All-male fry are preferred to prevent uncontrolled reproduction before harvest in intensive Nile tilapia (Oreochromis niloticus) aquaculture. Males also grow faster than females. An alternative approach to direct hormonal masculinisation of tilapia fry is to produce fry that are genetically male. However, sex determination system in tilapia is fairly complex. Recent developments have resulted in a linkage map and genetic markers that can be used to analyse the sex determination system. To analyse the genetic sex determination mechanism and to develop marker-assisted selection in the Stirling Nile tilapia population, a fully inbred line of clonal females (XX) was verified using test crosses and DNA markers (mostly microsatellites) to use as a standard reference line in sex determination studies. A series of crosses were performed involving this line of females and a range of males. Three groups of crosses were selected (each group consisted of three families) from progeny sex ratio distributions, and designated as type ‘A’ (normal XY males x clonal XX females), type ‘B’ (putative YY males x clonal XX females) and type ‘C’ (unknown groups of males x clonal XX females), for sex linkage study. For type ‘A’, inheritance of DNA markers and phenotypic sex was investigated using screened markers from tilapia linkage group 1 (LG1) to confirm the LG1-associated pattern of inheritance of phenotypic sex and the structure of LG1. Screened markers from LG1, LG3 and LG23 were used to investigate the association of markers with sex in families of type ‘B’ and ‘C’. In addition, a genome-wide scan of markers from the other 21 LGs was performed to investigate any association between markers and sex, in only families of cross type ‘B’. LG1 associated pattern of inheritance of phenotypic sex was confirmed by genotype and QTL analyses in families of cross type ‘A’. Analyses of genotypes in families of type ‘B’ and ‘C’ showed strong association with LG1 markers but no association with LG3 and LG23 markers. Genome wide scan of markers from all other LGs did not show any significant association between any markers and the sex. The allelic inheritance of two tightly linked LG1 markers (UNH995 and UNH104) in families of type ‘B’ and ‘C’ identified polymorphism in the sex determining locus: one of the alleles was associated mostly with male offspring whereas another allele was associated with both progeny (mostly males in type ‘B’ families, and approximately equal numbers in type ‘C’ families). This knowledge was used to identify and separate supermales (‘YY’ males) that should sire higher proportions of male progeny, reared to become sexually mature for use as broodstock. Two of them were crossed with XX females (one clonal and one outbred) to observe the phenotypic expression of the strongest male-associated allele in progeny sex. The observations of 98% male (99 males out of 101 progeny) and 100% male (N=75) from these two crosses respectively, suggest that a marker-assisted selection (MAS) programme for genetically male Nile tilapia production could be practical. This study also suggests that the departures from the sex ratios predicted using a “simple” XX/XY model (i.e., YY x XX should give all-male progeny) were strongly associated with the XX/XY system, due to multiple alleles, rather than being associated with loci in other LGs (e.g., LG3, LG23). This study also tentatively names the allele(s) giving intermediate sex ratios as “ambivalent” and emphasizes that the presence and actions of such allele(s) at the same sex-determining locus could explain departures from predicted sex ratios observed in some earlier studies in Nile tilapia.

Thesis (MScAgric)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: DNA micro satellite loci express extensive allelic variation making them convenient markers for research in many fields employing population genetic tools, including aquaculture and conservation genetics. Twelve Oreochromis mossambicus populations from wild, captive and introduced sources in Southern Africa were screened for genetic variation at ten CA repeat micro satellite loci. Three of the loci - UNHI04, UNHlll, and UNH123 - were sufficiently well resolved to screen extensively and were interpreted according to a model of Mendelian inheritance. Data was analyzed in terms of genetic structure and levels of genetic variation, the effect of management regime in captivity through successive generations on genetic diversity, and the nature of phylogenetic relationships present between populations. Exact tests, carried out using Monte Carlo type multiple resampling techniques, and F-Statistics were used to detect and quantify genetic structure among the twelve populations. The Exact test X2 (P < 0.001), a FST of 0.27 (P < 0.001), eST of 0.26, RsT of 0.28, and a <l>ST of 0.17 all indicated significant structuring among the populations. The evident genetic structuring endorsed the practice of maintaining the populations as separate genetic stocks, in separate tanks, in order to preserve unique genetic material for aquaculture strain development. Populations also exhibited some