Dissertations / Theses on the topic 'Abalone culture'

A physiological study was carried out to determine the effects of temperature, quantity of food fed, feeding activity and size class on the oxygen consumption and ammonia production of the South African abalone Haliotis midae L.. A set of intermittent flow respirometer chambers were used in a recirculating system to measure the oxygen consumption rates of fed (postprandial) and unfed (postabsorptive) abalone for three different size classes (15, 30, and 50mm) at three different temperatures (16°, 20° and 23°C) over a 24 hour period. Ammonia production rates and food consumption rates were simultaneously determined. Oxygen consumption and Ammonia production rates per gram abalone were linear functions of abalone size, temperature, and mass of food consumed. Oxygen consumption and ammonia production rates were found to increase proportionally to temperature and in inverse proportion to abalone size. These rates were significantly higher for fed as opposed to unfed abalone. Equations were derived to predict oxygen consumption and ammonia production for fed and unfed animals for a range of temperatures and size classes of abalone. A 96h LC50 lethal toxicity test for exposure to ammonia established 1.08mg.1⁻¹ of unionised ammonia as the lethal limit. A test on the long term effects on growth of acute exposure (12h) to various sublethal concentrations of ammonia showed no significant effects on growth for concentrations below 0.88 mg.1⁻¹. A safe chronic exposure level of 0.02 mg.1¹ NH₃-N was used to predict optimal flow rates required per kilogram of abalone in a rearing tank over a range of size classes and temperatures. The physiological and biological requirements of H. midae and the implications of this knowledge for rearing tank management and optimal design are discussed.

The available literature on abalone nutrition was synthesised and the prospects for developing a complete pelleted dry feed for Haliotis midae evaluated. The similar body compositions, digestive structures, enzyme activity, acceptance of a wide variety of feed ingredients and comparable growth performance on formulated diets of various abalone species suggests that they have similar nutritional requirements. Abalone also appear to be similar to other farmed monogastric animals in that digestion is primarily extracelluar and they possess a large, muscular crop and stomach. Abalone energy metabolism is carbohydrate based. They are capable of digesting high levels of dietary protein but their ability to utilise fat is limited. Abalone have been shown to consume predictable amounts of dry feed which is efficiently converted into body weight. Efficiency indices of utilisation of formulated feeds, such as feed conversion ratio, protein efficiency ratio, net protein utilisation and apparent digestibility have been successfully applied to abalone. It was concluded that the prospects for developing complete diets for H. midae using a conventional animal feed science model were good. The ability of H. midae to utilise a range of proteins which had potential for inclusion in practical diets was investigated. Five protein rich ingredients, namely, casein, fishmeal, soya oil cake, Spirulina, and torula yeast, were fed to H. midae in semi-purified diets. Two algal diets, fresh Plocamium corallorhiza and dried Ecklonia maxima were fed as controls. Abalone fed on fishmeal and Spirulina based diets displayed significantly higher growth rates than diets containing soya oil cake, torula yeast, casein and E. maxima. Growth rates of abalone fed with P. corallorhiza were significantly lower than all other diets. All artificial diets yield lower feed conversion ratios (0.7-1.8) and higher protein efficiency ratios (3.3-6.5) ratios than the seaweed control diets (FCR = 2.8-3.4; PER = 2.2-3.0). The results indicated that fishmeal and Spirulina were the most suitable proteins for inclusion in practical diets for H. midae. The effect of protein level on growth rate and nutritional indices was evaluated by feeding starch bound, fishmeal based diets containing 27, 32, 37, 42 and 47% protein to juvenile H. midae. Weight gain was positively related to the level of dietary protein, increasing by 18% between 27% and 47% protein. Protein efficiency ratio declining from 3.2 to 2.3 with an increasing dietary protein content. Feed consumption rate was approximately 1% of body weight per day for all diets. Post-weaning abalone (ca. lOmm shell length, 0.2g) differed from larger juveniles (ca. 35mm, 8g) in their response to varying proportions of dietary protein and energy. The smaller animals appeared to have a lower protein requirement and poorer ability to utilise lipid than the larger juveniles. Proximate analyses revealed that the levels of protein, lipid and carbohydrate in abalone soft tissue increased with increasing dietary levels of these nutrients. Larger juveniles contained significantly higher levels of protein and carbohydrate, but lower levels of lipid, ash and moisture, than the smaller post-weaning abalone. The assimilation efficiency of [U-¹⁴C]-arginine by H. midae fed diets enriched with [U-¹⁴C]-arginine was only 0.45%. Furthennore, supplementation of diets with graded levels of crystalline arginine did not have any effect on growth rates. It was concluded that the prospects for defining the quantitative amino acid requirements of H. midae using crystalline amino acids are not promising. Rates of gastric evacuation and enzyme secretion were monitored in juvenile H. midae fed an extruded, fishmeal based dry feed. Gut fullness peaked 6h after feed was offered and the bulk of feed consumed was digested within 24 h. Enzyme secretion appeared to begin with the onset of feeding and continued for at least 6h after peak gut fullness was attained. Protease activity increased significantly following ingestion but amylase activity was maintained at a more or less constant level. A low level of lipase activity was observed suggesting that the ability of H. midae to digest fat is limited. The growth rate of H. midae fed an extruded, fishmeal based feed increased with increasing temperature between 12°C and 20°C. Between 20°C and 24°C a marked decline in growth rate accompanied by a deterioration in feed conversion and increased mortality was observed. Consumption of a dry pelleted feed was shown to be a function of body size and temperature. Based on these data a model which predicts a daily ration for H. midae was developed. The present study showed that H. midae efficiently utilises extruded dry feeds containing conventional feed ingredients. Although technical difficulties were encountered in measuring apparent digestibility, it was concluded that the prospects for developing practical diets according to established nutritional principles are promising.

Thesis (MSc)--Stellenbosch University, 2002.
ENGLISH ABSTRACT: The species Haliotis midae is the only abalone species of commercial importance to the South African fisheries industry. The species is currently under substantial pressure due to commercial harvest and illegal poaching, to the extent that genetic and biodiversity may be under threat. The species is also being cultured in commercial systems for supply to international markets. The South African production for 2002 amounted to 350 tons. The commercial production of abalone may also have implications with regard to genetic diversity of natural and commercial populations. The aim of this project was to assess the genetic diversity of the natural and commercial populations of abalone at the I&J Abalone Farm, Danger Point, Gansbaai, in South Africa, in order to make recommendation with regard to possible impact on natural diversity as well as internal management systems. Distortion of natural genetic diversity or a loss of internal diversity will have detrimental consequences. Representative samples have been obtained from the surrounding natural population, as well as the commercial populations, including the broodstock and various progeny groups. Both mtDNA and AFLP molecular techniques were used to assess genetic diversity. Data analysis showed that the genetic profile of the commercial population display sufficient genetic variation. The genetic structure of the commercial population also displays no significant deviation from that of the surrounding natural population, i.e. the population of origin. The results through give indication of a small, though unsignificant loss of genetic variation from the broodstock to the subsequent progeny groups. The investigation conclude that the commercial populations of abalone at the I&J Abalone Farm, Danger Point, Gansbaai, in its current format holds no threat to the disruption of the genetic diversity of the surrounding natural population. Further, the commercial population possesses sufficient genetic variation in view of future genetic development. There is signs of a slight loss of genetic variation in the change over from the broodstock to the progeny groups. This would necessitate careful and controlled replacement of the original broodstock with new animals from the commercial progeny groups as part of an ongoing process of domestication.
AFRIKAANSE OPSOMMING: Die spesie Haliotis midae is die enigste perlemoen spesie van kommersiële belang vir Suid Afrikaanse visserye. Die spesie verkeer tans onder toenemende druk as gevolg van kommersiële en onwettige vangste, tot so 'n mate dat genetiese en biodiversiteit daardeur geaffekteer kan word. Die spesie word in toenemende mate kunsmatig gekweek vir voorsiening aan internasionale markte. Suid Afrikaanse produksie vir 2002 beloop reeds sowat 350 ton per jaar. Die kunsmatige produksie van perlemoen het ook moontlike implikasies op die genetiese diversiteit van natuurlike en kommersiële populasies van die perlemoen, Haliotis midae. Die doel van die projek was om die aard van genetiese diversiteit van die natuurlike en kommersiële populasies van perlemoen by die I&J Perlemoen Plaas, Danger Point, Gansbaai in Suid Afrika te ondersoek, ten einde aanbevelings te maak ten opsigte van moontlike impak op natuurlike diversiteit asook ten opsigte van interne genetiese bestuurstelsels. Versteuring van natuurlike diversiteit of verlies van interne diversiteit hou nadelige gevolge in. Verteenwoordigende monsters is versamel vanuit die omliggende natuurlike populasies, sowel as van die kommersiële populasie op die plaas, insluitend die teelmateriaal en verskillende nageslaggroepe. Beide mtDNA en AFLP-molekulêre tegnieke is gebruik tydens die ontleding van genetiese diversiteit. Dataontleding het aangetoon dat die genetiese profiel van die kommersiële populasie voldoende genetiese variasie demonstreer. Die genetiese struktuur van die kommersiële populasie toon verder geen betekenisvolle verskil met die omliggende natuurlike populasie nie, dit wil sê met die populasie van oorsprong nie. Die resultate toon verder 'n geringe, dog nie betekenisvolle verlies van genetiese variasie wat waargeneem word met oorgang van die teelmateriaal na die onderskeie nageslaggroepe. Die bevindinge van die ondersoek is dus dat die kommersiële populasie op die I&J Perlemoen Plaas, Danger Point, Gansbaai, in die huidige formaat, geen bedreiging inhou vir die versteuring van die omliggende natuurlike genetiese diversiteit nie. Verdere bevindinge is dat die kommersiële populasie oor voldoende genetiese diversiteit beskik met die oog op toekomstige genetiese ontwikkeling. Daar is tekens van 'n geringe afname in genetiese diversiteit met die oorgang vanaf die teelmateriaal na die nageslaggroepe. Dit beklemtoon die belang van 'n gekontroleerde vervanging van die oorspronklike teelmateriaal met nuwe teeldiere vanuit die kommersiële populasie as deel van die domestikasieproses.

In the late 1980's abalone culturalists noticed reduced growth rate and shell deformities in some abalone stocks. These problems were the result of infestations by a shell boring polychaete, Terebrasabella heterouncinata. Under intensive abalone culture conditions the level of infestation can reach epidemic proportions and there are often severe consequences for the host abalone. Heavy sabellid infestation levels have placed the economic viability of several South African farms under threat. This study formed part of an ongoing project that is aimed at investigating the basic biology of Terebrasabella heterouncinata. The majority of abalone farmers in South Africa feed their abalone either naturally occurring kelp (Ecklonia maxima) or the formulated abalone feed, Abfeed. Farmers have suggested that the use of Abfeed is associated with higher sabellid infestation levels and changing the abalone diet from Abfeed to kelp helps reduce sabellid infestation. Speculation has arisen indicating that older, slower growing abalone are more susceptible to sabellid infestation. The effect of host abalone diet history and their growth on sabellid settlement success, morphometries and reproduction was quantified. To better understand the plasticity of the expression of life history traits the variability of morphometric and reproductive characteristics was compared between different farm environments. And more...

ENGLISH ABSTRACT: The first successful captive spawning of the South African abalone Haliotis midae occurred in the 1980’s and subsequently the commercial abalone industry in South Africa has developed, with an estimated investment of US$ 12 million and annual output of 500 to 800 tons by 2001, making South Africa the biggest abalone producer outside of Asia. Natural kelp is currently the major feed and the development of a suitable substitute, and improved disease management in abalone culture are seen as the primary factors limiting expansion of the industry in South Africa. Further, abalone growth rates are very slow and improvements in growth rate will lead to shortened production times with benefits to producers. Diseases in aquaculture have traditionally been combated using antibiotics as treatment (therapeutic usage) and preventative measure (prophylactic usage). In terrestrial livestock management, antibiotics are also used as growth promoters. The use of antibiotics in aquaculture has recently sparked concerns about the development of antibiotic resistance in pathogens of humans and aquaculture organisms, and alternative strategies to using antibiotics mainly focus on manipulating the microbial composition in the host organism, in order to establish a beneficial microbial population to prevent disease. The role that organic acids and their salts can play as growth promoters in the South African abalone Haliotis midae, and as manipulators of the gut microflora of this species of abalone was investigated and compared to the effects of antibiotics. Three different treatments were tested against a negative control and a positive control containing 30ppm avilamycin, a commercial antibiotic growth promoter (AGP) used in the pig and poultry industry. The 3 treatments consisted of 1% acetic and 1% formic acid (treatment AF), 1% sodium benzoate and 1% potassium sorbate (treatment SBPS), and 1% benzoic and 1% sorbic acid (treatment BS). Three different experiments were conducted to test the effects of the different acids and salts. The first experiment was under controlled optimum water temperature conditions (16.5ºC), another at elevated water temperature (20.5ºC) in order to test response during temperature stress conditions, and the final trial was conducted under uncontrolled practical production conditions. In an attempt to establish the mechanism by which the treatments have their effects (if any), the composition of the gut microflora of the abalone was monitored. It was found that the organic acids and salts investigated can enhance the growth rate of Haliotis midae in the size class 23 mm to 33 mm mean length significantly when compared to both control treatments. It was further found that the tested AGP had no effect on growth rate. None of the treatments had a significant effect on feed conversion ratio (FCR), Incidence cost (IC) or feed intake. It could also not be shown that the treatments affected the intestinal microflora of the abalone, although this might be due to inadequate microbiological methods. The mechanism by which the acids and salts have their effects could not be established. It was found that the animals in the controlled system underwent an initial adaptation period, which led to improvement in specific growth rate (SGR), FCR and IC as the experiment progressed during the controlled optimal conditions experiment. Large differences in FCR and IC was seen for controlled optimal conditions and production conditions which means that there is still a large scope for developing methods to improve practical on-farm feed utilisation by abalone. SGR, FCR and IC were negatively influenced by raising water temperature from 16.5ºC to 20.5ºC. The composition of the gut microflora of the abalone also changed significantly after the water temperature was raised. It appears that animal weight gain and shell growth respond differently to changing water temperatures, which is reflected in a change in Fulton condition factor. A relationship between the length and weight of abalone between 15 mm and 47 mm was established and it was found that Haliotis midae does not follow an isometric growth relationship. This relation can be used as a tool to improve farm management and therefore also profitability. Various micro-organisms were isolated from Haliotis midae during the trial, but their relationship and interaction with abalone is not clear. Clear dominance by specific species of bacteria was observed during certain periods. The current research has clearly showed the potential of organic acids and their salts to act as growth promoters in the South African abalone Haliotis midae, with application in both the local aquaculture and feed manufacturing industries. The possibility further exists that some aspects of the current research can be adapted to be applicable in other abalone species and even in other aquaculture species.
AFRIKAANSE OPSOMMING: Die eerste suksesvolle aanteel van die Suid-Afrikaanse perlemoen Haliotis midae in gevangeskap is in die 1980’s gerapporteer, waarna ‘n suksesvolle akwakultuur industrie ontwikkel het met ‘n geskatte produksievermoë van 500 tot 800 ton en kapitaalbelegging van US$ 12 miljoen in 2001. Suid-Afrika is tans die grootste perlemoen-produserende land wat buite Asië geleë is. Die ontwikkeling van ‘n geskikte alternatiewe voedselbron vir natuurlike kelp (tans die algemeenste voedselbron wat gebruik word in die kweek van perlemoen), sowel as verbeterde siektebestryding word tans gesien as die hooffaktore wat verdere uitbreiding in die Suid-Afrikaanse industrie beperk. Perlemoen het verder baie stadige groeitempo’s en enige verbetering in hierdie verband sal produksietye verkort en dus produsente bevoordeel. Siektes in akwakultuur word tradisioneel bestry deur gebruik te maak van antibiotiese behandeling (terapeutiese bestryding) of van voorkomende behandeling (profilaktiese bestryding). In gewone diereproduksie-sisteme (bv. varke en hoenders) word antibiotika ook gebruik as groeistimulante. Die gebruik van antibiotika in akwakultuur het onlangs die bekommernis laat ontstaan dat sekere menslike en dierepatogene weerstand kan ontwikkel teen sommige middels, wat die behoefte laat ontstaan het om siektebestryding sonder die gebruik van antibiotika te ontwikkel. Alternatiewe strategieë fokus grootliks daarop om die samestelling van die mikrobiese bevolking van die gasheer te manipuleer en sodoende ‘n voordelige bevolking in die gasheer te vestig, wat dan siektes voorkom. Daar is ondersoek ingestel na die rol van organiese sure en hul soute as groeistimulante en manipuleerders van die mikrobiese bevolking in die Suid-Afrikaanse perlemoen Haliotis midae. Drie verskillende behandelings is getoets en vergelyk met beide ‘n negatiewe- en positiewe kontrole (wat 30 dele per miljoen van ‘n kommersiële antibiotiese groeistimulant bevat het). Die drie formulasies het onderskeidelik bestaan uit ‘n mengsel van 1% etanoëen 1% metanoësuur (behandeling AF), 1% bensoë- en 1% sorbiensuur (behandeling BS) en 1% natriumbensoaat en 1% kaliumsorbaat (behandeling SBPS). Om die effekte van hierdie formulasies te toets, is daar 3 proewe gedoen. Een proef is gedoen onder temperatuurbeheerde toestande teen ‘n optimum watertemperatuur van 16.5ºC terwyl ‘n ander gedoen is onder onbeheerde, praktiese produksie-omstandighede. ‘n Verdere beheerde proef is gedoen teen ‘n watertemperatuur van 20.5ºC om die effek van die verskillende formulasies te toets wanneer die diere aan temperatuur-spanning blootgestel word. Die samestelling van die mikrobiese bevolking in die dunderm van die perlemoen is deurentyd gemonitor in ‘n poging om die meganisme vas te stel waarvolgens die sure en soute hul effek het, indien daar enige effek waargeneem word. Daar is gevind dat die onderskeie sure en suursoute die groeitempo van Haliotis midae met ‘n gemiddelde lengte van 23 mm tot 33 mm beduidend kan verhoog in vergelyking met die groeitempo’s van beide kontroles. Daar is gevind dat die antibiotiese groeistimulant geen effek het op die groei van die diere nie en dat geen behandelings ‘n beduidende effek op voeromsetting, voerkoste of voerinname gehad het nie. Daar kon nie bewys word dat enige van die formulasies of die antibiotika ‘n effek gehad het op die mikrobes in die spysverteringskanaal van die perlemoene in die sisteem nie, alhoewel die gebrek aan ‘n effek moontlik toegeskryf kan word aan die onakkurate en onvoldoende mikrobiologiese metodes wat gebruik is tydens die studie. Die meganisme waarvolgens die sure werk kon nie vasgestel word nie. Daar is verder gevind dat die diere in die temperatuur-beheerde eksperiment aanvaklik deur ‘n aanpassingsperiode gegaan het, wat tot gevolg gehad het dat die spesifieke groeitempo, voeromsetting en voerkoste verbeter het met die verloop van die eksperiment. Daar is groot verskille gevind in die voeromsetting van beheerde optimale toestande en onbeheerde produksietoestande, wat impliseer dat daar nog baie ruimte en geleenthede is om metodes te ontwikkel wat beter voeromsetting bewerkstellig tydens perlemoenproduksie. Spesifieke groeitempo, voeromsetting en voerkoste is nadelig beïnvloed toe die watertemperatuur verhoog is vanaf 16.5ºC na 20.5ºC. Die samestelling van die mikrobiese bevolking in die spysverteringskanaal van die perlemoen het ook beduidende veranderinge ondergaan tydens hierdie temperatuur verhoging. Dit wil voorkom asof die lengtegroei van die dop en die toename in massa verskillend reageer op ‘n verandering in watertemperatuur en hierdie effek word weerspieël in die verandering in Fulton-kondisiefaktor. ‘n Verwantskap tussen totale doplengte en totale gewig van Haliotis midae kon vasgestel word vir diere tussen 15 mm en 47 mm en daar is gewys dat H. midae nie ‘n isometriese groeipatroon volg nie. Hierdie verwantskap kan aangewend word tydens produksiebestuur om produksie te verbeter en daardeur ook winsgewendheid te verhoog. Verskeie mikrobes is tydens die verloop van die proef geïsoleer, maar die rol van en interaksie tussen hierdie mikrobes en die Suid-Afrikaanse perlemoen is nie duidelik nie. Sekere bakterieë het die mikrobiese bevolking in die spysverteringskanaal van die perlemoen in hierdie proef oorheers tydens sekere groeiperiodes. Die huidige navorsing het duidelik aangetoon dat organiese sure en hul soute as groeistimulante kan optree in die Suid-Afrikaanse perlemoen Haliotis midae, met toepassings in die plaaslike akwakultuur- en voervervaardigins-industrieë. Dit beskik verder oor die potensiaal om aangepas te word sodat dit toepaslik is in ander perlemoenspesies en selfs ander akwakultuur organismes.

