The semen cryopreservation in cryotubes reduces the time needed for filling, freezing and thawing of the samples, while optimizing the procedures for artificial fertilization. However, no study has yet been performed with tambaqui semen in this container. The aim of this study was to evaluate the influence of cryotubes (1.6 and 4.5 mL) and thawing time (60ºC/70s and 60ºC/90 s) on the quality and fertility of tambaqui cryopreserved semen. For that, semen samples were diluted in freezing solution (1:9) composed with 75% glucose 290 mOsm , 10% methylglycol and 5% egg yolk, and frozen in liquid nitrogen vapor (in a dry shipper), and stored in liquid nitrogen (-196 °C). The semen samples was thawed at 60ºC in bath water during 70s or 90s and the semen quality was evaluated (Total motility - MT; Progressive motility - MP; Curvilinear velocity - VCL; Straight-line velocity - VSL and Average path velocity - VAP). In this study was also determined the time viability of spermatozoa thawed, maintained under refrigeration at 5°C, and assessed over 24 hours. Besides the kinetic parameters were evaluated sperm morphology and membrane integrity of spermatozoa. The fertilizing capacity of semen was evaluated from the samples thawed in the best time. All parameters of sperm kinetic showed higher values when the semen samples were thawed in 90 s compared to 70 s, independently of cryotube type. No significant differences were observed in sperm kinetic parameters after thawing between samples frozen in 1.6 ml and 4.5 mL cryotubes, except for total motility that was higher in 1.6 mL (47 ± 14% ) compared with 4.5 mL cryotubes (40 ± 11%), independently of thawing time. After activation, the spermatozoa significantly reduced the values of kinetic parameters within 37 seconds, except the MT which remained constant during this period. Relying on the sperm parameters evaluated (VCL, VSL, VAP and membrane integrity for both cryotubes and MT, MP and morphology only for 1.6 mL cryotube) the frozen semen maintained the quality during 3h after thawing. The fertilization rate obtained with fresh semen (74±6%) was higher than cryopreserved semen (1.6 mL - 45±9% and 4.5 mL - 41±12%). The two cryotubes did not differ in this parameter. A high correlation (p <0.05) was observed between fertility and sperm kinetics parameters (MT - 89%, MP - 86%; VCL - 79%; VSL - APV and 69% - 78%). It is concluded that 1.6 and 4.5 mL cryotubes can be used in the cryopreservation of tambaqui semen being recommended to be thawed at 60°C for 90s and their use in fertilization procedures within 3 hours after thawing since kept at 5°C. / A criopreservação de sêmen em criotubos reduz o tempo necessário para o envase, congelamento e descongelamento das amostras, além de otimizar os procedimentos de fertilização artificial. No entanto, nenhum estudo ainda foi realizado com o sêmen de tambaqui neste recipiente. Assim, o objetivo do presente trabalho foi avaliar a influência do tipo de criotubo (1,6 e 4,5 mL) e do tempo de descongelamento (60ºC/70s e 60ºC/90s) sobre a qualidade e fertilidade do sêmen de tambaqui criopreservado. Para isso, amostras de sêmen foram diluídas em solução de congelamento (1:9 v/v) composta por 75% de glicose 290 mOsm, 10% de metilglicol e 5% de gema de ovo, sendo envasadas em criotubos de 1,6 e 4,5 mL, congeladas em vapor de nitrogênio líquido no botijão dry-shipper (-175ºC) e armazenadas em botijão criogênico a -196°C. Para avaliação do tempo de descongelamento do sêmen, os criotubos foram imersas em água a 60°C durante 70 s ou 90 s e a qualidade espermática imediatamente avaliada (Motilidade total - MT; Motilidade progressiva - MP; Velocidade curvilinear - VCL; Velocidade em linha reta - VSL e Velocidade média da trajetória - VAP). Neste estudo foi determinado também o tempo de viabilidade dos espermatozoides descongelados, mantidos sob refrigeração a 5°C e avaliados durante 24 horas. Além dos parâmetros de cinética espermática foram avaliadas a morfologia e a integridade da membrana plasmática dos espermatozoides. A capacidade de fertilização do sêmen foi avaliada a partir das amostras descongeladas no melhor tempo. Todos os parâmetros de cinética espermática apresentaram valores superiores quando as amostras de sêmen foram descongeladas por 90s em relação ao tempo de 70s, independentemente do tipo de criotubo. Não foram observadas diferenças significativas nos parâmetros de cinética espermática pós-descongelamento entre as amostras congeladas nos criotubos de 1,6 e 4,5 mL, com exceção da MT que foi superior nos criotubos de 1,6 mL (47±14%) em comparação com os criotubos de 4,5 mL (40±11%), independentemente do tempo de