Typescript. / Thesis (Ph. D.)--University of Hawaii, 1967. / Bibliography: leaves 96-104. / xi, 104 l illus., tables

Thesis (Ph. D.)--Rhodes University, 2004. / Title from PDF t.p. (viewed on Apr. 23, 2006). Includes bibliographical references (leaves 222-257).

M.Sc. / Aquatic toxicology is the qualitative and quantitative study of toxic effects of pollutants on aquatic organisms. The main goal in toxicity testing is to predict, in combination with other environmental factors, with known accuracy, a concentration of a specific toxicant that will not harm a system and to make this prediction in a responsible and cost effective manner. There are a variety of unique toxicity tests, with fish being one of the most popular organisms to work with, due to being the best-understood organism in the aquatic environment and its commercial importance. Zinc, copper and cadmium are three biologically important heavy metals that are commonly used in various industries. Low concentrations zinc and copper are essential micronutrients for both plants and animals, but in higher concentrations they become toxic to the environment and its biota. Cadmium has a chemical structure similar to that of zinc and is often found in association with it, but it is a very toxic substance. The effects of zinc, copper and cadmium on the free-embryo life stage (yolk sac phase) of Oreochromis mossambicus were examined, for evaluation as a possible ¡§early life stage¡¨ fish lethality assessment. ¡§Fish early life stage tests¡¨ are considered to be relatively quick, comparable and inexpensive screening tools for testing effluents and chemicals. The yolk-sac stage is considered the most sensitive life stage in fish. O. mossambicus free-embryos feed endogenously and are indigenous to southern Africa and are therefore a good choice for lethality testing. The effects of the same metals on randomly selected Culicidae (mosquito) larvae were also tested. The reason for randomly selected larvae was to determine the possibility of doing toxicity testing without a species-specific culture. Mosquitoes are very common and well known due to being vectors of various human diseases. Recommendations towards future studies, to determine the usefulness of both O. mossambicus embryos and Culicidae larvae as biological indicator organisms, were also looked at. / Prof. G.J. Steyn

M.Sc. / Please refer to full text to view abstract

A histological assessment allows aquatic scientists to assess fish health in polluted aquatic ecosystems at tissue and cellular level. However, a firm knowledge and understanding of normal histological structure is essential to ensure accurate and objective results. Hence, the histologist must be able to distinguish between toxicant induced lesions and the range of histological characteristics considered to be normal for that specific species. However, limited histological reference material and data describing normal conditions are available for southern African fish species. The aim of this baseline study was to establish reference material, both qualitative and quantitative, for two southern African freshwater fish species used as indicator species in toxicity studies at the University of Johannesburg, Clarias gariepinus and Oreochromis mossambicus. The reference material includes descriptive histology of selected target organs, and the associated values and intervals for related quantitative health aspects including somatic indices, condition factor, blood parameters (haematocrit and plasma proteins) and quantitative histological results. Essentially, this project originated from a need to better understand the normal histology of C. gariepinus and O. mossambicus, as a range of histological characteristics were identified in control groups in previous toxicity studies which could not be confirmed to be associated with normal conditions. To be able to conduct a baseline study, with the purpose of establishing reference material, it was essential that the history of the specimens be known and documented including age, sexual maturity, nutritional status and physical and chemical water quality data. The project therefore involved the breeding of fish specimens under controlled conditions. A subsequent necropsy and qualitative and quantitative histological assessments were executed on five target organs (usually included in toxicity studies within the department): liver, gills, gonads, heart and kidney. Fish of both species were bred in an environmental room in reconstituted, reverse osmosis water and reared until sexually mature. Twenty specimens with an approximately 50:50% sex ratio were collected for both species. A necropsy was performed on each specimen and tissue samples of the selected target organs were processed using standard techniques, and prepared for the qualitative and quantitative histological assessments using light microscopy. The results The results showed that no macroscopic abnormalities were identified during the necropsy regarding external features or internal organs of the specimens within the sample groups, and all fish appeared to be in good health. The target organs were then examined microscopically, followed by a description of the normal histological structure (qualitative assessment). In addition, the target organs were assessed by means of a specific quantitative histological assessment protocol, which provides a standardised methodology of criteria to objectively assess fish health (quantitative assessment). Histological alterations identified during this assessment were quantified and subsequent reference index values could be calculated for each organ. / Dr. G.M. Pieterse