Thesis (MSc)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The species Haliotis midae is the only commercially exploitable abalone species of the six found in the South African coastal waters. This species is under substantial pressure from both legal and illegal harvesters, to such an extent that it could be commercially extinct within four years. Efforts to alleviate the pressures on the natural populations of both illegal and legal harvesting are being made. The genetic management systems for abalone farming and ranching activities should be carefully evaluated. The loss of genetic diversity and the risks of contaminating the gene pools of natural populations in the vicinity of a farm should be minimized. Genetic evaluation studies will be at great importance to acquire the necessary data needed for genetic diversity and differentiation analysis. The aim at this study was to develop species-specific microsatellite DNA markers to assess the genetic diversity and differentiation within and between the brood stock and commercial stock of the Sea Plant Products abalone farm (Hermanus, Republic of South Africa) and natural populations related to the brood stock. The species-specific DNA markers were also used for parentage assignments within the tarm population (first for abalone) and preliminary OTL (quantitative trait loci)-discovery analysis studying growth rate segregation. Samples were taken of the farm's brood stock and commercial stock (Rows 2, 3, 4) as well as from two natural populations (Saldanha Bay and Black Rock) related to the brood stock. Various statistical parameters and software packages were used to assess genetic diversity and differentiation, to infer parentage and to look for OTL's. Eight species-specific microsatellite DNA markers were designed and used for data analysis. Data analysis showed a loss at genetic diversity from the brood stock to the commercial stock caused by the subdivision of the original brood stock into rows and the differential contributions of parents to the offspring. No genetic differentiation (Fst) was detected between the farm and natural populations, except for the offspring of Row2. levels of inbreeding (ns) were high for all loci within the populations. Thirty-eight percent of all studied offspring were confidently assigned to a couple. The preliminary QTldiscovery suggested the segregation of a number of alleles and genotypes with growth rate. The study concluded that the commercial abalone population of the Sea Plant Products abalone farm holds no threat to the disruption of the genetic diversity of the natural populations. It is proposed that the farm implement a rotational breeding program to increase the genetic diversity of the commercial population. Any newly acquired brood stock must be profiled before their introduction into the breeding program to assess the influence of the animals on the current levels of genetic diversity within the farm. The accuracy and reliability of parentage assignments and QTl-discovery need to be optimised by adding more loci and sampling more animals or even by trying and developing new methods.
AFRIKAANSE OPSOMMING: Van die ses perlemoen spesies wat langs die Suid-Afrikaanse kus gevind word, is die spesie Haliotis midae die enigste een wat van kommersiële belang is. Wettige, sowel as onwettige versameling, plaas hierdie spesie onder sulke geweldige druk dat dit dalk binne vier jaar verlore kan wees vir die kommersiële bedryf. Verskeie strategieë word tans geïmplimenteer om hierdie druk te verlig. Die genetiese bestuurstrategieë binne perlemoen plase moet deeglik ondersoek word. Die verlies aan genetiese diversiteit en die moontlikheid vir die kontaminering van die natuurlike populasies in die omgewing van die plaas se geenpoel, moet uitgeskakel word. Genetiese evaluasies sal van groot belang wees om die nodige data vir genetiese diversiteit- en differensiasie-analises te verkry. Die doel van die studie was om spesies-spesifieke mikrosatelliet DNA merkers te ontwikkel wat gebruik sou word om die genetiese diversiteit en differensiasie binne en tussen die broei diere en die kommersiële diere van Sea Plant Products se perlemoen plaas (Hermanus, Republiek van Suid- Afrika) en die natuurlike populasies wat verwant is aan die broei diere, te bepaal. Die spesies-spesifieke DNA merkers is ook vir ouerskap-bepalings binne die plaas se populasie gebruik, asook vir voorlopige OTL (quantitative trait locI) - ontdekking met betrekking tot groeitempo segregasie. Monsters van die plaas se broei diere en kommersiële diere (Ry 2, 3, 4) asook van twee natuurlike populasies (Saldanha Baai en Black Rock) wat verwant is aan die broei diere, is geneem. 'n Verskeidenheid van statistiese parameters en sagteware pakette is vir die genetiese diversiteit- en differensiasie-analises, vir ouerskap-bepalings en vir die opspoor van OTL's gebruik. Agt spesies-spesifieke mikrosatelliet DNA merkers is ontwerp en toe gebruik vir die data analises. 'n Verlies aan genetiese diversiteit vanaf die broei diere na die kommersiële diere is deur die data analises uitgewys. Dit is veroorsaak deur die verdeling van die oorspronklike broei diere in rye en die differensiële bydraes deur die ouers na die nageslag. Geen genetiese differensiasie (Fst) is tussen die plaas se populasie en die natuurlike populasies gevind nie, maar die nageslag van Ry 2 het wel differensiasie getoon. Die vlakke van inteling (Fis) was hoog oor alle lokusse binne alle populasies. Agt-en-dertig persent van die nageslag wat bestudeer is, was suksesvol met 'n ouerpaartjie geassosieer. Die voorlopige OTL-ontdekking studie toon die potesiële segregasie van 'n aantal allele en genotipes saam met groeitempo. Die bevindinge van die studie is dat die kommersiële populasie op die Sea Plant Products perlemoen plaas, geen bedreiging vir die versteuring van die natuurlike populasies se genetiese diversiteit inhou nie. Daar is voorgestel dat die plaas 'n rotasie-basis broeiprogram moet implimenteer om sodoende die genetiese diversiteit van die kommersiële populasie te verhoog. Enige nuwe broei diere se genetiese profiel moet ook voor die tyd ondersoek word om te sien wat se effek hierdie diere op die huidige genetiese diversiteit van die kommersiële populasie sal hê. Die akkuraatheid en betroubaarheid van die ouerskap-bepalings en OTL-ontdekking moet optimiseer word deur of meer lokusse te bestudeer, of meer diere in analises te gebruik of selfs om nuwe tegnieke te probeer of te ontwikkel.

An adequate supply of diatoms during the weaning stage (generally 5 - 10 mm shell length (SL)) is one of the primary constraints to the commercial culture of the South African abalone, Haliotis midae. Because of the seriousness of the problem, a project aimed at the development of an artificial weaning diet was initiated. Initially, the chemical composition (proximate composition, amino acid, fatty acid and mineral element profile) of juvenile H. midae was analyzed, as a general lack of such information was identified in a review. Due to the lack of knowledge on the nutritional requirements of H. midae, the formulation of the weaning diet was based on the essential amino acid (EAA) pattern of the shucked tissue, and the known nutrient requirements of haliotids. Subsequently, a water stable gel and pellet form of the diet were developed. The best water stability of a gel was obtained with a 1:3 agar/gelatine mixture which retained 70.7 ± 2.7 % of its dry weight after 24 h. Starch based pellets, however, retained 89.0 ± 0.6 % of their dry weight after 24 h. In a comparative growth trial, pellets produced a significantly better increase in SL and weight than gels after only 15 days. This was probably due to the better water stability of pellets, which resulted in a better nutritional quality than in gels. The feeding behaviour on both forms of the diet did not differ. Activity patterns were exclusively nocturnal and feeding frequency was consistently low. The percentage composition of the pelleted weaning diet, on a dry weight basis, was 5 % casein, 15 % gelatine, 15 % fish meal, 10 % Spirulina spp., 2.5 % fish oil, 2.5 % sunflower oil, 21.0 % dextrin, 23.0 % starch, 4.0 % of a mineral and 2.0 % of a vitamin mixture. The correlation coefficient between the EAA pattern of H. midae and the dietary EAA pattern was r⁷= 0.8989. Pellets were fed to juveniles in a 30 day growth trial to study the effect of photoperiod (12, 16, 20 and 23 hours of darkness) on growth and general nutritional parameters. A comparative experiment feeding diatoms was conducted under a 12hL: 12hD light regime at the same time. The SL and weight of the juveniles did not increase significantly with an increase in hours of darkness. The growth of juveniles fed on pellets did not differ significantly from those fed on diatoms. Percentage feed consumption (PFC), percentage feeding rate (PFR), feed conversion ratio (FCR), protein efficiency ratio (PER) and percentage protein deposited (PPD) were determined for the animals fed on pellets. None of the parameters were significantly affected by photoperiod. However, there were trends in that PFC increased with longer periods of darkness, while PPD decreased. The FCRs (0.44 ± 0.04 to 0.60 ± 0.19) and PERs (5.06 ± 1.74 to 6.64 ± 0.77) indicated that juveniles used the feed, and in particular the protein, very efficiently. Photoperiod did not have an effect on the specific activity of the digestive enzymes amylase, protease and lipase. The specific activity of amylase in the juveniles fed on diatoms was significantly higher than in the pellet fed groups. This was surprising as the main carbohydrate of diatoms is the ß-(l-3) glucan chrysolaminarin, and not starch, a ß-(l-4) glucan. Protease specific activity, on the other hand, was significantly higher in the pellet fed groups, indicating an ability to adapt to the high protein content in the artificial diet (35.48 %), compared to diatoms which had a protein content of 5 %. The specific activity of lipase did not differ significantly between groups, probably because of a similar lipid concentration (5 - 10 %) in diatoms and pellets. Finally, the effect of stocking density, ranging from 1250 to 10,000 juveniles/m2, on the growth of juveniles was evaluated. A model of hatchery productivity was developed based on this investigation. Hatchery productivity was defined as the number of juveniles per unit space reared through to the grow-out stage per unit time. The model predicted that maximum productivity would be achieved at a stocking density of 10,000 juveniles/m2. The results have shown that H. midae can be successfully weaned on an artificial diet, as the growth on the diet was not significantly different to growth obtained on diatoms. Long-term growth trials are needed to confirm these results. The importance of standardized experiments on the nutritional requirements and digestibility of abalone was emphasized. The importance of improved artificial diets, optimal culture conditions, as well as the application of biotechnological techniques to further abalone aquaculture was highlighted.

The abalone (Haliotis midae) culture industry in South Africa is becoming increasingly dependent on the use of formulated feeds, due to limitations in the supply of kelp. The bulk of the feeds that are currently available were developed based on the requirements of juvenile abalone cultured within the optimal temperature range for growth (18 - 20 °C). However, most abalone farming facilities are land-based pump ashore operations and are thus mostly exposed to temperatures outside of this range. In addition, these feeds have been found to be unsuitable for abalone cultured at elevated water temperatures (> 20°C). The aim of the study was to develop size and temperature specific diets for H midoe through optimisation of dietary protein, energy and lipid levels. Abalone were cultured under farm-like conditions in three partially recirculating temperature controlled systems at either 18, 22 or 24°C and fed formulated diets containing graded levels of protein (18,22 and 26 %) and energy (11.6, 13.5 and 16.2 MJ.kg·I ). Abalone were stocked into baskets at 5 % of available of surface area (n=36) and each diet (n=9) was fed to four baskets of abalone at each of the three temperature regimes for ten weeks. Abalone growth was temperature dependent, with growth declining from 4.33 g.month-I for abalone cultured at 18°C to 0.77 g.month-I at 24°C. Dietary protein could be reduced from 26 to 18 % provided dietary energy levels were maintained at 13.5 MJ.kg- l • A dietary energy level of 11.6 MJ.kg-1 was insufficient to meet the energetic requirements of H midae regardless of the protein content of the diet. The effects of water temperature and body size on the protein requirements of H midae were investigated by culturing abalone at temperatures within the optimal range for abalone farming (i.e. 14, 16 and 18°C). Three size classes of abalone (15, 50 and 80 mm) were fed formulated feed containing graded levels of dietary protein (20, 26, 32, 38 and 44 %) under controlled laboratory conditions for 12 weeks, and, in a separate experiment, under commercial farm conditions for 24 weeks. It was not possible to convincingly define the optimal protein levels for abalone of different sizes in this experiment because growth rates fell below average commercial growth rates obtained on farms. Growth was temperature dependent in the laboratory trial, with the rate of weight gain of the 15 mm (ANOV A: p=0.002) and 50 mm abalone (ANOV A: p=0.02) increasing significantly with an increase in temperature from 14 to 18°C. In the farm trial, dietary protein content did not affect the growth rate of the 10-15 or 80 mm abalone (ANOVA: p>0.05), however, the 50 mm abalone displayed significantly higher weight gain on the 32 % (4.72±0.20 g.month-I ) and 38 % (5.01±0.34 g.month-I ) protein diets compared to those fed the 20 % protein diet (3.75±0.13 g.month-I ) (ANOVA: p=O.OI). Although definition of optimal dietary protein levels were not possible, the effects of dietary protein content and water temperature on the growth of H midae were independent signifying that the protein requirements of abalone are temperature independent. In addition, there was no evidence to indicate that abalone of the different sizes tested here had different dietary protein requirements. The size specific dietary lipid and protein requirements of H midae were investigated by feeding two size classes of abalone (30 and 60 mm initial shell length) diets containing graded levels of dietary lipid (4, 7, 10, 13 and 16 %) and protein (34 - 39 %) for 12 weeks. The 30 and 60 mm abalone were stocked at 7 (n=200) and 9 % (n=36) of the available basket surface area respectively and each diet was fed to four baskets of abalone of each size class. The protein requirements of H. midae are influenced by the amount of available dietary energy and thus it is possible that the ability of abalone to utilise lipids as a source of energy differs in the presence of varying levels of dietary protein. High levels of dietary lipid negatively affected the growth, condition factor and soft tissue glycogen content of both size classes of abalone. This negative effect was greater in the 30 mm size class compared to the 60 mm abalone. The corresponding increase in feed consumption and feed conversion ratio in response to increasing levels of dietary protein also provides evidence that abalone are unable to utilise dietary lipids as an energy source and high levels of dietary lipid probably inhibit the uptake of carbohydrates and protein. High dietary lipid levels did however appear to promote gonad maturation. It was possible to reduce dietary protein from 34 to 20 % without negatively affecting growth through the maintenance of dietary energy levels and thus it is recommended that future experiments on the energy content of formulated feeds should focus on the improved use of carbohydrates. Reductions in the protein portion of formulated feeds for H. midae are possible provided the diet contains sufficient levels of energy supplied from carbohydrates. As the ability of abalone to utilise dietary lipid is limited, lipids are unlikely to play a significant role as an energy source in abalone feeds. Further investigations should focus on the utilisation of various carbohydrate sources in abalone feeds.

Abalone Haliotis midae farming in South Africa is highly intensive, employing pump-ashore, flow-through systems. Despite the known sensitivity of abalone to water quality, there is only a rudimentary understanding of water quality dynamics on South African abalone farms and its effects on abalone production. Furthermore, the potential for reusing the relatively dilute abalone farm effluent to culture other animal species has not been investigated. This study investigated the dynamics of water quality and growth on a South African abalone farm and assessed the suitability of the effluent for the culture of silver kob Argyrosomus inodorus and bloodworm Arenicola loveni loveni. Monitoring of water quality and abalone growth in abalone tanks revealed that oxygen concentrations decreased, while H⁺ ion and free-ammonia (NH₃) concentration increased in a gradient between the inflow and outflow. Abalone growth was positively correlated with oxygen concentration and negatively correlated with free-ammonia and H⁺ ion concentration. The oxygen (O) concentration of the farm influent was dependent upon the influents’ temperature (T) and was described by the relationship O (mg L⁻¹) = 11.244 – 0.208T (r²=0.74). Linear regression analysis of data collected from abalone farm tanks revealed that the concentration of total ammonia at the outflow of abalone tanks (μg TAN L⁻¹) was dependant upon temperature (°C), flow-rate (L s⁻¹ kg⁻¹ H. midae), abalone size (g) and length of time since the tank was last cleaned (d) (n = 125, r² = 0.80). The production of total ammonia (μg TAN s⁻¹ kg⁻¹) was related to temperature, abalone size and days that the tanks remained un-cleaned (n = 125; r² = 0.81). A diurnal cycle of respiration was evident in abalone tanks with higher oxygen consumption and H+ ion production at night. The oxygen concentration of farm effluent was related to temperature, farm biomass and flow rate by means of a linear regression equation (n = 40; r² = 0.69). The results demonstrated the importance of optimising the flow-rate per unit of biomass for various temperatures and sizes of abalone. As abalone size and temperature cannot be controlled under farm conditions, the flow-rate per unit of biomass which the abalone culture system receives will determine the quality of the culture water. The specific growth rate (0.48 ± 0.01 % BW d⁻¹), mortality (1.8 ± 0.5 %), feed conversion ratio (3.0 ± 0.2) and protein efficiency ratio (1.0 ± 0.1) of silver kob kept in either abalone farm effluent or control seawater for 120 days did not differ significantly (t-test, P>0.05). A 90 day growth trial indicated that abalone farm effluent is a suitable culture medium for bloodworm. Bloodworm supplied with control seawater lost weight at 0.19 ± 0.04 % BW d-1, while those given abalone effluent grew at 0.39 ± 0.07 % BW d⁻¹. Mortality was 6 ± 3 % in effluent and 11 ± 8 % in seawater. The bloodworm were efficient at processing solid waste. Abalone farm effluent initially contained 7.7 ± 13 mg L⁻¹ more suspended solids than control seawater, which contained 3.5 ± 0.5 mg L⁻¹, but after passing through bloodworm systems the concentration in abalone effluent was reduced to only 1.4 ± 3.5 mg L⁻¹ above that in control seawater. Therefore, abalone farm effluent could be reused as a culture medium for both silver kob and bloodworm. Future work is needed to investigate aspects of the feasibility of such systems such as growth rates at different sizes and stocking densities.

Inadequate information of the nutritive physiology and the dietary requirements of abalone are the principle factors that currently limit the development of a formulated feed for the commercial culture of Haliotis midae. The need to develop a method to determine apparent digestibility co-efficient's for abalone in order to facilitate further applied nutritional research was identified. Animals between 50 and 80 mm were collected from natural stocks along the east Cape coast of South Africa at Port Alfred and Great Fish point, and acclimated to laboratory conditions. Initial trials demonstrated that H. midae accepted and preferred a semi-purified diet to the seaweed Plocamium corallorhiza, one of the main components of it's natural diet. A technique of determining apparent digestibility co-efficient's (ADC) using the indirect method with chromic oxide as an inert marker was developed. Digestibility trials yielded higher dry matter (DMADC) and crude protein apparent digestibility co-efficient's (CPADC) for the semi-purified diet than for two species of algae, Gelidium amanzii and P. corallorhiza (83.7% and 95.6%, 70.7% and 80.0%, and 29.9% and 57.3% respectively). The ability of the animals to utilize terrestrial animal and plant ingredients efficiently makes it feasible to use conventional feed ingredients in formulated feeds for H. midae. Trials to determine the effect of different temperatures (15°C, 18°C and 22°C) on DMADC and CPADC of the semipurified diet showed that peak digestibility occurred at 18°C. There was also a positive relationship between temperature and consumption rate. Although no enzyme studies with H. midae have been conducted, the peak ADC's at 18°C is attributed to an increase in enzyme activity at this temperature. Transit time, an inverse function of temperature and consumption, is considered to be responsible for the decrease in the ADC' s at 22°C in conjunction with a possible decrease in enzyme activity at this temperature. A photoperiod trial to investigate the effect of darkness on DMADC and CPADC of the semi-purified diet revealed that digestive efficiency decreased with increasing hours of darkness. There was also a positive relationship between duration of darkness and the rate of consumption. The decrease in ADC's is attributed to decreased transit times as the duration of darkness increased . The contribution of this project to the understanding of abalone nutrition, the development of a formulated abalone feed and systems design for abalone farms is discussed.