descongelamento. Após ativação, os espermatozoides reduziram significativamente os valores dos parâmetros de cinética dentro de 37 segundos, exceto a MT que se manteve constante neste período. Baseando-se na maior parte dos parâmetros espermáticos avaliados (VCL, VSL, VAP e Integridade da membrana plásmatica para ambos os criotubos e MT, MP e Morfologia espermática somente para o criotubo de 1,6 mL) o sêmen congelado manteve sua qualidade durante 3h após o descongelamento. A taxa de fertilização obtida com o sêmen in natura (74±6%) foi superior ao sêmen criopreservado (1,6 mL - 45±9% e 4,5 mL - 41±12%). Os dois criotubos não diferiram entre si neste parâmetro. Uma alta correlação significativa (p<0,05) foi observada entre a fertilização e a cinética espermática (MT - 89%; MP - 86%; VCL - 79%; VSL - 69% e VAP - 78%). Conclui-se que os criotubos de 1,6 e 4,5 mL podem ser utilizados na criopreservação do sêmen de tambaqui, sendo recomendado seu descongelamento a 60°C por 90s e seu uso em procedimentos de fertilização dentro de 3 horas após o descongelamento desde que mantido a 5°C.

M.Sc. (Aquatic Health) / This study took place in the Groot Marico River catchment situated in the North West Province of South Africa. The catchment holds many rivers and associated tributaries that are home to a variety fish species with some being endemic to the catchment. A small minnow species, the Marico barb Barbus motebensis was chosen as the study species due to it being endemic to the catchment with limited information available on them with regards to their reproductive status in terms of their reproductive timing and number of potential offspring. Two tributaries from the upper Groot Marico River were chosen as sampling sites for the collection of B. motebensis across the four seasons. These two tributaries presented two genetically different populations, which allowed for a comparison of the findings to ascertain whether any differences in reproductive aspects occurred between the two tributary populations during the different seasons. The Marico barb is under threat from predation pressure posed by the largemouth bass Micropterus salmoides introduced into the system. Micropterus salmoides is a known alien invasive fish that eats any animal food item it encounters, especially fish. Due to B. motebensis and M. salmoides sharing the same habitat preferences, B. motebensis stands little chance of survival in the same water inhabited by the predatory M. salmoides. Many studies have shown bass to drive other fish species to the point of extirpation from a system through predation pressure. Currently B. motebensis is classified as vulnerable on the Red List of Threatened Species, although this status may be further compromised with the presence of M. salmoides...

To contribute to the understanding of the invasion biology of common carp Cyprinus carpio in southern Africa, this thesis investigated the life history, relative abundance, long-term population demographics and trophic niche utilisations of non-native common carp C. carpio in relation to two endemic cyprinids, Orange River mudfish Labeo capensis and smallmouth yellowfish Labeobarbus aeneus in South Africa‟s largest impoundment, Lake Gariep. The growth zone deposition rates in astericus otoliths of the three species were validated as biannual for C. carpio and as annual for L. capensis and L. aeneus, which allowed for reliable estimation of lengths-at-age upon which growth, age-at-maturity and mortality rates could be estimated. Cyprinus carpio exhibited fast growth, matured relatively early at two years of age and attained a maximum age of seven years. Labeo capensis grew significantly slower, but attained older ages of up to 12 years. Females showed notably delayed maturation at approximately six years of age. The life history parameter estimates for L. aeneus were similar to those of L. capensis. These species-specific life history characteristics contributed to a substantially higher population growth potential of C. carpio compared to L. capensis and L. aeneus. Delta-lognormal and delta-gamma Generalized Linear Models (GLMs) were used to analyse patterns of relative abundance of L. capensis, L. aeneus and C. carpio. The application of these GLMs was necessary to account for large proportions of zeros and strong skewness in the catch-per-unit-effort (CPUE) from experimental gillnet and fisheries-dependent angler surveys. Confidence intervals around predicted abundance indices were obtained through the development of a generalised parametric bootstrap procedure. The resulting standardised abundance indices were coupled with results from analysis