significant deviations from Hardy Weinberg equilibrium characterised by an overall reduced heterozygosity across the loci. In microsatellite studies, null alleles are often suggested as major contributors to heterozygote deficits. To test for null alleles, two controlled crosses of 0. mossambicus were made. The progeny from each cross were examined for expected parental allelic ratios at the UNHI04, UNHlll and UNH123 loci. All three loci presented evidence of possible null alleles. Accelerated inbreeding and genetic drift through successive generations in captivity can reduce heterozygosity and gene diversity. To investigate loss of diversity a sample taken from the Bushmans population in 1999 (N = 25) was compared with a Bushmans 2000 sample (N = 36). The comparison highlighted altered allele frequencies, a significant increase in average observed heterozygosity and a non-significant change in average expected heterozygosity using the UNHI04 and UNH123 loci. Calculation of genetic distances and phylogenetic comparisons between the populations provided insight into the degree of management required in conserving genetic diversity in natural populations of Mozambique tilapia. UPGMA and Neighbour-Joining techniques were used to construct phylogenetic trees using Dm and ({)~)2 distance matrices. Clustering of populations appeared to reflect geographic locality of the source populations, however certain populations were not congruent with geography. Mantel tests were used to expose a possible association between genetic distance matrices generated from each individual locus. An association would support a geographic background to population genetic structure. The Mantel tests did not provide conclusive evidence. Mantel tests for association between the combined locus Dm and (81l)2 genetic distance matrices and a geographic distance matrix were similarly non-significant. Multi-dimensional scaling (MDS) plots of Euclidean distance values for Dm and (81l)2 matrices presented a two-dimensional view of the genetic distance data. The degree of similarity with the UPGMA and Neighbour-Joining tree-clustering pattern was higher for the (81l)2 than for the Dm MDS plots. Scatter plots indicated a reliable non-linear correlation between Euclidean distance and genetic distance for the two-dimensional MDS. The micro satellite markers employed in this research provided molecular information needed for complimenting a co-study on quantitative genetic evaluation of the twelve populations. The quantitative co-study provided measures of average length and weight gain indices for the populations based on progeny growth trials. No significant correlation of average heterozygosity (gene diversity) with either average weight or length gain was found. The significant genetic diversity and structure present between the twelve populations provided rationale for implementing strategies to conserve natural 0. mossambicus populations as genetic resources, and manage captive populations for long term maintenance of genetic diversity. / AFRIKAANSE OPSOMMING: Die verstaffing van groot alleliese variasie deur DNA mikrosateliete maak van hulle gerieflike merkers vir navorsing in 'n verskeidenheid van velde wat gebruik maak van populasie genetiese gereedskap, ingesluit akwakultuur en bewarings genetika. Twaalf 0. mossambicus populasies wat verkry was vanuit die natuur, in gevangeneskap en ingevoerdes, van Suidelike Afrika was getoets vir genetiese variasie by tien verskillende CA-herhalende mikrosateliet loci. Drie van die loci - UNHI04, UNHlll en UNH123 - is op grootskaal getoets en volgens In model van Mendeliese oorerwing geinterpreteer. Die data was ontleed volgens genetiese struktuur en vlakke van genetiese variasie, die effek wat bestuur strategie in gevangeneskap op genetiese diversiteit in opeenvolgende generasies uitgeoefen het, so wel as die aard van die filogenetiese verhoudings wat teenwoordig is tussen die populasies. "Exact" toetse is uitgevoer deur gebruik te maak van Monte Carlo tipe veelvuldige hermonsterinsamelings tegnieke en F-statistieke is gebruik vir die deteksie en kwantifisering van die genetiese struktuur tussen die twaalfpopulasies. Die Exact toets X2 (P < 0.001), 'n FST van 0.27 (P < 0.001), SST van 0.26, RsT van 0.28, en 'n <DST van 0.17 gee almal 'n indikasie van betekenisvolle strukturering tussen die populasies. Die genetiese struktuur bevestig die beleid dat die populasies behou moet word as aparte genetiese voorraad, in aparte tenke, om te verseker dat die unieke genetiese materiaal behoue bly om akwakultuur variante te ontwikkel. Populasies het ook betekenisvolle verskuiwings van die Hardy Weinberg ekwilibrium getoon, wat gekarakteriseer word deur 'n algemene verlaging van heterosigositeit oor die loci. Nul allele word dikwels aanbeveel om in mikrosateliet studies groot bydraes te maak tot hetersigotiese defekte. Om vir nul allele te toets was twee gekontroleerde kruisings van 0. mossambicus gemaak. Die nageslag van elke kruising was getoets vir verwagte ouer alleliese verhoudings by die UNHI04, UNHlll en UNH123 loci. Al drie loci het getoon dat dit moontlike nul allele