Lessons from established breeding programs: terrestrial and aquatic animals

Some relevant components of selection program theory and implementation are reviewed. This includes pedigree recording, genetic evaluation, balancing genetic gains and genetic diversity and tactical integration of key issues. Lessons learned are briefly described – illustrating how existing method and tools can be useful when launching a program in a novel species, and yet highlighting the importance of proper understanding and custom application according to the biology and environments of that species.

Perspectives from agriculture: advances in livestock breeding - implications for aquaculture genetics

In this paper we present livestock breeding developments that could be taken into consideration in the genetic improvement of farmed aquaculture species, especially in freshwater fish. Firstly, the current breeding objective in aquatic species has focused almost exclusively on the improvement of body weight at harvest or on growth related traits. This is unlikely to be sufficient to meet the future needs of the aquaculture industry. To meet future demands breeding programs will most likely have to include additional traits, such as fitness related ones (survival, disease resistance), feed efficiency, or flesh quality, rather than only growth performance. In order to select for a multi-trait breeding objective, genetic variation in traits of interest and the genetic relationships among them need to be estimated. In addition, economic values for these traits will be required. Generally, there is a paucity of data on variable and fixed production costs in aquaculture, and this could be a major constraint in the further expansion of the breeding objectives. Secondly, genetic evaluation systems using the restricted maximum likelihood method (REML) and best linear unbiased prediction (BLUP) in a framework of mixed model methodology could be widely adopted to replace the more commonly used method of mass selection based on phenotypic performance. The BLUP method increases the accuracy of selection and also allows the management of inbreeding and estimation of genetic trends. BLUP is an improvement over the classic selection index approach, which was used in the success story of the genetically improved farmed tilapia (GIFT) in the Philippines, with genetic gains from 10 to 20 per cent per generation of selection. In parallel with BLUP, optimal genetic contribution theory can be applied to maximize genetic gain while constraining inbreeding in the long run in selection programs. Thirdly, by using advanced statistical methods, genetic selection can be carried out not only at the nucleus level but also in lower tiers of the pyramid breeding structure. Large scale across population genetic evaluation through genetic connectedness using cryopreserved sperm enables the comparison and ranking of genetic merit of all animals across populations, countries or years, and thus the genetically superior brood stock can be identified and widely used and exchanged to increase the rate of genetic progress in the population as a whole. It is concluded that sound genetic programs need to be established for aquaculture species. In addition to being very effective, fully pedigreed breeding programs would also enable the exploration of possibilities of integrating molecular markers (e.g., genetic tagging using DNA fingerprinting, marker (gene) assisted selection) and reproductive technologies such as in-vitro fertilization using cryopreserved spermatozoa.

Genetic improvement of the herbivorous blunt snout bream (Megalobrama amblycephala)

Selection experiments with the herbivorous blunt snout bream or Wuchang bream (Megalobrama amblycephala) were started in 1985. Mass selection for size and length/depth ratio resulted in a significant increase in growth and better shape, while inbreeding led to a significant decrease in growth. The total selection ratio from fry to mature brooders was about 0.03 per cent per generation. In the grow out stage, the average daily body weight gains of two lines of fifth generation (F5) fish were 29 per cent and 20 per cent respectively more than the control group, with an average of 5.8 per cent and 4 per cent improvements per generation, respectively. The body was 4 per cent deeper in ratio of standard length/body depth. The effects of inbreeding were examined by crossing full-sibs, the offspring of which were kept without selection. The third generation inbred fish showed 17 per cent lower growth as compared to the control group, with an average of 7.5 per cent per generation. The results demonstrate that selection is a powerful tool to improve the economic traits of the blunt snout bream, but inbreeding can rapidly lead to a reduction in performance. In 2000, the 6th generation of selected bream was certified by the Chinese Ministry of Agriculture as a good breed for aquaculture.

Genetic and morphological identification of pelagic juvenile rockfish collected from the Gulf of Alaska

Pelagic juvenile rockfish (Sebastes spp.) collected in surveys designed to assess juvenile salmonids and other species in the Gulf of Alaska in 1998 and 2000–2003 provide an opportunity to document the occurrence of the pelagic juveniles of several species of rockfish. Often, species identification of rockfish is difficult or impossible at this stage of development (~20 to 60 mm), and few species indigenous to Alaska waters have been described. Use of mitochondrial DNA markers for rockfish species allowed unequivocal identification of ten species (S. aleutianus, S. alutus, S. borealis, S. entomelas, S. flavidus, S. melanops, S. pinniger, S. proriger, S. reedi, and S. ruberrimus) in subsamples from the collections. Other specimens were genetically assignable to groups of two or three species. Sebastes borealis, S. crameri, and S. reedi were identified using morphological data. Combining genetic and morphological data allowed successful resolution of the other species as S. emphaeus, probably S. ciliatus (although S. polyspinis cannot be totally ruled out), and S. polyspinis. Many specimens were initially morphologically indistinguishable from S. alutus, and several morphological groups included fish genetically identified as S. alutus. This paper details the characteristics of these pelagic juveniles to facilitate morphological identification of these species in future collections. (PDF file contains 32 pages.)

