Longitudinal analyses of correlated response efficiencies of fillet traits in Nile tilapia.

Recent studies with Nile tilapia have shown divergent results regarding the possibility of selecting on morphometric measurements to promote indirect genetic gains in fillet yield (FY). The use of indirect selection for fillet traits is important as these traits are only measurable after harvesting. Random regression models are a powerful tool in association studies to identify the best time point to measure and select animals. Random regression models can also be applied in a multiple trait approach to analyze indirect response to selection, which would avoid the need to sacrifice candidate fish. Therefore, the aim of this study was to investigate the genetic relationships between several body measurements, weight and fillet traits throughout the growth period and to evaluate the possibility of indirect selection for fillet traits in Nile tilapia. Data were collected from 2042 fish and was divided into two subsets. The first subset was used to estimate genetic parameters, including the permanent environmental effect for BW and body measurements (8758 records for each body measurement, as each fish was individually weighed and measured a maximum of six times). The second subset (2042 records for each trait) was used to estimate genetic correlations and heritabilities, which enabled the calculation of correlated response efficiencies between body measurements and the fillet traits. Heritability estimates across ages ranged from 0.05 to 0.5 for height, 0.02 to 0.48 for corrected length (CL), 0.05 to 0.68 for width, 0.08 to 0.57 for fillet weight (FW) and 0.12 to 0.42 for FY. All genetic correlation estimates between body measurements and FW were positive and strong (0.64 to 0.98). The estimates of genetic correlation between body measurements and FY were positive (except for CL at some ages), but weak to moderate (-0.08 to 0.68). These estimates resulted in strong and favorable correlated response efficiencies for FW and positive, but moderate for FY. These results indicate the possibility of achieving indirect genetic gains for FW and by selecting for morphometric traits, but low efficiency for FY when compared with direct selection.

Successful xenogeneic germ cell transplantation from Jundia catfish (Rhamdia quelen) into adult Nile tilapia (Oreochromis niloticus) testes.

Fish germ cell transplantation presents several important potential applications for aquaculture, including the preservation of germplasm from endangered fish species with high genetic and commercial values. Using this technique in studies developed in our laboratory with adult male Nile tilapias (Oreochromis niloticus), all the necessary procedures were successfully established, allowing the production of functional sperm and healthy progeny approximately 2months after allogeneic transplantation. In the present study, we evaluated the viability of the adult Nile tilapia testis to generate sperm after xenogeneic transplant of germ cells from sexually mature Jundia catfish (Rhamdia quelen) that belong to a different taxonomic order. Therefore, in order to investigate at different time-periods post-transplantation, the presence and development of donor PKH26 labeled catfish germ cells were followed in the tilapia seminiferous tubules. From 7 to 20days post-transplantation, only PKH26 labeled spermatogonia were observed, whereas spermatocytes at different stages of development were found at 70days. Germ cell transplantation success and progression of spermatogenesis were indicated by the presence of labeled PKH26 spermatids and sperm on days 90 and 120 post-transplantation, respectively. Confirming the presence of the catfish genetic material in the tilapia testis, all recipient tilapias evaluated (n=8) showed the genetic markers evaluated. Therefore, we demonstrated for the first time that the adult Nile tilapia testis offers the functional conditions for development of spermatogenesis with sperm production from a fish species belonging to a different order, which provides an important new venue for aquaculture advancement.

Modelagem das trajetórias médias do peso vivo e do rendimento de carcaça de tilápias do Nilo (Oreochromis niloticus) por meio de funções B-spline / Modeling the mean trajectories of live body weight and carcass yield of Nile tilapia (Oreochromis niloticus) by B-spline functions

Empregando o método dos quadrados mínimos e polinômios B-spline quadráticos, diferentes modelos estatísticos foram testados para identificar o mais apropriado para modelar as trajetórias médias do peso vivo e do rendimento de carcaça de tilápias do Nilo (Oreochromis niloticus). Dados de peso vivo (8.758) e de rendimento de carcaça (2.042) de tilápias com idades entre 106 e 245 dias foram obtidos de 72 famílias provenientes de 36 machos e 72 fêmeas. As variáveis sexo e tanque de criação foram consideradas como classificatórias, e os coeficientes dos polinômios B-spline quadráticos com dois a cinco intervalos de mesmo tamanho foram utilizados como covariáveis. Segundo a maioria dos critérios de ajuste utilizados, os modelos com polinômio B-spline quadrático com cinco intervalos de mesmo tamanho apresentaram os melhores ajustes. O aumento do número de intervalos do polinômio B-spline melhorou o ajuste dos polinômios aos dados. A inclusão dos efeitos classificatórios de sexo, tanque de criação, interação entre esses efeitos e polinômio B-spline quadrático aninhado a essa interação indicou que, com o decorrer do tempo, cada sexo, cultivado em diferente tanque, apresentou trajetória média diferente, sendo necessária a inclusão do aninhamento do tempo na interação sexo x tanque de criação para que, em programas de melhoramento genético da espécie, os valores genéticos dos candidatos à seleção não sejam sub ou superestimados.