kan wees. Versnelde inteling en genetiese drywing deur opeenvolgende generasies in gevangeneskap kan die heterosigositeit en diversiteit verminder. Om die vermindering van diversiteit te toets was 'n monster van die Busmans 1999 (N=25) populasie vergelyk met 'n monster van die Bushmans 2000 (N=36) populasie. Die vergelyking het veranderde alleel frekwensies, 'n betekenisvolle vermeerding in gemiddelde waargeneemde heterosigositeit en 'n onbetekenisvolle verandering in gemiddelde verwagte heterosigositeit getoon deur gebruik te maak van die UNHI04 en UNH123 loci. Berekening van genetiese afstande en filogenetiese vergelykings tussen die populasies het nuwe insig gegee oor die graad van bestuur wat nodig is om genetiese diversiteit in die natuurlike populasies van 0. mossambicus tilapia te behou.UPGMA en Neighbour-Joining tegnieke was gebruik om filogenetiese bome op te stel deur gebruik te maak van Dm en (OIl)2 afstand matrikse. Populasie bondeling het geblyk om geografiese lokaliteit van die bron populasies te toon, alhoewel van die populasies nie met die geografiese lokaliteit ooreengestem het nie. Mantel toetse is gebruik om 'n moontlike assosiasie tussen genetiese afstand matrikse wat verkry is van elke loci bloot te stel. 'n Assosiasie sou 'n geografiese agtergrond tot populasie genetiese struktuur steun. Oortuigende bewys is nie deur die Mantel toetse verskaf nie. Mantel toetse vir assosiasie tussen die gekombineerde loci Dm en (OJ..l)2 genetiese afstand matrikse en In geografiese afstand matriks was ook onbetekenisvol. 'n Tweedimensionele beskouing van die genetiese afstand data is voorgestel deur multidimensionele skaal (MDS) grafieke van Euclidean afstand waardes van die Dm en (OJ..l)2 matrikse te teken. Die graad van ooreenstemming met die UPGMA en Neighbour-Joining boom samevoeging patroon was hoër vir die (OJ..l)2 as vir die DmMDS grafieke. Verspreiding grafieke het 'n vertroubare nie-liniêre korrelasie tussen Euclidean afstande en genetiese afstande vir die twee-dimensionele MDS grafieke getoon. Die mikrosateliet merkers wat in die studie gebruik was het molekulêre informasie verskaf wat nodig is vir 'n komplimentêre studie oor die kwantitatiewe genetiese evalueering van dié twaalf populasies. Die kwantitatiewe studie het afmetings van gemiddelde lengte en gewig vermeerdering van die populasies verskaf gebaseer op nageslag proewe. Geen betekenisvolle korrelasie van gemiddelde hetersigositeit (geen diversiteit) was getoon met óf gemiddelde gewig óf lengte vermeerdering. Die betekenisvolle genetiese diversiteit en struktuur teenwoordig tussen die twaalf populasies het rede gegee om strategieë te implimenteer om natuurlike 0. mossambicus populasies te konserveer as genetiese bronne en om populasies in gevangeneskap te bestuur vir langtermyn instandhouding van genetiese diversiteit.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The effects of acute inbreeding on growth performance, yield and occurrence of deformity traits were studied in experimental full-sibling inbred populations of Mozambique tilapia Oreochromis mossambicus at three levels of inbreeding coefficients, namely F = 0.000, F = 0.250 and F = 0.375. The base population with F = 0.000 was established through the crossing of two geographically separated and genetically unrelated farm stocks. At each generation, the inbreeding depression indicated by body weight (BW), standard length (SL), specific growth rate (SGR) and yield were highly significant, but no linear relationship was found between level of inbreeding and inbreeding depression. Both condition factor (K) and the number of observed deformities appears not to be significantly affected by inbreeding at all three levels. Over all, the average inbreeding depression at F = 0.250 and F = 0.375 respectively was found to be 46.5 percent and 46.6 percent for body weight (BW); 18.2 percent and 18.0 percent for standard length (SL); 21.8 percent and 20.3 percent for specific growth rate (SGR) and 5.752 percent and 8.940 percent for flesh yield. The outbred Control group differed significantly (P<0.05) from the six inbred family groups in terms of body weight (BW), standard length (SL), specific growth rate (SGR) and yield at all levels of inbreeding studied (F = 0.000, F = 0.250 and F = 0.375). Average inbreeding depression for body weight (BW) amongst the six inbred families ranged from 39.6 to 54.2 percent at F = 0.250 (in Gen 2) and 45.6 to 47.3 percent at F = 0.375 (in Gen 3). The inbreeding depression coefficient for body weight (BW) per 10% increase in F, amongst the six inbred families, ranged from 15.9 to 21.7 percent at F = 0.250 and from 12.2 to 12.6 percent at F = 0.375. Average inbreeding depression for standard length (SL) amongst the six inbred families ranged from 14.0 to 22.3 percent at F = 0.250 and from 17.2 to 18.4 percent at F = 0.375. The inbreeding depression coefficient for standard length (SL) amongst the six inbred families ranged from 5.6 to 8.9 percent at F = 0.250 and from 4.6 to 4.9 percent at F = 0.375. Average inbreeding depression for specific growth rate (SGR) amongst the six inbred families ranged from 