The southern African shoreline is inhabited by a great diversity of patellid limpets of which most are endemic to South Africa. These limpets have evolved foraging mechanisms that partition ecological resources and reduce interspecific competition, resulting in ecological specialists and generalists. The evolution of ecological specialization or generalization remains poorly understood and there is no agreement on how such evolutionary transitions are correlated with levels of genetic diversity. This study investigated evolutionary correlations between territoriality in foraging and genetic structure of southern African patellid limpets (Cymbula and Scutellastra spp.) using stable δ13C and δ15N ratios, mitochondrial and nuclear DNA markers. The outcomes of the study were best rationalized and understood in the context of the scale of analysis in terms of both space and taxonomy. At biogeographic scales and the interspecific level, the stable isotope signatures and genetic structure of these limpets are determined by extrinsic factors such as biogeography and oceanography. However, at the smallest site scales and the intraspecific level, the isotope signatures and genetic diversity of these limpets are significantly correlated to their foraging traits. At large scales, there are no correlations between foraging behaviour and either genetic structure or isotope signature. At smaller scales, territorial Scutellastra foragers display both isotopic enrichment and greater haplotype diversity than congeneric non-territorial foragers. Thus, the isotope signatures and genetic structure of these limpets are determined by intrinsic species-specific response linked to their foraging behaviour. However, this pattern was intriguing as differences between territorial and non-territorial limpets in both isotopic signature and genetic diversity were observed only along the south coast when the same species were compared in other biogeographic regions. The significant interaction effect between foraging behaviour and stable isotope signatures was only observed from the sites within the Agulhas Bank or which are strongly influenced by the Agulhas current. This south enrichment in isotopic ratios is due to the mixing differences between onshore and offshore waters as the Agulhas current moves from east to south. At the generic level, the correlation between foraging behaviour and isotope signatures and genetic structure were particularly profound for Scutellastra species. The molecular phylogeny revealed deep evolutionary divergence between territorial and non-territorial Scutellastra spp. This divergence was concordant with morphological differences in shell shape and radula anatomy between territorial and non-territorial species. A taxonomic review of the scutellastrid spp. is proposed, suggesting possible re-consideration of the genus as two genera characterized by either territoriality or non-territoriality. The divergence between territorial and non-territorial species in both Scutellastra and Cymbula took place approximately in the early Oligocene. Major climatic cooling and decreases in sea level occurred during the Oligocene and this probably exposed much of the lower intertidal zone, increasing new potential habitats and algal availability. The Oligocene exposure of rocky shores and algal abundance in the lower intertidal zone probably elicited resource partitioning amongst these patellid limpets and subsequently, the evolution of territorial and non-territorial species. Analyses of the demographic history of these patellid limpets revealed evidence of post-glacial spatial expansion around the Pleistocene, implying these limpets were at population equilibrium during the dramatic LGM sea temperatures. Thus, these limpets managed to expand their range during dynamic oceanographic oscillations and dramatic sea-level changes in the Pleistocene. This study highlighted the importance of applying ecological traits as a subject to investigate and comprehend the evolutionary ecology of marine herbivores. The foraging traits of these true limpets are reflected in both their stable isotope ratios and genealogy, presumably as an evolutionary consequence of competition.

Dispersal rates are known to play a fundamental role in establishing the genetic structure found within a species. Dispersal in marine benthic invertebrates is largely dependent on life history strategy. The presence of pelagic larvae will potentially result in high dispersal within a species, while direct development limits dispersal. This study used two intertidal pulmonate limpet species to investigate the relationship between dispersal potential and genetic structure of populations. Siphonaria capensis produces benthic egg masses, which release pelagic larvae. These float in ocean currents before settling and metamorphosing into adults. S. serrata reproduces by direct development. The larvae hatch as completely metamorphosed juveniles after developing in an egg case not far from the parent. The high dispersal capacity of S. capensis pelagic larvae should result in high levels of within-population variation since individuals within a particular population may originate from several different populations. This should lead to high levels of gene flow along the coast and to low genetic differentiation among populations. On the other hand, the low dispersal potential of S. serrata should cause low levels of genetic variation within populations, as relatives will remain close to one another. Low levels of gene flow and high genetic differentiation among populations are expected in this species. In dealing with the relationship between life history strategy and population genetic structure in these two species, a series of indirect techniques was utilised. Multivariate analyses of the morphological variation within each species showed that variation in size was similar between the two species and largely governed by environmental factors. Variation in shell shape was largely under genetic control and supported the predictions made for each species. Variation in total proteins was relatively uninformative in terms of examining the differences between the two species. Allozyme analysis and mtDNA sequencing clearly showed differences between the species in terms of their population genetic structure. High levels of gene flow were found within S. capensis. This was strongly influenced by ocean currents, with the close inshore Agulhas current along the Transkei coast contributing to high levels of dispersal and hence gene flow. Low levels of gene flow occurred within S. serrata, resulting in low within-population variation and high among-population differentiation.

