Mind the gaps: investigating the cause of the current range disjunction in the Cape Platanna, Xenopus gilli (Anura: Pipidae)

Low-lying areas of the Cape at Africa’s south-westernmost tip have undergone dramatic marine-remodelling, with regular changes in sea-level following glacial cycles. Species for which marine barriers are impenetrable underwent concomitant radical distribution changes which may account for current range disjunctions. The Cape platanna, Xenopus gilli, is a frog distributed in only three disjunt areas within low-lying regions of the southwestern Cape. We determined the relationship between frogs from these three disjunct areas, by using a combination of morphometric analysis and mtDNA (ND2 and 16S fragments) sequences of 130 frogs from eight ponds. Coalescent analyses on molecular data dated the divergence in two major clades to around 4.6 Mya, a period during which major uplifting on the eastern side of the subcontinent caused climate changes throughout southern Africa. Principal components analysis showed significant morphometric differences between each clade on head and limb measurements. Consistent differences in ventral colouration and patterning were also observed. We report on increased levels of hybridisation with X. laevis throughout the range of X. gilli, which reaches at least 27% hybrids in some ponds. Urgent conservation actions are required to control habitat loss from alien invasive vegetation, and prevent introgression with the domestic-exotic, X. laevis

Molecular phylogenetics reveals a complex history underlying cryptic diversity in the Bush Squeaker Frog (Arthroleptis wahlbergii) in southern Africa

Throughout the Miocene, the African landscape underwent broad climatic shifts that profoundly influenced the distribution of fauna and flora. Since the late Miocene, these shifts have created a landscape in southern Africa that is strongly characterised by savanna and arid environments. Forests persist in small fragments, primarily in mountainous or heterogeneous landscapes. Arthroleptis wahlbergii is a small frog endemic to eastern South Africa that has presumed low dispersal ability. Because of its preference for forests, the dynamics of forests since the late Miocene in this region might have promoted diversification within A. wahlbergii. To investigate whether habitat fragmentation might have driven divergences among populations, we carried out species distribution modelling and population level and phylogenetic analyses using two genetic loci (16S, mitochondrial; RAG-1, nuclear) sequenced for 48 individuals from 14 forests across the c. 500 km range of this species. There is substantial population level structuring within A. wahlbergii, however the structure does not relate to forest types or catchments. We instead propose that the structure is a result of dynamic and idiosyncratic changes in forest connectivity over the Pleistocene. We identified two geographically circumscribed clades, the northern of which corresponds to true A. wahlbergii. The southern clade corresponds to populations from which Arthroleptis wageri FitzSimons, 1930 was described. This has long been considered a synonym of A. wahlbergii, but our molecular phylogenetic and distribution modelling supports recognising A. wageri as a distinct species

Occurrence and extent of hybridisation between the invasive Mallard Duck and native Yellow-billed Duck in South Africa

Hybridisation between invasive and native species represents a significant threat to biodiversity. The Mallard Duck (Anas platyrhynchos) is known to hybridise with numerous closely related Anas species in regions where they have been introduced, threatening the genetic integrity of native ducks and in some instances contributing to their extinction risk. Mallard Ducks were introduced into South Africa in the 1940s and are now naturalised and widespread in the country. It has been speculated that Mallard Ducks are hybridising with native Yellow-billed Ducks (A. undulata) in South Africa, but evidence for this remains observational or purely anecdotal. Here we use data from nuclear microsatellite markers and mitochondrial DNA sequencing to show that hybridisation is indeed occuring between these two species. We found evidence for the occurance of hybridisation, mostly as crosses between Mallard Duck hens and Yellow-billed Duck drakes. Surprisingly, our results suggest that introgressive hybridisation is primarily occuring into the invasive Mallard Duck population (mostly Mallard Duck backcrosses were detected), evidenced by directionally-skewed gene flow and sex-biased mating. Whether these findings reflect true assortative mating or a case of Haldane’s rule remains unknown. We also found evidence of high connectivity between Yellow-billed Duck populations, as far as 1000 km apart, in South Africa. Taken together these results suggest that hybrid genotypes can disperse over vast distances between populations and lead to genetic pollution, even in the absence of invasive Mallard Ducks. Active management of Mallard Duck populations has been met by public resistance in some areas in South Africa, partly because of a lack of evidence showing clear impacts by these birds. This study provides some of the first scientifically-documented evidence for such impacts

