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Thesis (M.S.)--Rhodes University, 2001. / Title from PDF t.p. (viewed on Apr. 23, 2006). Includes bibliographical references (leaves 100-110).

The South African hake resource faces divergent fishing pressures and management issues. Although the resource consists of two species, management was simplified because the resource was only subject to trawl effort and because of the similarity in population parameters, single species assessment models could be adopted. The impact of trawling on the stock is considered to be well understood and the resource has shown recovery since exclusion of foreign vessels. The 1990s were punctuated by major political change and the need for transformation has resulted in an expansion of the number of fishing rights holders. Longlining has been reintroduced and there is no clear understanding of how the combined fishing pressures will affect the population structure of either Merluccius capensis or M. paradoxus. Information is vital to successful management. Fisheries are complex and intricate, and at times appear impossible to control or monitor. An integrated information system provides easily understood graphical explanations of complex issues. This thesis assessed the dynamics of the trawl and longline fisheries between 1994 and 1999 using a geographical information system (GIS). The accessability of a GIS incorporates the needs of scientists, managers and fishing communities. The simple GIS developed in this study revealed shifts in effort, facilitated the calculation of spatially precise catches and biomasses and highlighted the inadequacy of current sampling coverage. Trawlers were shown to fish the same areas consistently during the years investigated, with highest fishing intensity and average CPUE achieved at depths between 300 and 500m. Analysis of the longline sector revealed several similarities to the trawl sector, fishing intensity was highest between 301 and 500m, suggesting that both sectors face a "friction of distance" dilemma. The distribution and abundance of hake, in particular the exploitable proportion of the population, was determined by a combination of depth and substrate type. The selectivity patterns of trawlers and longliners were briefly investigated with the results illustrating that gear selectivity of the M. capensis stock was depth dependent. The deeper fishing occurred, the larger the length-at-selection. As a result, the M. capensis parental stock faces unprecedented fishing pressure. In the absence of reliable species-specific catch data, logistic and linear regression models were developed to split the hake catch into its respective species components. Large discrepancies between the predicted M. capensis catch for the two models were noted. Regression estimates constructed at a finer spatial scale may provide a better fit than the current depth logistic employed by Marine and Coastal Management. A first attempt at assessing the M. capensis resource on the West Coast using an age-structured production model was presented. It was found that a lengthy and accurate M. capensis catch series is required before it is possible to successfully model the dynamics of the stock. It is necessary to incorporate finer spatial detail in the collation of catch data and collection of sampling data. It would be unadvisable to assume that the stock is stable or recovering. The implications of a size/sex relationship must be investigated and properly appraised.

The motivation for this study was to examine the exploitation patterns of the M. capensis and M. paradoxus hake fisheries on the Southeast Coast, and determine the size and species of hake caught in each of the hand-line, long-line, and trawl methods. The handline hake fishery has increased substantially over the last several years and concerns are beginning to emerge about the impact this will have on the inshore resource collected on the South Coast between August 1998 and July 1999 was used to describe the hand-line method and estimate annual landings. Data on the size and species in longline catches of hake caught during 1997 were already available for this study. Size distributions in trawl catches were determined from commercial category landing data reported by catch weight and depth. The species composition in these catches determined by comparison using RV Afrikana survey data collected in the same depth regions. Location plays a significant role in determining the sizes and species of hake caught by each gear. Hand-lines catch smaller sizes on average than do long-lines, inshore trawls target mainly M. capensis while offshore trawls catch both hake species. A substantial amount of the hand-line hake caught on the South Coast is not reported. Examination of the exploitation patterns reveal that intense trawling pressure is directed at the smaller sized M. paradoxus inhabiting the depth region between 160-400-meters. Inshore trawls discard a large amount of small sized M. capensis within the 100-meter isobath. A preliminary stock assessment on the status of each hake species found that M. paradoxus appears to be over-exploited while M. capensis was in better shape. However, length-based pseudo-cohort analysis, used in this assessment, is critically reliant on having length frequency data from a steady state population in equilibrium. This limits the application of this model for management purposes and this finding is purely theoretical at this stage. Results suggest that each hake species is under a different pattern and level of exploitation and the multi-species nature of hake stocks on the South Coast should be considered in developing optimum management policies. Future work should focus on developing appropriate age/length keys so that an age-based VPA, which is more powerful than the length-based approach, can be applied towards stock assessments on the South Coast. Alternatively, length-data covering a longer period should be compiled and the equilibrium assumption further investigated so that the results from length-based models can be used with more confidence.