17.9 to 27.9 percent at F = 0.250 and from 16.7 to 27.2 percent at F = 0.375. The inbreeding depression coefficient amongst the six inbred families ranged from 7.2 to 11.2 percent at F = 0.250 and from 4.5 to 7.3 percent at F = 0.375. Average inbreeding depression for yield amongst the six inbred families ranged from 0.4 to 7.7 percent at F = 0.250 and from 8.5 to 10.2 percent at F = 0.375. The inbreeding depression coefficient for yield amongst the six inbred families ranged between 0.2 and 3.1 percent at F = 0.250 and from 2.3 to 2.7 percent at F = 0.375. The condition factor (K) of the six inbred families showed no significant differences to the Control (P > 0.05) at all levels of inbreeding with K-values ranging from 1.42 to 2.85. The occurrence of morphological deformities in all seven family groups including the Control showed no noticeable trend, with a random, nonlinear occurrence of fluctuating asymmetry observed at different inbreeding levels in O. mossambicus. This study demonstrates that inbreeding has a significant negative effect on the production traits of Oreochromis mossambicus, especially growth. Results from this study emphasize the need to create awareness amongst small scale farmers of the importance of preventing uncontrolled inbreeding in production systems, as well as to monitor inbreeding levels during the process of dissemination of improved fish strains to small scale fish growers in developing countries, including Africa. / AFRIKAANSE OPSOMMING: Die gevolge van akute inteling op die groei prestasie, opbrengs en voorkoms van misvorming eienskappe bestudeer in eksperimentele full-broer ingeteelde bevolkings van Mosambiek tilapia Oreochromis mossambicus op drie vlakke van inteling koëffisiënte, naamlik F = 0,000, F = 0,250 en F = 0,375. Die basis bevolking F = 0,000 gestig deur die kruising van twee geografies geskei en geneties onverwant plaas aandele. Op elke generasie, die inteling depressie aangedui deur die liggaam gewig (BW), standaard lengte (SL), spesifieke groeitempo (SGR) en opbrengs was hoogs betekenisvol, maar geen lineêre verband is gevind tussen vlak van inteling en inteling depressie. Beide toestand faktor (K) en die aantal waargeneem deformiteite verskyn om nie beduidend beïnvloed deur inteling op al drie vlakke. Oor alles is, die gemiddelde inteling depressie by F = 0,250 en F = 0,375 onderskeidelik gevind om 46,5 persent en 46,6 persent vir die liggaam gewig (BW) wees; 18,2 persent en 18,0 persent vir standaard lengte (SL); 21,8 persent en 20,3 persent vir spesifieke groeitempo (SGR) en 5,752 persent en 8,940 persent vir vlees opbrengs. Die outbred beheer betekenisvol verskil (P <0,05) van die ses ingeteelde familie groepe in terme van die liggaam gewig (BW), standaard lengte (SL), spesifieke groeitempo (SGR) en opbrengs op alle vlakke van inteling bestudeer (F = 0,000 , F = 0,250 en F = 0,375). Gemiddeld inteling depressie vir die liggaam gewig (BW) onder die ses ingeteelde families gewissel 39,6-54,2 persent by F = 0,250 (in Gen 2) en 45,6-47,3 persent by F = 0,375 (in Gen 3). Die inteling depressie koëffisiënt vir die liggaam gewig (BW) per 10% toename in F, onder die ses ingeteelde families, het gewissel 15,9-21,7 persent by F = 0,250 en 12,2-12,6 persent by F = 0,375. Gemiddeld inteling depressie vir standaard lengte (SL) onder die ses ingeteelde families gewissel 14,0-22,3 persent by F = 0,250 en 17,2-18,4 persent by F = 0,375. Die inteling depressie koëffisiënt vir standaard lengte (SL) onder die ses ingeteelde families gewissel 5,6-8,9 persent by F = 0,250 en 4,6-4,9 persent by F = 0,375. Gemiddeld inteling depressie vir spesifieke groeitempo (SGR) onder die ses ingeteelde families gewissel 17,9-27,9 persent by F = 0,250 en 16,7-27,2 persent by F = 0,375. Die inteling depressie koëffisiënt onder die ses ingeteelde families gewissel 7,2-11,2 persent by F = 0,250 en 4,5-7,3 persent by F = 0,375. Gemiddeld inteling depressie vir opbrengs onder die ses ingeteelde families het gewissel van 0,4-7,7 persent by F = 0,250 en 8,5-10,2 persent by F = 0,375. Die inteling depressie koëffisiënt vir opbrengs onder die ses ingeteelde families gewissel tussen 0,2 en 3,1 persent by F = 0,250 en 2,3-2,7 persent by F = 0,375. Die toestand faktor (K) van die ses ingeteelde families het geen betekenisvolle verskille op die Beheer (P> 0,05) op alle vlakke van inteling met K-waardes wissel 1,42-2,85. Die voorkoms van morfologiese afwykings in al sewe familie groepe, insluitend die beheer het geen merkbare neiging, met 'n ewekansige, nie-lineêre voorkoms van wisselende asimmetrie waargeneem op verskillende vlakke in inteling O. mossambicus. Hierdie studie toon dat inteling het 'n beduidende negatiewe uitwerking op die produksie-eienskappe van Oreochromis mossambicus, veral groei. Resultate van hierdie studie beklemtoon die noodsaaklikheid om bewustheid onder kleinboere van die belangrikheid van die voorkoming van onbeheerde inteling in die produksie stelsels, sowel as om te monitor inteling vlakke tydens die proses van verspreiding van verbeterde vis stamme klein skaal vis produsente in ontwikkelende lande te skep, insluitend Afrika.