Aspects of the biology and ecology of Patella granularis were investigated along a 130km stretch of the south-east coast of South Africa. Distribution, biomass, density and population structure were investigated at seven localities. In addition, a more detailed study of the growth rate and reproductive biology of populations inhabiting three different substrata (aeolianite, quartzitic sandstone and mussel shells) was conducted. The genetic relationships between these three populations was also examined, as was the foraging behaviour of the limpets inhabiting an aeolianite and a quartzitic sandstone shore. Finally, differences in food availability on the different substrata were studied. On the south-east coast, P. granularis has a wide intertidal distribution, occurring from the upper Balanoid zone through to the Cochlear zone, where it is a common inhabitant of mussel shells. The mean shell length of P. granularis was found to decrease down the shore. The largest limpets (46.6 mm shell length) were found on an offshore island in Algoa Bay. At most localities investigated, the sex ratio deviated from a 1:1 ratio with more males than females being recorded on five shores. Both limpet density and biomass were lower on the south-east coast when compared to data published for west coast populations. On the south-east coast, both density and dry biomass were highest in the lower Balanoid zone. Allozyme electrophoresis indicated that P. granularis inhabiting aeolianite, quartzitic sandstone and mussel shells are all part of a single population. Extremely high genetic identity values (0.998), low levels of heterozygosities (0.035 - 0.061), low levels of polymorphisms (25% - 31%) and low FST values (0.021) all suggest that the three populations of P. granularis form a common breeding group, despite the high levels of phenotypic plasticity observed. On all shores, P. granularis was found to grow allometrically, increasing in shell height more rapidly than shell length. Estimation of the growth rate (determined by the Von Bertalanffy growth model) of P. granularis suggested that limpets inhabiting the mussel shells grew more slowly, and attained a smaller maximum size, than those inhabiting both the aeolianite and the quartzite (K = 0.25, 0.32 and 0.33 respectively; 27.12 mm, 31.89 mm and 32.96 mm respectively). Previous work has shown that west coast P. granularis grow more quickly (K = 0.7) and reach a greater size (. 40 mm). Translocation of limpets among sites suggested that limpet size in the mussel beds was spatially constrained. Shell microgrowth bands were deposited tidally, but could not be used for aging limpets due to shell erosion. Limpets from the aeolianite had the greatest reproductive fitness, producing more eggs (.366 000/limpet) than those inhabiting quartzite (.119 500/limpet) or mussel shell limpets (.85 800/limpet). Aeolianite limpets also spawned throughout the year, whereas those from the quartzite and mussel shells spawned twice a year (once in winter and once in summer) although a great deal of interannual variability was observed. The onset of sexual maturity occurred at a similar age in all limpets (1 - 2 years) and is probably genetically entrenched. P. granularis inhabiting both an aeolianite and a quartzitic shore were active during nocturnal low tides. All limpets returned to a home scar after foraging. Whilst foraging, limpets inhabiting the aeolianite shore moved shorter distances (.17 cm) at a slower rate than those from quartzite (.30 cm). Limpets that were translocated from one substratum to the other initially moved similar distances to the source group, but after a maximum period of one week, moved distances that mirrored those moved by the resident limpets. Neither season nor tidal phase influenced the distances foraged. No directionality in foraging was found. Wear of radula teeth, particularly the pluricuspid tooth, was greater in limpets from the quartzite. It is hypothesised that the observed differences in life-history parameters and foraging behaviour of limpets both within the south-east coast and between the west and south-east coasts are related to food abundance. Chlorophyll-a, and hence microalgal biomass, was consistently higher on aeolianite (.2.5 times) than on both quartzite and mussel shells. Estimates of chlorophyll-a were higher (although not significantly) in winter. Previous studies determined that primary productivity is also higher along the west coast. Finally, the lack of evidence for migration of limpets from the low-shore to high-shore in south-east coast P. granularis is discussed. It is suggested that this species settles opportunistically within its physiological tolerances and responds morphologically to localised environmental conditions.