Haliotis midae is the cornerstone of the South African abalone fishery. For more than a decade, the wild abalone stock of South Africa has suffered decline due to over-exploitation and illegal activities such as poaching. Prior to 1970, no regulations were in place concerning the annual landings. As a result the fishery was exploited as if it were an infinite resource. It is this initial uncontrolled harvesting (regardless of age) and poaching that has driven the abalone resource decline. Due to the slow growth rate exhibited by abalone as a species, natural replenishment of wild stock following exploitation and poaching was far below the rate of exploitation of this resource. Studies on the growth of abalone have mainly been conducted under laboratory conditions. The purpose of this study was to measure the growth of abalone, fed different diets, under commercial culture conditions. Three food types were used namely; commercial pellets, seaweed (Ulva spp.) and dried kelp bars (Ecklonia maxima). Four diets were obtained from the three food types namely; combination of commercial pellets and seaweed (Diet A), commercial pellets only (Diet B), seaweed only (Diet C) and dried kelp bars only (Diet D). The food types used in this study represent both artificial (Commercial pellets) and natural feeds (seaweed and kelp) used in commercial abalone culture. The growth of two cohorts (40-50 mm and 50-60 mm) was followed over a 426 day period, with data for the first 183 days being used for statistical analysis to determine performance of a given diet. The best growth rates were found in abalone fed Diet A (40-50 mm: 2.64 mm.month-1; 50-60 mm 2.78: mm.month-1) and B (40-50 mm: 2.20 mm.month-1; 50-60 mm: 2.35: mm.month-1). These (Diets A and B) gave higher growth rates when compared to Diets C and D (natural diets), whose growth rates ranged between 0.50 mm.month-1 and 1.71 mm.month-1 for both cohorts. Also observed in this study was that, the mixture of formulated diet and seaweed gave better growth than formulated diet given exclusively.

Thesis (MSc)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The indigenous abalone, Haliotis midae has been a successfully cultured aquaculture species in South Africa since 1990. It has a slow growth rate and takes from two to five years to reach market size. Like for most other commercially important abalone species, the slow growth rate of H. midae is a cause of concern with regard to the profitability of farming and global competitiveness of the species. Ploidy manipulation of the maternal genome, a universally growing practice in shellfish culture, is considered a promising method to improve the growth rate of abalone - a desirable trait in aquaculture organisms from a commercial perspective. This manipulation technique is employed to achieve sterility, which results in limited gonad development. The consequent re-allocation of resources to somatic growth results in improved growth. The purpose of this study was to establish a viable method for the induction and validation of triploidy, on a commercial scale, in the South African abalone, H. midae. The focus was on hydrostatic pressure as a method of induction and flow cytometry as the method of validation. The results obtained confirm hydrostatic pressure as an effective method for the induction of triploidy in H. midae, delivering high percentages of triploidy (>80%) over a wide range of pressures and times, in 48 hour-old larvae. Hydrostatic pressure had a negative effect on survival in 20 hour-old larvae. Flow cytometry was validated as a reliable, fast and accurate, though expensive, method for identification of triploidy in H. midae. As an outcome of this study a manual of “Procedures for the Induction and Validation of Triploidy in the abalone” is presented (Appendix 1) together with recommendations for further studies on triploidy in the South African abalone, H. midae.
AFRIKAANSE OPSOMMING: Die inheemse perlemoen, Haliotis midae, is sedert 1990 ‘n suksesvol gekweekte akwakultuur spesie in Suid-Afrika. ‘n Kenmerk van die spesie is die stadige groeitempo van tussen twee en vyf jaar ten einde bemarkbare grootte te bereik. Soos vir die meerderheid perlemoen van kommersiële belang, is hierdie stadige groeitempo rede tot kommer met betrekking tot die winsgewende kweek en wêreldwye mededingendheid van die spesie. Die manipulasie van ploïdie van die moederlike genoom is ‘n toenemende praktyk in skulpvisboerdery en word gereken as ‘n belowende metode om die groeitempo van perlemoen te verbeter. Hierdie manipulasietegniek word gebruik om steriliteit te verkry wat manifesteer as onderdrukte ontwikkeling van die geslagsklier. Die gevolg is die herkanalisering van bronne na somatiese groei. Die doel van hierdie studie was om ‘n lewensvatbare metode vir die induksie van triploïdie op ‘n kommersiële skaal in die Suid-Afrikaanse perlemoen, H. midae, te vestig. Daar is op hidrostatiese druk as metode vir die induksie en vloei-sitometrie as metode vir die geldigverklaring van triploïdie gefokus. Die resultate van hierdie studie bevestig dat hidrostatiese druk ‘n effektiewe metode vir die induksie van triploïdie in H. midae is. Hoë persentasies van triploïdie (>80%) is oor ‘n wye reeks van drukke en tye in 48 uur oue larwes verkry. Daar is gevind dat hidrostatiese drukbehandeling ‘n negatiewe effek op die oorlewing van 20 uur oue larwes het. Vloei-sitometrie is bevestig as ‘n betroubare, vinnig en akkurate, maar duur metode vir die identifikasie van triploïdie in H. midae. As ‘n uitvloeisel van die studie word ‘n handleiding “Procedures for the Induction and Validation of Triploidy in the abalone” (Appendix 1) aangebied tesame met aanbevelings vir verdere studies rakende triploïdie in die Suid-Afrikaanse perlemoen, H. midae.

According to farm records, cultured Haliotis midae (50-70 g.abalone⁻¹) were growing 10% slower in winter when compared to summer. This reduction in growth rate also coincided with enlarged gonads. Initial trials showed that there were differences in mean monthly growth rates ranging from 1.97 – 5.14 g abalone⁻¹ month⁻¹, and gonad bulk index (GBI) also varied between months (GBI range: 26.88 ± 12.87 to 51.03 ± 34.47). The investment of energy into gonad tissue growth did not compromise whole body growth as the abalone continued to gain weight throughout the reproductive periods, probably due to gonadal growth. Growth of this size class of abalone was not influenced by water temperature or day length, suggesting favourable on-farm culture conditions (regression analyses, p > 0.05). There is no need to implement a seasonal dietary regime. Cultured H. midae were fed artificial diets with different protein sources, including only soya, only fishmeal, a combination of soya and fishmeal, and these were compared to kelp-fed abalone. Kelp-fed abalone grew slower than those fed artificial feeds (p>0.05). Gonad growth was the greatest when soya meal was included in the diet (average GBI: 74.91 ± 23.31), while the average gonad size of abalone fed the fishmealbased diet had gonads which were 38% smaller, and kelp-fed abalone had gonads which were 75% smaller than those of the abalone fed on diets containing soya meal. The increased gonad mass in abalone fed on diets including soya meal could be attributed to phytoestrogenic activity, as a result of the presence of isoflavones found in the soya plant; this remains to be tested. The use of soya in brood stock diet development is advised. The influence of dietary protein to energy ratio (1.41 – 2.46 g MJ⁻¹) on growth and gonad size was tested. Protein and energy levels within the ranges tested (22 and 33% protein; 13.5 and 15.6 MJ kg⁻¹) did not interact to influence growth rates of cultured H. midae. GBI increased from 50.67 ± 4.16 to 83.93 ± 9.35 units as a function of dietary protein to energy ratio (y = 42.02 x⁰·⁸¹; r² = 0.19; regression analysis: F₁¸₃₈ = 8.9; p = 0.005). In addition, protein level influenced gonad size, with gonad growth being greater in abalone fed the high protein diet (factorial ANOVA: F₁¸₃₂ = 7.1, p = 0.012). Canning yields were reduced by 7% when the protein content was increased, while increasing the quantity of dietary energy improved canning yields by ~ 6% (one-way ANOVA: F₁¸₂₈ = 14.4, p= 0.001). The present study provided evidence that although growth rates are varying seasonally, reproductive investment is not hindering weight gain. Gonad growth can be influenced if desired by farms, depending on the level of soya inclusion, as well as the protein to energy ratio in the diet. Monthly variation in growth and gonad size, as well as the influence of diet on gonad growth were highlighted, and the implications for farm application and further research were discussed.

Terebrasabella heterouncinata is a small K-selected sabellid polychaete. It is a simultaneous hermaphrodite with a semi-continuous mode of reproduction, producing relatively few large eggs that are brooded within the parental burrow until the larvae emerge, to settle on the growing edge of the abalone shell. Despite its low fecundity, this worm has become problematic on abalone farms in South Africa. The present study was conducted to gain an understanding of the life history patterns of T. helerouncinata to determine how they contributed to the success of these worms under altered conditions. This study demonstrated that conditions prevalent on abalone farms were conducive to enhancing the reproductive success of this worm, and suggests that larger, more fecund worms may have been selected for in the decade that these worms have been present on the farms. Increased nutrient availability, and possibly the increased stability of the farm environment relative to its natural environment, has led to a 1.5-fold increase in the average size of the worms. Body size was found to be positively correlated with brood size, and this resulted in worms on farms brooding 3 to 4.5 times more offspring at a time than worms from wild abalone. The ability to increase the number of eggs produced at a time may have been limited by the fact that these worms have only two ovaries. Thus, the increase in fecundity may have been related primarily to the increase in the rate at which the eggs were laid by the worms on the farms, and the increase in the coelomic space available for the storage of these rapidly developing eggs. The ability to increase the rate at which oocytes develop may be related to the vitellogenic mechanisms employed by these worms. Vitellogenic oocytes are able to incorporate high molecular weight yolk precursors from the surrounding coelomic fluid through endocytotic activity. This may allow the oocytes to increase the rate at which they incorporate yolk material under conditions of nutrient enrichment. The increase in fecundity did not occur at the expense of offspring size and, presumably, quality. The increased reproductive output on the farms was compounded by a proportionate increase in the number of reproducing worms within the population. In addition, these worms are long-lived (worms from farmed abalone reached a maximum age of approximately 40 months) and exhibit negligible senescence. Thus, their reproductive output did not change significantly with an increase in age. Furthermore, the proportion of the reproductive worms did not decrease with an increase in age. Thus, within the age range tested, worms of all ages have the potential to make equal contributions to population growth. While diet and abalone stocking density could not be identified as having a significant effect on reproductive output and infestation rate under intensive culture conditions, it was demonstrated that in a naïve abalone population, the total intensity of infestation increased exponentially with time. This increase may be a consequence of an increase in fertilisation success. These worms continuously produce entaquasperm that are released into the water column. The sperm are collected by other individuals that then store the sperm in a single spermatheca. The ability to store sperm relieves individuals of a dependence on the synchronisation of spawning of eggs and sperm. As the population size and density increases, there could be more individuals releasing sperm into the water column, resulting in a continuous supply of sperm. The increased production of eggs would therefore not be constrained by a lack of sperm. The stored sperm are released into the brood chamber to fertilise eggs as they are laid, and this would probably increase the fertilisation success in the species. This study also provides evidence to suggest that reproduction in this worm has a seasonal component. Future studies should concentrate on measuring fertilisation success in greater detail, measuring the effect of season on reproduction, determining whether there are genetic differences between worms on farmed and wild abalone and determining whether wild worms have similar life-spans and age-related fecundity as worms on farms.

This study investigated the feasibility of ranching the abalone Haliotis rubra (Leach) and Haliotis laevigata (Donovan) on concrete artificial reefs at a site chosen by industry investors on silty bottom off Altona Beach, Port Phillip Bay, Victoria, Australia. The study started from the premise that artificial reef deployment combined with abalone stock enhancement may achieve the dual purpose of habitat and stock improvement. It evaluated the hydrodynamics of the site in relation to artificial reef structural stability and drift algal transport, the development and ecology of the artificial reef assemblage, and abalone enhancement by seeding the reefs with hatchery juvenile abalone and transplanting adult broodstock abalone. Despite a scarcity of reports on culturing abalone on artificial reef, a literature review examined broadly the functions of artificial reefs, particularly in the context of abalone enhancement, and the relationship between artificial reefs, hydrodynamics and ecology. The major finding was that the success of artificial reef projects is highly dependent on the environment of the chosen site, and that preliminary studies are essential to predict their likely success. This consideration alone provided strong justification to undertake this study. The topography at the Altona site was generally flat, with natural patches of low basalt boulder reef, offering low habitat complexity, yet supporting a diverse range of flora and fauna, including a low-density wild stock of H. rubra. Water depth was a mean of 3.3 m at low tide and 4.4 m at high tide. A single, uncomplicated, concrete artificial reef of H-shape design, was tested as abalone habitat. The hydrodynamic analysis confirmed previous studies of Port Phillip Bay, with mild current speeds of mean 0.045 m.s-1, and maximum-modelled wave height (H1/3) of 1.21 m and period (T1/3) of 4.51 sec. Water temperature ranged from 9.9ºC during July to 23.8ºC during January, with salinity averaging of 35.5 ppt. The site had a low probability of receiving drift algae, necessary as a food source for abalone, because of its geographic location, potentially affecting ranch productivity. Ecological monitoring of the three-year old artificial reef shows complex changes in the flora and fauna over time, particularly in respect of the sessile fauna. Key differences between the artificial reef and a nearby natural reef community were: lower cover of corallines and late colonisers, such as sponges. High levels of sedimentation were recorded at the Altona site. Hatchery juvenile H. laevigata, with mean survival of 15% after two years and a mean annual growth rate of 39 mm, showed the most promise for outplanting. In comparison, for hatchery juvenile H. rubra, mean survival was 9% after three years and mean annual growth rate was 22 mm. No natural recruitment of H. laevigata was recorded on artificial reef despite transplants of adult broodstock on the reefs. Natural recruitment of H. rubra was also low and insufficient to reliably contribute to abalone ranch stock. Having examined the hydrodynamic, ecological and enhancement attributes of the Altona site, the study concluded that the site was marginally viable for abalone ranching, and that an alternative site near Werribee, 20 km further southwest, had superior attributes for growth and survival of abalone.

An understanding of species specific water quality requirements is essential for efficient production of aquaculture products, an aspect not well documented for the land-based culture of the South African abalone, Haliotis midae. In order for the industry to remain competitive in international markets, efficient use of water supplies and the development of water reuse technology is needed. This study assessed the changes in water quality between tanks in a tiered serial-use raceway in relation to accumulated biomass and water flow and estimated the flow index (FI) (L h⁻¹ kg⁻¹) at which growth becomes significantly affected. The effect of dietary protein level, supplementation of pure oxygen and addition of sodium hydroxide (NaOH⁻) on water quality and fundamental production parameters in the serial-use raceways was also assessed. The serial-use raceways were used as a tool to create a range of water quality conditions at which the growth, feed conversion ratio (FCR) and condition factor (CF) of "cocktail" size (60 – 70 mm) H. midae could be monitored. The metabolic activity of the abalone resulted in a deterioration in water quality between tanks in series. pH (r² = 0.99; p < 0.001) and dissolved oxygen concentration (r² = 0.99; p < 0.001) were positively correlated with flow index (pH = 7.38 FI°·°² ; dissolved oxygen = 6.92 FI°·°⁴), while free ammonia nitrogen (FAN) (r² = 0.99, p < 0.001) and nitrite (NO²⁻ - N) (r² = 0.93, p < 0.001) were negatively correlated with flow index (FAN = 8.02 FI⁻°·⁷¹). Nitrite concentrations increased over time indicating colonisation of Nitrosomonas bacteria on the basket surfaces. A flow index of 7.2 – 9.0 L h⁻¹ kg⁻¹ was estimated as the minimum to avoid significant reductions in weight and shell length gain and increases in FCR values. Total ammonia nitrogen (TAN) and FAN concentrations were significantly correlated to dietary protein (P) (t = 6.63, p < 0.0001 and t = 6.41, p < 0.0001, respectively) and flow index (t = 5.42, p < 0.0001 and t = 3.9, p < 0.0002, respectively) and could be estimated using the models TAN = 9.73 P – 110.3 log (FI), and FAN = 0.132 P – 1.10 log (FI). Mean FAN concentrations were 67 and 41 % lower in tanks fed a diet containing 22 and 26 % protein respectively, when compared to tanks fed a 33 % protein diet. Supplementation with pure oxygen (103 ± 8 % saturation) improved shell length gain (t = 3.45, p = 0.026) in abalone exposed to high FAN (2.43 ± 1.1 μg L⁻¹) and low pH (7.6 ± 0.13), relative to a treatment with no oxygen supplementation (92 ± 6 % saturation). Addition of a sodium hydroxide solution resulted in elevated mean pH in treatment raceways when compared to control raceways. The increased pH resulted in significantly higher weight gain (g abalone⁻¹) (F₁·₁₂ = 4.51; p = 0.055) and shell length gain (mm abalone⁻¹) (F₁·₁₂ = 4.56; p = 0.054) at an α-error level of < 5.5 %. In two trials, weight gain and shell length gain were significantly correlated to pH (p < 0.001), and multiple regression of pH, dissolved oxygen and FAN consistently revealed pH to be the best predictor of growth. It is therefore suggested that decreasing pH is the first limiting water quality variable for abalone in serial-use raceways. As a decrease in water pH is linked to respiration by the abalone and subsequent increase in dissolved carbon dioxide (CO₂) concentration, future studies should examine the effects of CO₂ on H. midae metabolic rate, calcification rate and health. The results of this study will contribute toward our understanding of the specific water quality requirements for H. midae in commercial aquaculture systems, and influence the design and management procedures for abalone water reuse systems.