Employing the method of least squares and quadratic B-spline polynomials, different statistical models were tested to identify the most appropriate to model the mean trajectories of live weight and carcass yield of Nile tilapia (Oreochromis niloticus). Data of live weight (8,758) and carcass yield (2,042) of tilapias with ages between 106 and 245 days were obtained from 72 families derived from 36 males and 72 females. The sex and tank variables were considered as classificatory and the coefficients of quadratic polynomials B-spline of two to five intervals of the same size were used as covariables. According to most fit criteria used, the models with quadratic B-spline polynomial with five intervals of the same size presented the best adjustments. The increase in number of intervals of B-spline polynomial improved the fit of the polynomials to the data. The inclusion of classificatory effects from sex, tank, the interaction of these effects and the quadratic polynomial B-spline nested in this interaction indicated that, over time, each sex, grown in different tank, presented different mean trajectory, necessitating the inclusion of nesting time in the interaction sex x tank in order to avoid the under or overestimation of breeding values of the selection candidates in breeding programs of this species.

Production of oocytes of Nile tilapia (Oreochromis niloticus) for in vitro fertilization via hormonal treatments.

Only a few studies have described hormonal treatments for induction of synchronicity and gamete collection in Nile tilapia (Oreochromis niloticus), both important for assortative matings in breeding programmes and essential for polyploidy technologies. In this study, we compared the effectiveness of carp pituitary extract (CPE), Nile tilapia pituitary extract (TPE), human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) protocols on the induction of spawning and egg production in Nile tilapia. Among the hormonal treatments analysed, only hCG was effective for producing viable gametes for in vitro fertilization. To verify the viability of this hormonal treatment, hCG was tested using different doses (1000, 2000, 3000, 4000 and 5000 IU/kg) in a large number of females (208 animals) from two Nile tilapia lines. The results indicated that hCG doses between 1000 and 5000 IU/kg could be used to induce final oocyte maturation in Nile tilapia with collection of stripped oocytes. This is the first study to report differential reproductive responses to hormonal treatment between tilapia lines: line 1 was more efficient at producing eggs and post-hatching larvae after hCG induction than line 2. In conclusion, we demonstrated that the hCG protocol may be applied on a large scale to induce final oocyte maturation in Nile tilapia. The development of a protocol for in vitro fertilization in Nile tilapia may aid in breeding programmes and biotechnological assays for the development of genetically modified lines of Nile tilapia.

Molecular cloning and expression of epsilon toxin from Clostridium perfringens type D and tests of animal immunization.

Epsilon toxin produced by Clostridium perfringens types B and D causes enterotoxemia in sheep, goats and calves. Enterotoxemia can cause acute or superacute disease, with sudden death of the affected animal. It provokes huge economic losses when large numbers of livestock are affected. Therapeutic intervention is challenging, because the disease progresses very rapidly. However, it can be prevented by immunization with specific immunogenic vaccines. We cloned the etx gene, encoding epsilon toxin, into vector pET-11a; recombinant epsilon toxin (rec-epsilon) was expressed in inclusion bodies and was used for animal immunization. Serum protection was evaluated and cross-serum neutralization tests were used to characterize the recombinant toxin. To analyze the potency of the toxin (as an antigen), rabbits were immunized with 50, 100 or 200 microg recombinant toxin, using aluminum hydroxide gel as an adjuvant. Titers of 10, 30 and 40 IU/mL were obtained, respectively. These titers were higher than the minimum level required by the European Pharmacopoeia (5 IU/mL) and by the USA Code of Federal Regulation (2 IU/mL). This rec-epsilon is a good candidate for vaccine production against enterotoxemia caused by epsilon toxin of C. perfringens type D.