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.

Molluscs are an important component of South Africa’s biodiversity. The assessment of distribution patterns and factors influencing the biogeographic distribution are an integral part of assessing the conservation status of molluscs and their conservation management needs. The existing terrestrial mollusc data from South Africa were assessed in terms of their value to biodiversity conversation planning and management. Although the data on terrestrial molluscs are incomplete and would be misleading in terms of identifying specific areas for protection, the data do illustrate significant patterns and trends of mollusc endemism and diversity, which can be used to improve biodiversity conservation and management efforts. The distribution of molluscs across the South African landscape illustrated ten broad biogeographical patterns. Two of these patterns reflected ancient distribution patterns of molluscs and consisted of molluscs of the Gondwanaland/southern relict and Laurasian origins. Three biogeographic patterns occurred across the eastern regions. These patterns were defined as the tropical/subtropical east African, subtropical east of southern Africa and east African afromontane patterns. The biogeographic patterns in the west consisted of the characteristic temperate ‘Mediterranean’ Cape centre and the arid regions of northwestern Cape, Namibia and parts of Botswana. An additional biogeographic pattern identified as the nama karoo/central west was recognised. The final two biogeographical patterns described taxa that were widely distributed and taxa that exhibited disjunct distributions. Twenty-six families and forty-three genera were associated with more than one biogeographical pattern. The dominant biogeographic pattern was the tropical/subtropical east African component. Twenty-one families and forty-eight genera were associated with this biogeographical pattern. The east African Afromontane pattern was also a conspicuous biogeographic element in South Africa. Fewer families and genera were distributed in the western and central regions. The distributions of terrestrial molluscs were influenced by a combination of various factors, which included the presence of rivers, the escarpment, altitude, humidity, precipitation, temperature and biomes. Rivers could possibly restrict the distribution of certain mollusc taxa but did not appear to be the dominant factor that influenced the distribution of molluscs across the landscape. In terms of the effect of temperature on the distribution of molluscs, the mean daily and mean annual temperatures appeared to have more of an influence on the distribution patterns than the mean daily maximum and minimum temperatures. Mean annual temperatures influenced the distribution of all families and genera. The mean daily maximum temperature appeared to have little or no effect on the distribution of mollusc taxa. Humidity and biomes also appeared to influence the distribution of taxa. The least inhabited biome was the succulent biome. Many mollusc taxa occurred in the wetter, warmer areas with high humidity levels. Areas of high species richness and high endemic species richness in South Africa were identified using two systems of endemism, one based on distinctive gaps in the frequency distribution of terrestrial molluscs in South Africa and the other based on an existing classification of invertebrate endemism (Hamer & Slotow, 2002). Areas of high mollusc species richness and endemism were also compared to areas of high millipede species richness and endemism. The total number of South African mollusc endemics was 370 (83 % of 447 indigenous species). The dominant mollusc families in South Africa were Achatinidae, Charopidae, Streptaxidae, Subulinidae and Urocyclidae. The first system of endemism identified 56 site endemics (species with only one locality), 50 local endemics (0 < maximum distance < 60 km) and 145 regional endemics (60 km < maximum distance < 330 km). The Hamer & Slotow (2002) classification of endemism classed 67 species as site endemics (maximum distance between localities < 10 km), 47 as local (11 km < maximum distance < 70 km) endemics and 59 as regional endemics (71 km < maximum distance < 150 km). The analysis of mollusc data, with both systems of endemism, showed similar areas of high species and endemic species richness. Quarter-degree grid cells with highest species richness overlapped with grid cells with the highest number of endemic species. However these grid cells coincide with areas that have been intensively sampled and this bias limits the application of the data in conservation planning. The patterns of endemism for molluscs and millipedes within the provinces differed, indicating that the inclusion of a single taxon in conservation planning would inadequately reflect the diversity of invertebrates in South Africa. A preliminary list of specific priority endemic sites for terrestrial mollusc conservation was identified. It is essential that the existing data on invertebrates be evaluated and used to identify key patterns and trends in invertebrate diversity as this will allow for the inclusion of invertebrates in biodiversity conservation planning and management. The analysis of the existing mollusc data identified bio geographical patterns that are important to conservation planning both at the local and national level as well as commonalities and differences between molluscs and millipede distributions. The analysis also highlighted the importance of municipal areas for conservation of hotspots of diversity, particularly in the eastern coastal areas of South Africa. / Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2007.