Thesis (MScEng)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: Nearly all South African abalone farms function on an intensive pump-ashore, flow-through system. Large volumes of sea water that are pumped ashore flow through abalone or kelp harvesting tanks and finally gravitate back to the ocean. If the effluent from an abalone farm can be desalinated without permanent membrane fouling, then sea water reverse osmosis (SWRO) technology can be integrated effectively with established abalone farms without having to increase the farms’ intake system capacities. Without the need to construct and maintain an intake system, the overall cost of desalination can be reduced. Therefore, the aim of this study was to determine the feasibility and viability of integrating a SWRO desalination plant with a typical South African abalone farm. The project focused on four areas of concern, namely: - characterisation of typical South African abalone farm water - SWRO desalination plant pilot study and membrane fouling behaviour - general operation of a typical abalone farm and its implications for desalination - cost estimates and implications for the integration of an SWRO desalination plant with an abalone farm During a nine-month on-site investigation, sea water turbidity was reduced by up to 43% from a mean value of 0.82 NTU in the influent stream to 0.47 NTU in the combined effluent stream from the abalone tanks. Even with spikes in the influent turbidity, the turbidity of the combined effluent from all abalone tanks (excluding tank flush water) remained below 1 NTU. Dissolved organic carbon (DOC) in both the influent and combined effluent remained below 1 mg/litre. Ultrafiltration (UF) was selected as pre-treatment to the reverse osmosis (RO) in order to minimise potential fouling of the RO membranes. Membrane compaction of both the UF and RO membranes contributed significantly to initial flux losses – as much as 18% for the polyethersulfone (PESM) UF membranes and 20% for the thin film composite (TFC) polyamide RO membrane. However, this is comfortably in line with typical compaction values quoted in the literature. Without pre-flocculation, the UF was able to operate at a specific flux between 45 and 55 litre/m2/h (LMH) and recoveries ranging between 60 and 75%. Corresponding trans-membrane pressure (TMP) ranged between 0.59 and 0.76 bar. With ferric chloride pre-flocculation at a concentration of 3 mg/litre (as Fe3+) the UF could be operated at notably lower TMP values between 0.11 and 0.36 bar. These results indicate that provision should definitely be made for pre-flocculation when using UF as pre-treatment, despite the fact that the DOC concentrations and turbidity of the abalone farm effluent are quite low (DOC <1 mg/litre, NTU <1). It furthermore highlights the inability of DOC and turbidity alone to predict the membrane fouling potential of water. A better indicator of membrane fouling potential, albeit not perfect, is the modified fouling index (MFI0.45). This index follows a linear trend with foulant concentration and serves as a good indicator of the filterability of water. On-site measurements showed an increase in mean MFI0.45 values from 29 s/litre2 for the influent to 48 s/litre2 for the effluent from the abalone tanks, thereby confirming the need for pre-flocculation as part of UF pre-treatment. Chemically enhanced backwashing (CEB) of the UF membrane at least every 24 hours was found to be essential for its stable operation. Therefore, UF with pre-flocculation (3 mg/litre Fe3+) and regular CEB can be used effectively as pre-treatment method for the desalination of abalone farm effluent water. An RO ‘feed-and-bleed’ system was used to simulate the typical performance of the last membrane in a full-scale RO membrane bank. This RO membrane performed well with no signs of extreme fouling. The membrane produced a good quality permeate – for the last membrane in a membrane bank – reducing the TDS of the RO feed from 33 493 mg/litre to 969 mg/litre. These results compared well with simulated values by Reverse Osmosis System Analysis (ROSA; an RO simulator by DOW), indicating a TDS reduction from 33 271 mg/litre to 1 409 mg/litre at a feed pressure of 56 bar, and overall recovery of 44%. A steady performance of the RO membrane during the pilot study indicated that it is possible to desalinate abalone effluent water without notable permanent membrane fouling. A stable normalised flux rate of 8 LMH was achieved and RO membrane integrity remained intact with a salt rejection that ranged from 98.0 to 98.5%. No sudden reduction in permeate flux was observed as a result of fouling by unknown constituents present in the UF permeate. DBNPA (a non-oxidising disinfectant) was dosed once per week at a concentration of 10 – 30 mg/litre for 30 minutes. Scaling was controlled effectively by means of an antiscalant dosed at a concentration of 11 – 12 mg/litre in the feed stream. The CIP frequency was not optimised but a CIP frequency of once every 6 – 8 weeks appeared to be more than adequate to prevent permanent membrane fouling Advantages of integrating an SWRO desalination plant with a South African abalone farm include: - no lengthy and costly environmental impact assessment (EIA) is required to build a new intake system - shared capital and operational cost of intake system - dual incentive to keep constant good quality water flowing through the farm - early warnings regarding occurrences such as algal bloom and red bait - shared operational and management cost to keep pipelines clean - electricity saved (pumps for intake system) Disadvantages of integrating an SWRO desalination plant with a South African abalone farm include: - will require diverting of the abalone tank wash water from regular effluent - possible water ‘down-times’ due to maintenance operations on abalone farm Based on information from the literature the fixed capital cost depreciation rate (FCCDR) typically contributes approximately 40% and the operation and maintenance (O&M) cost typically contributes 60% to the unit production cost (UPC) of desalinated water. Furthermore, a SWRO desalination plant’s intake system can contribute between 5% and 33% to the FCCDR, depending on the nature and design of the plant. Consequently, the intake system can contribute between 2% and 13% of the UPC of desalinated sea water. This implies possible cost savings of between R0.15/m3 and R2.37/m3 for the production of fresh water (depending on site-specific design factors) when desalinating sea water effluent from on-shore abalone tanks. Integration of an SWRO desalination plant with a South African abalone farm is feasible and viable, provided that the necessary steps and precautions are taken to ensure a smooth and stable operation of the SWRO desalination plant. Cost savings on the part of all the stakeholders are possible if the correct contract can be negotiated.
AFRIKAANSE OPSOMMING: Byna alle Suid Afrikaanse perlemoenplase funksioneer op ‘n seewater deurvloeistelsel. Groot volumes seewater word aan wal gepomp en vloei deur die perlemoen of kelp-oes tenks. Hierdie water vloei dan uiteindelik terug na die oseaan as gevolg van swaartekrag. Indien die afvalwater van die perlemoenplase ontsout kan word sonder permanente membraanbevuiling kan seewater tru-osmose (SWTO)-tegnologie effektief met gevestigde perlemoenplase geïntegreer word sonder om die plase se water inname-stelsels se kapasiteite te vergroot. Sonder die behoefte aan uitbreiding en instandhouding van ‘n water inname-stelsel by so ‘n plaas behoort die totale koste van ontsouting aansienlik minder te wees. Die doel van hierdie studie was dus om die uitvoerbaarheid en lewensvatbaarheid van ‘n integrasie van ‘n SWTO ontsoutingsaanleg met ‘n tipiese Suid Afrikaanse perlemoenplaas te ondersoek. Ten einde dit te doen, het die projek op vier areas van belang gefokus, naamlik: - karakterisering van tipiese Suid-Afrikaanse perlemoenplaas water - SWTO ontsoutingsaanleg loodsstudie en membraan bevuilingsgedrag - algemene bedryf van ‘n tipiese perlemoenplaas en die implikasies vir ontsouting - kosteberamings en koste-implikasies met betrekking tot die integrasie van ‘n SWTO ontsoutingsaanleg met ‘n perlemoenplaas Gedurende ‘n nege maande op-perseel ondersoek is seewater troebelheid verminder met tot 43% van 'n gemiddelde waarde van 0.82 NTU in die invloeistroom tot 0.47 NTU in die gekombineerde afvalwaterstroom wat die tenks verlaat. Selfs met skerp wisseling in die invloeistroom troebelheid, bly afvalwaterstroom troebelheid deurgaans onder 1 NTU met die uitsondering van tenk spoelwater. Opgeloste organiese koolstof (OOK) in beide die invloeistroom en die gekombineerde afvalwaterstroom het deurgaans onder 1 mg/liter gebly. Ultrafiltrasie (UF) is gebruik as voorbehandeling vir die tru-osmose (TO) om sodoende potensiële bevuiling van TO membrane te minimaliseer. Membraan kompaksie van beide die UF en TO het merkbaar bygedra tot aanvanlike deurvloeiverliese – so veel as 18% vir die poli-etersulfoon (PESM) UF membrane en 20% vir die dun film saamgestelde (DFS) poli-amied TO membraan. Hierdie is egter gerieflik binne die tipiese kompaksiewaardes soos aangehaal in die literatuur. Sonder flokkulasie was die UF in staat tot temperatuur aangepaste deurvloeitempo van tussen 45 en 55 liter/m2/h (LMH) teen herwinningstempo’s tussen 60 en 75%. Ooreenstemmende trans-membraandrukkings (TMD) het gewissel tussen 0.59 en 0.76 bar. Met ysterchloried voor-flokkulasie teen 'n konsentrasie van 3 mg/liter (as Fe3+) kon die UF teen merkbaar laer TMD waardes bedryf word – tussen 0.11 en 0.36 bar. Hierdie resultate dui daarop dat daar beslis voorsiening vir pre-flokkulasie gemaak moet word wanneer UF as voorbehandeling gebruik word, ten spyte van die feit dat die OOK konsentrasie en troebelheid van die afvalwater van die perlemoenplaas redelik laag is (OOK <1 mg / liter, troebelheid <1 NTU). Verder lig dit die onvermoë uit om OOK en troebelheid alleen te gebruik om membraanbevuilingspotensiaal van water te voorspel. ‘n Beter aanwyser van membraanbevuilingspotensiaal, alhoewel nie perfek nie, is die aangepaste bevuilingsindeks (MFI0.45). Hierdie bevuilingsindeks volg ‘n lineêre neiging met die konsentrasie van onsuiwerhede en dien as ‘n goeie aanwyser van die filtreerbaarheid van water. Op-perseel metings het getoon dat ‘n toename in gemiddelde MFI0.45 waardes van 29 s/litre2 vir die invloeistroom tot 48 s/litre2 vir die afvalstroom van die perlemoentenks die behoefte vir voor-flokkulasie as deel van UF voorbehandeling bevestig. Chemies versterkte terugspoeling (CVT) van die UF membrane ten minste elke 24 uur is noodsaaklik gevind ten einde bestendige werking te verseker. Dus kan UF met voor-flokkulasie (3 mg/liter Fe3 +) en gereelde CVT effektief as voorbehandeling metode vir die ontsouting van perlemoenplaas afvalwater gebruik word. ‘n TO ‘voer-en-bloeistelsel’ is gebruik om die tipiese prestasie van die laaste membraan in ‘n volskaalse TO membraanbank te simuleer. Hierdie TO membraan het goed presteer sonder tekens van buitensporige membraanbevuiling. Vir die laaste membraan in ‘n membraanbank het die membraan goeie gehalte finale water gelewer – ‘n vermindering van die totaal opgeloste stowwe (TOS) van die TO voerwater van 33 493 mg/liter tot 969 mg/liter is behaal. Hierdie resultate het goed vergelyk met gesimuleerde waardes deur Reverse Osmosis Analysis System (ROSA, ‘n TO simulator deur DOW) wat ‘n TOS vermindering van 33 271 mg/liter tot 1 409 mg/liter by ‘n voerdruk van 56 bar en ‘n algehele herwinningstempo van 44% aandui. ‘n Bestendige werking van die TO membraan tydens die loodsstudie het getoon dat dit moontlik is om perlemoenplaas afvalwater te ontsout sonder merkwaardige permanente membraanbevuiling. 'n Stabiele genormaliseerde deurvloeitempo van 8 LMH is bereik en TO membraan integriteit het ongeskonde gebly met 'n sout verwerping wat gewissel het van 98.0 tot 98.5%. Geen skielike afname in finale water deurvloeitempo is waargeneem as gevolg van bevuiling deur onbekende onsuiwerhede in die UF finale water nie. DBNPA (‘n nie-oksiderende ontsmettingsmiddel) is een keer per week teen ‘n ‘n konsentrasie van 10 – 30 mg / liter vir 30 minute gedoseer. Mineraal skaalvorming is effektief beheer deur die dosering van ‘n anti-skaalmiddel teen 11 – 12 mg/liter in die TO voerstroom. Die skoonmaak-in-plek (SIP) frekwensie is nie ge-optimeer nie, maar ‘n SIP een keer elke 6 – 8 weke is meer as voldoende gevind om mikrobiese bevuiling te voorkom. Voordele van die integrasie van 'n SWTO ontsoutingsaanleg met 'n Suid-Afrikaanse perlemoenplaas sluit die volgende in: - geen lang en duur omgewings impak ontleding (OIO) is nodig vir die bou van ‘n nuwe inname-stelsel nie - gedeelde kapitaal en operasionele koste van inname-stelsel - tweeledige aansporing om konstant goeie gehalte watervloei deur die plaas te verseker - vroegtydige waarskuwings ten opsigte van gebeurtenisse soos rooigety - gedeelde bedryfs- en bestuurskoste om voerpype skoon te hou Nadele van die integrasie van 'n SWTO ontsoutingsaanleg met 'n Suid-Afrikaanse perlemoenplaas sluit die volgende in: - vereis herleiding van perlemoentenk spoelwater weg van gereelde afvalwater - moontlike watervloei-aftye weens instandhoudingsbedrywighede op die perlemoenplaas Gebaseer op inligting uit die literatuur dra die vaste kapitaal koste waardeverminderings-koers (VKKWK) gewoonlik ongeveer 40% en die bedryfs- en instandhoudingskoste (B&I) ongeveer 60% by tot die produksiekoste per eenheid (PKE) van ontsoute water.Verder kan ’n SWTO ontsoutingsaanleg se inname-stelsel tussen 5% en 33% tot die VKKWK bydra afhangende van die aard en ontwerp van die aanleg. Gevolglik kan die inname-stelsel tussen 2% en 13% tot die PKE van ontsoute seewater bydra. Dit impliseer ‘n moontlike kostebesparing van tussen R0.15/m3 en R2.37/m3 vir die produksie van vars water wanneer die afvalwater van perlemoentenks ontsout word. Integrasie van 'n SWTO ontsoutingsaanleg met 'n Suid-Afrikaanse perlemoenplaas is uitvoerbaar en lewensvatbaar indien die nodige stappe en voorsorgmaatreëls geneem is om ‘n vlot en bestendige werking van die SWTO ontsoutingsaanleg te verseker. Kostebesparings vir alle belanghebbendes (beleggers) is moontlik indien daar oor die korrekte kontrak onderhandel kan word.

Thesis (MSc(Agric))--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: The pnmary alm of this study was to investigate the national competitive advantage of the South African abalone industry. A further aim was therefore to investigate the environment, in which the South-African abalone industry operates. Secondary aims included presenting a detailed description of the roleplayers in the South African abalone industry, and considering whether the South African abalone industry can be made more competitive and, if so, how this can be achieved. The achievement of these aims required a framework to establish which data is necessary for the task. The framework selected involved a combination of Porter's Five Forces analysis and Diamond Model. The study suggests that South African abalone firms should promote South African abalone more actively, invest more funds in human resources, and that the industry's faith in government needs to be restored, which can be achieved by improving the services offered by public institutions like the South African Bureau of Standards (SABS), the Department of Trade and Industry (DTI) and Marine and Coastal Management (M&CM).
AFRIKAANSE OPSOMMING: Die doel met die studie was om die nasionale kompeterende voordeel van die Suid Afrikaanse perlemoen industrie te ondersoek. Om dit te bereik, moes die omgewing waarin die industrie fuksioneer ontleed en die rolspelers in die industrie deeglik beskryf word. 'n Verdere doel was om te bepaal of dit moontlik is om die kompeterendheid van die industrie te verhoog, en indien wel, hoe dit bereik kan word. Die raamwerk wat gekies is om die doelstellings te bereik bestaan uit 'n kombinasie van Porter se "Five Farces" analise en "Diamond Model". Die resultate dui aan dat die industrie Suid-Afrikaanse perlemoen meer aktief moet bekendstel, individuele ondernemings meer in hul menslike hulpbronne moet investeer, en dat die industrie se vertroue in die nasionale regering herstel moet word, deur die dienste te verbeter wat deur publieke instellings soos die Suid-Afrikaanse Buro van Stanaarde (SABS), Departement Handel en Nywerheid en Mariene- en Kusbestuur verskaf word.

Thesis (MScAgric)--Stellenbosch University
ENGLISH ABSTRACT: Triploidy is the genetic state of containing three sets of chromosomes per cell in stead of two as in diploid organisms. The South African abalone (Haliotis midae) is naturally a diploid organism that sexually matures between four to eight years of age. Early sexual maturity is a disadvantage in cultured abalone stock, as the process of gonad development and spawning is energy demanding, causing energy to be diverted away from somatic growth. This same problem has been extensively experienced in diploid bivalve molluscs, where triploidy has since been applied as a means to prevent sexual maturation from occurring, thereby speeding up the growth process and shortening the time to marketing. Because triploidy was effective in bivalves, it was thought that it could contribute to faster growth in abalone as well. A procedure for the induction of triploidy in the abalone, Haliotis midae, was developed by De Beer (2004) and yielded up to 100 percent triploidy in treated abalone larvae. The next step was to compare the growth of the diploids and triploids to establish whether there was indeed a growth advantage on the part of the triploids, in view of commercial application. By using the same techniques as described by De Beer (2004), three groups consisting of triploid and diploid siblings were produced and subscribed to a comparative growth trial. The groups were spawned in three different seasons. The main objective was to establish whether there was in fact a difference in growth between diploid and triploid siblings, and whether seasonal effects were associated with growth advantages for either triploids or diploids. The two growth parameters measured were shell length and body weight. Measurements commenced at eight months of age, when the abalone could be individually tagged and continued up to the age of 24 months. The over-all results provided no convincing evidence of statistically significant faster growth of triploid juveniles compared to that of diploids up to two years of age. Growth differences were detected between seasons, but could not confidently be ascribed to seasonal environmental effects. The regression of shell length to body weight was similar for diploids and triploids.
AFRIKAANSE OPSOMMING: Triploiede organismes bevat drie stelle chromosome per sel in plaas van twee soos dit normaalweg in diploiede diere voorkom. Die Suid Afrikaanse perlemoen (Haliotis midae) is van nature ‘n diploiede organisme wat tussen die ouderdom van vier tot agt jaar seksueel aktief word. Vroeë seksuele aktiwiteit is ongewens in kommersiële akwakultuur aangesien energie spandeer word aan gonade ontwikkeling in plaas van somatiese groei. Dieselfde probleem is vroeër in die oester bedryf ondervind waar dit deur middel van triploiede induksie aangespreek is. Triploiedie veroorsaak steriliteit en kan gebruik word as ’n metode om steriliteit op groot skaal te induseer. Steriliteit sou dan meebring dat meer energie beskikbaar is vir somatiese ontwikkeling, wat verhoogde groeitempo en n verkorte tyd tot bemarking beteken. Op soortgelyke wyse is dus gepostuleer dat triploiedie in perlemoen ook tot steriliteit kon lei. ‘n Triploiede induksie metode was ontwikkel deur Mathilde de Beer (2004) wat ‘n hoë persentasie triploidie in geinduseerde perlemoen opgelewer het. Die volgende logiese stap was om die groei van diploiede diere met die van triploiede diere te vergelyk om te bepaal of triploiedie wel ’n groei voordeel tot gevolg het met die oog op kommersiële toepassing. Deur van dieselfde tegnieke as De Beer (2004) gebruik te maak, is drie groepe, elk bestaande uit verwante diploiede en triploiede diere, geproduseer en ingeskryf aan n vergelykende groei proef. Die groepe was in drie verskillende seisoene geproduseer. Die hoof doelstelling van die proef was om groeitempo van diploiede en triploiede diere te vergelyk, asook om die invloed van seisoen op groei van diploide en triploide te bepaal. Twee groei eienskappe naamlik skulp lengte en liggaamsmassa is gemeet vanaf ‘n ouderdom van agt maande (wanneer die diere individueel gemerk kon word) tot ‘n ouderdom van 24 maande. Die algehele resultate het gedui op geen betekenisvolle verskil tussen die groei van triploiede en diploiede perlemoen tot op die ouderdom van twee jaar. Verskille het voorgekom in die groei tussen seisoene, maar daar kon nie bewys word dat die verskille die gevolg van seisoenale omgewingseffekte was nie. Diploiede en triploiede het dieselfde skulp lengte tot liggaamsmassa verhouding getoon tot op twee jaar ouderdom.

Thesis (MPhil (Animal Sciences. Aquaculture))--University of Stellenbosch, 2006.
T. heterouncinata is a sabellid polychaete endemic to South Africa and found primarily in the shells of the abalone Haliotis midae. With the intensification of abalone aquaculture around the world, T. heterouncinata has become a problematic pest by causing shell deformities, reducing abalone growth rates and, in some instances, high abalone mortalities. The problem of this sabellid was first noticed in Californian in the early 1980’s in Red abalone (Haliotis rufescens) production facilities. Many mitigation methods have been tested over the years and this paper investigates another two methods; a reduction in particulate load in the tank to reduce the food source of the sabellid which perhaps will reduce fecundity, and to use ultrasound as a possible mitigation method. This study found that filtration and reduction in suspended particles did not have a significant effect, but that ultrasound did have a significant effect in reducing T. heterouncinata populations.

