History and status of oyster exploitation and culture in South Africa, and the role of oysters as vectors for marine alien species

Includes bibliographical references (leaves 69-86).In South Africa, both wild and cultivated oysters are consumed. Edible wild oysters include Striostrea margaritacea, Saccostrea cucullata, Ostrea atherstonei and 0. algoensis and all occur along the South and East coasts. These oysters were, or are, exploited commercially, recreationally and via subsistence fishers with S. margaritacea being the most targeted species. The commercial harvesting areas are along the Southern Cape coast and in KwaZulu-Natal. The Southern Cape coast is the largest harvesting area with 102 of the 145 pickers employed in the region. Commercial and recreational harvesting is managed by the Marine and Coastal Management Branch of the Department of Environmental Affairs and Tourism. Data on the total annual catch of oysters in these provinces are minimum estimates, as collectors do not always comply with the harvesting regulations. Subsistence harvesting is largely unmanaged, except in KZN, and is particularly rife in the Eastern Cape Province. The culture of oysters is dependent on importing Crassostrea gigas spat mostly from Chile. Oyster production statistics are only available since 1985, but approximately two million Crassostrea gigas oysters were produced annually throughout the seventies and early eighties. Since then, production has fluctuated over the years with an approximate increase of six million between 1985 and 1991, a decrease of five million between 1991 and 1998, and is presently stable. The establishment and closure of a highly productive farm in the late eighties and early nineties respectively, as well as improved production in recent years, has resulted in these trends. Although the market for oysters has grown, production has not kept up with demand, due to a lack of suitable locations for mariculture purposes. Finding suitable sites for oyster cultivation along the Northern Cape coast and establishing local oyster hatcheries for C. gigas oysters is suggested as the way forward. The latter would also prevent associated marine alien species from being imported with spat. Globally, oysters are well known vectors of marine alien species and despite oyster imports as early as 1894 into South Africa, this topic has been afforded little or no local attention. A visit to various oyster farms in South Africa resulted in the discovery of four newly-recorded alien species: the black sea urchin Tetrapygus niger, from Chile, the European flat oyster Ostrea edulis, thought to be locally extinct following its intentional introduction into South Africa in 1946, Montagu's crab Xantho incisus, from Europe, and the brachiopod Discinisca tenuis, from neighbouring Namibia. Oyster imports are suggested as their most likely vector into South Africa and the biological attributes of some emphasizes the possible threat and the need to limit or prevent their spread. Local or intraregional translocation of C. gigas and associated species, including aliens colonizing the area, may aid in this spread. Oysters host a diverse community of epi-and infaunal fouling taxa, which can be accidentally translocated along with their hosts in the course of commercial oyster trade. Thus, the types and quantities of fouling taxa occurring on farmed Crassostrea gigas were examined. How effectively these taxa are removed by standard cleansing techniques and whether those that persist after washing, survived intraregional translocation, were also examined. Cleaning and translocating oysters significantly reduced both the quantity (by more than 30 and 40 times respectively) and variety of fouling taxa. Although the mean abundance (A) or biomass (B) of taxa in uncleansed oysters (A: 79.48±233.10 (SD), B: 0.034±0.314 (SD)) were greatly reduced in cleansed oysters (A: 2.30±7.65 (SD), B: 0.0003±0.002 (SD)), small quantities still managed to survive translocation (A: 1.87± 7.43 (SD), B: 0.006±0.020 (SD)). Thus, the effectiveness of exposing oysters to freshwater or heated seawater as a more thorough cleansing regimen, to prevent the translocation of such taxa, were examined. Results indicated that oysters were able to survive for a longer time in freshwater (0% mortalities after 18 h) than in heated seawater (26.7% mortalities after 40 sec), but most taxa were eliminated more effectively by the latter treatment (e.g. 88.5% of the mudworm Polydora hoplura died after 20 sec compared to 97.5% after 18 h in freshwater). However, only a single reproductive individual of an alien species may be required for a successful introduction, and soaking for 20 sec in heated seawater would still be ineffective. An alternative treatment of 18 h in freshwater and 20 sec in heated seawater or freshwater, is suggested as a more effective treatment

Thermal preference and performance in a sub-Antarctic caterpillar: A test of the coadaptation hypothesis and its alternatives.

