Many studies addressing the toxicity of silver nanomaterials have found that smaller sized silver nanoparticles are usually more toxic to organisms and in cell culture than particles of larger sizes yet it is not entirely clear why. We investigated the size dependent toxicity of silver nanoparticles by measuring the response of embryonic zebrafish (Danio rerio) following exposure to a library of thirteen distinct silver nanoparticle size distributions with mean diameters between 8.9 nm and 112.6 nm. Data analysis using dose���response modeling revealed that silver nanoparticles (AgNP) induced embryo toxicity that is dependent on the total surface area and not on the mass or particle number in solution. Included in this study is a comparison between embryo toxicity induced by silver nitrate (AgNO���) and AgNPs for cardiovascular endpoints, as well as an investigation into the influence of the chorion on AgNP toxicity. This study demonstrates the importance of using alternative dose metrics in nanotoxicology, and highlights the value of using the embryonic zebrafish to explore nanomaterial structure activity relationships. / Graduation date: 2013

No description available.

The inclusion of soya as a dietary protein source in the formulated feed, Abfeed® S34 (Marifeed Pty (Ltd), Hermanus) for farmed abalone, Haliotis midae has resulted in larger gonads during reproductive seasons compared to the gonads of abalone fed kelp or diets that included fishmeal as the only main protein source. The aim of this study was to determine if the isoflavones present in the soya were responsible for this increase in gonad size and the subsequent effects on farmed abalone growth. Animals weighing between 40-50 g were fed one of seven isonitrogenous and isoenergetic diets containing either 0, 25, 50 or 100 percent of the soya component of the commercial feed (Abfeed® S34, Marifeed Pty (Ltd), Hermanus) from September 2013 to March 2014. An additional three diets were formulated to include crystalline isoflavone (ISO). These diets were identical to the 0 percent soya diet (i.e. the fishmeal only diet - FM), only ISO was included at the same rate that ISO occurred in the three soya diets. Data were analysed using a multiple forward stepwise regression analysis (MSR) to test the effects of ISO concentration, soya concentration, time, sex, time by concentration interaction and sex by concentration interaction on growth and gonad development and to identify those variables that most contributed to the model. The inclusion of crystalline ISO failed to promote larger gonads and had no effect on abalone growth, while growth and gonad development was dose dependent on soya inclusion rates with sex and time contributing to the models. Mean monthly weight gain in males correlated with increasing soya concentrations (c) (MSR, y = 3.24 + 0.002c, r2 = 0.23, p = 0.03), ranging from 3.11 ± 0.55 g abalone-1 month-1 to 4.43 ± 0.46 g abalone-1 month-1, while both male and female monthly length gain was not influenced by soya concentration with an overall mean of 1.62 ± 0.05 mm abalone-1 month-1 (MSR, p = 0.05 and p = 0.81, respectively). By December, the whole body mass, meat mass and visceral mass in both males and females decreased with increasing soya levels. However, by February, female whole body mass, meat mass and visceral mass positively correlated with soya levels. At the end of the study, male abalone fed FM with soya equivalent to the commercial feed had the highest whole body mass (69.00 ± 2.48 g abalone-1), meat mass (41.80 ± 1.12 g abalone-1), visceral mass (9.00 ± 2.47 g abalone-1) and gonad bulk index (42.70 ± 9.82 g abalone-1), while females were not influenced by soya concentrations with an overall whole body mass of 63.46 ± 0.79 g abalone-1. Weight loss was observed in all treatments between February and March, probably due to a spawning event. The moisture content in the meat was not influenced by treatment, however, visceral water loss was effected by both ISO and soya concentration with time and sex contributing to the model. The visceral water loss of females fed graded levels of soya decreased as a function of soya from December to March, and from December to February for males, whereas females fed ISO-enriched diets decreased as a function of ISO concentration (c) at the end of the study from 74.98 ± 0.88 to 73.10 ± 0.75 percent (MSR, y = 74.97 – 0.0025c, r2 = 0.20, p = 0.048). The inclusion of crystalline ISO had no significant effect on oogenesis in female farmed Haliotis midae, while the distribution of the predominant oocyte stage, stage 7 (second last stage prior to spawning) was dose-dependent in abalone fed increasing soya concentration (c) (MSR, y = 33.38 + 0.03c, r2 = 0.32, F(1, 18) = 8.52, p = 0.01). The increase in stage 7 oocytes in abalone fed FM with soya did not reduce the number of oocytes (44.96 ± 3.01 oocytes mm-2) present within the lumen, while the number of oocytes (o) in abalone fed the FM-only based diets decreased with increasing abundance of stage 7 oocytes (MSR, y = 58.28 – 0.48c, r2 = 0.38, F(1, 18) = 12.51, p = 0.002), possibly due to the increase in size of the oocytes with thicker jelly coats. This study provided evidence that crystalline isoflavone had no influence on abalone gonad development over five months, while soya had a dose-dependent effect on growth, gonad mass and oogenesis in farmed Haliotis midae. Formulated abalone feed could be manipulated at certain times of the year to obtain maximum growth. These implications and further studies were discussed.