of stable isotope ratios of fish tissues and potential food resources and revealed that C. carpio was mainly confined to soft-bottom habitats, where it predominantly foraged on benthic invertebrates. Labeo capensis was abundant in a wide range of benthic habitats and was consumed basal food resources such as detritus. Labeobarbus aeneus was found to feed mostly on pelagic zooplankton. There were no significant interspecific differences in trophic niche space, suggesting limited resource competition among the three species. Standardised historical and contemporary gillnet CPUE data indicated slow population growth rates of L. capensis and L. aeneus during the first ten years postimpoundment, but showed high biomass levels some four decades after impoundment. These results could be corroborated by stochastic age-structured production model (ASPM) simulations. In contrast to the two endemic species, the gillnet CPUE of C. carpio showed a clear „boom and bust‟ pattern, which, based on ASPM simulations, could be best explained by increased food availability during the first five years postimpoundment, followed by suboptimal conditions thereafter. Together, these results provided evidence that the establishment of the C. carpio population did not prevent the slow but successful long-term establishment of the two large endemic cyprinids. Both endemic fishes revealed specialised feeding within the impoundment.

Freshwater fish are the most imperilled vertebrate group with a high projected extinction rate. In general, the world's freshwater ichthyofauna is in a poor state and is threatened by habitat destruction, pollution, the effects of alien species, damming, water extraction and overfishing. Within South Africa, legislation supporting the conservation of aquatic habitats and its associated fauna is strong, but there is a lack of capacity and poor co-ordination between agencies responsible for the conservation and management. The Greater Addo Elephant National Park (GAENP) includes the upper catchments of five of the Sundays River tributaries. Since the optimal method of conserving freshwater aquatic biota is to conserve the headwaters of either the river or its tributaries, there is an opportunity to conserve the native ichthyofauna of the Sundays River system. The principal objective of this thesis was, therefore, to provide information pertinent to the conservation and management of the fishes in the Sundays River system. Sixteen species of freshwater and estuarine fish were sampled, of which eight were alien. Species composition in each of the rivers, with the exception of the Kabouga River, had changed significantly from previous studies. The distribution ranges of several indigenous species had reduced, while those of some alien species was increased. Two species not previously recorded, Labeo capensis and Gambusia affinis, were sampled. Alien species were sampled from all the tributaries. Diversity indices and community assemblage models were identified as approaches for monitoring the ichthyofauna in this species depauperate system. Diversity indices were not recommended for use as a monitoring tool, as they provided insufficient detail on community composition and structure. A modified depletion estimator was applied to multiple pass electrofishing data to determine the effects of various physico-chemical variables on the absolute abundance of Pseudobarbus afer, an endangered species. Pseudobarbus afer density was found to be positively correlated to the presence of boulders. Where P. afer were present, they were abundant. Of the total sampled catch, 21.8% was comprised of alien fishes. Clarias gariepinus was the most successful invader having penetrated all but one of the tributaries. Micropterus salmoides changed the species composition of the Wit River significantly through predation, and is potentially the most dangerous of the alien species in the Sundays River system, capable of extirpating a population of P. afer in as little as three days. Only one man-made barrier to the upstream migration of alien fishes was present on the tributaries, and several addition barriers are required to safeguard the populations of indigenous fish in the headwaters of these tributaries. Several management and conservation tools are presented and discussed, including river rehabilitation, translocation, eradication of alien species, erection of barrier weirs and captive breeding programmes. Public awareness and education is stressed because without support from the communities surrounding the GAENP, management plans for conservation of the indigenous ichthyofauna will, in all likelihood, fail.

