v. vittata and S. v. sanctipauli. Stable isotope values indicated that juveniles originated from at least three populations. Irrespective of their morphological and genetic characteristics, most immatures moulted in Antarctic waters, and adults moulted in various habitats. Their colony of origin could not therefore be inferred from stable isotope values from feathers. Results indicate that morphological groupings may reflect a north–south cline across the Indian Ocean. Adequate conservation strategies require rigorous reassessment of the currently accepted subspecies, including DNA analyses of samples from the breeding grounds, particularly on Amsterdam and St Paul Islands.

Broadcast-spawning marine organisms with long pelagic larval duration are often expected to be genetically homogeneous throughout their ranges. When genetic structure is found in such taxa, it may be in the form of chaotic genetic patchiness: i.e. patterns that might seem independent of any un- derlying environmental variation. The joint analysis of population genetic data and marine environmental data can elucidate factors driving such spatial genetic diversity patterns. Using meso-scale sampling (at a scale of 10s to 100s of km), microsatellite data and advection connectivity simulations, we studied the effect of temperate southern Australian ocean circu - lation on the genetic structure of the snail Nerita atramentosa. This species has a long pelagic larval duration and is represented as a single metapopula- tion throughout its ~3000 km range, but even so, we found that its dispersal potential is lower than ex- pected. Connectivity simulations indicate that this is a result of the larvae that remain on the continental shelf (where currents are erratic and often shoreward) re- turning to the coast in much larger numbers than lar- vae that become entrained in the region's shelf-edge boundary currents. Our study contributes to the growing evidence that departures from the expecta- tions of panmixia along continuous and environ - mentally homogeneous coastlines are not limited to low-dispersal species, and it identifies on-shelf larval retention as an important factor limiting dispersal.

Estuaries, at the confluence of marine and freshwater systems, are mostly of geolog- ically recent origin and as such make excellent models for understanding recent speciation events. Using molecular approaches, we compared genetic diversity and demographic histories in 2 closely related southern African klipfish species, the marine Clinus superciliosus and the estuar- ine C. spatulatus. Strong genetic differentiation was identified using both mtDNA control region and nDNA S7 sequencing, despite some haplotype sharing. Coalescent-based modelling suggests that species divergence occurred during the Late Pleistocene or, more likely, during the Early Holocene, when present-day estuaries formed. Analyses of population demography suggest that C. superciliosus has undergone historical population expansion, whereas C. spatulatus is charac- terized by a population decline, potentially driven by repeated cycles of population crashes linked to the opening and closing of estuarine systems. This is also reflected in values of genetic diversity, which are almost an order of magnitude lower in the estuarine than in the marine species. Given the unique evolutionary history of C. spatulatus, a species that is restricted to only 2 South African estuaries, we highlight the need for a better understanding of the processes that have shaped the evolution of estuarine populations. The identification of unique genetic lineages in estuaries can help to better guide conservation and management efforts for some of South Africa's most fragile habitats.

Global climatic oscillations have shaped the contemporary genetic structure of marine taxa in different ways. Previous demographic studies have indicated that various intertidal marine species display genetic signatures of demographic expansion that either pre- or postdate the Last Glacial Maximum. Such expansions and the ability of species to colonise new habitats will influence their genetic structure, but the link between scales of larval dispersal and the strength of phylogeographic structure is not always clear. We analysed a fragment of the mitochondrial COI gene of 11 sympatric species of intertidal southern African patellid limpets to investigate how ancient oceanographic dynamics have shaped and maintained their contemporary spatial genetic variation. Our data show that the patellid limpets investigated display congruent evidence of spatial expansion during the Late Pleistocene or Early Holocene, which corresponds with the establishment of the contemporary southern African shoreline. We argue tha...