D.Phil. / The endocrine system regulates important physiological processes in an organism. Numerous natural and synthetic chemicals are released into the environment and can disrupt the normal functioning of the endocrine system, influencing physiological processes even at relatively low concentrations. These chemicals are known as endocrine disrupting chemicals (EDCs). These compounds are highly persistent in nature due to a resistance to biodegradation, and because of their hydrophobicity. EDCs bio-accumulate especially in the fatty tissue of aquatic and terrestrial organisms. Developmental and reproductive disorders are potentially harmful effects of exposure. EDC sources include agricultural and urban runoff, and industrial effluent discharges that eventually pass through sewage- and wastewater treatment plants. These chemicals degrade at various rates and to varying extents during treatment processes, forming many new chemicals. Methods to predict the occurrence of endocrine disruption include the occurrence of intersex, and changes in spermatogenesis and secondary sexual characteristics (SSCs) of supposedly exposed organisms. The urogenital papilla (UGP) is a SSC whose development and morphology is hormonally controlled and may be influenced when exposed to EDCs. The sharptooth catfish (Ciarias gariepinus) is often used in environmental toxicity studies. Sex is determined by the external dimorphic UGP found only in males. Intersex has been observed in this species in South African freshwater systems. A field study was carried out in the Rietvlei (RVD) and Marais (MD) dams in the Rietvlei Nature Reserve (RNR). MD acts as a sedimentation dam before water enters RVD, and is theoretically more polluted. The RNR system receives effluent from various domestic and industrial sources including agriculture, industries, informal settlements and municipal sewage treatment plants. Four sampling surveys, two low and two high flow, were carried out over a period of two years. Supposedly male catfish (n=97) were collected and identified as male or possible intersex based on the UGP. Water and sediment was collected for target EDC and endocrine disrupting metal (EDM) analyses. Fish length and mass, and UGP length and width were measured. Laparotomy was performed and gonads were inspected macroscopically for alterations. A section of the gonad and the entire UGP was removed and processed according to standard techniques for histological analysis. The gonadosomatic- (GSI), hepatosomatic- (HSI) and UGP length (UGPLI) indices were calculated. Fish fat was removed and analysed for target EDCs.

N,N'-methylenebisacrylamide (MBA) was studied because of its effectiveness in inducing heritable translocations in germ cells of male mice. The health impact of translocations was studied through anatomical analysis of the progeny of semisterile translocation carriers. As expected, the semisterility of translocation carriers resulted primarily from embryonic death during periimplantation stages due to unbalanced chromosome sperm segregants. Among conceptuses that survived to mid- and late-gestation stages, there was an increased incidence of developmental anomalies including fetal death and phenotypic defects. These abnormalities are associated with unbalanced chromosome complements that allow survival to the later stages of development.