Solen cylindraceus is an infaunal filter-feeding bivalve inhabiting the intertidal mud banks of many southern African estuaries. It is particularly abundant in the Kariega estuary (33°41'S; 26°42'E) where it reaches densities of 400m⁻² (192g shell-free dry wt. m⁻²). The Kariega is a permanently open, marine dominated estuary about 18km in length, and S. cylindraceus is most abundant in its middle and upper reaches. Some physical characteristics of the estuary (temperature, salinity, sediment and water turbidity) are described, and the possible role of these factors in determining the density and distribution of S. cylindraceus within the Kariega estuary, is discussed. The structure of the alimentary system, gills and labial palps of S. cylintfraceus is described, all of which showed no major variation from the "typical" eulamellibranchiate form. Solen cylintfraceus was found to be a euryhaline osmoconformer with a salinity tolerance range of 15-65%. When animals were removed from their burrows, osmotic equilibration of the haemolymph was rapid (1-2 hours). By contrast, in animals left undisturbed in their burrows, osmotic equilibration was retarded (72-204 hours). It is suggested that the observed decrease in the rate of change of haemolymph osmolarity for animals in their burrows is linked to the stability of the interstitial salinity. A temperature tolerance range of 5-44°C was determined for S. cylintfraceus (in situ), in which prolonged exposure to 5°C and 40-45°C (12-36 hours respectively) resulted in a decreased burrowing ability, coma and death. Animal burrowing responses were not affected by temperatures in the range 15-35°C. Field experiments were carried out over several tidal cycles, in which the measurement of crystalline style volume was used as a means of assessing extracellular digestive activity. No major variation in style volume was recorded and it appeared that S. cylindraceus did not exhibit any cyclical pattern of style dissolution and regeneration. It is suggested that S. cylindraceus feeds continuously from the water column during high tide and possibly within its burrow, at or below the water table, during low tide. At a suspensoid concentration of 5Omg l⁻¹, S. cylindraceus was found to filter water almost continuously (90-95% of the time). Time spent filtering dropped to 68% at 100mg l⁻¹ and 32% at 500mg l⁻¹. Filtration rates for summer collected animals (25°C) were 22.86 ± 4.36ml min.⁻¹, some 3ml min.⁻¹ greater than that recorded for winter (16°C) collected animals. Filtration rate may be expressed as a function of shell length by the equations: y=0.247x¹̇⁰⁶⁶ (winter) and y=0.758x⁰̇⁸²⁶ (summer). Solen cylindraceus was capable of acclimating its filtration rate to both high and low temperatures under laboratory conditions. Filtration rate exhibited a thermal optimum in the range 15-35°C, declining at higher and lower temperatures. Q₁₀ values of filtration decreased rapidly from greater than 4 to less than 2, when the thermal optimum was reached. Maximum rates generally occurred at approximately 5°C above the temperature to which the animal had been acclimated. Optimal filtration rates (19-23ml min.⁻¹) were recorded in the salinity range 15-45%. When subjected to abrupt changes in salinity, filtration rates were immediately depressed. The extent and duration of these decreased filtration rates were dependent upon the magnitude and direction of salinity change, and were always less in animals exposed to hyper- than hyposaline conditions. Animals exposed to increased temperature and simultaneous elevated or unchanged salinity, showed a slight increase in filtration rate followed by rapid acclimation. A decrease in both temperature and salinity resulted in an initial decrease in filtration rate and a longer acclimation period. The ability of S. cylindraceus to acclimate fully within a wide temperature and salinity range, and to filter maximally in hypersaline conditions may, in part, explain its unusually high abundance in the Kariega estuary, despite it being close to the southernmost limit of the animal's geographical distribution. No significant difference in flItration rate was recorded at suspensoid concentrations of 5-100mg 1⁻¹. However, at 250 and 500mg l⁻¹ filtration rates decreased significantly, and coincided with increased levels of pseudofaecal production. Solen cylindraceus retained particles down to 2.5-3.0µm with great efficiency (ca. 60-90% efficiency). Below this particle size, retention efficiency decreased rapidly and a net production of particles was recorded below 1.51µm. Particle retention was independent of temperature (15 and 25°C) and salinity (15 and 35%). Use was made of stable carbon isotope analyses (¹³C/¹²C ratios) in an attempt to determine the important food sources of S. cylindraceus within the Kariega estuary. The results obtained demonstrated an enrichment in δ¹³ values for S. cylindraceus from the upper (-27.9%) to the middle (-25%c) and lower (-21.6%o ) reaches of the estuary, with no seasonal variation apparent. The bivalve was substantially more depleted in ¹³C relative to the dominant aquatic macrophytes Zostera capensis (-9.1 to -15.6%o) and Spartina maritima (-12.5%o). The use of δ¹³ alone, however, to unequivocally "pin point" specific food sources of a filter feeder in a predominantly detritus based food web, is limited. It is suggested that in the Kariega estuary, riparian litter and other terrestrially derived vegetation contribute to the carbon pool. A possible contribution of ¹³C depleted food sources via chemoautotrophic and/or anaerobic pathways, to the diet of S. cylindraceus, is suggested.