Bibliography: leaves 122-135.
The successful aquaculture of the abalone Haliotis midae requires a multi-disciplinary approach. Experiments were designed to provide insight into various aspects of abalone biology and seawater system design. A comparative evaluation of a closed and open seawater system for an H. midae hatchery was performed. Monthly seawater temperatures and nitrite levels were higher in the closed system. The salinity of the closed system seawater varied more than that of the open system. The pH of seawater in the closed system varied between 7.7 and 8.2. This was lower than the 7.9 to 8.35 pH range of the open system seawater. An open system is thus clearly preferable to a closed system. Haliotis midae reproductive condition can be assessed visually by examining the shape and colour of the broodstock gonads. Spawning experiments showed that abalone should be starved for at least 24 hours prior to spawning induction. Haliotis midae can be induced to spawn by treatment with hydrogen peroxide when exposed to seawater at a pH of between 9.0 and 9.9 (males and females) Spawning can reliably be induced by using final hydrogen peroxide concentrations in the range of 7 to 25 mM for male and female H. midae. Gravid broodstock should be exposed to hydrogen peroxide for 100 to 300 minutes during spawning induction. There was no significant impact observed for prior conditioning of the brood-stock to the spawning tanks, lunar phase and timing of spawning induction relative to sunset. Haliotis midae larval toxicity to chlorine, copper and ammonia was investigated. The LT50 to a chlorine concentration of 0.06 mg/l was 170 minutes. For copper the LT50 of larvae exposed to 0.12 mg/l was 53 minutes. The LT50 of ammonia at 5 mg/l was 600 minutes. A brief synopsis of the major findings is presented in the last chapter. Some discussion on the future prospects of the abalone industry is also provided.

Abalone, Haliotis midae, farmers in South Africa that feed formulated diets reported a periodic drop in abalone growth during periods of increased gonad development. A large drop in abalone biomass was noticed after presumed spawning events. This study was aimed to determine the effect of diet and sex-sorting on gonad development in abalone. Experiments were conducted on a commercial abalone farm from July 2012 to the end of June 2013. Isonitrogenous and isoenergetic diets were formulated with two protein sources. A fishmeal and soybean meal (S-diet) diet and a fishmeal only (F-diet) diet were fed to abalone (50 - 70 g abalone⁻¹) over 12 months. Weight and length gain, gonad bulk index (GBI), visceral index (%) and meat mass index (%) were determined monthly and seasonally. A histological study on the female gonads was conducted. This study also included an experiment to test the effect of sex-sorting (70 - 80 g abalone⁻¹) on growth and body composition with treatments including males (M), females (F) and equal numbers of males and females (MF). Weight gain and length gain were faster in S-diet-fed abalone (RM-ANOVA, F ₍₁, ₁₆₎ = 7.77, p = 0.01; F ₍₁, ₆₉₎ = 49.9, p < 0.001, respectively). Gonad development was significantly affected by the inclusion of soybean meal with S-diet-fed abalone showing higher GBI-values than F-diet-fed abalone (RM-ANOVA, F ₍₁, ₃₃)= 16.22, p = 0.0003). Male abalone had higher GBI-values than females (RM-ANOVA, F ₍₁, ₃₃₎ = 39.87, p < 0.0001). There was no significant difference in average feed conversion ratio (FCR) between diets over time (RM-ANOVA, F ₍₁, ₂₁₎ = 0.008, p = 0.97). However, average FCR-values were significantly highest between November 2012 and March 2013, the presumed spawning season. The visceral mass (gut and gonad) as a proportion of whole mass (visceral index, %) was significantly higher in abalone fed the S-diet (RM-ANOVA; F ₍₁, ₆₉₎ = 68.06, p < 0.0001). There was no difference in meat mass index (%) between diets for both male and female abalone (RM-ANOVA; F ₍₇, ₂₄₈₎ = 0.80, p = 0.60; F ₍₇, ₂₄₁₎ = 1.7, p = 0.11,respectively). Meat mass index significantly decreased from September 2012 to February 2013 coinciding with the period of high GBI-values. The distribution of oocyte maturity stages differed between diets. The majority of oocytes within S-diet-fed abalone were fully mature stage 8 oocytes compared to a majority of stage 7 oocytes in F-diet-fed abalone. Histology corroborated peaks in GBI-values for abalone fed both diets. There was no significant difference in growth, GBI, visceral index (%) and meat mass index (%) between abalone sorted into monosex and mixed-sex populations. Thus, the presence of the opposite sex did not have an effect on growth and gonad mass in H. midae. The phytoestrogens daidzin, glycitin, genistin, daidzein, glycitein and genistein were present in soybean meal and only traceable amounts were found in the F-diet. This study provided evidence that soybean meal present in formulated feed affected growth and gonad development in H.midae. The difference in the distribution of the maturity stages of oocytes was affected by diet. Sex-sorting abalone into monosex and mixed-sex populations had no influence on weight and length gain and gonad development.

Thesis (MSc)--University of Stellenbosch, 2007.
ENGLISH ABSTRACT: The indigenous abalone Haliotis midae is one of the most remarkable and highly exploited species of marine molluscs in South Africa. It is the only species of southern African Haliotidae to be commercially reared and has been successfully cultured for almost two decades. Its short history of domestication along with market demands and the need to develop efficiency in the production process has resulted in an increased interest in the possible genetic improvement of this species. The unhurried growth rate associated with H. midae is a cause of particular concern to the industry, predominantly with regards to profitability and competitiveness in the market place. A modest amount of work has so far been directed at establishing a means of enhancement for selective breeding on the commercial level. Genetics plays a key role in the establishment of successful improvement programmes in various aquaculture species. The aim of this study was to develop species-specific microsatellite markers for the abalone and subsequently perform parentage assignment on farm produced animals entered into a growth performance recording scheme. Animals were obtained from the hatcheries of three commercial abalone farms situated in the Walker Bay region in the Western Cape. Microsatellites were isolated using the enrichment-based FIASCO method, and characterised into perfect, imperfect and compound repeats according to the structural nature of their repetitive units. From the partial gDNA libraries obtained and 365 screened colonies, a total of 54 loci were located. PCR primers were designed for 36 markers and the 15 primer pairs that displayed loci with the highest level of polymorphism were subsequently chosen for fluorescent labelling. The markers were tested on a subset of 32 wild H. midae individuals to determine their usefulness and efficiency in genotyping. Five markers, along with five others that were previously designed, were chosen for assigning parentage to the animals submitted to the performance recording scheme. Three thousand offspring from each of the three participating farms were equally divided and reared at five different locations. From each location 20 fast growing and 20 slow growing juveniles, as well as the broodstocks, were sampled and genotyped using the ten chosen microsatellite loci. Two farms had 60% of offspring unambiguously assigned to a single parental couple. Assignments showed patterns of dominant male and female brooders, but no trend in brooders specifically contributing to fast or slow growing offspring. Parentage assignment for the third farm was, however, unsuccessful due to lack of broodstock data. In future, screening of all available broodstock will ensure acquisition of relevant pedigree information. The results obtained in this study are an initial step in the development of a genetic improvement programme for commercial Haliotis midae.
AFRIKAANSE OPSOMMING: Die inheemse skulpvis Haliotis midae is een van die mees merkwaardige en hoogs oorbenutte mariene slakspesies in Suid-Afrika. Dit is die enigste suidelike Afrika Haliotidae spesie wat kommersieel benut word en dit word al meer as twee dekades suksesvol geteel. Die spesie se kort domestiseringsgeskiedenis, toenemende mark aanvraag en die behoefte om meer effektiewe produksie daar te stel, het gelei tot toenemende belangstelling in die moontlike genetiese verbetering van die spesie. Die stadige groeitempo geassosieer met H. midae is veral ‘n punt van kommer vir die industrie, veral in terme van winsgewendheid en kompetering in die markplek. Minimale werk is sover gedoen in die daarstelling van verbetering deur selektiewe teling op ‘n kommersiële skaal. Genetika speel ’n sleutelrol in die daarstelling van suksesvolle verbeteringsprogramme van verskeie akwakultuur spesies. Die doel van hierdie studie was om spesie-spesifieke mikrosatelliet merkers vir perlemoen te ontwikkel en vervolgens ouerskapsbepaling van kommersiële diere, wat deelneem aan ‘n groeiprestasie aantekenstelsel, uit te voer. Diere is voorsien deur die teelstasies van drie kommersiële perlemoenplase geleë in die Walker Bay omgewing in die Wes-Kaap. Mikrosatelliete is geïsoleer deur die verrykings-gebaseerde FIASCO metode, en gekarakteriseer as perfekte, onderbroke of saamgestelde herhalings gebaseer op die strukturele aard van die herhalings eenhede. Vanaf die gedeeltelik gDNA biblioteke wat bekom is en 365 gesifte kolonies, is ‘n totaal van 54 loki opgespoor. PKR inleiers is ontwerp vir 36 merkers en die 15 inleierpare, wat loki met die hoogste polimorfisme geamplifiseer het, is vervolgens geselekteer vir fluoreserende merking. Die merkers is getoets op ’n kleiner groep van 32 natuurlike H. midae individue om hulle bruikbaarheid en genotiperingseffektiwiteit te bepaal. Vyf merkers is saam met vyf reeds ontwikkelde merkers gekies vir ouerskapsbepaling van die diere in die prestasie aantekenstelsel. Drieduisend nageslag diere vanaf elkeen van die drie deelnemde plase is gelykop verdeel en grootgemaak op die vyf verskillende lokaliteite. ‘n Monster van 20 vinnig groeiende en 20 stadig groeiende jong perlemoen, sowel as broeidiere, is vanaf elke lokaliteit geneem en gegenotipeer deur middel van die 10 geselekteerde mikrosatelliet loki. Sestig persent van twee van die plase se nageslag is onteenseglik toegesê aan ‘n enkele ouerpaar. Ouerskapstoekenning het patrone van dominante vroulike en manlike broeidiere getoon, maar geen tendens in terme van bydrae tot vinnig en stadig groeiende nageslag kon gevind word nie. Ouerskapstoekenning vir die derde plaas was onsuksesvol as gevolg van ’n gebrek aan data vir die broeidiere. In die toekoms sal genotipering van alle beskikbare broeidiere die daarstelling van relevante stamboominligting verseker. Die resultate verkry in hierdie studie verteenwoordig ‘n eerste stap in die ontwikkeling van ’n genetiese verbeteringsprogram vir kommersiële Haliotis midae.

Thesis (MSc (Genetics))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: The South African abalone, Haliotis midae, is the only endemic species of commercial value. Aquaculture remains the only avenue for expanding the industry, since the closure of the fishery. The current focus is on implementing a molecular breeding programme; thus the development of molecular markers for linkage mapping and QTL analysis is a priority. Various markers, mainly anonymous, have been developed for H. midae; however emphasis is being placed on the development of gene-linked type I molecular markers. The present study investigates and demonstrates the use of public sequence collections to develop type I markers for a species with limited genomic resources, via three strategies: Surveying anonymous H. midae microsatellite markers’ flanking regions to find homology to gene sequences in public databases, cross-species marker transfer of anonymous markers from H. rubra and H. discus hannai demonstrating putative gene associations and lastly EST marker mining (SNP and microsatellites) from various Haliotids and testing transfer to the target species. Approximately 17% of H. midae anonymous markers showed significant similarity to genes. The current study also reports higher cross-species transferability from both H. rubra and H. discus hannai to H. midae (39% and 20.5%, respectively) than previously demonstrated and 15 EST-microsatellites and 16 EST-SNPs were successfully mined. Furthermore, the non-random distribution of microsatellites and high nucleotide diversity in the H. midae genome was confirmed. This is a low cost and time effective method for marker development and presents a continuous and dynamic resource that could be used for future marker development and characterisation as sequence information in public databases grow exponentially.
AFRIKAANSE OPSOMMING: Die Suid-Afrikaanse perlemoen, Haliotis midae, is die enigste van vyf inheemse spesies van kommersiële waarde. Na die noodgedwonge sluiting van die vissery, is akwakultuur die mees praktiese oplossing om die perlemoen industrie uit te brei. Die huidige fokus is gerig op die implementering van ‘n molekulêre teel-program en dus is die ontwikkeling van molekulêre merkers vir genetiese kartering en kwantitatiewe kenmerk lokus analise, van uiterste belang. Tipe II merkers is voorheen vir die perlemoen ontwikkel, maar huidige tendense lê klem op die ontwikkeling van geen-gekoppelde tipe I merkers. Die huidige studie ondersoek die gebruik van publieke databasisse vir die ontwikkeling van tipe I molekulêre merkers vir ‘n spesie met beperkte genomiese bronne. Drie strategieë is geïmplementeer: Eerstens is ‘n opname gemaak van die homologie van perlemoen tipe II merker-vleuelende volgordes met geen volgordes in databasisse. Verder is die oordraagbaarheid van tipe II merkers vanaf H. rubra en H. discus hannai wat assosiasie met gene toon ondersoek. Laastens is ‘n Uitgedrukte Volgorde Merk (UVM) (Expressed Sequence Tag, EST) merker-ontginnings metode vanaf verskeie Haliotis spesies en toetsing van oordraagbaarheid na die teiken spesie uitgevoer. Ongeveer 17% van die tipe II H. midae merkers het geniese assosiasie getoon. ‘n Hoër tussen-spesie oordraagbaarheid vanaf beide H. rubra en H. discus hannai na H. midae (39% en 20.5%, onderskeidelik) word gerapporteer in vergelyking met vorige studies en 15 UVM-mikrosatelliete en 16 UVM-enkel nukleotied polimorfismes (single nucleotide polimorphism, SNP) is ontwikkel. Verder bevestig die studie die nie-lukrake verspreiding van mikrosatelliete en hoë nukleotied diversiteit in die perlemoen genoom. Die gebruik van publieke databasise vir die ontwikkeling en karakterisering van tipe I molekulêre merkers is tyd- en koste-besparend en bied ‘n volgehoue en dinamiese bron vir toekomstige gebruik.

Abalone poaching is a major problem in South Africa. The South African abalone, Haliotis midae, rates as an extreme example of high levels of illegal harvesting. The research aimed at examining the role of the different role players in preventing poaching of the species in the East London area of the Eastern Cape Province of South Africa and determining the challenges the officers faced as there was an increase in poaching in the area. To achieve this aim, the research techniques including questionnaires, interviews and focus groups were used. Fisheries compliance and enforcement faces challenges of the illegal harvesting of abalone since 1994. In the past 18 years (1994-2012), and more specifically in the past nine years (2004-2012), poaching of abalone has increased at an alarming rate along the East London coastline. The failure of the state to issue fishing rights and conduct effective sea-based compliance, combined with the incentives to fish abalone created the conditions for rapid emergence of illegal harvesting. The uncontrolled fishing had a dramatic effect on the stock, and the average size of abalone decreased significantly. The Eastern Cape Province abalone cultivation industries were developed due to the decline in harvesting of abalone. Government departments such as the Department of Agriculture, Forestry and Fisheries; the Department of Economic Development, Environmental Affairs and Tourism; the South African Police Services together with the Eastern Cape Parks and Tourism Agency have conducted joint operations to combat the illegal harvesting of abalone. These operations have led to many arrests of abalone poachers along the East London coastline. The quantity of confiscated abalone has increased from 2007 to 2011. The positive results achieved by the departments during joint operations showcase robust efforts to eradicate the environmental transgression in the East London Coastline. Joint operations are encouraged by all the departments to save the species for future generations. Workshops involving different stakeholders had to take place and the policies in place must have a bottom-up approach where communities are involved.

Thesis (PhD (Genetics))--University of Stellenbosch, 2009.
ENGLISH ABSTRACT: South African abalone, Haliotis midae, has been the subject of major concern regarding its survival and conservation over the last decade or more. Being the only one of five endemic species with commercial value, there is considerable interest and urgency in genetic management and improvement of this species. Limited genetic information and the increasing conservation concern of this species are considered the key motivations for generating information on the micro- and macro-evolutionary processes of H. midae, the overall objective of this study. This study reported the first microsatellite and Single Nucleotide Polymorphism (SNP) markers developed specifically for Haliotis midae. Both these marker types were applied to elucidate the degree of gene flow in nine natural abalone populations whilst testing for two contrasting hypotheses; panmixia versus restricted gene flow. Data was analysed using a series of methodological approaches ranging from traditional summary statistics to more advanced MCMC based Bayesian clustering methods with and without including spatial information. Using only microsatellite data, the historical demography of the species was also examined in terms of effective population size and population size fluctuations. Finally, the evolutionary positioning and origin of Haliotis midae with regards to other Haliotis species was investigated based on mitochondrial and nuclear sequence data. Both microsatellite and SNP data gave evidence for subtle differentiation between West and East coast populations that correlates with a hydrogeographic barrier in the vicinity of Cape Agulhas. Population substructure was supported by AMOVA, FCA and Bayesian clustering analysis. Clustering utilizing spatial information further indicated clinal variation on both sides of the proposed barrier with a region in the middle coinciding with a secondary contact zone, indicating possible historical isolation during glacial periods. Overall, the similar degree of substructure observed with both microsatellites and SNPs supported the existence of contemporary and/or historical factors with genome-wide effect on gene flow. The population expansion measured with the microsatellites was inconsistent with the known recent decline but taking the species’ life cycle and large effective population size into account, a shrinkage in population size will probably only be apparent in a few generations time. On a macro-evolutionary scale, this study presents the first classification of South African abalone as a monophyletic group within the Haliotidae family. The topology based on the combined mitochondrial and nuclear dataset is highly suggestive of a relatively recent radiation of the SA species from the Indo-Pacific basin. The study concludes by describing the most likely factors that could have affected overall population structure and makes suggestions on how the given genetic information should be incorporated into strategies aimed towards the effective management and conservation of Haliotis midae.
AFRIKAANSE OPSOMMING: Die Suid-Afrikaanse perlemoen, Haliotis midae, is oor die laaste dekade of meer die onderwerp van groot bekommernis betreffende die spesie se oorlewing en bewaring. Aangesien dit die enigste van vyf endemiese SA spesies is met kommersiёle waarde, is daar besonderse belang en erns in die genetiese beheer en verbetering van die spesie. Beperkte genetiese inligting en ‘n toenemende behoefte om die spesie te bewaar is die hoof motivering agter die generering van informasie rakende mikro- en makro-evolusionêre prosesse in Haliotis midae en is die oorhoofse doel van hierdie studie. Hierdie studie beskryf die eerste mikrosatelliete en enkel basispaar polimorfismes wat ontwikkel is spesifiek vir Haliotis midae. Beide tipe merkers is aangewend om die mate van gene vloei in nege wilde perlemoen populasies te ondersoek terwyl twee hipoteses ondersoek is; panmiksie versus beperkte gene vloei. Data is geanaliseer deur gebruik te maak van ‘n reeks metodieke benaderings wat wissel van tradisionele opsommings statistieke tot meer gevorderde MCMC gebasseerde groeperings metodes met of sonder die gebruik van geografiese data. Mikrosatelliet data is ook aangewend om die historiese demografie van die spesie te bepaal in terme van effektiewe populasie grootte asook veranderinge in populasie groottes. Laastens is die evolusionêre posisionering en oorsprong van Haliotis midae teenoor ander Haliotis spesies ondersoek deur gebruik te maak van mitokondriale en nukleêre DNA volgorde data. Beide mikrosatelliet en enkel basispaar polimorfisme data lewer bewys van ‘n subtiele genetiese verskil tussen wes en ooskus populasies wat verband hou met ‘n hidrografiese skeiding in die omgewing van Kaap Agulhas. Populasie struktuur is ondersteun deur die analise van molekulêre variansie (AMOVA), faktoriale komponente analise asook Bayesiese groeperings analise. Groeperings analise wat geografiese informasie insluit dui klinale genetiese variasie aan beide kante van die skeiding aan met ‘n area in die middel wat ooreenstem met ‘n sekondêre kontak gebied. In totaal, ondersteun die soortgelyke mate van struktuur verkry met beide die mikrosatelliete en enkel basispaar polimorfismes die bestaan van hedendaagse en/of historiese faktore met genoom wye invloed op gene vloei. Die toename in populasie grootte vasgestel deur die mikrosatelliet data stem nie ooreen met die onlangse afname waargeneem in die spesie nie, maar met inagneming van Haliotis midae se lewenssiklus en groot effektiewe populasie grootte, sal die afname in populasie grootte moontlik eers oor ‘n paar generasies na vore kom. Op ‘n makro-evolusionêre skaal lewer hierdie studie die eerste klassifikasie van Suid-Afrikaanse perlemoen as ‘n monofiletiese groep binne die Haliotidae familie. Die topologie gebaseer op ‘n gesamentlike mitkondriale en nukleêre datastel is hoogs aanduidend van ‘n relatiewe onlangse verspreiding van die Suid-Afrikaanse spesies uit die Stille-Indiese Oseaan. Die studie sluit af deur die mees algemene faktore te bespreek wat populasie struktuur kon beïnvloed het en maak voorstelle op watter wyse hierdie genetiese inligting aangewend kan word vir die effekiewe beheer en bewaring van Haliotis midae.