Sertraline is a selective serotonin reuptake inhibitor (SSRI) that is widely used for the treatment of depression and anxiety. Due to the abundant therapeutic use of sertraline, low levels have been detected in municipal wastewater effluents suggesting that aquatic organisms may be exposed. The purpose of this study was to evaluate the steroidogenic effects of sertraline on larval (FHM) and adult female fathead minnows (FFHM), Pimephales promelas. Larval FHM were exposed to 0.1, 1, and 10 µg/L sertraline for 28 days and analyzed via RT-qPCR for differential expression of 11β-Hydroxysteroid dehydrogenase (11β-HSD), 20β-Hydroxysteroid dehydrogenase (20β-HSD), aromatase (CYP19), and nuclear thyroid receptor alpha (TRα). FFHM were exposed to 3 or 10 µg/L sertraline for 7 days with the brain and ovary excised at exposure termination. Juvenile FHM exposed to 0.1 μg/L sertraline had a significant upregulation of both 20β-HSD and TRα. FFHM exposed to 10 µg/L sertraline had a significant upregulation of 11β-HSD expression in brain tissue, while no steroidogenic changes were observed in the FFHM ovary. Similarly, in FFHM brain tissue, CYP19 and 20β-HSD expression levels were significantly higher in fish exposed to 10µg/L sertraline compared to control. The significance of these findings with respect to survival, growth and reproduction are currently unknown, but represent future research needs.

Thesis (MPhil)--Stellenbosch University, 2003. / ENGLISH ABSTRACT: A farm in Zimbabwe (Farm A) obtains its water for tbe poultry operations from tbe Makavusi River that is heavily contaminated with bacteria and performance results are thereby reduced. Anolyte is a Russian invention that claims to have bactericidal effects that could be beneficial to broilers. The aim of this research is to investigate the effect of Anolyte on production profits of broilers given water of poor quality by carrying out a number of trials. Trials that were conducted are basic in vitro tests on Anolyte within the laboratory, water analysis on water supply to the broiler section (both chemical and bacteriological), and a detailed trial within an experimental trial site comparing broiler performance results with different chemical water treatments and different dilutions of Anolyte. Broiler chicks from different hatcheries (Hatchery A and Hatchery B) were also compared. Performance results were based on growtb rate, mortality, feed conversion ratios (FCR), performance efficiency factors (PEF) and profit margin comparisons. Results showed that in tbe laboratory, Anolyte had an antibacterial activity. In the broiler performance trial on Hatchery B broilers, the live weights at forty two days of age in broilers given 15% Anolyte differed from other treatments (P<0.05) other than 10% Anolyte (P > 0.05). However, the FCR and mortalities did not differ (P > 0.05) between treatments. In the trial on Hatchery A broilers to 42 days of age, growth rates in broilers given 15% Anolyte differed from tbe control, Chematron and 20% Anolyte (P < 0.05). However, the FCR and mortalities did not differ between (P>0.05) treatments. In comparing chicks from different hatcheries, Hatchery B broilers differed (P < 0.05) from Hatchery A broilers in live weights at 42 days on 10% Anolyte, 15% Anolyte and on Chematron treatments. However, tbe cumulative mortalities and FCR did not differ (P > 0.05) between the Hatchery B and Hatchery A on any treatment. When comparing profitability, tbe treatment that had the greatest margin in comparison to tbe control (untreated drinking water) was 15% Anolyte for both tbe Hatchery B and Hatchery A trials. / AFRIKAANSE OPSOMMING: 'n Plaas in Zimbabwe, (Plaas A) verkry water, vir hul pluimvee bedryf vannuit die Makavusi rivier. Die rivier is swaar besmet met bakterië en as gevolg daarvan verhoed dit dat hulle hul volle opbrengs potential bereik. Anolyte is 'n Russiese uitvindsel wat aanspraak maak daarop dat dit bakteriëdodende effekte besit wat tot die bevordering van slaghoenders kan lei. Die doel van die navorsing is om Anolyte op proef te stel aangaande produksie profyt op slaghoenders waaraan swak kwaliteit water verskaf is. Verskeie toetse is uitgevoer n1. 'n basiese 'in vitro' toets in samewerking met die laboratorium, water ontleding op water wat aan die slaghoender seksies verskaf is (beide chemikalies and bakterieologies) asook 'n intensiewe proefnemeing waarin die verskeie obrengs resultate, van chemikaliese behandelde water en verskillende verdunnings van Anolyte, met mekaar vergely is, in 'n experimetele navorsings eenheid. Slaghoender kuikens van twee verskillende broeihuise (n1. Broeihuis A en Broeihuis B) is ook met mekaar vergelyk. Die resultate is gabasseer op groei tempo, sterftes, voer omsettings verhouding, opbrengs geskikthied faktore asook vergelykings op wins. Laboratorium navorsing het bevestig dat Anolyte wel antibakteriese aktiviteit getoon het. Terwyl in die proefneming op Broeihuis B slaghoenders, het die lewende gewig van hoenders, op 42 dae van ouderdom, waaraan 15% Anolyte verskaf is, verskil van ander behandelings (P<0.05) met die uitsondering van 10% Annolyte (P>0.05). Alhoewel die voer omsetting verhoudings en sterftes nie in die verskillende behandelings verskil het nie. In die proefnemeing op Broeihuis A slaghoenders tot 42 dae van ouderdom, het die ontwikkeling van slaghoenders, waaraan 15% Anolyte verskaf is, verskil van die kontiole, Chematron en 20% Anolyte. Die voer omsettings verhoudings en sterftes het egter geen verskil getoon tussen die verskeie behandelings nie. Resultate van die vergelyking tussen die kuikens uit die twee verskillende broeihuise, het getoon dat die Broeihuis A en Broeihuis B lewensgewigte, op 42 dae van ouderdom, wel met mekaar verskil het met 10% Anolyte, 15% Anolyte sowel as die Chematron behandelings. Daar was egter geen verskil, (P>O.05) tussen die twee tipes aangaande elke tipe se saamgestelde sterftes en die voer omsettings verhoudings, in welke behandeling nie. Die vergelykings op wins het getoon dat die behandeling met die grootste brulo marge, die was van 15% Anolyte in vergelyking met die kontiole (onbehandelde drinkwater) vir beide die Broeihuis A en die Broeihuis B proefnemings.