The need to monitor fresh water ecosystems for pollution is increasing, as is the need to develop a biomarker sensitive to a range of environmental insults. Recently, heat shock proteins have been identified as possible biomarkers of environmental contamination. However, evaluation as to their use as a biomarker of metal contamination in fish species endemic to Southern Africa is limited. The purpose of this study was to identify what members of the 70 kDa family of heat shock proteins (Hsp70) were present in the liver of Oreochromis mossambicus, and if their accumulation was altered after short-term (96 hour) exposure to aqueous copper. In addition to copper exposure, the effect of acclimation media was also examined. Tissue-level analysis was done by means of histological examination so as to determine if alterations in the accumulation of the Hsp70 family had a marked effect on the structural integrity of the liver. Specimens of Oreochromis mossambicus acclimated in either aged tap water or borehole water were placed in flow-through systems and exposed to either 10% or 20% of the LC50 value of cupric chloride for a duration of 96 hours. Control groups were run in conjunction with the exposure groups so as to set control values by which to compare. Heat shock protein analysis was done by Western blotting after separation of hepatic proteins by SDS-PAGE. For the purpose of histological analysis, representative samples were randomly selected. Analysis of the hepatic heat shock protein 70 family identified the presence of three (3) members, each of a different molecular weight. These included members of 70 kDa (Hsp70), 74 kDa (Hsp74) and 76 kDa (Hsp76). In addition to these findings, it was found that Oreochromis mossambicus accumulated high levels of particular members of the heat shock protein 70 family under unstressed conditions, affording the fish adaptability to environmental extremes. Furthermore, individuals acclimated in aged tap water showed decreased Hsp76 accumulation after exposure to sub-lethal copper concentrations, whereas those individuals acclimated in borehole water retained relatively high levels of Hsp76. Additionally, it was shown that the hepatic structure deteriorated in those individuals acclimated to the aged tap water after copper exposure, with observed increases in vacuolation, number of macrophage centres present and the occurrence of intracellular golden-brown granules. However, there was little change from the already-altered hepatic structure of those individuals acclimated in borehole water, with conspicuous golden-brown granules the most obvious histopathological condition present. Histological examination therefore proved to supplement the heat shock protein results obtained. This study thus concluded that a decrease in the accumulation of the Hsp70 family resulted in a negative organismal response, initiating deleterious alterations in the hepatic structure. Additionally, this study concluded that past water quality has a marked effect on a given biomarker response, and should be taken into careful consideration when conducting biomarker studies. / Prof. J.H.J. Van Vuren