The two most frequently exploited species along the south east coast of South Africa are the gastropods, Haliotis midae (abalone) and Turbo sarmaticus (alikreukel). H. midae is a high valued commercial species, and suffers intense levels of illegal fishing. T. sarmaticus however, has no commercial value but is the preferred food item for impoverished subsistence communities. Owing to the fact that no legal commercial fishery exists for either species along the south coast, very few studies have been undertaken, especially in the heavily exploited infralittoral. Infralittoral size frequency distributions for both species revealed significant variation in density and size among sites of varying exploitation pressure. Densities ranged between 0 – 2.23 m⁻² (H. midae) and 0.03 – 4.93 m⁻² (T. sarmaticus) and maximum shell lengths ranged from 49.4 – 153.5 mm (H. midae) and 28.3 – 104.4 mm (T. sarmaticus). Relatively high densities and large sizes were found in marine reserves and secluded areas, and low densities and small sizes at sites near to large population centres and within the former Ciskei homeland region. Mean size of the largest 10% of the population, total density and sexually mature density were significantly related to exploitation predictors for both species. In addition, densities of H. midae juveniles were significantly related to exploitation predictors, suggesting that recruitment may be suppressed at the most exploited sites. Exploitation of T. sarmaticus tended to be localized with refuge and subtidal populations persisting. H. midae exploitation was however, far more extensive and intense. Growth of H. midae was investigated using three methods; mark-recapture, cohort analysis and growth banding analysis at Kowie Rocks, Port Alfred. The most useful of these methods for determining growth was a new technique described for growth banding analysis; which was validated using cohort analysis and measurements of shells of known age. This technique was less time consuming and labour intensive than previously described methods. Abalone growth was best described by the Schnute (1981) growth function. Systematic geographic variation in growth was observed for 10 sites along the South African coastline. Significant differences in growth among sites existed for animals between 0-4 years (P < 0.0001) and 4-6 years (P < 0.0001), and in the mean maximum sizes attained (P < 0.001). In general, abalone from the south east/east coast were found to have faster growth rates, smaller mean maximum sizes and attained sexual maturity earlier than those along the south west/ west coast. Haliotis midae recruit and juvenile densities were found to differ significantly among sites of varying exploitation pressure (P < 0.0001) and among months for recruit densities (P < 0.001). Exploited sites had low recruit and juvenile densities compared to unexploited sites and peak recruitment occurred during October/ November 2005. Recruit densities were significantly related to infralittoral adult densities during two of the three sampling months (P<0.05), when recruitment was low. No relationship was observed during the period of high recruitment, with all sites receiving high recruit densities. It was concluded that variation in recruit densities was the result of a combination of both density-dependent relationships (i.e. local spawner density and temporal variability in recruitment intensity) and the possible dispersal capabilities of H. midae. In addition, it was concluded that at present recruitment overfishing was not occurring along the south east coast. Post-recruitment mortality rates were variable but relatively constant, with hypothetical percentage survival and density curves revealing high rates and similar mortality curves among sites. Variation in juvenile densities was consequently a result of initial recruit densities and not variation in post-recruitment mortality. T. sarmaticus populations were found to be regionally sustainable and persisted along the south east coast due to adjacent intertidal and subtidal refuge populations. However, H. midae populations are becoming decimated along the south east coast. From the information obtained in this study new management proposals were suggested and discussed, such as closed areas and region-based management fisheries together with stock enhancement. These suggestions may prove to be feasible alternatives to present management strategies.