Thesis (MSc Food Sc )--Stellenbosch University, 2002. / ENGLISH ABSTRACT: The focus of this study was the development of food products produced from the neck flesh of Cape hake (Merluccius capensis) as a means of adding value to hake heads. The product prototypes that have been developed include curried fish chowder (packaged in stand-up pouches), fish spread (packaged in plastic casings) and Chakalaka hake (packaged in both cans and retortabie pouches). A formula for fish stock, which was used as a base ingredient in the three product prototypes, has also been standardised. Shelf life testing was regarded an essential part of the development processes for the refrigerated product prototypes i.e. the curried fish chowder and the fish spread. Shelf life determinations involved microbiological testing based on set microbiological standards as well as sensory monitoring and pH testing. Proximate chemical- and mineral analyses were performed on freeze-dried samples of the developed product prototypes. The efficiency of the antimicrobial peptides enterocin 1071 A and 1071 B, as biological preservatives, versus that of conventional artificial preservatives was evaluated in the fish spread prototype. Three batches of fish spread were prepared: one containing the enterocin crude extract; the second a combination of sodium benzoate and potassium sorbate, while the third batch containing no preservatives served as the control. Microbiological- and histamine tests coupled with organoleptic monitoring and pH testing were carried out over a 21-day period. It was concluded that although enterocins 1071A and 1071B had some preserving effect in the fish spread, the artificial preservative combination was the superior preserving agent. However, neither the biological preservatives nor the artificial preservative combination succeeded in providing a satisfactory shelf life. The preserving agents used in this study were however not necessarily included at optimum levels and higher levels could possibly lead to an improved shelf life. The influence of two processing temperatures [121°C (249.8°F) and 116°C (240.8°F)] and two food container types (retortable pouch and can) on the sensory quality characteristics of the Chakalaka hake-prototype was investigated. The results indicated that the shorter processing time obtained with a higher processing temperature resulted in a product of better sensory quality. The sensory quality characteristics of Chakalaka hake processed in cans at 121°C were closest to that considered desirable for the product. / AFRIKAANSE OPSOMMING: Die fokus van hierdie studie was die ontwikkeling van voedselprodukte geproduseer uit die nekvleis van stokvis (Merluccius capensis) met die doeI om waarde by stokviskoppe te voeg. Die volgende produk-prototipes is ontwikkel: 'n dik vissop met 'n kerriegeur (verpak in regopstaande sakkies), 'n vissmeer (verpak in plastiekomhulsels) en Chakalaka hake (verpak beide in blikkies en retortbestande sakkies). 'n Formule vir visaftreksel, wat as 'n basis-bestanddeel in die drie produk-prototipes gebruik is, is ook gestandaardiseer. Rakleeftydstudies het 'n belangrike deel van die ontwikkelingsprosesse van die verkoelde produk-prototipes, d.i. die dik vissop en die vissmeer, uitgemaak, Rakleeftydbepalings het mikrobiologiese toetsing, gebaseer op vasgestelde mikrobiologiese standaarde, sowel as die monitor van sensoriese eienskappe en pH metings behels. Proksimale chemiese- en mineraal analises is uitgevoer op gevriesdroogde monsters van die ontwikkelde produk-prototipes. Die effektiwiteit van die antimikrobiese peptiede enterosien 1071 A en 1071 B, as biologiese preserveermiddels, is ondersoek in vergelyking met dit van konvensionele kunsmatige preserveermiddels in die vissmeer-prototipe. Drie mengsels vissmeer is berei waarvan die eerste enterosien kru-ekstrak bevat het; die tweede 'n kombinasie van natrium bensoaat en kalium sorbaat, terwyl 'n derde mengsel geen preserveermiddels bevat het nie en gedien het as kontrole. Mikrobiologiese- en histamien toetse is gelyklopend met organoleptiese monitering en pH metings oor 'n tydperk van 21 dae op monsters van die drie vissmeermengsels uitgevoer. Die ondersoek het getoon dat enterosien 1071 A en 1071 B wel 'n mate van bederfwering in die vissmeer meegebring het, maar dat die kunsmatige preserveermiddelkombinasie 'n beter preserveringseffek gehad het in die produk. Nie die biologiese preserveermiddels óf die kunsmatige preserveermiddelkombinasie kon 'n bevredigende rakleeftyd teweegbring nie. Die preserveermiddels in hierdie studie is egter nie noodwendig in optimale hoeveelhede gebruik nie en hoër vlakke kan moontlik tot 'n verbeterde rakleeftyd lei. Die effek van twee prosesseringstemperature [121 °C (249.8°F) en 116°C (240.8°F)], sowel as twee verpakkingstipes (retortbestande sakkie en blik), op die sensoriese kwaliteitseienskappe van die 'Chakalaka hake' -prototipe is ondersoek. Die resultate het aangedui dat die korter prosesseringstyd verkry met 'n hoër prosesseringstemperatuur, gelei het tot 'n produk van beter sensoriese gehalte. Die sensoriese kwaliteitseienskappe van 'Chakalaka hake' geprosesseer in blikke by 121°C was die naaste aan dit wat beskou word as gewens vir die produk.

Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Cape hake (Merluccius capensis and M. paradoxus) is commercially the most important trawl-caught fish off the South African, coastline and due to current intensive fish processing procedures Cape hake contributes the most to the total fishwaste production. Besides its commercial importance fish is also regarded as one of the single most important consumable natural resources, either in the raw or frozen form. Most of South Africa's commercially trawled demersal fish has already been partially cleaned (i.e. headed and gutted) before landing with non-marketable bycatch and hake-waste normally disposed of as discards, resulting in a waste of a potential protein source. This study was thus aimed at fulfilling several objectives namely: observing the current large-scale commercial Cape hake harvesting procedure; constructing prediction models for several morphological parameters (whole hake mass, headed & gutted hake mass, hake head mass, hake head length, hake head breadth and hake head height) of Cape hake (Merluccius ssp.), using whole hake length as the independent variable; and determining the chemical composition (moisture, protein, fat, ash, macro and trace elements) of several hake head sections (clean head, neck flesh, tongue, tongue cartilage, jaw, gills, heart, intestines, gut, kidney, kidney & kidney bone and gut & gall); determining the effect that storage has on the fatty acid profile of both the clean head and neck flesh sections. The results obtained would supply necessary data required for techno-economic investigations in the use of hake heads. For each of the six prediction models constructed, there was an increase in the variance of the data points of categories 3 (64-80 cm) and 4 (>80 cm) as opposed to categories 1 (30-46 cm) and 2 (47-63 cm). This could be attributed to a smaller sample set for both categories 3 and 4 or due to an expected increase in the variance when investigating larger biological samples. There was also a clustering of data in the three areas for each prediction model namely, within category 1 and across categories 2 and 3 and 3 and 4. This emphasised the latitudinal stratification of the Cape hake population by age, hence their stratification by size. The prediction models constructed for both boat trips 2 and 3 differed significantly (p<0.01) from that of boat trip 1, with the exception of the hake head length (cm) prediction model. The constructed prediction models, for each of the three respective boat trips, showed good predictive abilities as was indicated by the low Mean Square Error (MSE) values for the test sets, and high Pearson's correlation coefficient (r) values. These prediction models can be used in the fishing industry with confidence for Cape hake within the time frame each respective boat trip was carried out. The neck flesh could be regarded as the most important concerning chemical composition whereas the jaw could be seen as the most important when one considers mineral content. This therefore means that the jaw section, once appropriately processed is a potential Ca, Na and Fe source for supplementing diets of people suffering from a Ca, Na or Fe deficient diet. With regard to chemical status the neck flesh section is seen as a good potential source of both protein and fat, which could be attributed to the fact that hake muscle constitutes a major portion of this section. This section could thus be used to supplement the protein and fat of an existing food product, which is protein and fat deficient for people suffering from a protein and fat deficient diet. Similarly, a market could be created for the production of an economical food product with the neck flesh section being the main ingredient. Once this have been accomplished, fishing vessels may be persuaded to retain their Cape hake fish-waste for further processing due to the value of the prepared food products and thereby maintain profitability while abiding to governmental law. In conclusion non-government scientists should have more input in the decision-making process concerning matters affecting South Africa's marine biodiversity in order for future key policy and legislation drafts to be effective. Improvement of current fish preservation techniques and the known chemical composition of currently discarded material will result in informed