Yucca (Yucca schidigera) and Soapbark (Quillaja saponaria), both native desert plants, are major commercial sources of saponin extracts. Yucca schidigera is native to the southwestern United States and to the arid Mexican desert, and Quillaja saponaria is found in arid areas of Chile. Saponins have detergent or surfactant properties with both water-soluble and fat-soluble components.The use of natural saponins from yucca and soapbark as an additive in the diet, has improved production in aquatic organisms. The purpose of this study was to evaluate the effect of inclusion of Nutrafito plus (NTF+), which is a mixture of Yucca and Soapbark extracts, on growth, survival, and development of Nile tilapia Oreochromis niloticus and the effect on growth, survival, development, and digestive enzyme activities for juvenile Pacific white shrimp Litopenaeus vannamei.The extracts were included at different levels in the diets of tilapia and shrimp in four experiments. In experiment 1, at the end of 6 weeks there was no significant difference (p > 0.05) between growth rates of tilapia fed 6 different diets, with no mortality nor abnormal behavior in any of the treatments. Water quality parameters were determined weekly and remained within recommended limits for Nile tilapia culture.In the second trial, at the end of 40 days, there was no significant difference (p > 0.05) between growth rates of fish fed 7 different diets with various levels of Nutrafito. There were no mortalities during the experiment.In the third trial, 20 tanks (140 L each) were stocked with 10 shrimp each. Tanks were divided into 5 treatments with 4 replicates each. There was significant difference (p < 0.05) between growth rates of shrimp fed 5 different diets, with higher growth rate at higher levels of inclusion.In the fourth trial, 15 tanks (140 L each) were stocked with 10 shrimp each. Tanks were divided into 5 treatments with 3 replicates each. There was significant difference between growth rates and feed conversion ratio of shrimp fed 5 different diets. In addition, an analysis for digestive enzymes activity was done and no significant difference (p > 0.05) was found between treatments.

Orientador: Joaquim Gonçalves Machado Neto / Banca: Flávio Ruas de Moraes / Banca: Marcello Pardi de Castro / Resumo: Objetivou-se determinar a CL50-24h e classificar o malathion (Komvektor 44% EW) pela toxicidade aguda para tilápias (Oreochromis niloticus); determinar as concentrações na água que resultam em aproximadamente 10, 30, 50 e 70% de mortalidade das tilápias após 24h de exposição aguda; observar os sinais clínicos e alterações histológicas causadas pelo malathion nos peixes sobreviventes em 15 e 30 dias de recuperação; registrar as alterações nas variáveis de qualidade de água; e determinar a porcentagem de mortalidade dos peixes do lote em que os animais sobreviventes se recuperam da intoxicação aguda com o malathion. Os valores da CL50-24h do malathion e as concentrações que causam entorno de 10, 30, 50 e 70% de mortalidade dos peixes foram calculados com os dados de ensaios de toxicidade aguda em laboratório. O ensaio de recuperação dos peixes expostos por 24h às concentrações parcialmente letais foi realizado com lotes de animais em mesocosmos de 1.000 L e o de recuperação dos animais sobreviventes às intoxicações em mesocosmos de 150 L. Os sinais clínicos da intoxicação nos sobreviventes foram observados e as análises histopatológicas feitas em fragmentos de brânquias, fígado e rim coletados 24h após intoxicação aguda e aos 15º e 30º dias de recuperação. As variáveis de qualidade da água foram aferidas. A CL50-24h do malathion foi 5,15 mg.L-1, que o classifica como moderadamente e muito tóxico. As concentrações de 3,0; 4,5; 5,0 e 7,0 mg.L-1 de malathion causam,respectivamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim of this study was to determine the LC50-24h and to classify the malathion (Komvektor 44% EW) according with acute toxicity to tilapias (Oreochromis niloticus); to determine the concentrations in the water wich caused 10, 30, 50 and 70% of mortality in tilapia after 24h of acute exposition; to observe the clinical signs and histological changes caused by malathion in surviving fish in 15 and 30 days of recovery; to register the changes in the parameters of water quality caused by malathion; and to determine the percentage of mortality in the surviving groups recover from acute intoxication by malathion. The LC50-24h values of the malathion to tilapias and the concentrations that cause about 10, 30, 50 and 70% of fish mortality was calculated with the acute toxicity dates in laboratory. The recovery test of exposed fish for 24h at partially lethal concentrations was carried out with groups of animals in mesocosm of 1000L and the recovery of the animals surviving to intoxication in mesocosm of 150L. Clinical signs of intoxication in survival fish was observed and the histologic analysis in tissue fragments of gills, liver and kidney collected after 24h acute intoxication at 15 and 30 days of recovery. The water quality parameters were recorded. The LC50-24h of malathion was 5,15 mg.L-1, and this value is classified in moderate and very toxic. The concentrations of 3,0; 4,5; 5,0 and 7,0 mg.L-1 caused, respectively, mortalities of 17,5%, 27,5%, 45% and 55% of fish in labor... (Complete abstract click electronic access below) / Mestre