This thesis investigates the variability occurring in the secondary metabolites produced by three South African marine molluscs. Chapter Two discusses the isolation and spectroscopic structure elucidation of the metabolites isolated from two Siphonaria species. The re-investigation of Siphonaria capensis yielded siphonarienfuranone (2.2) as the only common polypropionate isolated from both the 1998 and 2009 collections of S. capensis from the same areas suggesting possible seasonal or genetic variation in polypropionate production. The sterol cholest-7-en-3,5,7- triol (2.33) was also isolated form the 2009 collection of S. capensis and this is the first time this compound has been isolated from a Siphonaria species. The second species, Siphonaria oculus is closely related to S. capensis and the investigation into the former’s secondary metaboliteproduction revealed 2.2 as a major metabolite suggesting an inter-species overlap in polypropionate production. Three new polypropionate metabolites, 2.35, 2.36 and 2.37 were also isolated from S. oculus. An unsuccessful attempt was made to establish the absolute configuration of 2.37 using the modified Mosher’s method and the limited amount of 2.37 available prevented any further attempts at resolving the absolute configuration of this compound. The 1H NMR analysis of the defensive mucus collected directly from S. oculus revealed the presence of the acyclic polypropionate 2.37 as a minor metabolite. The absence of characteristic signals for the furanone containing compounds 2.2, 2.35 and 2.36, might suggest that these compounds cyclise from a hypothetical acyclic precursor (2.38) during standard work up of bulk acetone extracts of Siphonaria species. Chapter Three discusses the re-isolation and spectroscopic structure elucidation of the metabolites isolated from the nudibranch, Leminda millecra. Three known natural products, millecrone A (3.1), 8-hydroxycalamenene (3.6) and cubebenone (3.8) were re-isolated from our 2010 collection of L. millecra, as well as the new minor metabolite 8-acetoxycalamenene (3.16). The cytotoxic prenylated toluquinones and toluhydroquinones (3.9-3.15) initially isolated from the 1998 collection of L. millecra were not found in the 2010 collection supporting the hypothesis that these compounds may be of fungal origin. L. millecra clearly shows variability in the compounds sequestered by this species with millecrone A (3.1) being the only common metabolite in the three investigations of L. millecra to date. An unsuccessful attempt was made to establish the absolute configuration of 3.1, 3.6 and 3.8 through initial LAH reduction of the ketone moiety contained in 3.1 and 3.8 and esterification of the resultant diastereomeric alcohol mixtures and the phenol functionality in 3.6 with (1S)-camphanic chloride. Crystallisation of the (S)- camphanate esters of 3.6 and 3.8 for X-ray analysis were unsuccessful, while the unexpected conjugate addition of a hydride in 3.1 resulted in complex diastereomeric mixtures which could not be separated by HPLC.

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.