Ph.D. / Heavy metals like copper are toxicants commonly found in mining, industrial, agricultural and waste effluents in South Africa¡¯s riverine systems. In high concentrations these metals could have a negative effect not only on river systems, but also on fish populations. It is therefore important to determine the effect of these heavy metals on the fish at an early stage, before the fish show any sign of being exposed to the pollutants and before any bodily functions are affected. It is known that polluted water, especially heavy metal pollution like copper, can cause physiological and biochemical alterations in fish. Histopathology is a mechanism, which can provide an indication of fish health by determining early injury to cells and can therefore be considered an important tool to determine the effect of pollutants like copper on fish tissue. At present very little is known about the effect of toxicants on the histology of fish tissue of freshwater fishes in South Africa. This is important in gonads, where many variables not only in different fish species, but also in the number of developing germ cells in the testes exist. The large number of fish species and the variation in their anatomy and reproductive patterns make knowledge of the anatomy of all fish species difficult. In order to perform a histopathological analysis, it is important to be familiar with the normal histology of the fish organs that are being investigated. It was therefore necessary to describe the histology of selected organs in South Africa¡¯s freshwater fish species. The Mozambique tilapia, O. mossambicus was chosen as test organism and copper was selected as toxicant. The normal morphology and histology of the testes of O. mossambicus were described in order to serve as a baseline study and to provide histological parameters for further toxicological work on the gonadal histopathology of fish. Fish were exposed to sub-lethal concentrations of copper (CuCl2) for a short-term (96 hours) in experimental flow-through systems under controlled laboratory conditions (23 ¡¾ 1¨¬C). An external investigation of each of the 30 control and 40 copper exposed fish was performed. Standard length, body mass and gonadal mass were recorded and the maturity of each fish was obtained histologically. A light and electron microscopic examination was performed in order to describe the histology and histopathological changes. The histopathological changes were obtained by comparing the histology and ultrastructure of the testis of the control group with the information obtained for the testes that were exposed to copper. Fish obtained from the control group that were run simultaneously with every exposure group were used to describe the normal histology of the testes of O. mossambicus. The aim was to describe the normal morphology and histology of the testes of O. mossambicus so that the information obtained during this investigation could serve as a baseline study when dealing with toxicity studies. The different germ cells of the spermatogenesis process were also described with a view to assess not only the reproductive pattern, but also to provide histological parameters which might form a basis for any toxicological work on the gonadal histopathology of fish. Identification of reproductive biomarkers requires an understanding of all the principal cellular components of the testes and their normal range of variation. The cellular components that were described are the Sertoli and Leydig cells as well as the primary and secondary spermatogonia, primary and secondary spermatocytes, spermatids and spermatozoa. The information on the GSI, gonad reproductive stages as well as the histological examination provided the baseline which was needed for comparison of the testes of fish that were exposed to copper. The cellular and sub-cellular responses at light and electron microscopic level were examined to determine early cell changes and damage in the testes which were caused by different sublethal exposures of copper. There were no differences in the gross structure of the testes of the control group and the group that were exposed to copper. The testes of the fish that were exposed to 0.16, 0.40 and 2.0 mg Cu/§¤ all revealed disorganisation of the lobules, disintegration of the spermatogonia as well as disintegration of the interstitial tissue. The exposure of fish to sublethal concentrations of copper induced histopathological changes in the testes of all three groups of fish that were exposed to copper. The results also showed that the intensity of the histopathological changes increased with an increase in the concentration of copper. As a biomarker of exposure to toxicants, histopathology represents a useful tool to asses the degree of pollution. The results from this investigation showed a decrease in the GSI after being exposed to copper, when compared with the control groups, which were not exposed to copper. The information on the effect of copper on the GSI, gonad reproductive stages as well as the histological examination also provides information on gonadal health and maturational stage. This information however provides structural, rather than functional information regarding gonadal health. The results showed that the GSI and gonadal staging should be used together with the gonadal histopathology in order to provide a baseline for comparison when dealing with other toxicological studies. The information obtained during this investigation showed that fish histology and histopathology could be used as a biomarker to provide information on the effect of heavy metals on fish health. It is however suggested that the information obtained during this investigation should be extended to field investigation in order to be validated as a reliable histopathological biomarker. / Prof. J.H.J. van Vuren Prof. G.J. Steyn