This thesis presents an evaluation of the dispersive solid-phase extraction (d-SPE) method referred to as the quick, easy, cheap, effective, rugged and safe (QuEChERS) method for the determination of four organochlorine pesticide residues in fish samples. The pesticides investigated in this study were o, p′-DDT, p, p′-DDE, aldrin and dieldrin. The combined use of Gas Chromatography with an Electron Capture Detector (GC-ECD) and sensitive Time of Flight (TOF) mass detector facilitated the identification of the target analytes. In the absence of certified reference material, the overall analytical procedure was validated by systematic recovery experiments on spiked samples at three levels of 2, 5 and 10 ng/g. The targeted compounds were successfully extracted and their recovery ranged from 76 to 96% with relative standard deviations of less than 13%. The optimum QuEChERS conditions were 2g of fish powder, 10 ml acetonitrile and 1 min shaking time. The optimal conditions were applied to assess the levels of chlorinated pesticides in blunt-tooth catfish (Clarias ngamensis), tigerfish (Hydrocynus vittatus), Oreochromis andersonii and red-breasted tilapia (Tilapia rendalli) from the Okavango Delta, Botswana. Dieldrin, p, p′-DDE and aldrin were detected in all the analysed samples with a concentration range of 0.04 – 0.29, 0.07 – 0.33, 0.04 – 0.28 and 0.03 – 0.24 ng/g per dry weight in O. andersonii, C. ngamensis, T. rendalli and H. vittatus respectively. These concentrations were below the US-EPA 0.1 μg/g allowable limit in edible fish and the Australian Maximum Residue Limit (MRL) of 50 - 1000 ng/g in fresh fish. DDT was not detected in all the fish species investigated. The mean lipid content recorded in the fish samples were 1.24, 2.16, 2.18 and 4.21% for H. vittatus, T. rendalli, O. andersonni and C. ngamensis respectively. No systematic trend was observed between fish age and pesticide levels in fish. Acetylcholinesterase (AChE) activity assays were performed to assess the effects of organochlorine pesticides in Clarias ngamensis. The enzyme activity recorded in Clarias ngamensis from the Okavango Delta and the reference site was 12.31 μmol of acetylcholine iodide hydrolysed/min/g brain tissue. The enzyme activity remained the same indicating no enzyme inhibition. The conclusions drawn from this study are that the QuEChERS method is applicable for the determination of organochlorine pesticide residues in fish matrices. The fish from the Okavango Delta are safe for human consumption.

Grand Coulee Dam created Franklin D. Roosevelt Lake as part of the Columbia Basin Project. Located in northeastern Washington State, the Project provides economically important hydropower (19 billion kilowatt hours per year), irrigation (225,000 ha), flood control, and sport fishing ($5 to 20 million annually). A good system understanding aids in balancing these beneficial uses for the 230 km long reservoir. The reservoir's atypical 45-day mean residence time is much shorter than a typical lake, and much longer than for a riverine dam. The spring freshet requires drawdowns of 15 to 20 m for flood control—the driving characteristic of reservoir operations. A physically based two-dimensional hydrodynamic and water quality model, CE-QUAL-W2 Version 3.5 (Cole and Wells, 2006), is coupled with a fish bioenergetics model based on the Stockwell and Johnson model (1997, 1999) to examine the effects of hydrodynamics on the reservoir algae-zooplankton-kokanee food web. This model was applied and calibrated to Lake Roosevelt with model improvements of multiple zooplankton compartments and zooplankton omnivory. Calibration parameters included temperature, dissolved oxygen, nutrients, algae, and zooplankton. The fish bioenergetics model is applied over the entire reservoir model space to generate a spatial and temporal fish growth potential distribution. The fish model refinements include sub-daily time-steps and an optimized vertical foraging strategy. The linked model suggests that kokanee fish growth potential is seasonally limited by both warm water and prey densities. While the lake ecology is significantly affected by the reservoir operations in general, the pelagic fish growth potential did not appear sensitive to minor changes in reservoir operations. However, the model suggests that the advantageous foraging locations shift seasonally and that optimal foraging strategies are dependent on fish size.