Locomotor performance constrained by morphology and habitat in a diverse clade of African frogs (Anura: Pyxicephalidae)

Understanding how morphology relates to function and how the two change across species in different environments and regions can provide clues to the drivers of speciation and diversity within a clade. Locomotion is a vital trait for most animals to function, and locomotor adaptations should involve morphological changes that maximize fitness in specific environments. We focus on the Pyxicephalidae, a speciose sub-Saharan anuran family with remarkable morphological and ecological diversity, and hypothesize that morphological differences among these species affect locomotor performance and that these represent adaptations to their surrounding environment. We tested 215 individuals across 25 species for various aspects of locomotor performance by filming frogs jumping and swimming, inducing sustained movement around a circular track and rotating a non-stick surface until adhesion failure. We then used Ornstein–Uhlenbeck models to determine whether species showed trait selection between different environments. We found that some morphological traits were significantly correlated with aspects of locomotor performance between species. Microhabitats showed separate selective optima for both morphology and locomotor performance, specifically between combinations of semi-aquatic, fossorial, semi-arboreal and terrestrial species. Our study shows how ecological conditions coincide with morphological and locomotor differences within a continental arena, generating a remarkably diverse African frog family

Unfolded, misfolded, and self-organized short alanine-rich peptides: implications for fundamental science, human disease, and biotechnology

Protein folding is the reversible transition by which an unordered polypeptide chain attains its functional 3-D native structure. A detailed understanding of the principles which govern the protein folding process, such as how sequence codes for structure, remains elusive. Achieving a complete picture of the folding process requires information regarding structural preferences in the unfolded state. Moreover, understanding the principles which govern protein aggregation is of significant biomedical and biotechnological importance. Herein, short alanine-based peptides are used as model systems for studying both the structural preferences in the unfolded state as well as protein aggregation in relation to human disease, and exploitation of the self-assembly process for various biotechnological applications.It is now a central dogma of protein science that the unfolded state is not conformationally random, as was originally believed, but that, instead, residual structure exists. Here, we elucidate the conformational propensities of alanine in the unfolded state using short alanine-rich peptides as model systems. The intrinsic conformational propensities of alanine, as well as nearest neighbor effects are illuminated using various vibrational spectroscopic methods, combined with NMR results.Protein and peptide aggregation is affiliated with various seemingly unrelated diseases, including several neurodegenerative diseases and the systemic amyloidoses. It is of current belief that aggregation is a general feature of the protein energy landscape, suggesting that the various unrelated human pathologies linked to protein aggregation are linked by common principles. Herein, fibril formation of a short alanine-based peptide with no known disease affiliation is probed by vibrational circular dichroism (VCD) spectroscopy. In particular, it is demonstrated that peptide fibrils give rise to VCD intensity enhancement, illustrating the use of the technique as a novel means to probe aggregation kinetics.In addition to the biomedical relevance, protein and peptide self-assembly can be exploited as a means of constructing biomaterials with inherent biofunctionality. In this regard, oligopeptide-based hydrogels have shown potential as drug delivery systems and tissue engineering scaffolds. Herein, the unique properties of a novel class of self-assembling alanine-rich oligoopeptides are presented. In particular, it is demonstrated that conformational instability can be exploited to tune the physicochemical properties of hydrogels formed by such systems, for the potential use in various biotechnological applications.Ph.D., Physical Chemistry -- Drexel University, 201

Cannibalism or congeneric predation? The African clawed frog, Xenopus laevis(Daudin), preferentially predates on larvae of Cape platannas, Xenopus gilli Rose & Hewitt