decisions of future matters concerning its disposal. / AFRIKAANSE OPSOMMING: Kaapse stokvis (Merluccius capensis en M. paradoxus) is kommersieel Suid-Afrika se belangrikste vis spesie. Aangesien die Suid-Afrikaanse visprosesseringsbedryf baie intensief is, dra die Kaapse stokvis verwerkingsindustrie die grootste gedeelte by tot die totale visafval produksie. Die meeste van Suid-Afrika se visvangste word gedeeltelik skoongemaak voor landing terwyl nie-kommersiële byvangste en visafval gewoonlik oorboord gegooi word tydens die vangproses. Dit lei tot die vermorsing van 'n potensïele proteïen bron. Hierdie studie was dus gemik om: die huidige grootskaalse kommersiële Kaapse stokvis visvangsproses waar te neem; voorspellingsmodelle vir verskeie morfologiese parameters (heel vis massa, vis massa sonder kop en binnedele, stokvis kop massa, stokvis kop lengte, stokvis kop breedte en stokvis kop hoogte) vir Kaapse stokvis (Merluccius ssp.) te ontwikkel deur die hele lengte van die vis te gebruik as die onafhanklike veranderlike; die chemiese samestelling (vog, proteïen, vet, as, makro en spoor elemente) van verskillende dele van die viskop (skoonkop, nekweefsel, tong, tong kraakbeen, kaak, kiewe, hart, ingewand, derm, nier, nier & nierbeen en derm & gal); sowel as die effek van opberging op die vetsuurprofiel van beide die skoonkop en nekweefsel dele van die Kaapse stokvis kop. Hierdie resultate sal dan gebruik word vir die tegnies-ekonomies ondersoek in die gebruik van Kaapse stokvis koppe. Vir elk van die ses voorspellingsmodelle ontwikkel, was daar 'n vermeerdering in die variansie van die datapunte vir kategorieë 3 (64-80 cm) en 4 (>80 cm) teenoor kategorieë 1 (30-46 cm) en 2 (47-63 cm). Dit kan moontlik wees as gevolg van die kleiner monster trekking vir beide kategorieë 3 en 4 of as gevolg van verwagte toename in variansie wanneer groter biologiese monsters ondersoek word. Daar was ook 'n groepering van data in drie plekke vir elke voorspellingsmodel naamlik; binne in kategorieë 1 en oor kategorieë 2 en 3 en 3 en 4. Dit beklemtoon die geografiese breedte van die Kaapse stokvis populasie op grond van ouderdom, en dus die geografiese breedte op grond van grootte. Die voorspellingsmodelle ontwikkel vir beide die tweede en derde bootvangs het betekenisvol verskil (p<0.01) van die eerste bootvangs, behalwe die vir die stokvis kop lengte (cm) voorspellingsmodel. Die voorspellingsmodelle vir elk van die bootvangste het goeie voorspellingsvermoë getoon wat bewys is deur die lae Gemiddelde Kwadraat Fout waardes vir toetsgroepe en hoë Pearson's korrelasie koeffisiënt (r) waardes. Hierdie voorspellingsmodelle wat ontwikkel is, kan dus met vertroue in die Kaapse stokvis visvangsbedryf gebruik word mits dit ooreenstem met die periode waarin elke bootvangs uitgevoer was. Die nekweefsel gedeelte is die mees belangrikste met betrekking tot chemiese samestelling en die kaak die belangrikste in terme van minerale samestelling van die verskeie viskop dele. Die kaak is dus, as dit voldoende geprosesseer word, 'n goeie potensïele bron van Ca, Na en Fe en kan dus gebruik word om die dieet van mense wat 'n gebrek het aan hierdie minerale aan te vul. Met betrekking tot die chemiese samestelling van die nekweefsel gedeelte kan dit beskou word as 'n goeie potensiële bron van beide proteïen en vet, wat toegeskryf kan word aan die feit dat spierweefsel 'n groot deel uitmaak van hierdie viskop gedeelte. Hierdie viskop gedeelte sal dus uitstekend wees om die proteïen- en vetinhoud van 'n voedselproduk wat van nature 'n lae proteïen- en vetinhoud het te verhoog en hierdie produk sou dan geteiken word op daardie gedeelte van die gemeenskap wat 'n proteïen en vet tekort in hul dieet het. As dit eers alles in plek is, dan sal die visvangs bedryf hul Kaapse stokvis afval behou vir verdere prosessering deurdat dit gebruik word om die voedingsinhoud van bestaande voedsel soorte sal verbeter en terselfdertyd sal hulle aan wetgewing voldoen. Gevolglik sal nie-regerings navorsers meer betrokke moet wees by die besluitnemingsproses met betrekking tot sake wat die Suid-Afrikaanse mariene lewe affekteer en wat toekomstige wetgewing meer effektief sal maak. Die verbetering van huidige vis preserveringstegnieke gepaardgaande met die kennis van die chemiese samestelling van die Kaapse stokvis koppe sal lei na beter toekomstige besluite oor die afset daarvan.