Investigations were carried out into aspects of the bio!ogy and ecology of Helcion pectunculus along the coait of South Africa. These included studies of the distribution, density and biomass of the limpet at six sites along the east coast plus one site on the west coast; the growth of H pectunculus on both west and east coasts; a comparison of the reproductive biology of the east and west coast populations; the foraging activity and feeding behaviour of H pectunculus and the driving forces behind the rhythmic behaviour of this limpet; the importance of the crevice environment in the biology and ecology of H pectunculus. Helcion pectunculus has a restricted zonation, with the majority of animals residing in crevices in the upper Balanoid zone during diurnallowtides, although individuals were occasionally found in the lower Balanoid zone on shores with a gently sloping aspect. This limpet occurs in higher densities (50-lO0 individuals/m2) on shores which have large numbers of crevices and boulders i.e. quartzitic sandstone shores. On most shores, the ratio of males to females differed significantly from a 1: 1 ratio with the highest ratio being obtained on the west .. coast (3 males: 1 female). At all sites, the populations of H pectunculus exhibited strong sexual dimorphism. Males and females were always found to differ in size, with individuals of < 20 mm shell length generally being male whilst limpets with a shell length of> 22 mm were generally female. Helcion pectunculus grows allometrically, increasing in height faster than length, which is expected of a high shore gastropod mollusc attempting to reduce evaporative water loss. Growth rates were similar on both east and west coasts regardless of the differing oceanographic conditions. The theoretical values of Lmax were also similar being 30.86 mm and 30.71 mm respectively. Micro-growth bands are laid down within its' shell which have the same periodicity as the tidal cycle and these enabled age estimates to be made. Younger individuals were male whilst older animals were female, suggesting that H pectunculus is a protandrous hermaphrodite. Histological examination proved, unequivocally, that this limpet undergoes a protandric sex change, changing from male to female when they are about 2 years old. Both east and west coast popUlations had a marked reproductive cycle, exhibiting two spawning periods a year, one in April and another in November. The possibility that the reproductive pattern exhibited is now phylogeneticallyconstrained is discussed. It is suggested that H pectunculus has evolved a reproductive cycle which will allow its planktonic larvae to utilise the valuable phytoplankton bloom food source whilst using onshore winds to ensure that larvae are not transported out to sea and lost. The number of foraging excursions carried out by individuals of H pectunculus was found to have a significant effect on Gonad Index and hence potential reproductive output. The activity pattern of H pectunculus varied depending upon micro-habitat; animals inhabiting both east and west facing rock surfaces are active during nocturnal low tides whilst animals on west facing rock surfaces are also active during daytime low tides whilst in the shade. Limpets travel further during foraging excursions in winter (X = 85.53 cm) than in either spring (x = 55.7 cm) or summer (X = 48.8 cm) and also during spring low tides (x = 89.8 cm) compared with neaps (x = 40.9 cm). This limpet exhibits rigid homing to a fixed scar within a-crevice and feeding excursions were found to cons.is.t. of three distinct phases, a rapid outward phase, a slower foraging phase and a rapid homeward phase. Foraging was always highly directional, with a mean vector which took limpets onto an area of the rockface with the highest microalgal biomass and also the smoothest rock surface. Helcion pectunculus exhibits a free-running endogenous rhythm of locomotor activity with both circadian and circatidal components and it is suggested that this rhythm plays a role in allowing the limpet to avoid unfavourable environmental conditions. The exogenous entrainment factor of this endogenous rhythm was the time of exposure to air in the field. There was found to be an organized distribution of limpets within crevices with smaller, younger limpets being towards the back of the crevice and larger, older limpets towards the crevice mouth. It is hypothesized that juvenile limpets of this species actively select and settle at the backs of crevices responding to chemical cues of adult conspecifics. The crevice refuge supplies the limpets with a stable and buffered environment with higher relative humidities (X = 72.3%) and lower rock surface temperatures (X = 19.7°C) than adjacent exposed rock surfaces (X = 64.5%; x = 22.9°C). Limpet body temperatures were significantly lower in