Thesis (MSc (Genetics))--University of Stellenbosch, 2010. / Includes bibliography. / Title page: Dept. of Genetics, Faculty of Science. / ENGLISH ABSTRACT: Haliotis midae, or Perlemoen, is the only cultured species of abalone in South Africa and is under great international demand. This species is considered endangered, making sustainable farming practises and law enforcement against poaching essential for maintaining wild stocks. A limited amount of broodstock animals are provided to each farm from which thousands of offspring are grown and exported. The prevention of inbreeding and preservation of genetic diversity within farmed stocks is necessary for future sustainable farming and production of genetically stable offspring. Further research into the genetic dynamics of Perlemoen will provide the knowledge for advanced management programs for optimal farming practises and essentially sustainable production. This study focuses on genetic linkage map development with the intention of future identification of markers associated with genes of economic importance, such as growth rate. Identification of markers linked to genes responsible for such phenotypic traits will ultimately allow farming practises to select naturally genetically superior animals for breeding, thereby enhancing production. For the construction of a genetic linkage map of H. midae, microsatellite markers were developed using two strategies: FIASCO and screening of next generation sequence-bysynthesis contig data. The FIASCO-derived markers were characterised by genotype screening in 32 individuals from a full-sib family and analysed using Mendelian segregation expectations. The Illumina-derived markers were characterised by genotype screening in 32 individuals from wild populations and analysed against Hardy-Weinberg expectations. Forty four microsatellite-family combinations were obtained from FIASCO of which 28 provided informative genotype results (32% success). Twenty two markers were developed from sequence-by-synthesis screening. Fourteen provided reliable genotypes (37%) and six conformed to Hardy-Weinberg expectations. These markers were used, in addition to 156 previously developed markers, to develop sex-specific and sex-average linkage maps in two full-sib families consisting of approximately 100 offspring each. One hundred and six polymorphic loci were used for linkage analysis (LOD>3) in both families. The number of linkage groups obtained from sex-specific maps ranged from 13-16. The average genome length ranged from 500 cM to 800 cM with an average marker spacing of 10 cM. The sex-average linkage map provided 18 linkage groups with an average genome length calculation of 1800 cM and average marker spacing of approximately 13 cM. The linkage maps created in this study are preliminary but provide a stepping stone towards a high density map incorporating high throughput markers. This also provides a base for QTL mapping studies, in which phenotypic traits of interest can be identified and associated to specific locations in the H. midae genome for marker-assisted selection. / AFRIKAANSE OPSOMMING: Haliotis midae, ook bekend as Perlemoen, is in groot internasionale aanvraag en is ook die enigste klipkous spesie waarmee in Suid Afrika geboer word. Hierdie spesie word as bedreig beskou en daarom is volhoubare boerdery bedrywe en wetstoepassing teen stroping noodsaaklik om wilde populasies te beskerm. Elke perlemoenplaas word met ‘n beperkte aantal broeidiere verskaf, waarvan die nageslag dan gekweek en uitgevoer word. Voorkoming van inteling en handhawing van genetiese diversiteit binne gekweekte populasies is noodsaakllik vir toekomstige volhoubare kweking en produksie van ń geneties stabiele nageslag. Verdere ondersoeke na die genetiese dinamika van Perlemoen sal die nodige kennis verskaf om sodoende gevorderde bestuursprogramme te ontwikkel, wat tot optimale kweek praktyke en effektiewe volhoubare produksie sal lei. Hierdie studie fokus op die ontwikkeling van ‘n genetiese koppelingskaart met die voorneme om toekomstige merkers te identifiseer wat met gene van ekonomiese belang, soos byvoorbeeld groei tempo geassosieerd is. Identifisering van merkers wat vir sulke fenotipiese eienskappe verantwoordelik is sal sodoende toelaat dat boerdery praktyke kan selekteer vir diere vir verbeterde teling en produksie. Mikrosatelliet merkers is ontwikkel om die genetiese koppelingskaart saam te stel. Die volgende twee strategieë is benut: FIASCO en sifting van volgende generasie volgordebepaling-deur-sintese “contig” data. Die FIASCO-afgeleide merkers is gekarakteriseer deur genotipiese sifting in 32 individue van ‘n volsib familie en is deur Mendeliese segregasie verwagtinge ge-analiseer. Die Illumina-afgeleide merkers is gekarakteriseer deur genotipiese sifting in 32 individue van wilde populasies en is met Hardy-Weinberg ewewig ge-analiseer. Vier en veertig mikrosatelliet-familie kombinasies is deur FIASCO verky, waarvan 28 informatiewe genotipiese resultate gelewer het (32% sukses). Twee en twintig merkers is vanaf volgordebepaling-deur-sintese sifting ontwikkel. Veertien van hierdie merkers het betroubare genotipes (37%) verskaf en ses het aan Hardy-Weinberg verwagtinge voldoen. Hierbenewens is 156 voorheen ontwikkelde merkers gebruik om geslagspesifieke en geslagsgemiddelde koppelingskaarte in twee volsib families saam te stel. Hierdie volsib families het uit ń naslag van 100 elk bestaan. Een honderd en ses polimorfiese lokusse is vir koppelingsanalise gebruik, waar ‘n LOD waarde groter as drie statisties betekenisvol geag was. Die aantal koppelingsgroepe verkry van geslagspesifieke kaarte het tussen 13 en 16 gewissel. Die gemiddelde genoom lengte het van 500 cM tot 800 cM met ‘n gemiddelde merker spasiëring van 10 cM. Die geslagsgemiddelde koppelingskaart het 18 koppelingsgroepe gehad met ‘n gemiddelde genoom lengte berekening van 1800 cM en ‘n gemiddelde merker spasiëring van ongeveer 13 cM. Die koppelingskaarte wat in hierdie studie geskep is, is voorlopig en verskaf ‘n grondslag vir die ontwikkeling van ‘n hoër digtheidskaart, wat hoë deurset merkers inkorporeer. Dit verskaf ook ‘n basis vir kwantitatiewe kenmerk lokus karteringstudies. Hierdie karteringstudies kan fenotipiese eienskappe van belang identifiseer en assosieer met spesifieke posisies binne die H. midae genoom vir merker bemiddelde seleksie.