En los últimos años se han desarrollados numerosas investigaciones con el fin de mejorar el cultivo del camarón gigante de Malasia, investigaciones en policultivo, siembra de juveniles graduados y la adición de substrato han dado muy buenos resultados. La mayoría de estas investigaciones han tenido como objetivos la maximización de los niveles de producción, obtener mejoras en los pesos finales e incrementar la resistencia a enfermedades. El objetivo del presente estudio fue evaluar el crecimiento y la supervivencia del camarón gigante de Malasia Macrobrachium rosenbergii con 2 densidades de siembra (D1 y D2) y 2 porcentajes de incremento de substratos (S1 y S2) en policultivo con Oreochromis niloticus tilapia roja. Para alcanzar este objetivo se utilizó un diseño factorial 2 x 2, es decir, se evaluaron 4 tratamientos, 2 niveles por cada uno de los 2 factores (D1S1, D1S2, D2S1 y D2S2), cada tratamiento con tres repeticiones. Los datos de talla, peso, supervivencia y biomasa, obtenidos de las biometrías fueron analizados utilizando el software estadístico Statgrapfhic v.7,0 analizado al 95% de confianza (α es menor que 0,05). Después de 80 días de cultivo se observó que el tratamiento T3 (D2S1) obtuvo los mejores crecimientos (7,05 ± 2,13 g) y los mejores porcentajes de supervivencia (97,5%) para el camarón gigante de malasia. Los dos niveles de adición de substrato fueron igualmente efectivos para el crecimiento del camarón, se encontró una relación inversa entre el incremento de los substratos y los porcentajes de supervivencia. El crecimiento en talla de la tilapia solo fue afectado por la interacción de los factores densidad y substrato donde los tratamientos T2 (D1S2) y T3 (D2S1) presentaron crecimientos significativamente mayores; el crecimiento en peso de la tilapia solo fue afectado por el factor substrato, encontrándose una relación directa; no se encontró diferencia significativa (p es menor que 0,05) para los porcentajes de supervivencia. Se encontró una relación directa entre el incremento de la densidad y la biomasa final del camarón gigante de Malasia, siendo el nivel D2 significativamente mayor que D1. El tratamiento T3 (690,1 Kg ha-1) presentó una biomasa significativamente mayor que T2 (420,9 Kg ha-1) y T1 (387,1 Kg ha-1), T3 y T4 no fueron significativamente diferentes (p es mayor que 0.05). Para la tilapia se encontró una relación directa entre en incremento del substrato y la biomasa final, siendo el nivel S2 significativamente mayor que S1. La biomasa del tratamiento T2 (629,7 Kg ha-1) fue significativamente mayor respecto a T1 (453,4 Kg ha-1). En el análisis de producción los tratamientos T2 y T3 alcanzaron los mejores valores, el análisis de proyección mostró que el policultivo camarón tilapia, con adición de substrato, es una buena alternativa para obtener mejores rendimientos y ganancias. / In recent years, several researches have been developed with the objective of improve the culture of the fresh water prawn Macrobrachium rosenbergii, for instance polyculture, size grading juveniles and adding artificial substrate to the ponds have achieve results, in addition the majority of those investigations in the fresh water prawn have had different objectives, such as increasing the productions levels, increasing the harvest weight and increasing the resistance of different illnesses. This study was designed to evaluate the growth and supervivence of the freshwater prawn Macrobrachium rosenbergii with 2 different densities (D1 y D2) and 2 percentages of added substrates (S1 y S2) in polyculture with Oreochromis niloticus “red tilapia”. In order to achieve this objective, the experiment was designed such as 2 x 2 factorial arrangement consisting in 4 treatment combinations with 2 levels for each of 2 factors (D1S1, D1S2, D2S1 y D2S2) and which 3 replicate for each treatment. The size, supervivence and total production obtained from biometries were analyzed using the software Statgrapfhic v.7,0 at 95 % confidence (α =0,05). After 80 days of culture, T3 (D2S1) treatment had the best weight (7,05 ± 2,13g) and percentage of supervivence (97,5%) to freshwater prawn, in addition the different levels of substrates had similar effect on the harvest weight of freshwater prawn, moreover an inverse relation was found between added substrate and percentage of supervivence. The body length of tilapia was only affected by density and substrate interaction, where T2 and T3 treatment had the best growth. The harvest weight of tilapia was only affected by the substrate and a direct relation was found between them. A significantly effect (p is less than0,05) was not found between the different percentages of supervivence. A