Abstract not available

Ph.D. (Zoology) / Histomorphology as a tool to assess the health status of five economically important fish species in the Okavango Delta Panhandle, Botswana the Okavango Delta, and an inland freshwater wetland of national and international importance was designated by the Ramsar Convention in 1996. This site is of global importance because of a wide diversity of fish species found in this freshwater ecosystem. The Okavango Delta is also one of the last 'near pristine' sites on the African continent and therefore serves as an ideal reference site to describe the normal histomorphology of selected organs in sentinel fish species which could be used as reference for future toxicological studies. The aim of this study was to assess the health status of five economically important fish species namely, Clarias gariepinus (Burchell, 1822), Clarias ngamensis (Castelnau, 1861), Serranochromis robustus jallae (Gunther, 1864), Serranochromis angusticeps (Boulenger, 1907) and Hydrocynus vittatus (Castelnau, 1861) sampled from the Okavango Delta Panhandle, by describing the normal histomorphology of the liver, gills and gonads of the sentinel species. The sentinel fish species are of significant importance because they form an integral part of the small-scale commercial, subsistence and recreational fisheries in this ecosystem. In the past few decades, fish-health research has focused almost exclusively on histopathological alterations resulting from pollution caused by exposure to chemicals, parasitesand pathogens. Limited attention has been devoted to the normal histomorphology and health status of fish species in various aquatic ecosystems including the Okavango Delta Panhandle. Also, histopathological evaluations noted only the presence of abnormalities without quantitative assessment of the extent of the abnormalities within each tissue section. The absence of baseline normal histomorphology and numerical data however, makes it difficult to establish cause-and-affect relationships between pathology and contamination patterns in various fish species. It is imperative therefore, that the normal histomorphology of organs in fish species occurring in this region should be known so that histopathological conditions maybe accurately diagnosed…

The principal objective of this thesis was to gain an understanding of the movement patterns of spotted grunter Pomadasys commersonnii, an estuarine-dependent fishery species, in the turbid, freshwater dominated Great Fish Estuary. Both manual and automated telemetry methods were used to monitor the movements of spotted grunter during two separate studies conducted in summer and spring 2003 and 2004. Acoustic transmitters were surgically implanted into twenty spotted grunter with lengths between 263 and 387 mm TL in the first study and twenty spotted grunter ranging between 362 and 698 mm TL in the second study. The specific objectives were to gain an understanding of (i) the time spent in the estuarine environment (ii) the space use and home range size, and (iii) the abiotic factors governing the movement patterns of spotted grunter in the estuary. The nursery function of estuarine environments was highlighted in this study as adolescent spotted grunter spent a significantly larger proportion of their time in the estuary than adult fish (p < 0.0001; R² = 0.62). The increased frequency of sea trips, with the onset of sexual maturity, provided testimony of the end of the estuarinedependent phase of their life-cycle. Although considered to be predominantly marine, the adult spotted grunter in the Great Fish Estuary utilised the estuary for considerable periods. Adults are thought to frequent estuaries to forage, seek shelter and to possibly rid themselves of parasites. During this study, the number of sea trips made by tagged fish ranged from 0 to 53, and the duration ranged from 6 hours to 28 days. The tidal phase and time of day had a significant effect (p < 0.05) on the sea trips undertaken by fish. Most tagged spotted grunter left the estuary during the night (84%) on the outgoing tide, and most returned in the evening (77%) during the incoming tide. Sea temperature (p < 0.0001; R² = 0.34), barometric pressure (p = 0.004; R² = 0.19) and wind (p = 0.01) had a significant effect on the number of spotted grunter recorded in the estuary. Spotted grunter were more prone to return to the estuary after high barometric pressure, when low sea temperatures (upwelling events) prevailed. There was a significant positive relationship between home range size and fish length (p = 0.004; R² = 0.20). Small spotted grunter (< 450 mm TL) appeared to be highly resident, with a small home range (mean size = 129 167 m²), that was generally confined to a single core area. Larger individuals (> 450 mm TL) occupied larger home ranges (mean size = 218 435 m²) with numerous core areas. The home ranges of small and large spotted grunter overlapped considerably yielding evidence of two high use areas, situated 1.2 km and 7 km from the mouth of the Great Fish Estuary. Tagged spotted grunter were located in a wide range of salinity, turbidity and temperature, but were found to avoid temperatures below 16 ºC. The daily change in environmental variables (salinity, temperature and turbidity) had a significant effect on the change in fish position in the estuary (p < 0.0001; R² = 0.38). The distribution of tagged spotted grunter, particularly the larger individuals, in the Great Fish Estuary was influenced by the tidal phase (p < 0.05); they moved upriver on the incoming tide and downriver on the outgoing tide. This study provides an understanding of the movement patterns of spotted grunter in the estuary and between the estuarine and marine environments. Consequently, it provides information that will assist in the design of a management plan to promote sustainability of this important fishery species. The techniques used and developed in this study also have direct application for further studies on other important estuarinedependent fishery species.