A merluza, Merluccius hubbsi Marini, 1933, é uma espécie demerso-pelágica típica da região Sudeste-Sul do Brasil, presente entre 21°S e 34°S. Capturada em pequenas quantidades pelas frotas de arrasto, a partir de 2001 tornou-se alvo de pescarias na plataforma externa e talude superior e, desde então, tem apresentado claros sinais de sobrepesca. Estudos sobre o crescimento, a mortalidade e a reprodução apontavam a necessidade de identificar os estoques de merluza na região Sudeste-Sul, visando o adequado ordenamento de sua pescaria, sendo este o objetivo do presente estudo. Com esta finalidade, exemplares da coleção do Museu de Zoologia da USP, otólitos de exemplares juvenis da coleção do Laboratório de Ictiofauna do Instituto Oceanográfico e exemplares coletados em 2004, junto às frotas pesqueiras no Rio de Janeiro, em Santos, em Itajaí e em Rio Grande, no verão e no inverno, foram utilizados para procurando identificar variações geográficas. Foram analisados dados sobre caracteres morfométricos e merísticos, sobre o crescimento de juvenis e de adultos e sobre a reprodução. Os caracteres morfométricos não variaram; os caracteres merísticos apresentaram maiores valores ao sul de 33°S; o crescimento de juvenis e adultos é diferencial entre as regiões, associado à diferentes épocas de desova, com pico no verão na região Sudeste e, no inverno na região Sul. Foram identificados dois estoques: um no sudeste, entre 21°S e 29°S, e outro no sul, entre 29°S e 34°S, este último compartilhado com o Uruguai e a Argentina. Para a gestão imediata da pescaria de M. hubbsi, sugere-se para os dois estoques, restringir a pesca de arrasto durante quatro meses, sendo nos dois primeiros entre 250 e 500 m e nos dois últimos entre 100 e 250 m. Para o estoque sudeste esta restrição deve ser aplicada, para a área entre 21°S e 25°S, de outubro a janeiro e, para a área entre 25°S e 29°S, de novembro a fevereiro. Para o estoque sul a restrição deve abranger a área entre 29°S e 34°S, e vigorar entre abril e julho. / The Argentine hake, Merluccius hubbsi Marini, 1933, is a demersal-pelagic fish inhabiting the South and the Southeast Brazilian area, between 21°S and 34°S. Since 2001, this species has been strongly targeted by trawlers operating at the external continental shelf and the slope. Previous studies about growth, mortality and reproduction pointed the necessity to identify stocks of this species which is the objective of this paper. Samples from the Museu de Zoologia USP collection, otoliths from juveniles fishes from the Ictiofauna Laboratory - IOUSP collection, and specimens collected during Summer and Winter of 2004 from fishing boats operanting in Rio de Janeiro, Santos, Itajaí and Rio Grande were used to analyze geographic variations. Morphometric and meristic data, juveniles and adults growth and reproduction characteristics were analysed. The morphometric characters did not present any variation, instead meristic counts tended to be larger in direction to south of 33°S. Juveniles and adults growth are different between the south and the southeast regions, and associated to different spawning periods, which is in the Summer and in the Winter, respectively. Two stocks were identified: one in the Southeast, between 21°S and 29°S, the other one in the south between 29°S and 34°S, the last one shared with Uruguay and Argentina. In order to manage these stocks, it is suggested the prohibition of the fishery activity during four months, the first two at depth between 250 and 500 m and the last two between 100 and 250 m. For the Southeast stock this restriction would be applied between 21°S and 25°S from October to January and between 25°S and 29°S from November to February. For the south stock the prohibition would be adopted in the area between 29°S and 34°S from April to July.