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.

Le développement de l’halioticulture est récent et a été très rapide au niveau mondial. Il s’explique par une demande supérieure à l’offre et par un épuisement des stocks dans le milieu naturel. Pour soutenir leur développement, la plupart des filières de production halioticole ont récemment mis en place des programmes de sélection génétique. En France, la filière ormeaux peine à se développer. Cette thèse CIFRE, cofinancée par France Haliotis, seule écloserie d’ormeau en France, a donc été initiée afin de définir les bases d’un programme de sélection pour améliorer la rentabilité de son élevage. La thèse s’est déroulée en deux étapes : l’étude des effets du processus de domestication observés sur les ormeaux issus de cette écloserie, et l’estimation de paramètres génétiques dans son système de production. Deux expérimentations comparant une population sauvage et une population d’élevage ont permis de montrer que les ormeaux produits sur l’élevage jusqu’en 2013 présentaient des performances égales à celles des ormeaux sauvages. A l’âge adulte, les ormeaux d’élevage ont néanmoins montré une moins grande sensibilité aux manipulations d’élevage et un comportement d’évitement des prédateurs produit par rapport aux ormeaux sauvages. L’estimation de paramètres génétiques a pu être effectuée grâce à la constitution d’un jeu de 123 marqueurs SNP permettant la reconstruction du pedigree de familles élevées en mélange pendant 4 ans. L’estimation de l’héritabilité de différents caractères associés à la croissance montre que des progrès importants pourraient être obtenus par sélection sur le poids à 42 mois. En revanche, les caractères comportementaux et physiologiques étudiés dans cette thèse n’apparaissent pas ou peu héritable. Un programme de sélection adapté au système de France Haliotis a donc été proposé pour améliorer le poids des animaux à 4 ans. L’analyse des corrélations génétiques suggère qu’une sélection multicritères pour améliorer conjointement la croissance, le taux de chair et contrôler l’évolution du développement gonadique, pourrait également être envisagée. / In the context of an increasing demand and a declining supply from fisheries, worldwide aquaculture production of abalone has greatly increased in the last decades. Selective breeding has been initiated to improve growth in major abalone species. In France, the abalone production was initiated 30 years ago and struggles to develop. This CIFRE PhD co-financed by France Haliotis, the only French hatchery in activity, was initiated to study early domestication process of farmed abalone population and to provide knowledge required to design a selective breeding program adapted to this company. Wild and farmed populations were compared, showing that, until 2013, farmed and wild abalone presented similar performances. However, lower physiological and behavioural responses to farm stressor were observed, as well as less response to predator contact.Estimations of genetic parameters were performed using a novel set of 123 SNP markers in order to successfully assign parentage in a single cohort resulting from the reproduction of 40 broodstock reared under common conditions over 4 years. Heritability estimates were obtained for a large number of production traits, indicating that individual selection of live weight when 42-month-old would lead to significant gains. A selective breeding program suitable for France Haliotis production system has been proposed to improve the weight at 4 years. The analysis of genetic correlations suggest that a multi-criteria and selective breeding program to improve growth, meat yield and to control reproductive allocation could also be established, but would be more complex to implement