Thesis (Msc Food Sc (Food Science))--University of Stellenbosch, 2009.
An 84-day study was conducted to find a suitable diet and feeding level for the culture of South African abalone (Haliotis midae) in Namibia. Two experimental diets, namely, a seaweed diet (SWD) Laminaria pallida (macrophytic algae) and a formulated diet (FD) (macro-algae), for use in abalone (Haliotis midae) feed development, were evaluated. The animals used in this study were juveniles (24.33 ± 3.14 mm shell length; 2.72 ± 0.83 g live weight, mean ± SE) and sub-adults (58.07 ± 10.33 mm shell length and 41.96 ± 20.61 g live weight, mean ± SE). The nutrient profile of the SWD and FD displayed no differences in the protein and carbohydrate levels. Crude protein levels ranged from 4.91 to 17.68% (dry matter (DM) basis). The lipid levels in the FD (0.25%) were almost 0.56% lower than that in the SWD (0.76%). The feed conversion ratio (FCR) and protein efficiency ratio (PER) for the sub-adult abalone ranged from 2.80 to 10.90 and 0.10 to 0.40, respectively. The juvenile abalone fed on the FD yielded significantly lower (P < 0.05) FCRs (0.8) and higher PERs (1.20) than their counterparts fed on the SWD. A similar trend was observed for the sub-adult abalone although the differences were not significant (P > 0.05). The relative growth rate (RGR) of juvenile fed on the FD was 25% lower compared to those fed on the SWD, while that of the sub-adult abalone fed on the FD was 29% lower compared to the abalone fed on the SWD. From the daily growth rate (DGR) in terms of daily body weight (DGRBW) calculated after the 84-day period, repeated-measures ANOVA (RANOVA) indicated no interaction between time period and diet. Although slightly lower, the DGRBW for the juvenile abalone fed on the SWD diet (0.033 g/day) did not differ significantly from the DGRBW of abalone fed on the FD (0.079 g/day). In contrast, sub-adult abalone fed on the SWD exhibited significantly higher DGRBW compared to those fed on the FD. Although the abalone fed on the FD was slightly higher in nutritional content, there was no significantly difference (P > 0.05) in the nutritional profile of the abalone soft body tissue fed on either the SWD or FD. There was no significant (P > 0.05) difference in preference when comparing the aroma of the abalone meat samples fed on either the SWD or FD. However, there was a significant difference (P < 0.05) in the consumers’ preference in terms of flavour for the abalone sample fed on the FD. The trained taste panel results indicated that there was no difference in the aroma and flavour of the abalone fed on the different diets (P > 0.05). This study showed that cultured juvenile H. midae, readily accepted a FD, producing high consumption and survival rates. The FD still warrants further refinement and testing for it to become a more effective mariculture feed with commercial potential.

Thesis (MSc)--University of Stellenbosch, 2008.
ENGLISH ABSTRACT: The five leading abalone producers in South Africa have initiated a genetic enhancement program for Haliotis midae in a collaborative effort to improve economically valuable traits. Several independent objective-specific studies were initiated, including the establishment of a Performance Recording Scheme (PRS), utilised in this study, and necessary to monitor the ongoing performance of individuals as the move from mass-selection to marker assisted selection (MAS) is implemented. The primary objective of this study was parentage assignment of F1 offspring mass-selected for size at approximately one year and allocated to either a “faster” or a “slower” growth group. Nine microsatellite markers were used to genotype juveniles and potential parents, with assignment completed using CERVUS 2.0. Average growth results for Abagold and HIK were comparable for both growth groups. Slight environmental effects, although not statistically significant, were evident as growth advantages for juveniles within the faster growth group at two of the five locations and for juveniles within the slower growth group at one of the five rearing locations. Despite measures to standardise environmental influences, variables are difficult to control within the reality of a production environment; and potential genotype x environment interactions may require further investigation and factoring into future breeding programs. The additional costs associated with MAS often make the technology prohibitive to most aquaculture operations, despite the significant genetic gains to be realised from its implementation. Cost-optimising routine processes such as DNA extractions may be one approach to reduce these additional costs. Chelex®100 appears to be a suitable alternative to the CTAB method – being quick and cost-effective to perform. Applying this method in combination with the high throughput of a robotic platform warrants further evaluation. For the microsatellite development, 50% of positive recombinant clones contained inserts. Sequencing of these clones produced 16% perfect repeats and 47% imperfect repeats for which 52 primer sets were designed and tested. In total, 31 polymorphic microsatellite loci of different motifs and composition were developed. Sixty-one percent of sequenced clones were deemed redundant and pre-screening for both uniqueness and the presence of microsatellites would reduce unnecessary sequencing thus improving the efficiency of the FIASCO method and reducing costs. Nine loci were selected for parentage assignments. Null alleles were present for all the selected markers; however, frequencies were below the critical level of 5%. Parentage yielded 91% and 90% successful assignment for Abagold and HIK respectively; however, observations indicate that a measure of relatedness may exist between breeders. Recommendations with regards to future family breeding include, for both Abagold and HIK, retaining selected breeders based on their respective contributions to the F1 progeny while reassessing the potential of remaining breeding stock under more controlled breeding conditions. No obvious trends were observed for growth with most individuals producing both faster and slower growing offspring. Juveniles will be reassessed at two years to determine whether the size advantage or disadvantages were maintained and to ascertain whether growth advantages/disadvantages may be gender specific.
AFRIKAANSE OPSOMMING: Die vyf mees toonaangewende perlemoen produseerders in Suid Afrika het „n genetiese verbeteringsprogram vir Haliotis midae geinisieer in „n gesamentlike poging om ekonomiese belangrike eienskappe te verbeter. Verskeie onafhanklike fokus-spesifieke studies is geinisieer, insluitend die totstandkoming van „n groeiprestasie aantekenstelsel, soos gebruik in hierdie studie, en wat noodsaaklik is om die aaneenlopende prestasie van individue te moniteer soos daar beweeg word van massa seleksie tot merker bemiddelde seleksie. Die primêre fokus van hierdie studie was die ouerskapsbepaling van F1 nageslag wat massa geselekteer is op ouderdom 1 jaar vir grootte en as of “vinniger” of “stadiger” groeiers geklassifiseer is. Nege mikrosatelliet merkers is gebruik om jong perlemoen individue en moontlike ouers te genotipeer, met die ouerskapstoekenning bereken deur CERVUS 2.0. Groei resultate vir Abagold en HIK was vergelykbaar vir beide groei groepe op drie van die lokaliteite. Geringe omgewingseffekte, alhoewel nie statisties betekenisvol nie, was sigbaar as „n groei voordeel vir jong individue op twee van die vyf lokaliteite. Ongeag maatstawe om omgewingsinvloede te standardiseer, is varieerbares moeilik om te beheer in die produksie omgewing en genotipe x omgewings interaksies mag verdere navorsing vereis en behoort in ag geneem te word in toekomstige telingsprogramme. Die onkoste wat met merker bemiddelde seleksie geassosieer word, maak die tegniek soms onaantreklik vir die meeste akwakultuur operasies; nie teen staande die genetiese voordele wat die gebruik daarvan veroorsaak. Die koste-optimiseering van roetine prosesse, soos byvoorbeeld, DNA ekstraksies, is dalk een aanslag om die addisionele koste te verminder. Chelex®100 blyk „n geskikte alternatief tot die CTAB metode te wees – die tegniek is vinnig en koste-effektief om uit te voer. Die gebruik van hierdie metode in kombinasie met die hoë deurvloei van ‟n robotiese sisteem behoort verder ondersoek te word. Vir die mikrosatelliet ontwikkeling het slegs 50% van die positiewe rekombinante klone invoegings bevat. Nukleotiedvolgorde bepaling van hierdie klone het 16% perfekte herhalings en 47% onderbroke herhalings bevat waaruit 52 inleierstelle ontwikkel en getoets is. In totaal is 31 polimorfiese mikrosatelliet loki van verskillende motiewe en samestelling ontwikkel. Een-en-sestig persent van die volgorde bepaalde klone is oortollig geag en vooraf sifting vir beide uniekheid en die teenwoordigheid van mikrosatelliete sal onnodige volgorde bepaling verhoed, die effektiwiteit van die FIASCO tegniek verhoog sowel as addisionele koste verminder. Nege loki is geselekteer vir ouerskapsbepaling. Nul allele was teenwoordig vir al die geselekteerde merkers, maar die frekwensies was egter laer as die 5% kritieke waarde. Ouerskap is 91% en 90% suksesvol bepaal vir Abagold en HIK onderskeidelik. Waarnemings dui egter daarop dat daar verwantskappe mag wees tussen van die broeidiere. Voorstelle in terme van toekomstige familie teling sluit is, vir beide Abagold en HIK, om geselekteerde broei diere te behou gebaseer op hulle onderskeie bydraes tot die F1 nageslag asook die herevaluaring van die potensiaal van die oorblywende broei diere onder meer beheerde teling toestande. Geen voor-die-handliggende tendense is waargeneem vir groei nie met die meeste individue wat beide vinniger en stadiger groeiende nageslag geproduseer het. Jong individue moet geherevalueer word op tweejarige ouderdom om te bepaal of die groei voordeel of nadele behou is en om te bepaal om groei voordele/nadele geslagspesifiek is.

Includes bibliographical references (leaves 91-111).
The feasibility of growing red seaweed, Gracilaria gracilis (Gigartinales, Rhodophyta) in land-based culture systems was investigated in two Western Cape abalone farms i.e. Jacobsbaai Sea Products (JSP) and Irvin & Johnson (I&J) and large size culture tanks (I&J). All the tanks were stocked with at 2 kg m⁻² supplied with unfiltered seawater (24 and 4 volume exchanges d⁻¹) JSP and I&J, respectively. This study also tested the efficiency of G. gracilis as a biofiltering species as well as investigating the effectivenss and influence of nutrient-enriched seawater i.e. turbot and abalone effluent (JSP) and fertilized seawater and abalone effluent (I&J) on the growht of Gracilaria.

Thesis (PhD (Genetics))--University of Stellenbosch, 2010.
Includes bibliography.
ENGLISH ABSTRACT: The slow growth rate of Haliotis midae impedes the optimal commercial production of this most profitable South African aquaculture species. To date, no comprehensive effort has been made to identify genes associated with growth variation in farmed H. midae. The aim of this study was therefore to investigate growth variation in H. midae and to identify and quantify the expression of selected growth-related genes. Towards this aim, molecular methodologies and cell cultures were combined as a time-efficient and economical way of studying abalone transcriptomics and cell biology. Modern Illumina sequencing-by-synthesis technology and subsequent sequence annotation were used to elucidate differential gene expression between two sibling groups of abalone demonstrating significant growth variation. Following transcriptome sequencing, genes involved in growth and metabolism, previously unknown in H. midae, were identified. The expression of selected target genes involved in growth was subsequently analyzed by quantitative real-time PCR (qPCR). The feasibility of primary cell cultures for H. midae was furthermore investigated by targeting embryo, larval and haemolymph tissues for the initiation of primary cell culture. Larval cells and haemocytes could be successfully maintained in vitro for limited periods. Primary haemocyte cultures demonstrated to be a suitable in vitro system for studying gene expression and were subsequently used for RNA extraction and qPCR, to evaluate differential growth induced by bovine insulin and epidermal growth factor (EGF). Gene expression was thus quantified in fast and slow growing abalone and in in vitro primary haemocyte cultures treated with different growth stimulating factors. The results obtained from transcriptome analysis and qPCR revealed significant differences in gene expression between large and small abalone, and between treated and untreated haemocyte cell cultures. Throughout in vivo and in vitro qPCR experiments, the up-regulation of genes involved in the insulin signaling pathway provides evidence for the involvement of insulin in enhanced growth rate for various H. midae tissues. Besides the regulation of target genes, valuable knowledge was also gained in terms of reference genes, during qPCR experimentation. By quantifying the stable expression of two genes (8629, ribosomal protein S9 and 12621, ornithine decarboxylase) in various tissues and under various conditions, suitable reference genes, that can also be used in future H. midae qPCR studies, were identified. By providing evidence at the transcriptional level for the involvement of insulin, insulin-like growth factors (IGFs) and insulin-like growth factor binding proteins (IGFBPs) in improved growth rate of H. midae, the relevance of investigating ways to stimulate insulin/IGF release in aquaculture species was again emphasized. As nutritional administration remains the most probable route of introducing agents that can stimulate the release of insulin-related peptides, continuous endeavours to stimulate abalone growth through a nutritional approach is encouraged. This is the first time next generation sequencing is used towards the large scale transcriptome sequencing of any haliotid species and also the first time a comprehensive investigation is launched towards the establishment of primary cell cultures for H. midae. A considerable amount of sequence data was furthermore annotated for the first time in H. midae. The results obtained here provide a foundation for future genetic studies exploring ways to optimise the commercial production of H. midae.
AFRIKAANSE OPSOMMING: Die stadige groeitempo van Haliotis midae belemmer die optimale kommersiele produksie van hierdie mees winsgewende Suid-Afrikaanse akwakultuur spesie. Tot op hede is geen omvattende poging aangewend om gene verwant aan groeivariasie in H. midae te identifiseer nie. Die doel van hierdie studie was dus om groeivariasie in H. midae te ondersoek en om spesifieke groei-gekoppelde gene te identifiseer en hul uitdrukking te kwantifiseer. Ter bereiking van hierdie doel is molekulêre metodes en selkulture gekombineer as 'n en tydsbesparende en ekonomiese manier om perlemoen transkriptomika en selbiologie te bestudeer. Moderne Illumina volgordebepaling-deur-sintese tegnologie en daaropvolgende annotasie is gebruik om verskille in geenuitdrukking tussen naby-verwante groepe perlemoen, wat noemenswaardige groeivariasie vertoon, toe te lig. Na afloop van die transkriptoom volgordebepaling is gene betrokke by groei en metabolisme, vantevore onbekend in H. midae, geïdentifiseer. Die uitdrukking van uitgesoekte teikengene betrokke by groei is vervolgens ge-analiseer deur kwantitatiewe "real-time PCR" (qPCR). die lewensvatbaarheid van 'n primêre selkulture vir H. midae is ook ondersoek deur embrio, larwe en hemolimf weefsels te teiken vir die daarstelling van primêre selkulture. Larweselle en hemosiete kon in vitro suksesvol onderhou word vir beperkte periodes. Primêre hemosietkulture het geblyk 'n gepaste in vitro sisteem te wees om geenuitdrukking te bestudeer en dit is vervolgens gebruik vir RNS ekstraksie en qPCR, om differensiële groei, geïnduseer deur insulien en epidermale groeifaktor (EGF), te evalueer. Geenuitdrukking is dus gekwantifiseer in vinnig- en stadiggroeiende perlemoen en in in vitro primêre hemosiet selkulture wat behandel is met verskillende groei stimulante. Die resultate wat verkry is van transkriptoomanalise en qPCR het noemenswaardige verskille in geenuitdrukking tussen groot en klein perlemoen, en tussen behandelde en onbehandelde hemosiet selkulture uitgelig. Die op-regulering van gene betrokke by die insulien sein-padweg, tydens in vivo en in vitro qPCR eksperimente, bied getuienis vir die betrokkenheid van insulien in die verhoogde groeitempo van verskeie H. midae weefsels. Benewens die regulering van teikengene is waardevolle kennis ook ingewin in terme van verwysingsgene tydens qPCR eksperimentering. Deur die stabiele uitdrukking van twee gene (8629, ribosomale proteien S9 en 12621, ornitien dekarboksilase) te kwantifiseer in verskeie weefsels en onder verskeie kondisies is gepaste verwysingsgene, wat ook in toekomstige H. midae qPCR eksperimente aangewend kan word, geïdentifiseer. Deur getuienis vir die betrokkenheid van insulien, insuliensoortige groeifaktor en insuliensoortige groeifaktor-bindingsproteïene by verbeterde groei van H. midae op transkripsievlak te bied, is die toepaslikheid van bestudering van maniere om insulienvrystelling in akwakultuurspesies te stimuleer, beklemtoon. Aangesien voeding die mees waarskynlike roete is om middele wat insuliensoortige peptiedvrystelling stimuleer daar te stel, word vogehoue pogings om perlemoengroei deur die regte voeding te stimuleer, aangemoedig. Hierdie is die eerste studie wat volgende generasie volgordebepaling (“next generation sequencing”) gebruik vir die grootskaalse transkriptoom volgordebepaling van enige haliotied spesie. Dit is ook die eerste keer dat ‘n omvattende ondersoek geloods word na die daarstelling van primêre selkulture vir H.midae. ‘n Aansienlike hoeveelheid volgorde data is ook vir die eerste keer geannoteer in H. midae. Die resultate wat hier verkry is bied ‘n basis vir toekomstige genetiese studies wat maniere ondersoek om die kommersiële produksie van perlemoen te optimiseer.