Thesis (MPhil)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Tilapia is presumed to have a well-defined genetic mechanism of sex determination, but not all sex ratios are compatible with a monofactorial sex determination model. A theory of autosomal gene influence, as well as temperature sex determination (TSD) has been proposed in order to explain large variations in sex ratios. This study assessed the variation in progeny sex ratio in O. mossambicus as a basis for the application of YY male technology in the production of all-male progeny groups. Three populations of O. mossambicus were sampled that are representative of the geographical distribution of the species in Southern Africa. Progeny groups were produced from randomly selected parents and maintained at constant temperature during the labile period of sex differentiation. Variation in sex ratios between different families of the same strain as well as between different strains was calculated. No significant differences were observed in sex ratio between strains, though a significant intra-group variation was identified. This study identified both male and female-biased sex ratios. The data in general conform to a monofactorial sex determination model. Male-biased sex ratio observed in one strain can possibly be ascribed to temperature sex determination (TSD). This strain of O. mossambicus originated from an area with different annual temperature patterns and the possibility of TSD having an adaptive advantage is discussed. This thesis also presents the results of a program to produce monosex male tilapia through the application of the YY male technology in O. mossambicus. Viable XY female and YY male genotypes were produced. XY females sired progenies ranging from 68-94% male ofspring, while YY males sired a mean progeny of 94% male phenotypes. From these results it is concluded that YY male technology provide a viable method for the production of all-male progeny in O. mossambicus. Once available on a commercial scale, the technology can be made more reliable through the application of the appropriate selection methods. / AFRIKAANSE OPSOMMING: Tilapia spesies is oorwegend onderworpe aan ‘n goed gedefinieerde enkelfaktor model van genetiese geslagsbepaling, hoewel afwykings van sodanige geslagsbepalingsmodel soms waargeneem word. Die invloed van outosomale gene, sowel as omgewings temperatuur word voorgehou as verklaring van die waargenome variasies in geslagsverhoudings. Hierdie studie behels die evaluasie van variasie in geslagsdifferensiasie van O. mossambicus as basis vir die implementering van die YY manlike tegnologie vir die produksie van slegs-manlike nageslag groepe. Monsters is bekom van drie populasies van O. mossambicus verteenwoordigend van die geografiese verspreiding van die spesie in Suider Afrika. Nageslag groepe is geproduseer vanaf ewekansig gekose ouerpare en by ‘n konstante temperatuur gehuisves tydens die sensitiewe tydperk van geslagsdifferentiasie. Variasie in geslagsverhouding tussen verskillende families binne dieselfde subpopulasie sowel as binne verskillende populasies is bepaal. Geen betekenisvolle verskille is waargeneem in die geslagsverhouding tussen die verskillende populasies nie terwyl betekenisvolle verskille tussen families binne ‘n populasie waargeneem is. Die data voldoen aan ‘n enkelfaktor genetiese geslagsbepalingsmodel, met die uitsondering van een populasies waar ‘n moontlike omgewingsverwante geslagsbepalingseffek waargeneem is. Die tesis sluit ook in die resultate in van ’n program om YY-tegnologie in O. mossambicus te vestig ten einde manlike nageslag groepe te produseer. Funksionele vroulike XY- en manlike YY genotipes is ontwikkel. Die vroulike XY-genotipes het manlike nageslag groepe opgelewer wat wissel van 68%-94% manlik, terwyl manlike YY-genotipes nageslag groepe gelewer het wat gemiddeld 94% manlik was. Die resultate bevestig die lewensvatbaarheid van YY-tegnologie vir die kommersiële produksie van manlike nageslaggroepe in O. mossambicus.