A merluza, Merluccius hubbsi Marini, 1933, é uma espécie demerso-pelágica típica da região Sudeste-Sul do Brasil, presente entre 21°S e 34°S. Capturada em pequenas quantidades pelas frotas de arrasto, a partir de 2001 tornou-se alvo de pescarias na plataforma externa e talude superior e, desde então, tem apresentado claros sinais de sobrepesca. Estudos sobre o crescimento, a mortalidade e a reprodução apontavam a necessidade de identificar os estoques de merluza na região Sudeste-Sul, visando o adequado ordenamento de sua pescaria, sendo este o objetivo do presente estudo. Com esta finalidade, exemplares da coleção do Museu de Zoologia da USP, otólitos de exemplares juvenis da coleção do Laboratório de Ictiofauna do Instituto Oceanográfico e exemplares coletados em 2004, junto às frotas pesqueiras no Rio de Janeiro, em Santos, em Itajaí e em Rio Grande, no verão e no inverno, foram utilizados para procurando identificar variações geográficas. Foram analisados dados sobre caracteres morfométricos e merísticos, sobre o crescimento de juvenis e de adultos e sobre a reprodução. Os caracteres morfométricos não variaram; os caracteres merísticos apresentaram maiores valores ao sul de 33°S; o crescimento de juvenis e adultos é diferencial entre as regiões, associado à diferentes épocas de desova, com pico no verão na região Sudeste e, no inverno na região Sul. Foram identificados dois estoques: um no sudeste, entre 21°S e 29°S, e outro no sul, entre 29°S e 34°S, este último compartilhado com o Uruguai e a Argentina. Para a gestão imediata da pescaria de M. hubbsi, sugere-se para os dois estoques, restringir a pesca de arrasto durante quatro meses, sendo nos dois primeiros entre 250 e 500 m e nos dois últimos entre 100 e 250 m. Para o estoque sudeste esta restrição deve ser aplicada, para a área entre 21°S e 25°S, de outubro a janeiro e, para a área entre 25°S e 29°S, de novembro a fevereiro. Para o estoque sul a restrição deve abranger a área entre 29°S e 34°S, e vigorar entre abril e julho. / The Argentine hake, Merluccius hubbsi Marini, 1933, is a demersal-pelagic fish inhabiting the South and the Southeast Brazilian area, between 21°S and 34°S. Since 2001, this species has been strongly targeted by trawlers operating at the external continental shelf and the slope. Previous studies about growth, mortality and reproduction pointed the necessity to identify stocks of this species which is the objective of this paper. Samples from the Museu de Zoologia USP collection, otoliths from juveniles fishes from the Ictiofauna Laboratory - IOUSP collection, and specimens collected during Summer and Winter of 2004 from fishing boats operanting in Rio de Janeiro, Santos, Itajaí and Rio Grande were used to analyze geographic variations. Morphometric and meristic data, juveniles and adults growth and reproduction characteristics were analysed. The morphometric characters did not present any variation, instead meristic counts tended to be larger in direction to south of 33°S. Juveniles and adults growth are different between the south and the southeast regions, and associated to different spawning periods, which is in the Summer and in the Winter, respectively. Two stocks were identified: one in the Southeast, between 21°S and 29°S, the other one in the south between 29°S and 34°S, the last one shared with Uruguay and Argentina. In order to manage these stocks, it is suggested the prohibition of the fishery activity during four months, the first two at depth between 250 and 500 m and the last two between 100 and 250 m. For the Southeast stock this restriction would be applied between 21°S and 25°S from October to January and between 25°S and 29°S from November to February. For the south stock the prohibition would be adopted in the area between 29°S and 34°S from April to July.