Thesis (MSc)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: Haliotis midae (locally also known as perlemoen) is the largest of five endemic species found along the coast of South Africa. It is the only species with commercial value contributing to the exploitation of these animals. Due to declines of natural stocks, farming practices were established during the early 1990s in order to supply the international demand. To facilitate efficient breeding methods and ensure the sustainability of these commercial populations, genetic management, which can be accomplished with the use of molecular markers such as single nucleotide polymorphisms (SNPs), is necessary. Single nucleotide polymorphisms have become the markers of choice in various applications in aquaculture genetics due to their abundance in genomes, reduction in developmental costs and increased throughput of genotyping assays. Identification of SNPs in non-model species such as H. midae can be achieved by in silico approaches. In silico methods are suitable for de novo SNP identification and are both cost- and time-efficient. It is based on the analysis of multiple alignments where mismatches may be reported as candidate SNPs. Various medium-throughput genotyping methods are available to confirm putative SNPs, but the ideal method depends on factors such as cost, accuracy and multiplexing capacity. Although SNP markers can have various applications within the aquaculture environment the focus for this current study was saturating the linkage map of H. midae with additional markers. This would assist in the identification of quantitative trait loci associated with economically important traits, which in turn could ultimately be employed for marker-assisted selection and improved molecular breeding programs. In order to identify in silico SNPs, sequenced transcriptome data from a previous study was used and subjected to a series of criteria: minor allele frequency 10%, minimum coverage 80, 60 bp flanking regions. Selected loci were genotyped using a 192-plex assay with the Illumina GoldenGate genotyping assay with the VeraCode technology on the BeadXpress platform, in individuals from six mapping families. A conversion rate of 69.35% and global success rate of 76.34% was achieved. Polymorphic loci were subjected to linkage analysis using JoinMap® v.4.1 to create sex-average and sex-specific maps and to saturate the current linkage map for H. midae. Along with previously developed markers, 54% of the newly developed SNPs could be successfully incorporated into the linkage map of H. midae. A total of 18 linkage groups were observed with an average marker spacing of 6.9 cM and genome coverage of 79.1%. Bioinformatic analyses and setting stringent criteria to identify SNPs from sequenced transcriptomic data proved to be an efficient way for SNP discovery in the current study. Genotyping of the identified loci with the GoldenGate genotyping assay demonstrated a high success rate; providing a genotyping assay adequate for species with little genomic information. The linkage map created in this study illustrated the utility of SNP markers in conjunction with microsatellite markers for linkage map construction and the adequate marker spacing obtained provides a step closer to quantitative trait loci mapping in this species.
AFRIKAANSE OPSOMMING: Haliotis midae (plaaslik ook bekend as perlemoen) is die grootste van vyf inheemse spesies wat langs die kus van Suid-Afrika aangetref word. Dit is die enigste spesie van kommersiële waarde wat bydraend is tot die uitbuiting van hierdie diere. As gevolg van die afname in hierdie natuurlike hulpbron het boerdery praktyke gedurende die vroeë 1990's ontstaan om in die internasionale aanvraag te voorsien. Ten einde doeltreffende teelmetodes te beoefen en die volhoubaarheid van hierdie kommersiële populasies te verseker is genetiese bestuur, wat bewerkstellig kan word deur die gebruik van molekulêre merkers soos enkel nukleotied polimorfismes (ENPs), baie belangrik. Enkel nukleotied polimorfismes is gewilde merkers in verskeie toepassings in akwakultuur genetika as gevolg van hul oorvloed in genome, verlaagde ontwikkelingskoste en verhoogde deurset van ENP-genotiperingstoetse. Identifisering van ENPs in nie-model spesies soos H. midae kan uitgevoer word deur in siliko benaderings te gebruik wat geskik is vir de novo ENP identifisering en ook tyd- en koste-effektief is. Dit word gebaseer op die analise van veelvuldige inlynstellings waar nukleotiedes wat nie ooreenstem nie as kandidaat ENPs gerapporteer kan word. Om kandidaat ENPs te bevestig, kan verskeie medium-deurset genotiperingsmetodes uitgevoer word, maar die ideale metode word bepaal deur faktore soos koste, akkuraatheid en multipleks kapasiteit. Alhoewel ENP merkers in verskeie toepassing binne die akwakultuur omgewing gebruik kan word was die fokus van die huidige studie om die koppelingskaart van H. midae te versadig. Dit sal bydrae tot die identifisering van kwantitatiewe eienskap lokusse wat gekoppel kan word aan ekonomies belangrike eienskappe wat dan op die beurt weer vir merkerbemiddelde seleksie gebruik kan word en uiteindelik ten opsigte van die verbetering van molekulêre teelprogramme aangewend kan word. Ten einde in siliko ENPs te identifiseer is transkriptoomdata van 'n vorige studie gebruik en onderwerp aan 'n reeks kriteria: geringste alleelfrekwensie 10%, minimum dekking 80, 60 bp gebiede weerskante van polimorfisme. Geïdentifiseerde lokus-genotipering is met behulp van 'n 192-pleks toets uitgevoer met die Illumina GoldenGate genotiperingstoets met die VeraCode tegnologie op die BeadXpress-platform, in individue afkomsitg vanaf ses karteringsfamilies. 'n Omskakelingskoers van 69.35% en 'n algehele sukseskoers van 76.34% is bereik. Polimorfiese lokusse is onderwerp aan koppelings-analise met behulp van JoinMap® v.4.1 om geslags-gemiddelde en geslags-spesifieke kaarte te skep asook om die kaart wat beskikbaar is vir H. midae te versadig. Saam met voorheen ontwikkelde merkers is 54% van die nuut ontwikkelde ENPs suksesvol opgeneem in die kaart van H. midae. 'n Totaal van 18 koppelingsgroepe is verkry met 'n gemiddelde merker-spasiëring van 6.9 cM en 'n genoomdekking van 79.1%. Die gebruik van bioinformatiese analises en streng kriteria om ENPs vanaf transkriptoomdata te identifiseer blyk doeltreffend te wees in hierdie studie. Genotipering van die geïdentifiseerde lokusse met die GoldenGate genotiperingstoets dui op 'n hoë suksessyfer en verskaf 'n voldoende genotiperingstoets aan spesies met min genomiese inligting. Die koppelingskaart in hierdie studie het geïllustreer dat die ENP merkers suksesvol saam met mikrosatelliet merkers gebruik kan word vir koppelingskaart konstruksie en dat die voldoende merker-spasiëring verkry 'n stap nader aan kwantitatiewe eienskap lokus kartering in hierdie spesie bied.

Thesis (MScAgric)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: The species Haliotis midae is of great commercial value to the South African abalone industry and is mainly exported to Asian markets, specifically China. Up to 50% is sold as canned products with H. midae registering an average canning yield of approximately 35%. The species is presently genetically undomesticated and breeding programmes are being introduced to improve a range of production traits of which growth and yield is of primary importance. The objective of the study was to determine genetic parameters such as heritability, genotypic and phenotypic correlations of yield-related traits to assess the potential genetic improvement through selective breeding. A series of yield-related parameters were identified that is of relevance to the standard abalone canning procedure. Low to moderate heritabilities where recorded for most traits, including pre-shuck/live weight (0.20 ± 0.06), post-shuck weight (0.15 ± 0.05), post-gut weight (0.15 ± 0.05), post-brine weight (0.19 ± 0.06), pre-canning weight (0.19 ± 0.06), post-canning weight (0.21 ± 0.06), shell weight (0.16 ± 0.05), canning yield percentage (0.08 ± 0.03) and shell weight to post-gut weight ratio (SW: PGW) (0.09 ± 0.04). Weight related parameters are phenotypically highly correlated (0.86 ≤ r ≤ 0.99) but show negative correlation with canning yield percentage (-0.38 ≤ r ≤ 0.04). The nett yield of abalone shows a relatively strong positive correlation with the live weight (r = 0.66). Shell length is highly heritable (h2 ≈ 0.48) and show a strong positive correlation with live weight (r = 0.94). Shell weight is also highly correlated with live weight (r = 0.80) and the SW: PGW ratio does not show a significant correlate with live weight (r = 0.03). Weight-related traits show heritability values ranging from 0.15 to 0.20 that could allow a positive genetic response. Shell length (as a linear growth parameter) shows a high heritability (h2 ≈ 0.48) and a strong positive correlation with live weight (r = 0.94) which also makes it suitable for use as a selection criterion in breeding programmes for improved growth rate. Direct selection for canning yield is compromised by the destructive nature of measurement and the low heritability (h2 < 0.10). The negative correlations between yield as a percentage and growth traits (-0.38 ≤ r ≤ 0.04) further complicate its use as a direct breeding objective. Although the canning yield as a percentage shows a decrease with an increase in live weight, the nett canning yield increases (r = 0.66) with the live weight. It is therefore recommended to use shell length as a criterion for selection for increased growth rate and nett yield, thereby optimising profitability.
AFRIKAANSE OPSOMMING: Die spesie Haliotis midae is van groot kommersiёle waarde tot die Suid-Afrikaanse perlemoenindustrie en word meestal uitgevoer na markte in Asiё, spesifiek China. Tot 50% van die perlemoen wat in Suid-Afrika geproduseer en uitgevoer word, word verblik en huidiglik is die verblikkingsopbrengspersentasie ongeveer 35%. Haliotis midae is tans geneties onderontwikkeld en die gebruik van teelprogramme word nou geimplementeer met die doel om 'n verskeidenheid eienskappe te verbeter, waarvan groei en opbrengs van primêre belang is. Die doelwit van die studie was om genetiese parameters soos oorerflikheid en ook die genotipiese en fenotipiese korrelasies van obrengsverwante eienskappe te bepaal om sodoende die potensiёle genetiese verbetering as gevolg van selektiewe teeling te assesseer. 'n Reeks obrengsverwante eienskappe is geidentifiseer wat relevant is binne bestaande en standaard kommersiёle perlemoenverblikkingsprotokolle. Lae tot matige oorerflikheidswaardes is waargeneem en sluit in lewende, of voor-ontskulpingsgewig (0.20 ± 0.06), na-ontskulpingsgewig (0.15 ± 0.05), na-oopvlekkingsgewig (0.15 ± 0.05), na-pekelgewig (0.19 ± 0.06), voor-verblikkingsgewig (0.19 ± 0.06), na-verblikkingsgewig (0.21 ± 0.06), skulpgewig (0.16 ± 0.05), verblikkingsopbrengspersentasie (0.08 ± 0.03) en 'n skulpgewig tot na-oopvlekkingsgewig verhouding (SW: PGW) (0.09 ± 0.04). Gewigsverwante parameters is fenotipies hoogs gekorreleerd met mekaar (0.86 ≤ r ≤ 0.99) maar toon 'n negatiewe korrelasie met die verblikkingsopbrengspersentasie (-0.38 ≤ r ≤ 0.04). Die netto opbrengs van perlemoen dui op 'n relatiewe sterk positiewe korrelasie met lewende gewig (r = 0.66). Skulplengte is hoogs oorerflik (h2 ≈ 0.48) en toon 'n sterk positiewe korrelasie met lewende gewig (r = 0.94). Skulpgewig is ook hoogs gekorreleerd met lewende gewig (r = 0.80) en die SW: PGW verhouding toon geen beduidende korrelasie met lewende gewig nie (r = 0.03). Gewigsverwante eienskappe toon oorerflikheidswaardes wat varieer tussen 0.15 en 0.20 en kan 'n moontlike genetiese respons lewer. Skulplengte (as 'n liniêre groeiparameter) toon 'n hoё oorerflikheid (h2 ≈ 0.48) en 'n sterk positiewe korrelasie met lewende gewig (r = 0.94) wat dit gepas maak vir gebruik as 'n seleksiekriterium in 'n teelprogram met verbeterde groeitempo as doel. Direkte seleksie in terme van verblikkingsopbrengs word ingeboet danksy die destruktiewe natuur van die metingsmetodiek asook 'n lae oorerflikheid (h2 < 0.10). Die negatiewe korrelasies tussen verblikkingsopbrengs (uitgedruk as 'n persentasie) en groeieienskappe (-0.38 ≤ r ≤ 0.04) dien as 'n verdere komplikasie in die gebruik van dié eienskap as direkte teeldoelwit. Alhoewel die verblikkingopbrengs 'n afname toon soos lewende gewig toeneem, is daar steeds 'n positiewe korrelasie tussen die netto verblikkingsopbrengs en die lewende gewig (r = 0.66). Dit word dus aangeraai om skulplengte as seleksiekriterium vir verbeterde groeitempo en netto opbrengs te gebruik om sodoende wins te maksimaliseer.

Thesis (PhD) -- tellenbosch University, 2014.
ENGLISH ABSTRACT: This study is part of the continuing global research effort dedicated to finding alternative aquafeed ingredients, which not only replace fish meal and fish oil as sources of essential nutrients in aquafeeds, but also provide specialist functional properties when included in feeds. Due to constraints in supply of fish meal and fish oil originating from wild capture fisheries, the continually growing aquafeed industry requires new sources of raw materials for the production of high quality feeds. The aim of the study was to investigate the specialist functional properties of feed ingredients (with emphasis on immune-stimulation potential) derived from fish processing waste after enzymatic hydrolysis by endogenous proteolytic enzymes (autolysis). Further aims were to (i) quantify effects of these feed ingredients on production performance of two species relevant to the South African aquaculture industry, namely Mozambique tilapia Oreochromis mossambicus and South African abalone Haliotis midae, (ii) compare performance to commercially available enzymatically produced feed ingredients, and (iii) separate the functional effects of these fish processing waste derived feed ingredients from the acid used to preserve them against bacterial spoilage during the autolysis process. Oil recovered after autolysis of rainbow trout viscera proved to be an attractive feed ingredient due to favourable effects on the non-specific cellular immune function of both Mozambique tilapia and South African abalone. However, in South African abalone, increased immune function due to inclusion of fish oil was accompanied by a significant decrease in production performance. The inclusion of hydrolysed proteins, obtained by autolysis of fish waste, in aquaculture feeds also improved non-specific immunity and survival of Mozambique tilapia significantly – independently of the preserving acid – although the same was not observed for South African abalone. Production performance was dependent on dietary hydrolysed protein inclusion levels in both species; excessive inclusion resulted in decreased production performance. The performance of dietary hydrolysed protein from autolysis and those from commercial production processes were significantly different, possibly as a result of different raw material origins and production processes. It is further shown that formic acid can contribute to improved water stability in abalone feeds, a novel mode of action not previously described. The study concludes that the simple autolysis process for processing of fish waste can provide aquafeed ingredients with immune stimulatory potential, which can contribute to improved production performance in the Mozambique tilapia and the South African abalone. The result can contribute to improved sustainability of the aquafeed industry, through substitution of fish meal and fish oil derived from capture fisheries with processed fish waste components.
AFRIKAANSE OPSOMMING: Hierdie studie vorm deel van die voortdurende en wêreldwye soeke na nuwe akwakultuur voerbestanddele wat nie net vismeel en –olie kan vervang as bron van noodsaaklike voedingstowwe nie, maar wat ook gespesialiseerde funksionele eienskappe openbaar wanneer dit in akwavoere ingesluit word. As gevolg van beperkings in die voorsiening van vismeel en –olie afkomstig vanaf wilde visserye, word die voortdurend groeiende akwavoer bedryf genoodsaak om nuwe grondstowwe te benut vir die vervaardiging van hoë kwaliteit voere. Die doelwit van die studie was om moontlike gespesialiseerde funksionele eienskappe (met spesifieke klem op potensiële immuun-stimulasie) van voerbestanddele te ondersoek wat herwin is vanaf reënboogforel proseseringsafval, na ensiematiese hidrolise d.m.v. endogene ensieme teenwoordig in die afval (outolise). Verdere doelwitte was om (i) effekte op produksie prestasie van hierdie bestanddele te kwantifiseer in twee akwakultuur spesies relevant tot die Suid- Afrikaanse bedryf (naamlik die Mosambiekse kurper Oreochromis mossambicus en die Suid- Afrikaanse perlemoen Haliotis midae), (ii) om produksie prestasie te vergelyk met kommersieel beskikbare voerbestanddele voorberei d.m.v. ensiemtegnologie, en (iii) om die moontlike effekte van die voerbestanddele te skei van die van die sure gebruik om die bestanddele te preserveer tydens die outolitiese proses. Daar is bevind dat olie herwin na outolise van reënboogforel ingewande ‘n goeie voerbestanddeel is wat gunstige effekte op die nie-spesifieke, sellulêre immuniteit van beide Mosambiekse kurper en Suid-Afrikaanse perlemoen gehad het. In die Suid-Afrikaanse perlemoen het verbeterde immuunfunksie gepaard gegaan met ‘n verswakking in produksie prestasie. Die gehidroliseerde proteïen komponent van ge-outoliseerde prosesseringsafval het ook beduidende verbetering in nie spesifieke immuniteit en oorlewing van Mosambiekse kurper tot gevolg gehad - onafhanklik van die preserverende suur - maar dieselfde is nie in die Suid-Afrikaanse perlemoen waargeneem nie. Produksie prestasie was afhanklik van die insluitingsvlakke van gehidroliseerde proteïen in beide spesies en dit is bevind dat oormatige insluiting produksie nadelig beïnvloed. Prestasie van proteïen afkomstig van outolise en die van ‘n kommersiële produksieproses het beduidend verskil, moontlik as gevolg van verskillende grondstowwe en prosesseringstegnieke. Daar is verder vir die eerste keer getoon dat mieresuur ‘n beduidende verbetering in waterstabiliteit in sekere perlemoenvoere teweeg kan bring. Die studie kom tot die slotsom dat die eenvoudige outolise proses funksionele akwavoerbestanddele kan produseer wat kan bydra tot ‘n verbetering in produksie prestasie in die Mosambiekse kurper en Suid-Afrikaanse perlemoen. Die resultaat kan bydra tot verbeterde volhoubaarheid van die akwavoer bedryf, deur vismeel en –olie afkomstig van wilde visserye, te vervang.

Thesis (PhD)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: The South African abalone, Haliotis midae, commonly known as perlemoen, is an economically important gastropod mollusc. Historically, this species maintained a lucrative fisheries sector; however with increasingly lower landings there has now been a shift to aquaculture. Efforts to conserve natural populations and to improve abalone aquaculture production are thus running in parallel. Previous studies reported significant disparities in parental contributions in aquaculture populations that could explain the rapid divergence of commercial stocks from wild populations. Furthermore, subtle, but significant, population differentiation has also been reported for wild populations on the west-, south-, and east coast of the South African coastline. This study therefore aimed to investigate the evolutionary forces, in particularly selection, facilitating population divergence in wild and cultured H. midae populations using a population genomics approach. By using both microsatellite- and single nucleotide polymorphism (SNP) markers it was found that approximately 10% to 27% of the H. midae genome may be influenced by selection. When incorporating these loci into analyses of population differentiation (e.g. AMOVA, factorial correspondence analysis and estimates of genetic distance) there was a marked increase in genetic divergence between wild and cultured populations (especially when using microsatellite loci) and amongst populations from different geographic regions (particularly supported by the SNP loci). The differences in population clustering as highlighted by microsatellite- and SNP markers can most likely be attributed to the genomic distribution of the respective loci: The SNP markers were developed from EST sequences and therefore mostly represents protein structural variation; whereas the microsatellite markers, found to be putatively under selection, were mainly located in regulatory motifs. The results of this study therefore confirmed previous observations of divergence amongst wild- and cultured populations, but more importantly demonstrated that selection is an important factor driving this divergence. In wild populations selection probably facilitates adaptation to local environmental conditions, whilst amongst aquaculture population adaptation to captivity, husbandry practices and artificial selection may be important determinants. There is evidence for population bottlenecks in wild- and cultured populations; nonetheless long-term effective population sizes seem to be large. Amongst the wild populations, however, short-term population sizes appear to be small most likely due to differential spawning rates amongst reproductively active animals leading to temporal fluctuation in genetic diversity. The results indicate that contact between wild and cultured abalone should be minimised to prevent any adverse effects due to outbreeding depression. With regards to conservation, an emphasis on maintaining adaptive diversity of the wild stocks might be warranted. Continued genetic monitoring is advisable for both wild and cultured abalone populations as to optimally manage the abalone resource for both conservation and commercial viability and sustainability.
AFRIKAANSE OPSOMMING: Die Suid-Afrikaanse perlemoen, Haliotis midae, is 'n ekonomies belangrike buikpotige weekdier. Histories het hierdie spesie 'n winsgewende vissery gehandhaaf, maar met steeds dalende vangste is daar nou 'n verskuiwing na akwakultuur. Pogings om natuurlike populasies te bewaar en perlemoen te verbeter vir verhoogde akwakultuur produksie loop dus in parallel. Vorige studies het bevind dat beduidende verskille in ouerlike bydraes tot die nageslag, in akwakultuur populasies, kan verduidelik hoekom die populasies so vinnig divergeer van die wilde voorouers. Verder, is subtiele, maar betekenisvolle genetiese differensiasie tussen wilde populasies aan die wes-, suid-en ooskus van die land gevind. Hierdie studie is dus daarop gemik om ondersoek in te stel na die mate waartoe verskeie evolusionêre prosesse, in besonder seleksie, die populasie divergensie in beide wilde en gekweekte H. midae teweegbring deur gebruik te maak van ‘n populasie genomika benadering. Deur gebruik te maak van beide mikrosatelliet- en enkel nukleotied polimorfisme (ENP) merkers is dit bevind dat ongeveer 10% tot 27% van die H. midae genoom moontlik beïnvloed word deur seleksie. Met die gebruik van loki onder seleksie tydens die ontleding van populasie differensiasie (bv. AMOVA, faktoriaal korrespondensie analise en genetiese afstand ramings) was daar 'n merkbare toename in genetiese divergensie tussen wilde- en gekweekte populasies (veral wanneer mikrosatelliet loki gebruik is) en onder die populasies vanuit verskillende geografiese gebiede (veral ondersteun deur die ENP loki). Die verskille in die populasie groeperings soos uitgelig deur die mikrosatelliet- en ENP-merkers kan waarskynlik toegeskryf word aan die genomiese verspreiding van die onderskeie loki: Die ENP-merkers is ontwikkel vanaf uitgedrukte volgorde merker (UVM) volgordes en daarom verteenwoordig dit meestal proteïen strukturele veranderinge, terwyl mikrosatelliet merkers eerder in regulatoriese motiewe geleë is. Die resultate van hierdie studie steun dus vorige waarnemings, maar meer belangrik, het dit getoon dat seleksie ‘n betekenisvolle faktor in populasie divergensie in beide wilde en gekweekte populasies is. In wilde populasies fasiliteer seleksie waarskynlik die aanpassing tot plaaslike omgewingstoestande terwyl seleksie onder die gekweekte populasies teweeggebring kan word as gevolg van aanpassing tot aanhouding, boerdery praktyke en kunsmatige seleksie. Daar is bewyse vir populasie bottelnekke in wilde- en gekweekte populasies; tog blyk langtermyn effektiewe populasiegroottes om redelik groot te wees. Onder die wilde populasies is egter gevind dat kort-termyn populasiegroottes klein kan wees, waarskynlik as gevolg van differensiële broeikoerse onder reproduktiewe diere. Dit het tot gevolg dat daar beduidende fluktuasies is in temporale genetiese diversiteit. Die resultate dui daarop dat kontak tussen wilde en gekweekte perlemoen tot 'n minimum beperk moet word om enige nadelige effekte weens uitteling depressie te voorkom. Verder, met betrekking tot bewaring, is ‘n klem op die handhawing van aangepaste genetiese diversitiet dalk geregverdig. Voortgesette genetiese monitering word aanbeveel vir beide wilde- en gekweekte perlemoen populasies ter wille van die optimale bestuur van die perlemoen hulpbron vir beide bewaring en kommersiële lewensvatbaarheid en volhoubaarheid.
International Foundation for Science
National Research Foundation of South Africa
Stellenbosch University