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.

Magister Philosophiae - MPhil / The principle aim of this study was to gain insights into the perceptions of the people living in the Cape Agulhas Area of South Africa on the issues of small-scale fishing and the historical claims to fishing rights of the communities living in the fishing villages of Struisbaai and Waenhuiskrans. A further aim was to identify the gap between knowledge and the implementation of fishing rights policies by analysing existing policies on small-scale fishing and thereby identifying possible lessons and guidelines for policy formulation. Key focus areas were the historical recollections of people, their knowledge of past, current and proposed fishing legislation and their opinions, recommendations and guidelines regarding new and proposed policies. A qualitative framework was used for this study, using key informants and the taking of life histories. The two fishing communities served as case studies and life histories were documented using semi-structured interviews. The study drew on previous research in historical practices and indigenous knowledge systems and special attention was given to scientific and archaeological research. The policy processes from 1994 to the present were described and themes were identified in the literature and the life histories. The study found that many of the methods used in earlier years is still used today, passed over from generation to generation. / South Africa

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.

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.

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.

Recruitment, the addition of new individuals to a population, must be understood to make predictions about population growth of marine invertebrates. Red abalone (Haliotis rufescens) represent a former important commercial fishery in California, and until recently, supported a major recreational fishery. However, there have been statewide declines since the 1960s due to overfishing, disease, and climatic factors. Thus, understanding population dynamics to inform management and population restoration is critical. Recruitment dynamics of red abalone are poorly understood, with no prior knowledge of seasonal trends. To address this knowledge gap, I assessed monthly (July 2016-June 2017) and annual (2012-2016) settlement rates of red abalone in the Monterey Bay, which has low density abalone populations due to sea otter predation. I evaluated associations between abalone recruitment and oceanographic factors (temperature, wave forces, and upwelling index) and food availability (kelp density) to understand potential predictors of recruitment. Abalone recruitment occurred year round, with generally higher recruitment in late summer to early fall (July-October) and peaks in August and October. This is the first demonstration of year-round abalone recruitment in the field. On a monthly basis, there were no statistically significant relationships between recruitment and oceanographic factors or food availability. Annual abalone recruitment was consistent in all years, with the exception of 2015 when recruitment majorly decreased during the second year of the North Pacific marine heatwave (i.e., warm blob and El Niño Southern Oscillation (ENSO) events). The failure of recruitment during only the second year of warm temperature suggests that prolonged extreme temperatures lead to reproductive failure. The consistent annual recruitment in the Monterey Bay contrasts with sporadic recruitment observed in Sonoma and Mendocino Counties in northern California. This finding was unexpected because red abalone in northern California were twice as dense as those in Monterey Bay at the time of the study. Possible hypotheses behind the observed consistent recruitment in Monterey Bay despite low densities include that: sheltered embayments retain larvae and promote recruitment, predation by sea otters aggregates abalone in crevices and promotes fertilization success, and the perennially present Macrocystis pyrifera kelp forests better support abalone growth and fecundity than northern California forests dominated by annual Nereocystis leutkeana.