Southern Africa is a biodiversity hotspot of patellid limpets, with three genera (Helcion, Cymbula and Scutellastra) identified and described in the region. Scutellastra is the most diverse and most frequently studied of these and, along with Cymbula, includes species with territorial and non-territorial foraging behaviours. We used three mitochondrial markers (12S rRNA, 16S rRNA and COI) and one nuclear marker (ATPSβ intron) to assess evolutionary relationships among species of Cymbula and Scutellastra with these two foraging behaviours and to identify which foraging mode is the more ancient. Maximum Likelihood and Bayesian Inference phylogenetic analyses revealed that the species sharing a foraging type are monophyletic in both genera. Territoriality is a derived character, as the clades with this foraging type are nested within a tree that otherwise comprises non-territorial taxa. These include Helcion, which was recovered as sister to the Cymbula/Scutellastra clade, and the next basal genus, Patella, which is ancestral to all southern African patellogastropods. Deep genetic divergence between the two foraging traits reflects strong adaptive effects of resource partitioning in the evolution of southern African patellid limpets.

Ascidians are considered to have lower dispersal potential than most other sessile marine invertebrates with planktonic propagules by virtue of a very brief propagule duration. The larvae of colonial forms remain in the water column for only a few minutes, whereas most solitary forms settle in less than 24 h. This difference in propagule duration has been used to explain why allozyme data from colonial ascidians on the Australian east coast were genetically distinct at different sampling sites, whereas a solitary species exhibited no genetic structure. Spatial homogeneity in solitary species is surprising because genetic structure of species with much higher dispersal potential can be characterised by isolation by geographic distance, suggesting that these disperse by means of a stepping-stone pattern of dispersal. I reassessed the dispersal potential of solitary ascidians using DNA sequence data from the mitochondrial cytochrome oxidase subunit 1 gene and the intron of the nuclear adenine nucleotide transporter gene of a common south-eastern Australian solitary ascidian, Pyura praeputialis, using samples that span the species’ distribution range. Congruent with earlier findings, there was no evidence for stepping-stone dispersal, but it must be conceded that these results could be strongly affected by frequent adult dispersal, particularly by means of anthropogenic vectors, as well as insufficient marker resolution.

The southern African crab Hymenosoma orbiculare was recently split into five distinct species, of which three are estuarine/coastal and have peripatric distributions that are linked to temperature-defined marine bioregions. This suggests that the species’ ranges may be limited by physiological adaptations to their thermal environment. We explored this hypothesis by rearing the larvae of the warm-temperate lineage of H. orbiculare and the warm-temperate/subtropical H. longicrure at a range of water temperatures, and found clear temperature-dependent differences in the duration of larval development. Our study contributes to the growing body of evidence that stresses the importance of adaptation to regional environmental conditions, rather than physical dispersal barriers on their own, in limiting the mixing of marine species between temperature-defined biogeographic regions.Keywords: crown crab, Hymenosoma longicrure, Hymenosoma orbiculare, planktonic larval duration, range limits, temperature stress, thermal adaptation, zoeaAfrican Journal of Aquatic Science 2014, 39(3): 347–350

Aim Based on mitochondrial DNA (mtDNA) data, many organisms with ranges spanning multiple biogeographical regions exhibit genetic structure across the transition zones between these regions, while others appear to be genetically homogenous. No clear link has been found between the presence or absence of such spatial genetic discontinuities and species’ dispersal potential, confounding the formulation of general predictions concerning genetic structure. The fact that discrepancies between mtDNA and nuclear markers are common across semi-permeable barriers suggests that a lack of structure could be attributable to mtDNA-specific properties of inheritance. We re-examined genetic structure in the coastal crab Hymenosoma orbiculare, a species that is represented by a single mtDNA lineage across the Atlantic/Indian Ocean biogeographical transition zone, by comparing mtDNA data with nuclear DNA data. Location South Africa’s cool-temperate and warm-temperate marine biogeographical provinces. Methods DNA sequence data from the mitochondrial COI gene and the intron of the nuclear ANT gene were generated for 150 individuals of H. orbiculare. For each locus, we determined whether the sharing of alleles between provinces was the result of either the retention of ancestral polymorphism or of secondary contact. Results We recovered two nuclear intron lineages whose spatial genetic structure reflects contemporary biogeographical and oceanographical conditions, indicating that the existence of a single mtDNA lineage is not a function of unexpectedly high levels of dispersal. Main conclusions MtDNA-based genetic homogeneity is increasingly being reported in coastal organisms with ranges spanning biogeographical transition zones that define distinct evolutionary lineages in other species. Our results stress the importance of revisiting single-locus data sets by means of multilocus genetic approaches before any conclusions can be drawn about the role of biogeographical transition zones in driving genetic structure.