Dissertation (PhD)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: Concern has increased that widespread adverse effects are occurring in humans and wildlife populations as a result of exposure to environmental chemicals (mostly man-made) that disrupt the normal functioning of the endocrine system. Many pharmaceutical, agricultural and industrial chemicals, acting as endocrine modulating compounds (EDCs), have been detected in an accumulated form in food, in drinking water and in the environment. Although the levels of these chemicals can be measured analytically, it is important to evaluate biological activity. For this, animal models are used and relevant bioassays developed. These assays are based on biological markers, which are specific xenobiotically-induced physiological responses and are usually deviations in cellular or biochemical components, processes or structures. Vitellogenin is a large protein complex, produced in the liver under estrogen control and circulates in the plasma, destined for incorporation into the developing oocyte in female oviparous vertebrates. Since vitellogenin production is low or nonexistent in males, its presence may therefore be interpreted as evidence of exposure to estrogenic environmental contaminants. In this study the African Clawed Frog, Xenopus laevis was used as model to study the effects of estrogens on biochemical and physiological parameters associated with vitellogenesis. As a starting point the seasonal female reproductive cycle in a natural Xenopus laevis population in terms of ovarian state, plasma vitellogenin and plasma estrogen levels was studied. It was shown that plasma vitellogenin and estrogen levels were seasonal and correlated to a seasonal ovarian cycle, which peaked during spring. However, although seasonality existed, there were reproductively active individuals at any time during the year. Increases in plasma estrogen levels predated increases in plasma vitellogenin levels and ovarian maturation. Lipoprotein profiles, as well as plasma cholesterol, triglyceride and phospholipid concentrations were determined and it was shown that estrogen affected these in such a way that these parameters could be used as additional markers in estrogen contamination studies. In order to develop an in vitro bioassay to screen for estrogenic activity, the use of hepatic tissue cultures was investigated. Optimal culture conditions were established and increased sensitivity in the estrogenic response was obtained by using liver slices from male frogs that were pre-treated with estrogen. Validation studies proved that this bioassay could be employed for routine screening of water and chemical samples. In order to refine the Xenopus laevis vitellogenin ELISA and liver slice bioassay, existing polyclonal anti-vitellogenin antibodies were replaced by culturing monoclonal antibodies. Selected antibodies were characterised and ELISAs developed and validated. This study showed that the newly developed Xenopus laevis vitellogenin ELISA and liver slice bioassay have the potential to be employed in environmental monitoring programmes. / AFRIKAANSE OPSOMMING: Daar is toenemende besorgdheid dat afwykings in mens- en dierbevolkings voorkom as gevolg van blootstelling aan chemikalieë (hoofsaaklik mensgemaak) in die omgewing wat die normale werking van die endokrienstelsel versteur. Verskeie farmaseutiese, landbou- en industriële chemikalieë, wat as endokrienversteurders optree, is in die omgewing gevind. AI kan die