direct relation was found between density and total production of the freshwater prawn, where D2 obtained better results than D1. T3 treatment (690,1 kg ha-1) had better yield than T2 (420,9 kg ha-1) and T1 (387,1 kg ha-1), T3 and T4 were not different. Tilapia had a direct relation between added substrate and total production, where S2 was significantly higher than S1. The production from T2 treatment (629,7 kg ha-1) was significantly higher than T1 (453,4 kg ha-1). The production analysis showed that polyculture of prawn-tilapia with added substrate, is a good alternative in order to obtain best productions and weights. / In recent years, several researches have been developed with the objective of improve the culture of the fresh water prawn Macrobrachium rosenbergii, for instance polyculture, size grading juveniles and adding artificial substrate to the ponds have achieve results, in addition the majority of those investigations in the fresh water prawn have had different objectives, such as increasing the productions levels, increasing the harvest weight and increasing the resistance of different illnesses. This study was designed to evaluate the growth and supervivence of the freshwater prawn Macrobrachium rosenbergii with 2 different densities (D1 y D2) and 2 percentages of added substrates (S1 y S2) in polyculture with Oreochromis niloticus “red tilapia”. In order to achieve this objective, the experiment was designed such as 2 x 2 factorial arrangement consisting in 4 treatment combinations with 2 levels for each of 2 factors (D1S1, D1S2, D2S1 y D2S2) and which 3 replicate for each treatment. The size, supervivence and total production obtained from biometries were analyzed using the software Statgrapfhic v.7,0 at 95 % confidence ( =0,05). After 80 days of culture, T3 (D2S1) treatment had the best weight (7,05 ± 2,13g) and percentage of supervivence (97,5%) to freshwater prawn, in addition the different levels of substrates had similar effect on the harvest weight of freshwater prawn, moreover an inverse relation was found between added substrate and percentage of supervivence. The body length of tilapia was only affected by density and substrate interaction, where T2 and T3 treatment had the best growth. The harvest weight of tilapia was only affected by the substrate and a direct relation was found between them. A significantly effect (p<0,05) was not found between the different percentages of supervivence. A direct relation was found between density and total production of the freshwater prawn, where D2 obtained better results than D1. T3 treatment (690,1 kg ha-1) had better yield than T2 (420,9 kg ha-1) and T1 (387,1 kg ha-1), T3 and T4 were not different. Tilapia had a direct relation between added substrate and total production, where S2 was significantly higher than S1. The production from T2 treatment (629,7 kg ha-1) was significantly higher than T1 (453,4 kg ha-1). The production analysis showed that polyculture of prawn-tilapia with added substrate, is a good alternative in order to obtain best productions and weights. / Tesis

This thesis explores the feasibility of using melanophore-based biosensors from Oreochromis niloticus. Melanophores are one type of pigmented cell of the scales and fins of fish that respond in a motile fashion to a diverse range of stimuli. Fish scales were employed as the first step in determining the utility of melanophores as biosensors. Responsiveness of melanophores in scales was quantitated with several bioactive agents. Experiments with depolarizing potassium ion, guanethidine, yohimbine (an adrenergic antagonist), and capsaicin (a sensory stimulant) provided evidence that melanophores are under nervous regulation. Conditions were developed to allow simplification of intact scale preparations, entailing epidermal removal and dennervation of scales. This simplification resulted in increased sensitivity and responsiveness to a larger array of bioactive agents, including a cyclic AMP analog. The simplified preparation was successfully tested for its ability to function as a biosensor using pharmaceutical eye drops; the response observed was determined to be due to naphazoline, an adrenergic agonist. Methods were developed that enabled culturing of chromatophores independent from scales. Cultured chromatophores were found to be responsive to bioactive agents with a comparable degree of sensitivity as simplified scale preparations. Attempts were undertaken to develop co-cultures of chromatophores with other cell types and with further development melanophore-based biosensors can be exploited. / Graduation date: 1997