Globalization has seen an unprecedented dispersal of exotic and alien species worldwide resulting in worldwide homogenization and sometimes extinction of indigenous or endemic taxa. When an exotic species becomes established in a new habitat the invasive organisms are capable of having an impact on indigenous community dynamics and the overall structure and function of ecosystems. Furthermore, the impact of invasion is determined by the geographical range, abundance and the per-capita or per-biomass effect of the invader. However, the success of the introduced organisms is reliant on their ability to acclimate to the physiochemical conditions of the newly invaded environment.Freshwater ecosystems are especially vulnerable to invasions because there are numerous potential routes of introduction including intentional pathways such as stocking, and unintentional pathways such as the release of ballast water and aquarium releases. Efforts to limit the introduction of invasive species or to manage established exotic populations are often hindered by insufficient understanding of the natural history of problematic species. Relatively little is known regarding the physiological tolerances of many taxa. Knowledge about specific species ecophysiological constraints allows for the prediction of future patterns of invasion more accurately, including where an introduced organism would probably survive, thrive and disperse. Furthermore, data on the physiological tolerances of an introduced exotic organism may provide data necessary for effective management and control. This studyinvestigated three invasive species in the Nseleni River system in a protected area in KwaZulu-Natal. The species studied were, Tarebia granifera (Quilted melania – Lamarck, 1822), Pterygoplichthys disjunctivus (Suckermouth armoured catfish - Weber, 1991) and Eichhornia crassipes (water hyacinth – (Martius) Solms-Laubach,). The Nseleni River flows into Lake Nsezi which is responsible for providing potable water to the surrounding towns and industry, as well as the surrounding rural communities. The Enseleni Nature reserve has become the centre for biodiversity dispersal in the immediate area, due to the change in landscape surrounding the protected area.An important step in developing alien invasive species management strategies in protected areas is determining their extent and invasive traits. Tarebia granifera is a prosobranch gastropod originally from South-East Asia that has become invasive in several countries around the world including South Africa. Snail populations were sampled at nine sites throughout the Nseleni/Mposa river system every six weeks over a twelve month period. The snail was abundant throughout the system, especially in shallow waters of less than 1m in depth.The first positive identification the loricariid catfish Pterygoplichthys disjunctivus for the Nseleni River was in 2006. The original introduction is believed to have been via the aquarium trade. The aim of the study was to assess the usefulness of the unified framework with regard to management of fish invasions by assessing the invasion stage of the loricariid population and identifying appropriate management actions using the Blackburn et al. (2011) framework. The fish were sampled at nine different sites and three different depths over a period of twelve months, as well as when two ichthyological surveys were carried out on the Nseleni River system. This invasive fish has been located throughout the system and both male and female fish were collected. The smallest fish sampled was a fingerling of a day or two old and the smallest pregnant female was a mere 270mm TL. This is a clear indication that this fish is breeding in the river system.Although T. granifera and P. disjunctivus were abundant in the Nseleni/Mposa river system, it was not clear what their role in the system was, and in particular if they were competing with any of the indigenous species. Therefore, isotope samples were collected from numerous taxa over a two week period, with the exception of Pterygoplichthys disjunctivus samples, which were collected over 12 months. The δ13C and δ15N signatures of all samples were determined. The niche overlap between the invasive and indigenous snails was effectively zero (1.02E-13%), indicating no shared food resources. The medium ranges of dNRb (7.14) and dCRb (9.07) for the invasive fish indicate that it utilizes a wider range of food resources and trophic levels than the majority of indigenous fish. A medium CDb value (2.34) for the invasive fish species, P. disjunctivus, describes medium trophic diversity, with three indigenous species possessing higher diversity and three possessing lower diversity. Furtherresults indicated that there was no direct dietary competition between P. disjunctivus and indigenous species. Eichhornia crassipes was first recorded on the Nseleni River in 1978, and has been shown to have a significant negative impact on the biodiversity of the Nseleni/Mposa River system and therefore required a control intervention. Although biological control using the two weevil species Neochetina eichhornia (Warner) and N. bruchi (Hustache) has been credited with affecting a good level of control, the lack of a manipulated post-release evaluation experiments has undermined this statement. Five experimental plots of water hyacinth of 20m2 were sprayed with an insecticide to control weevils. After ten months the plants in the sprayed plots were significantly bigger and heavier than those in the control plots that had natural populations of the biological control agents. This study has shown unequivocally that biological control has contributed significantly to the control of water hyacinth on the Nseleni/Mposa River system.The management plan for the Enseleni Nature Reserve identifies the need to control invasive and/or exotic organisms within the boundary of the protected area. In addition, set guidelines have been implemented on how to control these organisms, so that indigenous organisms are least affected. Lack of control of exotic organisms can have serious consequences for indigenous species. It is therefore of utmost importance that the population dynamics of the invading organism be understood, what the potential impact could be and how to control them. Furthermore, it has also acknowledged the threat of possible exotic species invasions from outside of the protected area that might result in threats to the protected area and that these must be investigated, researched and managed.This thesis has identified Tarebia granifera, Pterygoplichthys disjunctivus and Eichhornia crassipes as being a threat to indigenous biodiversity within the protected area, as well as in adjacent areas to the protected area. The thesis will therefore investigate the hypothesis that both Tarebia granifera and Pterygoplichthys disjunctivus are having a direct negative effect on available food resources for indigenous species of organisms. In addition, this thesis will investigate if theNeochetina species that have previously been introduced onto E. crassipes are having any negative effect on this invasive alien aquatic plant.