Thesis (MScAgric)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: The abalone Haliotis midae is the most important aquaculture species in South Africa. The industry is dependent on export to Far Eastern markets in a variety of forms, including live, frozen, canned or dried. The species is considered undomesticated in the sense that the current commercial broodstock has been obtained from natural populations through a process of random collection. Global competition has necessitated the South African industry to introduce a genetic improvement program to increase biological productivity and financial profitability. The objective of this study was to assess the genetic variation and to estimate key parameters in terms of growth and yield related traits, between family groups that form part of the breeding program. The study reports on heritability estimates of growth rate (0.14 ± 0.05), canning yield (0.08 ± 0.03), and drip loss during live export (0.03 ± 0.02). The high genetic correlation (0.94 ± 0.34) between shell length and live weight enables industry to utilise either weight or shell length as a criteria during operational practices such as sorting, grading and harvesting. The correlation of 0.85 ± 0.01 between live weight and canning loss indicates that animals that weigh more have a lower dressing percentage. Based on these low heritability values obtained for yield related traits it is recommended not to include these traits in the selection program at this stage. The findings of the study were however, compromised by the availability of a limited number of family groups, the age differences between families and the effect of different locations on the variance in phenotypes. Further investigation is needed to confirm the credibility of the results.
AFRIKAANSE OPSOMMING: Die perlemoen Haliotis midae is die belangrikste akwakultuurspesie in Suid Afrika. Die industrie is afhanklik van uitvoere na markte in die Verre Ooste. ‘n Verskeidenheid van produkte word uitgevoer, insluitend lewendige, gevriesde, gedroogte en verblikte perlemoen. Die spesie word as ongedomestikeerd beskou aangesien die huidige teeldiere op ‘n lukrake wyse uit natuurlike populasies versamel is. Globale kompetisie het die Suid Afrikaanse industrie genoodsaak om ‘n genetiese verbeteringsprogram in werking te stel om sodoende die biologiese produktiwiteit en finansiële winsgewendheid te verbeter. Die studie poog om genetiese variasie in groei en opbrengsverwante kenmerke tussen familiegroepe wat deel uitmaak van die teelprogram te ondersoek. Oorerflikheid van groeitempo (0.14 ± 0.05), opbrengs na verblikking (0.08 ± 0.03), en vogverlies na lewendige uitvoer (0.03 ± 0.02) is beraam. Die hoë korrelasie (0.94 ± 0.34) tussen gewig en skulplengte stel die industrie in staat om beide massa en skulplengte as kriteria te geruik tydens operasionele praktyke van sortering, groepering en oes. Die korrelasie van 0.85 ± 0.01 tussen gewig en verlies na verblikking dui aan dat swaarder diere ‘n laer uitslagpersentasie het. As gevolg van die lae oorerflikheidswaardes vir opbrengsverwante kenmerke word daar aanbeveel dat hierdie kenmerke nie op hierdie stadium ingesluit word in die seleksieprogram nie. Resultate is egter beïnvloed deur ‘n beperkte aantal familie groepe, ouderdomsverskille tussen families en die effek van verskillende lokaliteite op die variansie in die fenotipes. Verdere ondersoeke is nodig om die geloofwaardigheid van die bevindinge te bevestig.

Abalone culture is becoming increasingly important as over-exploitation and poaching worldwide are depleting wild stocks, yet demand for abalone products continues to increase. Although techniques for nursery production of juveniles and grow-out to market size are well established, optimization of growth rates and maintaining stock health requires a better understanding of factors influencing growth, water quality and health status during the grow-out phase. Previous studies on abalone at the University of Tasmania have investigated a range of husbandry, nutritional and water quality issues. The research covered in the current study used a range of experimental scale grow-out systems, in conjunction with a bioassay system, to investigate a number of factors previously identified as potentially affecting growth and water quality in commercial grow-out systems for greenlip (Haliotis laevigata) and blacklip (H rubra) abalone. Previous work has investigated the effect of environmental ammonia and low dissolved oxygen levels on abalone growth as separate exposures (Harris 1999). For the present study, this work was expanded to investigate the effect on growth of low dissolved oxygen levels combined with elevated ammonia levels as both chronic exposure and as intermittent, 8-hour exposures in two separate trials.

The Australian greenlip abalone Haliotis laeviqata is of commercial importance in the abalone diving industry. The species is also believed to have culture potential and accordingly control over reproduction is considered valuable. The major purpose of the study was to accelerate gonad development resulting in spawning outside the natural spawning season. Abalone were collected from Franklin sound in the Furneaux group of islands off the north-east tip of Tasmania. The important features of the conditioning tank were elevated water temperature, the provision of water movement within the tank to distribute feed to sedentary abalone and a diet of preferred red algae. Animals collected on 27 April 1990 were induced to spawn on 21 August 1990, 112 days or 1750 degree days following commencement of gonad conditioning. The natural spawning season of the source population was found to be November to March. A variety of methods for measuring reproductive development of abalone were used. This allowed the utility of individual methods to be examined and comparisons made between methods. Two gonad indices, the gonad bulk index (GBI) and the modified gonad bulk index (MGBI) were used as were a number of assessment methods with a histological component: oocyte size/frequency distribution, mean oocyte diameter, an ovarian phase method and percentage mature spermatozoa. The MGBI was considered more sensitive than the GBI, detecting first significant gonad growth following six weeks of gonad conditioning, compared to nine weeks for the GBI. The gonad indices increased from initial values of 14.1 ± 4.4, n=10 and 0.4 ± 0.2, n=10 for GBI and MGBI respectively to 72.3 ± 9.2, n=10 and 7.0 ± 2.0, n=10 following 105 days of gonad conditioning. Mean oocyte diameter (Am) increased from 30.7 ± 2.0, n=5 initially to 109.7 ± 6.0, n=5 during the same time period. The percentage of male germ cells present as mature spermatozoa increased from zero to 90.6 ± 16.3, n=5 following 24 weeks of conditioning.

A series of experiments were conducted to investigate the protein, energy and lysine requirements of greenlip abalone Haliotis laevigata. An initial experiment estimated the optimal crude protein requirement. Digestibility coefficients of the protein and energy components of feed ingredients were then estimated and used to formulate diets to investigate the optimal dietary ratio of digestible protein to digestible energy and the effect these parameters have on tissue quality in abalone. Finally, the lysine requirement of greenlip abalone was estimated using two methods of lysine supplementation. The first experiment involved feeding a series of semi-purified diets in which the essential amino acid profile was similar to that of the soft tissue of abalone. Abalone grew fastest at a crude protein level of 20.1%. Excess dietary protein reduced the growth rate. The optimal digestible protein:digestible energy ratio was estimated at 14.4 gDP.M.VDE. Leaching of essential amino acids over 24 hours immersion ranged from 26-54%. The apparent and true digestibility coefficients of the protein and energy components of five feedstuffs were estimated. Defatted soyflour provided the most digestible protein (96.7% digestible). Pre-gelled starch provided the most digestible energy (80.6%). Cost analysis indicated that defatted soyflour was the most costeffective source of protein and energy of the feedstuffs studied. In a factorial experiment, abalone were fed diets at ten levels of protein and two levels of energy. The optimal dietary digestible protein level was estimated at only 13%, and the optimal ratio of digestible protein:digestible energy between `12.3-17.9 g.M.J^-1`. Abalone grew poorly on diets with fish oil levels of 6.0 - 9.0%, although their growth rate increased when returned to a commercial diet containing less oil. The composition of abalone foot muscle and, to a lesser extent, viscera, was affected by dietary protein level and, in some cases, by dietary energy level. Carcass partitioning revealed that meat yield was also affected by dietary protein level. The lysine requirement of greenlip abalone was determined using two methods of lysine supplementation; the addition of free L-lysine-hydrochloride to diets and the use of gluten enriched with covalently-bound lysine. Abalone showed a linear response to free-lysine, and a curvilinear response to covalently-bound lysine. The former method was therefore unable to produce an estimate of dietary lysine requirement. Use of covalently-bound lysine predicted that maximal growth occurred when lysine accounted for 3.9% of crude protein.

Stock losses in the range 50-90% were recorded at experimental and pilot scale sea-based abalone farms in southern Tasmania in the mid-1990's. These were associated with spionid polychaete "mud worm" infestation, especially Boccardia knoxi. The overall aim of the multi faceted research presented here was to minimise the effects of spionid infestation. Studies of reproductive biology indicated initial infestation with B. knoxi could be delayed by placement of stock after the spring planktotrophic dispersal phase. This would also reduce infestation by a second species Polydora hoplura. Fieldwork during 1998-2001 indicated that large settlements of spionids might be relatively uncommon. Testing of 16 chemical/drug agents and freshwater bathing as a treatment for mud worm infestation failed to yield a useful candidate. Agents lacked penetration into shell burrows or were harmful to abalone at levels sufficient to kill spionids in situ. Air exposure of abalone for 2-4 hours at humidity less than approximately 63% was highly effective as a spionid treatment, especially in the first 6 months post infestation. Assessment of risk factors associated with spionid settlement found that elevated levels of spirorbid polychaetes enhanced mud worm colonisation. Stock size, rearing vessel design and position in the water column also led to differential spionid settlement outcomes. Spionid infestation was associated with a reduction in: growth, % flesh weight,, protein and carbohydrate reserves and with an increased respiration rate. Major histological changes were elevated levels of brown pigment in the right kidney and digestive gland consistent with mobilisation and consumption of energy reserves. The ranges in levels of sodium, potassium, calcium, magnesium, chloride, copper, glucose and protein in haemolymph were established for apparently healthy abalone and spionid infested animals.

碩士
國立臺灣海洋大學
生物科技研究所
95
Taiwan abalone (Haliotis diversicolor) is one of the important economical aquaculture shellfish in Taiwan. In 2002-2003, a massive death break out occurred in north-east of Taiwan, and virus was found to be the causative pathogen. The objective of this study is to establish tissue culture system of Taiwan abalone as the platform for studying virus infection in the future. In the preliminary data, the adhesion rate of Taiwan abalone cells was augmented by rising the concentration of L-15 and Grace’s medium. Therefore, the medium osmolarity is probably the key factor for Taiwan abalone cells adhesion. We also observed that high osmolarity (735~1,110 mOsmkg-1) of L-15 medium can promote abalone ovarian cells adhesion, and the proliferation can also be stimulated by the supplement of salt solution. In order to search better condition of abalone cell culture, we have developed a new medium MAH by modifying the formula of L-15 and Grace’s medium and also from the results of component assay of Taiwan abalone hemolymph extract. Comparing the growth of Taiwan abalone ovarian and hemocyte cells in modified LS20, GLS15 and MAH media, the best medium is GLS15. Although the adhesion cells are unable to attach after subculture, the suspension cells can be passaged. Till now, the suspension ovarian cells can be subcultured for 8 times, alive for 85 days; the suspension hemocyte can be subcultured for 11 times, alive for 96 days. In the medium supplement study, the supplement of abalone hemolymph extract can promote adhesion and proliferation of hemocyte, but not of insulin.

This study of land based abalone farming was conducted to investigate the industry's environmental impact and long-term sustainability. There is a paucity of information surrounding the environmental impacts of land based abalone farming despite the increase in the growth of the industry. Information which forms the basis of abalone farm environmental management and monitoring was developed as part of this project. This information is likely to provide a great deal of perspective to all existing and planned Australian abalone farms. Three preliminary experiments were conducted to determine how nutrients were being produced within the abalone farm and how they were varying on a diurnal scale. ; Production ofnutrients is clearly related to the formulated feed, however two possible means of nutrient production were deemed likely. Firstly from the formulated feed leaching or secondly from the degradation of the waste produced on farm. Finally a experiment was conducted to determine the pattern in nutrients exiting the abalone farm's effluent pipe. The nutrient stability of a commercially available formulated feed was assessed. This is of importance as on most Australian abalone farms there is usually a delay between the time the formulated feed enters the abalone culture tanks, and the time of consumption by the abalone. This leads us to believe that there may be nutrients from the formulated feed. The formulated feed leaching appears to be significant in terms of phosphorus (30% of total P content), whilst it appears to be relatively insignificant in terms of all other measured components. This research indicates that there may be a need for further research into the binding of phosphorus in Adams and Amos formulated feed. An examination of the tank waste was conducted to examine if the waste within the abalone culture tanks was a major source of dissolved nutrients that are produced as the waste degrades. Results show that approximately 50% of the C,N,P and organics of the formulated feed was collected in the abalone culture tank waste. Further the settlement pond waste contained approximately 30% of the C,N,P and organics of the formulated feed. This equates to approximately 40- 50% of the abalone tank waste nutrients (i.e. particulate waste) being degraded and remineralised as dissolved nutrients. The temporal variation in nutrient export for a single farm was characterised on a diurnal basis. Maximum concentrations of ammonium and phosphate were recorded at approximately sunrise at the farm outflow on the day of sampling. Therefore the time of day that water sampling takes place is important in the accurate assessment of environmental impact and may explain some of the variability in current monitoring program data sets. Results indicate that of the nutrients measured from a 20 tonne abalone farm, dissolved nitrogen and phosphorus were predominantly exported with exports of, up to, 1000g and 280g per day respectively. The source of these nutrients is the formulated feed. Daily feed rate can predict dissolved nitrogen and phosphate loads exported from the farm (P<0.05, r2 = 0.98 and 0.42 for Nand P, respectively). The relationship between daily feed rate and nutrient export was found to hold at three abalone farms around Tasmania (P<0.05, r2 = 0.71 for Nand r2 = 0.62 for P). These relationships can assist in the environmental management of the abalone industry and provide a consistent basis to judge the impact of abalone farming relative to other sources of nutrient inputs into our coastal environment. This study showed a 10-50 fold increase in biomass of nutrient scavenging seaweeds in the intertidal region in the vicinity of the abalone farm's discharge point. This was relative to both control sites and the discharge site prior to the commencement ofthe effluent discharge. Despite this the farm associated seaweed proliferation is unlikely to occur beyond 50 metres from point of discharge (i.e. at the end-of-pipe). Grazing and particulate feeding intertidal faunal communities were highly variable in time and space making it impossible to detect any effects of the farm's discharge. In the subtidal zone, there was no evidence of an impact on the macroalgal canopy assemblages within 50m of the end-of-pipe. The importance of solid separation devices was also highlighted in this study. Output ofparticulates from farms without Solid Separation Devices (SSD's) is likely to be significantly greater than farms with SSD's (i.e. approximately 30-50% of feed input is lost as particulate waste). Relative to farm inflow waters, Abalone Farms Australia (AFA) did not increase particulate loads into the marine environment (measured by weight); however, even with farms with SSD's, the composition of the particulates in the discharge is likely to change. For example, AFA the discharged particulates had a slightly greater (i.e. 3% greater) organic content relative to intake particulates. This study also showed that effluent nitrogen loads may be reduced by an average of 34% by a novel seaweed raft system held within the tanks. The seaweed raft system not only provided shade but most importantly a nutritious source of supplemental feed for the abalone. The present study intensively examined a single farm, and extensively examined a number of farms to gain a perspective on the environmental impact of the Tasmanian abalone industry relative to other industries that discharge nutrients into the marine environment. The study found that the entire Tasmanian abalone industry as at 2005 was likely to have little more impact than a small town of 600 people's sewage treatment plant (based on total nitrogen discharge). Additionally detailed characterization of abalone farm effluent and the subsequent environmental impacts of that effluent was determined to be negligible for AFA at current production rates (i.e. 20 tonnes of abalone biomass).

The Haliotidae is a family of marine gastropod molluscs, of the Order Archaeogastropoda (Schremp 1981). The world-wide family consists of 56 currently described species from temperate and tropical waters of both hemispheres. Approximately 25 abalone species are currently harvested commercially from at least 15 countries, with many more taken by recreational fisherman. This thesis examines the utility of molecular genetic markers in abalone research, with particular emphasis to the Australian fishery and aquaculture industries. The development of microsatellite markers in temperate abalone, and their use in stock structure studies of abalone fisheries in Australia and South Africa are presented. Twenty-two microsatellite loci were isolated from a Haliotis rubra partial genomic library, and their conservation in 12 other Haliotis species is presented. A maximum of 15 of these markers were retained in the most closely related species, Haliotis conicopora, but the species status of this group is questioned. Only 3 of the microsatellite loci examined were retained in each of two North American species, Haliotis corrugata and H. fulgens. Methods for the detection of microsatellite loci in other species is also discussed. A study of genetic variability in H. rubra at eight microsatellite loci in seven Tasmanian, one Victorian and two New South Wales locations is presented. These locations represent both fine (5 km) and broad (> 500 km) scale separation, and reveal a mean He of 0.552, with significant genetic differentiation between Tasmanian samples and those from mainland Australia (F cT = 0.003; P < 0.0001). No significant differentiation was identified between Tasmanian samples, or between mainland samples. Significant departures from Hardy-Weinberg equilibrium were common, in all instances due to an excess of homozygotes. Evidence for the existence of null alleles at two loci is presented as a factor in the departures from He.