Predators are not limited to prey from other species as they can cannibalise vulnerable individuals within their own population. The African clawed frog, Xenopus laevis (Daudin), is a predator with a broad diet, known to consume multiple prey species, including congeners and conspecifics. African clawed frogs occur in sympatry with the Endangered Cape platanna, Xenopus gilli Rose & Hewitt, which are under threat through competition and predation from X. laevis. We investigated the threat of X. laevis predation on X. gilli using choice and no‐choice experiments to evaluate the relative vulnerability of X. laevis and X. gilli larvae. Results showed that large X. gilli larvae had a significantly higher vulnerability to X. laevis predation compared to small X. gilli larvae. However, the same discrimination was not discerned when offered large and small X. laevis larvae, or mixed larvae of the same size. We report ontogenic shifts in behaviour of X. gilli larvae that may be a factor in contributing to the vulnerability of large X. gilli larvae to adult X. leavis predation. Congeneric predation likely has negative implications for the population structure of the Endangered X. gilli. Our study underlines the call for the removal of X. laevis to conserve populations of X. gilli

In a Pinch: Mechanisms Behind Potential Biotic Resistance Toward Two Invasive Crayfish by Native African Freshwater Crabs

It is essential to incorporate mechanisms of biotic resistance into predictions of ecological impact conferred by invasive species. Trophically and functionally analogous native species have high potential to confer biotic resistance or to be susceptible to competitive exclusion by the invading species. In species with dominance hierarchies and high aggression the role of weapons, such as chelae, is emphasised and selected for. Differences in traits such as crushing capacity can indicate prey handling capabilities, while correlations between closing force and morphology can be used to understand the role of signaling in agonistic contests. Closing force strength can be used to infer the outcomes of both direct (predation) and indirect (competition) trophic interactions. Southern Africa has been invaded by two freshwater crayfish species (Cherax quadricarinatus and Procambarus clarkii). Biotic resistance of freshwater crabs toward crayfish invasions varies between geographic location and co-evolutionary history, thus comparing invasion histories without incorporating geographic context can produce unequivocal conclusions. We compared the closing force and chelae morphology of both crayfish species with a native trophically analogous freshwater crab, Potamonautes perlatus. Closing force increased significantly with mass for all species. There was significant interaction between sex and species on closing force. Potamonautes perlatus females showed significantly stronger maximum chela closing force than male P. perlatus, both sexes of P. clarkii and female C. quadricarinatus. Contrastingly, male C. quadricarinatus had significantly higher closing forces than both sexes of P. clarkii and female C. quadricarinatus, however, there was no difference between female P. perlatus. Native P. perlatus has the capacity to hold a competitive mechanical advantage over both invaders, but this varies with sex. Chelae length was not a significant predictor for closing strength in any of the species, which may be related to dishonest signaling in decapod species. This makes it imperative to assess whether factors such as closing force actually translate to resource holding potential in a contest scenario. We thus provide evidence that African freshwater crabs may exhibit biotic resistance toward invasion and the first measurements for C. quadricarinatus closing force strength

Frog eat frog: exploring variables influencing anurophagy

Background. Frogs are generalist predators of a wide range of typically small prey items. But descriptions of dietary items regularly include other anurans, such that frogs are considered to be among the most important of anuran predators. However, the only existing hypothesis for the inclusion of anurans in the diet of post-metamorphic frogs postulates that it happens more often in bigger frogs. Moreover, this hypothesis has yet to be tested

Environmental constraints influencing survival of an African parasite in a north temperate habitat: effects of temperature on development within the host

The monogenean Protopolystoma xenopodis has been established in Wales for >40 years following introduction with Xenopus laevis from South Africa. This provides an experimental system for determining constraints affecting introduced species in novel environments. Parasite development post-infection was followed at 15, 20 and 25 °C for 15 weeks and at 10 °C for51 year and correlated with temperatures recorded inWales. Development was slowed/arrested at410 °C which reflects habitat conditions for >6 months/year. There was wide variation in growth at constant temperature (body size differing by >10 times) potentially attributable in part to genotype-specific host-parasite interactions. Parasite density had no effect on size but host sex did: worms in males were 1·8 times larger than in females. Minimum time to patency was 51 days at 25 °C and 73 days at 20 °C although some infections were still not patent at both temperatures by 105 days p.i. In Wales, fastest developing infections may mature within one summer (about 12 weeks), possibly accelerated by movements of hosts into warmer surface waters. Otherwise, development slows/stops in October–April, delaying patency to about 1 year p.i., while wide variation in developmental rates may impose delays of 2 years in some primary infections and even longer in secondary infections