The large cosmopolitan sea hare Bursatella leachii is a common resident in Eastern Cape river mouths during summer and late autumn where they congregate in beds of Zostera capensis to breed. In this thesis, the previously known toxic formamide marine secondary metabolite (-)-bursatellin (2.2), which may deter predators of South African specimens of the globally distributed sea hare Bursatella leachii, was isolated and identified (Chapter 2). There have been no previous chemical ecology studies of B. leachii and the latter half of this thesis is devoted to chemical ecology studies of this organism. Interestingly, the isolation of the (-)-diastereomer of 2.2 from specimens of B. leachii collected from the Kariega River mouth (near Kenton-on-Sea) suggests that the South African specimens of this species are similar to specimens collected from Puerto Rico and from the Mediterranean Sea. Two different chromatographic techniques for isolating 2.2 were compared in order to maximize the amount of 2.2 isolated from the Kariega River mouth sea hares. The doubling of selected resonances observed in both the ¹H and ¹³C NMR spectra of the bursatellin isolated in this study suggest one of three possibilities; either firstly, the presence of closely related compound(s), secondly, the presence of diastereomers or thirdly the presence of rotamers. Through NMR kinetic studies, we were able to establish that the presence of rotamers was very unlikely due to no change in the relative ratio (3:1) of the ¹H NMR signals with an increase in temperature. Although the attempted synthesis of the acetate derivative (2.28), as a means of separating a diastereomeric mixture was successful, the chromatographic separation of the proposed acetylated diastereomers was not successful. Preparation of the camphanate ester derivatives (e.g. 2.30) proved to be unsuccessful. Five B. leachii specimens were dissected, their organs separated and individually extracted with methanol. The methanol extracts were individually chromatographed on HP-20 media, and the distribution of bursatellin determined by isolation and NMR. It was evident from this investigation that the distribution of 2.2 within individual B. leachii specimens was found to be highest within the B. leachii ink gland. The lower amounts of 2.2 contained in the digestive system, relative to other organs, was hypothesized to occur because 2.2 is sequestered from the diet of the sea hare and efficiently moved from the gut to various organs around the body where it is stored. The absence of 2.2 from the skin was surprising and may be a result of a smaller mass of skin relative to other organs coupled with the limitations of the chromatographic separation techniques employed. Surprisingly, no bursatellin was found within juvenile sea hares. Chapter three discusses the isolation of ilimaquinone (3.1) and pelorol (3.19) from the sponge Hippospongia metachroma and the structure elucidation of each compound using computer modeling to illustrate the conformation. It was deemed necessary to isolate these well known and abundant bioactive marine natural products from a sponge as standard compounds in the bioassays given the paucity of 2.2 available for this study. Chapter four describes the assays used to test the biological activity of the bursatellin 2.2 compared to the generally bioactive ilimaquinone and the structurally related and commercially available broad spectrum antibiotic chloramphenicol. B. leachii, a shell-less marine mollusc inhabits a variety of intertidal habitats and, therefore, is exposed to several different predators, yet does not appear to have any specific predators. Potential predators of this sea hare in the Kariega Estuary could be fish and amphipods which are found in close proximity to these sea hares. Results of the assays showed that at roughly natural concentrations, (calculated from the isolated chromatographic yield) feeding was deterred by the fish and amphipods, which implied that 2.2 may confer a defensive role within the organism. The relatively high concentration present within the ink gland of B. leachii may support this hypothesis. Surprisingly, given its structural similarity to chloramphenicol, 2.3 did not show any antimicrobial action against five of the six bacterial strains against which it was screened [chloramphenicol inhibited the growth of all the bacterial strains at very low concentrations (0.25 mg/mL)]. Bursatellin was found to be only active against Staphylococus aureus at high concentrations ca. 2 mg/mL when compared to chloramphenicol. Neither bursatellin nor chloramphenicol showed anti-fungal activity. Although this study suggests that the sea hares may use chemical defences in addition to opaline ink to defend themselves, they also live within the seagrass Z. capensis, which possibly provides the sea hare with a cryptic form of physical defence against several predators that are unable to swim freely within the weed beds in the littoral zone of the estuary. / Adobe Acrobat 9.53 Paper Capture Plug-in