The effect of fractionating flax, peas and canola on the digestibility of these ingredients in <i>Nile tilapia</i> was determined. Dehulling of flax, and processing peas and canola to pea protein concentrate and canola protein concentrate, resulted in significant increases in the energy and dry matter digestibilities of these products (P < 0.05). Protein digestibility was significantly improved by dehulling flax but there were no differences in the protein digestibilities of pea, canola and their protein concentrates. The ability of the most digestible ingredients from Experiment 1 to replace fish meal in tilapia diets was examined in a 9 week growth trial. The diet formulations were arranged in a 2 x 4 factorial design with 2 types of plant protein mixtures used to replace fish meal (simple: soyabean meal and maize gluten meal or complex: soyabean meal, maize gluten meal, dehulled flax, pea protein concentrate and canola protein concentrate) and 4 levels of protein originating from fish meal (100%, 67%, 33% and 0%). Diets were formulated to contain equal digestible protein (380 g kg-1) and digestible energy (17.63 MJ kg-1). Fifty six tanks containing 10 male <i>Nile tilapia</i> were used in this experiment. Fish were fed to apparent satiation twice daily for a total of 63 days and growth and feed intake was measured for the entire experimental period. On day 64 of the experiment, one fish per tank was euthanized and a 1 cm segment of small intestine was prepared for measurement of villus length. The average daily gains, specific growth rates and feed efficiencies of fish fed diets with 0% fish meal were significantly lower than fish fed diets with the 33, 67 or 100% fish meal levels. Fish fed the complex diets had significantly higher average daily gains, specific growth rates, feed:gain ratios and protein efficiency ratios than those fed the simple diets. Villus length decreased with decreasing levels of fish meal and increased with increased diet complexity but the effects were not significant. The results indicated that replacement of fish meal with a complex mixture of plant ingredients may allow a greater replacement of fish meal in diets fed to <i> Nile tilapia</i>.

The effect of fractionating flax, peas and canola on the digestibility of these ingredients in <i>Nile tilapia</i> was determined. Dehulling of flax, and processing peas and canola to pea protein concentrate and canola protein concentrate, resulted in significant increases in the energy and dry matter digestibilities of these products (P < 0.05). Protein digestibility was significantly improved by dehulling flax but there were no differences in the protein digestibilities of pea, canola and their protein concentrates. The ability of the most digestible ingredients from Experiment 1 to replace fish meal in tilapia diets was examined in a 9 week growth trial. The diet formulations were arranged in a 2 x 4 factorial design with 2 types of plant protein mixtures used to replace fish meal (simple: soyabean meal and maize gluten meal or complex: soyabean meal, maize gluten meal, dehulled flax, pea protein concentrate and canola protein concentrate) and 4 levels of protein originating from fish meal (100%, 67%, 33% and 0%). Diets were formulated to contain equal digestible protein (380 g kg-1) and digestible energy (17.63 MJ kg-1). Fifty six tanks containing 10 male <i>Nile tilapia</i> were used in this experiment. Fish were fed to apparent satiation twice daily for a total of 63 days and growth and feed intake was measured for the entire experimental period. On day 64 of the experiment, one fish per tank was euthanized and a 1 cm segment of small intestine was prepared for measurement of villus length. The average daily gains, specific growth rates and feed efficiencies of fish fed diets with 0% fish meal were significantly lower than fish fed diets with the 33, 67 or 100% fish meal levels. Fish fed the complex diets had significantly higher average daily gains, specific growth rates, feed:gain ratios and protein efficiency ratios than those fed the simple diets. Villus length decreased with decreasing levels of fish meal and increased with increased diet complexity but the effects were not significant. The results indicated that replacement of fish meal with a complex mixture of plant ingredients may allow a greater replacement of fish meal in diets fed to <i> Nile tilapia</i>.

The structure and function of brain shows sexual dimorphism in the vertebrates. Sexual differentiation is divided into brain sexual differentiation and gonad sexual differentiation. Brain sexual differentiation is resulted from the neural development. In the present study, the neurosphere cloned from tilapia, Oreochromis mossambicus, was used. The characteristics of neurosphere derived from both primary- and sub- culture, were studied. The effects of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF, FGF2), and temperature on the neurosphere cloned from both primary culture and subculture, were investigated. These results show that the neurospheres, cloned form both primary- and sub- culture, is consist of the nestin-immunoreactive cell. Furthermore, the cell of the neurosphere shows an ability of differentiation. And the diameter of neurosphere in the subculture is significantly larger than that of primary culture. On the other hand, both FGF and temperature have an effect to increase the diameter of neurosphere in the primary culture.