Sewage effluents disposed into the marine environment from De Beers Marine Namibia diamond mining vessels have the potential to cause endocrine disruptive effects in marine organisms. Endocrine disruption refers to the alteration of the normal functioning of the endocrine system and various chemicals have the ability to mimic hormones, effecting endogenous hormone synthesis, transport, receptor interaction and intracellular signaling. The potential endocrine disruptive effects, caused by the release of different types of sewage effluents into the ocean, on fish species is a concern due to the commercial importance of fish species found in the mining area e.g. hake, sole, horse mackerel. Increased awareness of marine environmental degradation due to the presence of chemical contaminants has resulted in research being done on early warning systems, in the form of biomarkers. Cytochrome P450 monooxygenase 1A (CYP1A) and vitellogenin (vtg) are important proteins found in fish liver and blood, that have been used as biomarkers for the detection of pollutants in fish. CYP1A is a subfamily of the P450 superfamily of enzymes and catalyzes the oxidation, hydrolysis and reduction of exogenous and endogenous compounds (phase I reactions) and thus has the capacity to regulate the metabolism of several organic contaminants. CYP1A expression is altered by exposure to planar xenobiotic compounds e.g. polyaromatic hydrocarbons. Vtg is an important precursor for egg yolk proteins and plays a role in the growth and development of an oocyte. Expression of this protein is altered upon exposure to estrogenic compounds. The aim of this project was to isolate CYP1A from fish liver by differential centrifugation and optimize conditions for the CYP1A-mediated ethoxyresorufin-Odeethylase (EROD) assay and western blot analysis (to assess CYP1A expression). Another aim of this study was to evaluate the potential effects of biologically disruptive chemicals from sewage effluents, discharged into the marine environment, on the expression of CYP1A in two species of hake, Merluccius capensis and M. paradoxus (Cape hake). CYP1A in Cape hake is approximately a 60 kDa protein and the highest EROD activity was detected in the microsomal fraction after differential centrifugation. Optimal EROD assay conditions were observed at pH 7.5, a temperature of 25 °C, 10 μl of sample and a reaction time of 30 seconds. Enzyme stability assays indicated a drastic decrease in enzyme activity after 30 seconds. The EROD assay was not NADPH dependent but was limited by NADPH supply, with an increase of 300% in EROD activity being observed with the addition of 0.1 M exogenous NADPH. The addition of dicumarol (40 μM), a phase II enzyme inhibitor, showed a 232% increase in EROD activity. This is because dicumarol inhibited enzymes with the capacity to metabolize the product (resorufin) of the EROD reaction. With regard to western blot analysis, the optimal primary (rabbit antifish CYP1A peptide) and secondary (anti-mouse/rabbit antibody-horseradish peroxidase conjugate (POD)) antibody dilutions were determined to be 1:1000 and 1:5000, respectively. The comparison of CYP1A expression in Cape hake samples from De Beers Marine mining area and reference sites showed higher EROD activity (16.29 ± 0.91 pmol/min) in fish samples from the mining area in comparison to the reference site (10.42 ± 2.65 pmol/min). Western blot analysis was in agreement with the EROD assay results and a higher CYP1A expression was observed in fish from the mining sites. The increased CYP1A expression observed in fish from the mining area is not definitively an indication of a pollutant effect in the environment, as several environmental and biological factors (e.g. photoperiod and age) must also be considered before reaching this conclusion. Another aim of this study was to purify vtg from Cape hake blood samples. Cape hake vtg was purified from fish plasma by selective precipitation with MgCl2 and EDTA. Precipitated sample was subjected to anion exchange chromatography using fast protein liquid chromatography (FPLC). Vtg eluted as two broad peaks and had a molecular weight above 200 kDa. SDS-PAGE analysis also resolved smaller molecular weight proteins below 70 kDa, which were thought to be vitellogenin cleavage proteins, lipovitellin and phosphovitins. Western blot analysis was performed; however, it did not produce any conclusive results. The purification of vtg enables further studies in characterizing this protein and developing assay aimed at detecting estrogenic pollutants in the marine environment

Thesis (Ph. D. (Ichthyology and Fisheries Science))--Rhodes University, 2006.