Cryptic diversity represents a major challenge to the accurate assessment of biodiversity, but the combined use of genetic and morphological analyses has proven to be a powerful approach to detect it. This is especially important for groups for which genetic information is not yet available. Here, we studied the highly conspicuous habitat-forming Pyura stolonifera species complex (Tunicata), which, as has recently been revealed, shows surprising levels of cryptic diversity, but whose systematics and biogeographical patterns in Australasia nonetheless remain poorly understood. We first present detailed taxonomic information of all the species associated with the P.?stolonifera species complex. We then proceed to describe the results of an exhaustive survey that included south-east Australia, Tasmania, and New Zealand. Subsequently, we present morphological and mitonuclear genetic analysis of two unresolved lineages that comprise the species Pyura praeputialis and a species that is formally described here (Pyura doppelgangera sp. nov.). Although the ranges of these two species overlap on mainland Australia, we found no sites at which both species live in sympatry, and there was no morphological or genetic evidence of hybridization. Taken together, the present study illustrates the usefulness of a combined morphogenetic approach in unravelling overlooked marine diversity in a relatively well-studied region.

Aim To investigate how marine barriers shaped the demographic history of Atlantic seahorses (Syngnathidae: Hippocampus). Location Atlantic Ocean. Methods Range-wide sampling (n = 390) at mitochondrial and up to five nuclear DNA loci was carried out across the Hippocampus erectus species complex (H. erectus from the Caribbean/North America, H. patagonicus from South America and H. hippocampus from Europe and West Africa). Multi-species coalescent and approximate Bayesian computation (ABC) frameworks were used to estimate support of competing biogeographical hypotheses and demographic parameters, including lineage divergence times, effective population sizes and magnitudes of population size change. Results We identified four distinct lineages within the H. erectus complex. A posterior probability of 0.626 and corresponding Bayes factors ranging from 3.68 to 11.38 gave moderate to strong support for a basal divergence between South American populations of H. patagonicus and Caribbean/North American populations of H. erectus coincident with the inter-regional freshwater outflow of the Amazon River Barrier (ARB). Estimates of historical effective population sizes and divergence times indicate that European and West African populations of H. hippocampus expanded after colonization from a more demographically stable Caribbean/North American H. erectus. Main conclusions Our findings of trans-Atlantic colonization followed by isolation across a deep oceanic divide, and isolation across a freshwater barrier, may demonstrate a contrast in marine divide permeability for this group of rafters. Demographic inference supports the establishment of an ancestral population of the H. erectus complex in the Americas, followed by the ARB splitting it into Caribbean/North and South American lineages at a time of increased sedimentation and outflow. Our estimates suggest that following this split, colonization occurred across the Atlantic via the Gulf Stream currents with subsequent trans-Atlantic isolation. These results illustrate that rafting can be a means of range expansion over large distances, but may be insufficient for sustaining genetic connectivity across major barriers, thereby resulting in lineage divergence.

Small-scale genetic heterogeneity in marine broadcast spawners is often attributed either to physical factors that constrain larval dispersal or to stochasticity in reproductive success. In females of the mussel Perna perna, it has been attributed to asymmetrical levels of gene flow between bays and the open coast, with bays acting as sources of propagules. If nearshore currents are an important feature constraining dispersal, then genetic heterogeneity should also be identi- fied in other coastal invertebrates with similar dispersal potential, and the amount of genetic struc- ture in adults and juveniles should be similar, whereas temporal changes in reproductive success should manifest themselves in lower genetic diversity of juveniles. We compared sequence data of female P. perna with that of males, juveniles and 3 sympatric marine invertebrates. Congruent genetic structure was only found in a direct developer, suggesting that the region's oceanography does not have a strong structuring effect on species that, like female P. perna, have a planktonic dispersal phase. Furthermore, lack of genetic structure in male and juvenile P. perna indicates that there are no physical barriers that reduce larval exchange. Stochastic reproductive success is also an unlikely explanation for genetic structure in P. perna because levels of genetic diversity are similar in adults and juveniles. Together with the recent finding that the sex ratio in P. perna is skewed toward males, particularly at exposed coastal sites, these results point to a role for selec- tion in driving genetic structure between bays and coastal habitats by eliminating a large propor- tion of adult females from the open coast.