crevic~refuges compared to limpets on exposed rock surfaces. Body temperatures never exceeded the rock surface temperatures. It is suggested that this is the result of morphological adaptations such as shell ornamentation and allometric growth. Light levels above 1000 J.1E.m-2.s-1 inhibited foraging activity in H. pectunculuswhilst limpets subjected to 30-50% shade foraged even during daytime lowtides. This limpet is one of the least tenacious (2.75 ± 0.13 kg.cm-2 ) of all South African limpets and the possibility that wave activity governs both the activity patterns and homing behaviour of this limpet is discussed. Limpets deprived of a crevice refuge experienced extremely high mortalities, with 45% of the limpets being lost during the first high tide period. A hypothetical model of the hierarchy of exogenous factors controlling limpet foraging activity is introduced and discussed in relation to the results of this study. Finally, it is suggested that in addition to the "migratory" and "non-migratory" groups of limpets present on southern African shores a third group of limpets seem to be present which may be classed as "specialized non-migratory" species. These are species that do not migrate, garden or aggressively fight off like conspecifics. They have overcome the competition for space and food on intertidal rocky shores by adapting to a particular habitat which is exclusive to them alone. From the combined results of these studies, it can be stated that H. pectunculus has adapted physiologically, morphologically and behaviourally to successfully survive the extreme conditions in the upper Balanoid zone.

This thesis is divided into two parts. The first part (Chapter Two) documents a bioassay guided investigation of the ethyl acetate extracts of four marine invertebrates from Mozambique (an Irciniid sponge, a Haliclona sp. sponge, an ascidian tentatively identified as Diplosoma sp., and the soft coral Cladiella kashmani). Eight known compounds [ilimaquinone (2.1), renierone (2.7), N-formyl-1,2-dihydrorenierone (2.8), 1,6-dimethyl-7-methoxy-5,8-dihydroisoquinoline-5,8-dione (2.9), mimosamycin (2.10) 7Z-allylidene-5-hydroxy-7,7a-dihydro-2H-cyclopenta[b]pyran-6-one (2.11), flaccidoxide (2.18) and 11S,12S-epoxycembra-1Z,3E,7E-trien-14S-ol (2.19)] and a new diterpene [13S,14R-diacetoxy-11S,12R-epoxycembra-1Z,3E,7E-triene (2.20)] were isolated and identified using standard spectroscopic techniques. Anomalies in the published spectral data of 2.1 and 2.8 were exposed and corrected, and the absolute stereochemistry of the cembrane diterpenes 2.18 and 2.20 established using the modified Mosher’s method. The comparative activities of the nine natural products against four cancer cell lines (A549, LOX, OVCAR3, SNB19) are reported. The second part of the thesis (Chapter Three – Chapter Six) is concerned with an ecological, structural and synthetic study of diterpenes from the endemic South African pulmonate limpet Trimusculus costatus. Two new labdane diterpenes [6b,7a-diacetoxylabda-8,13E-dien-15-ol (3.10) and 2a,6b,7a-triacetoxylabda-8,13E-dien-15-ol (3.11)] were isolated from T. costatus and evaluated for anti-feeding activity against the common predatory fish Pomadasys commersonnii. A strategy for the semi-synthesis of 3.10 from rhinocerotinoic acid (4.14), a diterpene reportedly present in the ubiquitous South African shrub Elytropappus rhinocerotis, was devised in order to allow further bioactivity tests to be performed and unequivocally assign the unknown absolute stereochemistry of the T. costatus metabolites. Attempts to isolate rhinocerotinoic acid from local specimens of Elytropappus rhinocerotis were unsuccessful, and as the repetition of a published synthesis of 4.14 from (-)-sclareol (4.15) gave rhinocerotinoic acid in unacceptably low yields with poor stereoselectivity, an improved synthesis of 4.14 is presented. Comprehensive studies using hispanone (5.1) as a model compound showed that 6,7-dioxygenated labda-8-enes could be prepared from compounds possessing a 7-oxo-labda-8-ene skeleton with some degree of stereocontrol. In the process, fourteen new hispanone analogues were prepared and most of these were tested for activity in a suite of ten agro-chemical assays. The novel compound 7b-hydroxy-9a-carbonitrile-15,16-epoxylabda-13(16),14-dien-6-one (5.34) exhibited significant activity against the crop fungus Phytophthora infestans and is currently being subjected to further agro-chemical tests. Unfortunately, the results from the oxygenation study performed on the model compound 5.1 could not be directly extrapolated to rhinocerotinoic acid. Attempts to prepare the naturally occurring 3.10 from 4.14 via an alternative route were unsuccessful but yielded an analogue of 3.10 in which the substituents at C-6 and C-7 are in a diequatorial rather than a diaxial configuration.