Physiological ecologists have long assumed that thermoregulatory behaviour will evolve to optimise physiological performance. The coadaptation hypothesis predicts that an animal\u27s preferred body temperature will correspond to the temperature at which its performance is optimal. Here we use a strong inference approach to examine the relationship between thermal preference and locomotor performance in the caterpillars of a wingless sub-Antarctic moth, Pringleophaga marioni Viette (Tineidae). The coadaptation hypothesis and its alternatives (suboptimal is optimal, thermodynamic effect, trait variation) are tested. Compared to the optimal movement temperature (22.5°C for field-fresh caterpillars and 25, 20, 22.5, 25 and 20°C following seven day acclimations to 0, 5, 10, 15 and 5-15°C respectively), caterpillar thermal preference was significantly lower (9.2°C for field-fresh individuals and 9.4, 8.8, 8.1, 5.2 and 4.6°C following acclimation to 0, 5, 10, 15 and 5-15°C, respectively). Together with the low degree of asymmetry observed in the performance curves, and the finding that acclimation to high temperatures did not result in maximal performance, all, but one of the above hypotheses (i.e. \u27trait variation\u27) was rejected. The thermal preference of P. marioni caterpillars more closely resembles temperatures at which survival is high (5-10°C), or where feeding is optimal (10°C), than where locomotion speed is maximal, suggesting that thermal preference may be optimised for overall fitness rather than for a given trait

Similar metabolic rate-temperature relationships after acclimation at constant and fluctuating temperatures in caterpillars of a sub-Antarctic moth.

Temperature compensation in whole-animal metabolic rate is one of the responses thought, controversially, to characterize insects from low temperature environments. Temperature compensation may either involve a change in absolute values of metabolic rates or a change in the slope of the metabolic rate - temperature relationship. Moreover, assessments of compensation may be complicated by animal responses to fluctuating temperatures. Here we examined whole animal metabolic rates, at 0 °C, 5 °C, 10 °C and 15 °C, in caterpillars of the sub-Antarctic moth, Pringleophaga marioni Viette (Tineidae), following one week acclimations to 5 °C, 10 °C and 15 °C, and fluctuating temperatures of 0-10 °C, 5-15 °C, and 10-20 °C. Over the short term, temperature compensation was found following acclimation to 5 °C, but the effect size was small (3-14%). By comparison with caterpillars of 13 other lepidopteran species, no effect of temperature compensation was present, with the relationship between metabolic rate and temperature having a Q10 of 2 among species, and no effect of latitude on temperature-corrected metabolic rate. Fluctuating temperature acclimations for the most part had little effect compared with constant temperatures of the same mean value. Nonetheless, fluctuating temperatures of 5-15 °C resulted in lower metabolic rates at all test temperatures compared with constant 10 °C acclimation, in keeping with expectations from the literature. Absence of significant responses, or those of large effect, in metabolic rates in response to acclimation, may be a consequence of the unpredictable temperature variation over the short-term on sub-Antarctic Marion Island, to which P. marioni is endemic

Fluorescent Gene Tagging of Transcriptionally Silent Genes in hiPSCs

Summary: We describe a multistep method for endogenous tagging of transcriptionally silent genes in human induced pluripotent stem cells (hiPSCs). A monomeric EGFP (mEGFP) fusion tag and a constitutively expressed mCherry fluorescence selection cassette were delivered in tandem via homology-directed repair to five genes not expressed in hiPSCs but important for cardiomyocyte sarcomere function: TTN, MYL7, MYL2, TNNI1, and ACTN2. CRISPR/Cas9 was used to deliver the selection cassette and subsequently mediate its excision via microhomology-mediated end-joining and non-homologous end-joining. Most excised clones were effectively tagged, and all properly tagged clones expressed the mEGFP fusion protein upon differentiation into cardiomyocytes, allowing live visualization of these cardiac proteins at the sarcomere. This methodology provides a broadly applicable strategy for endogenously tagging transcriptionally silent genes in hiPSCs, potentially enabling their systematic and dynamic study during differentiation and morphogenesis. : Gunawardane and colleagues use CRISPR/Cas9 to deliver an excisable cassette to transcriptionally silent loci in hiPSCs, then accomplish excision of the cassette in a second step utilizing Cas9/CRISPR and the MMEJ and NHEJ DNA-repair pathways. Excision results in mEGFP tagging of the targeted loci. Upon differentiation, each of five tagged cell lines appropriately expresses a unique fluorescent fusion protein localized to the sarcomere in live cardiomyocytes. Keywords: CRISPR/Cas9, genome editing, cardiomyocyte differentiation, stem cells, iPSCs, MMEJ, live imaging, endogenous fluorescent tagging, mEGFP, HD