Thesis (MPhil)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: 60-day growth experiment was conducted to investigate the effect of dietary Lcarnitine supplementation on the production performance parameters of Mozambique tilapia, Oreochromis mosambicus. A number of approximately 140 tilapia fry with average weight of l.4g ± 0.71g were stocked in each of 40 fine-meshed hapas (I mx 1mx 1.5m) submerged within a complete recirculation pond system. During the first 30 days of the experiment water temperatures ranged from 19 to 23°C where after it decreased to 16-20°C for the consecutive 30-day period. Dietary treatments consisted of 8 replicates of 5 levels of L-carnitine supplementation labelled as Co, C250,C500, C750 and C 1000represented Omg, 250mg, 500mg, 750mg and 1000mg L-carnitine supplementation per kg feed respectively. Results were analyzed for significant differences using one-way analysis of variance (ANOVA) and Tukey's pairwise comparison test for growth rate, feed intake (FI) and feed conversion efficiency. After completion of the trial 8 fish from each hapa were sacrificed and analyzed for cephalosomatic index (CSI), dress out percentage (viscera, gills and head excluded), viscerosomatic index (VSI) and hepatosomatic index (HSl). Poor production performance results were generally observed as water temperatures were sub-optimal, especially during the second 30-days period. Results from the trial indicate no significant differences (P>0.05) between treatments for weight gain, FCR, FI and VS!. A negative trend was observed for FCR with increasing level of L-carnitine supplementation for both the first 30-day period (1.50±0.07, 1.53±0.08, 1.58±0.09 and 1.61±0.17 for C250,C50Q,C750and C 1000)as well as for the consecutive lower temperature 30-day period (2.22±0.10, 2.25±0.ll, 2.27±0.28 and 2.29±0.2l for C250, C500, C750 and C 10(0)'Although statistically not significant, fish fed the C250showed better performance in dress out percentage weight either than the control or the higher levels. The increasing trend for head weight with increasing level of L-carnitine supplementation were significant (P<0.05) from Co and C250with and above C500.The decreasing trend for liver weight with increasing level of L-carnitine supplementation became significant (P<0.05) with and above C750.The results of the current study showed a trend in the improvement of L-carnitine on the production performance parameters. However, the natural content of L-carnitine in the basal diet impaired with the inclusion levels, thus further research at lower inclusion levels is recommended. / AFRIKAANSE OPSOMMING: 'n Proef oor 'n tydperk van 60-dae is onderneem om die effek van L-karniten aanvulling op produksie prestasie parameters van Mosambiek tilapia (0. mosambicusi te ondersoek. 140 tilapia vingerlinge met 'n gemiddelde massa van lAg ± 0.7lg is ewekansig uitgeplaas in 40 eksperimentele hapa-hokkies (lmxlmx1.5m) in "n hersirkulasie sementdam-stelsel. Gedurende die eerste 30 dae van die proef het water temperatuur gewissel tussen 19 to 23°C waarna dit gedaal het na tussen l6-20°C vir die opeenvolgende 30-dag periode. Proef-rantsoen behandelings het bestaan uit 8 herhalings van 5 vlakke van L-karnitien aanvulling, naamlik Co,C250, C500, C750 en CIOOOvir Omg, 250mg, 500mg, 750mg en 1000mg L-karnitien aanvulling per kg voer afsonderlik. Resultate was ontleed vir betekenisvolle verskille deur gebruik te maak van analise van variansie (ANOVA) ontleding en die Tukey se vergelykende toets vir groeitempo, voerinname en voeromsettingsverhouding. Aan die einde van die proefperiode is 8 visse van elke hapa ontleed vir liggaamskomponent-samestelling (kop-, ingewande- en hepatosomatiese indekse. Ondergemiddelde produksie resultate is waargeneem wat toegeskryf kan word aan onder-optimale water temperature, veral gedurende die tweede 30-dag periode van die proef. Proef resultate het geen betekenisvolle verskille (P>0.05) in massatoename, voeromsettingsverhouding (VOV) of visserosomatiese indeks tussen behandelings getoon nie. 'n Negatiewe neiging is waargeneem vir VOV met toenemende vlakke van L-kamitien insluiting vir beide die eerste 30 dag periode (1.50±0.07, 1.53±0.08, 1.58±0.09 and 1.61±0.17 for C250, C500, C750 and CIOOO) sowel as vir die opvolgende 30-day periode nie (2.22±0.10, 2.25±0.11, 2.27±0.28 and 2.29±0.21 for C250, C50o, C750 and CIOOO). 'n Toenemende neiging vir kop-massa met toenemende L-kamitien insluiting was betekenisvol (P<0.05) vanaf Co en C250 met en hoër as C500. 'n Dalende neiging vir lewermassa met toenemde L-kinsluiting was betekenisvol (P<0.05) met en hoër as C750. Resultate van die proef dui oor die algemeen op 'n neiging tot verbeterde produksie prestasie parameters van tilapia vingerlinge met toenemde insluiting van Lkamitien. Verdere navorsing word aanbeveel om die invloed van natuurlike Lkamitien in die proteïen-bronne van die basaalrantsoen te op die gebrek aan betekenisvolheid van hierdie neiging te verklaar.

Thesis (M. Sc. (Ichthyology and Fisheries Science))--Rhodes University, 2006.