The aim of the present study was to determine the potential impact of intertidal grazers on the distribution of macro algae on the south coast of South Africa. Particular attention was paid to the large patellid limpet, Patella oculus, which is found thoughout the intertidal zone. Studies of gut contents revealed that Patella oculus was capable of ingesting not only the thallus of foliose (eg. Ulva spp.) and encrusting coralline macroalgae, but also spores and diatoms. The inclusion of these relatively small particles in the diet was surprising, as electron micrographs of the radula of P.oculus revealed that it is typically docoglossan in structure. Such radulae are thought to be poorly suited for collecting small food particles. Sand made up a significantly higher proportion of the gut contents than other particles at all shore heights, which suggests that P.oculus might be capable of excavating the rocky substratum, or of sweeping up sand, while searching for food. Analysis of the gut contents of other local herbivorous molluscs, was also carried out. These species included the winkles, Oxystele variegata and O.sinensis, and the small pulmonate limpets, Siphonaria concinna, S.capensis, and S.serrata. The guts of all species contained mainly spores and diatoms, although small fragments of Ulva sp. were found. The population structure of Patella oculus was investigated at two sites, Cannon Rocks and Old Woman's River. At Cannon Rocks, mean shell length of low-shore animals was significantly lower than that of both mid- and high-shore animals, while at Old Woman's River, no significant difference was found among shore heights. A regression equation for In (shell length) vs In (dry weight) was calculated, and based on length data, the biomass density (g dry mass.m⁻²) of P.oculus at Old Woman's River was estimated. Values ranged from 2.8 on the low- and midshore to 0.37 on the high-shore. A manipulative field experiment was used to determine the impact of mesograzers and macrograzers (such as Patella oculus) on the distribution of intertidal macro algae on the mid- and low-shore at Old Woman's River. Grazers were excluded using mesh cages (mesh size = 3mm), in two separate experiments, one in winter and the other in spring. Percentage cover of macroalgal species and sessile invertebrates was estimated at approximately 6 week intervals for up to 3 months. MANOV A showed that treatments did not significantly affect cover of macroalgae or barnacles during winter. However, towards the end of the spring experiment (midshore only) cover of barnacles and green foliose turfs did increase in those plots from which mesograzers and/or macro grazers were excluded. The failure of the statistical tests to detect significant differences at some time intervals may have been caused by high levels of variation among replicates. This suggests that factors other than grazing are of overriding importance in determining the distribution of local macroalgae. The existence of a possible symbiotic relationship between Patella oculus and the red foliose alga, Gelidium pristoides, was investigated. The availability of various substratum types, including rock, limpet shells, barnacles etc., and the proportion of the total cover of G.pristoides on each, was calculated. It was shown that a significantly higher proportion of the alga grew on limpet shells, although the availability of this substratum type was low. It is thought that the aggressive behaviour of P.oculus prevents all but juvenile Patella longicosta from grazing on its shell, thus providing a refuge from grazing for G.pristoides.