vlakke van hierdie stowwe analities bepaal word, is dit belangrik om hulle biologiese aktiwiteit te evalueer. Hiervoor word diermodelle aangewend om toepaslike toetse daarvoor te ontwikkel. Hierdie toetse word baseer op biologiese merkers, spesifieke xenobioties-geïnduseerde fisiologiese reaksies, en is gewoonlik afwykings van sellulêre- of biochemiese komponente, -prosesse of - strukture. Vitellogeen ('n dooiervoorloper) is 'n lipoproteïenkompleks wat, onder estrogeenbeheer, in die lewer vervaardig word en in die plasma sirkuleer vir uiteindelike inkorporasie in ontwikkelende oësiete van vroulike, ovipare werweldiere. Aangesien daar min of geen vitellogeen in manlike diere geproduseer word, is die teenwoordigheid daarvan 'n aanduiding dat die dier aan estrogeniese omgewingsbesoedeling blootgestel is. In hierdie studie is die Platanna, Xenopus laevis, as model gebruik om die gevolge van estrogene op biochemiese en fisiologiese veranderlikes, wat met vitellogenese geassosieer word, te bestudeer. As vertrekpunt is die seisoenale voortplantingsiklus van die wyfie, in terme van vitellogeen en -estrogeen vlakke in die plasma en aktiwiteit van die ovaria bepaal. Daar is aangetoon dat die estrogeen- en vitellogeenvlakke in die plasma met die ovariumsiklus, wat gedurende die lente hoogtepunte bereik, korreleer. Alhoewel daar seisoenaliteit bestaan, was daar dwarsdeur die jaar wyfies wat ovarium dooierneerlegging getoon het. Verhoging in estrogeenvlakke het vitellogeenpieke en rypwording van die ovaria voorafgegaan. Lipoproteïenprofiele, sowel as die cholesterol- , trigliseried- en fosfolipiedkonsentrasies in die plasma is bepaal en daar is aangetoon dat estrogeen hierdie medeveranderlikes in só 'n mate affekteer dat hulle as bykomende biomerkers vir estrogeenblootstelling in besoedelingstudies gebruik kan word. In vitro Xenopus laevis lewersnitte in die weefselkultuur omgewing is ondersoek om 'n biotoets te onwikkel vir die gebruik in vinnige estrogenisiteits-toetsing van watermonsters en chemise stowwe. Die mees gunstige kultuurtoestande is bepaal en die sensitiwiteit van estrogeenreaksies is verhoog deur lewer van mannetjies, wat vooraf met estrogeen behandel is, te gebruik. Hierdie biotoets se geldigheid is gestaaf en kan in roetine eerstevlaktoetsing van watermonsters gebruik word. Die gebruik van poliklonale teenliggaampies in 'n bestaande enzyme-linked immunosorbent assay (ELISA) is vervang deur spesiaal-ontwikkelde monoklonale anti-Xenopus laevis vitellogeen teenliggaampies. Uitgesoekte teenliggaampies, spesifiek teen die estrogeengeïnduseerde proteïene, is gekarakteriseer en ELISAs saamgestel en die geldigheid gestaaf. Hierdie studie het aangetoon dat die nuut-onwikkelde Xenopus laevis vitellogeen-ELISA en lewerkultuurbiotoets die potensiaal het om In omgewingsmoniteringprogramme gebruik te word.

Sucrose fading was used to initiate self-administration of ethanol on an FR4 schedule in male Fischer 344 rats. Rats showed low response rates for ethanol alone. After administration of liquid diet containing ethanol, ethanol intake increased over levels prior to administration of the liquid diet. In situ hybridization compared mRNA for the inducible or constitutive 70 kD heat shock proteins in ethanol and nonethanol rats. Both inducible and constitutive mRNAs were found in nonethanol and ethanol tissues. In peripheral organs, radiolableling was higher in ethanol tissue. In brain regions, nonethanol tissues showed higher radiolabeling.

Bibliography : leaves 102-106