Genetic improvement and effective dissemination: keys to prosperous and sustainable aquaculture industries

There is an increasing demand for fish in the world due to a growing population, better economic situation in some sectors, and greater awareness of health issues in relation to food. Since capture fisheries have stagnated, fish farming has become a very fast growing food production system. In this presentation, the author gives an overview of the technologies that are available for genetic improvement of fish, and briefly discuss their merit in the context of a sustainable development. He also discusses the essential prerequisites for effective dissemination of improved stock to farmers. It is concluded that genetic improvement programs based on selective breeding can substantially contribute to sustainable fish production systems. Furthermore, if such genetic improvement programs are followed up with effective dissemination strategies, they can result in a positive impact on farmers' incomes.

Potential applications of reproductive and molecular genetic technologies in the selective breeding of aquaculture species

The use of reproductive and genetic technologies can increase the efficiency of selective breeding programs for aquaculture species. Four technologies are considered, namely: marker-assisted selection, DNA fingerprinting, in-vitro fertilization, and cryopreservation. Marker-assisted selection can result in greater genetic gain, particularly for traits difficult or expensive to measure, than conventional selection methods, but its application is currently limited by lack of high density linkage maps and by the high cost of genotyping. DNA fingerprinting is most useful for genetic tagging and parentage verification. Both in-vitro fertilization and cryopreservation techniques can increase the accuracy of selection while controlling accumulation of inbreeding in long-term selection programs. Currently, the cost associated with the utilization of reproductive and genetic techniques is possibly the most important factor limiting their use in genetic improvement programs for aquatic species.

Manual for starch gel electrophoresis: A method for the detection of genetic variation

The procedure to conduct horizontal starch gel electrophoresis on enzymes is described in detail. Areas covered are (I) collection and storage of specimens, (2) preparation of tissues, (3) preparation of a starch gel, (4) application of enzyme extracts to a gel, (5) setting up a gel for electrophoresis, (6) slicing a gel, and (7) staining a gel. Recipes are also included for 47 enzyme stains and 3 selected gel buffers. (PDF file contains 26 pages.)

Genetic improvement and utilisation of indigenous tilapia in southern Africa: final technical report, December 1st 1998 to June 31st, 2002

Mozambique tilapia (Oreochromis mossambicus) is an indigenous tilapia species in southern Africa, until now the majority of genetic research has been carried out on Asian species of tilapia but this project aims to look at this African species. Those most suited to further development in aquaculture in southern Africa have now been identified. The genetic characterisation of strains has been completed. This information has aided the choice of strains for use in small scale aquaculture and for genetically male tilapia (GMT) production. They will form the basis of future strategies for further genetic improvement, and management of genetic diversity of Mozambique tilapia. The information will also contribute towards responsible management and development of genetic resources, particularly with regard to indigenous species of tilapia. Good progress has been made with the adaptation and implementation of producing the supermale fish required to produce all male offspring, resulting in faster growing populations of tilapia. The presence of the project and its associated activity has been a catalyst for a surge in interest in tilapia culture throughout southern Africa. [PDF contains 183 pages]

Genetic engineering of microorganisms: free release into the environment

Genetic engineering now makes possible the insertion of DNA from many organisms into other prokaryotic, eukaryotic and viral hosts. This technology has been used to construct a variety of such genetically engineered microorganisms (GEMs). The possibility of accidental or deliberate release of GEMs into the natural environment has recently raised much public concern. The prospect of deliberate release of these microorganisms has prompted an increased need to understand the processes of survival, expression, transfer and rearrangement of recombinant DNA molecules in microbial communities. The methodology which is being developed to investigate these processes will greatly enhance our ability to study microbial population ecology.

Genetic identification of four Malaysian mackerel species off Coast of Peninsular Malaysia based on molecular marker

Random Amplified Polymorphic DNA (RAPD) markers and cytochrome b (Cyt-b) gene sequences were utilized to fingerprint and construct phylogenetic relationships among four species of mackerel commonly found in the Straits of Malacca namely Rastrelliger kanagurta, R. brachysoma, Decapterus maruadsi and D. russelli. The UPGMA dendogram and genetic distance clearly showed that the individuals clustered into their own genus and species except for the Decapterus. These results were also supported by partial mtDNA cytochrome b gene sequences (279 bp) which found monotypic sequence for all Decapterus studied. Cytochrome b sequence phylogeny generated through Neighbor Joining (NJ) method was congruent with RAPD data. Results showed clear discrimination between both genera with average nucleotide divergence about 25.43%. This marker also demonstrated R. brachysoma and R. kanagurta as distinct species separated with average nucleotide divergence about 2.76%. However, based on BLAST analysis, this study indicated that the fish initially identified as D. maruadsi was actually D. russelli. The results highlighted the importance of genetic analysis for taxonomic validation, in addition to morphological traits.

Genetic differentiation between Atlantic salmon populations in the Windermere catchment

Genetic analysis, using single locus probes for genomic DNA, revealed that the juvenile Atlantic salmon populations in the Rivers Leven, Rothay and Troutbeck were related but genetically distinct. This genetic differentiation is greater than might be expected (by comparison with other salmon populations in the UK) and it is recommended that no action is taken which might promote genetic exchange between the three rivers. Thus, future fisheries management practices should treat the salmon from each site as separate genetic stocks. It is unlikely that any attempts to encourage fish currently spawning in the River Leven (downstream of Windermere) to utilize the upper catchment will be successful. The faster growth rate of juvenile salmon in the River Leven, compared with the River Rothay, probably results from a difference in temperature between the inflowing streams and the main outflow of Windermere. Precocious sexual maturation of some male parr was found in all three populations but the incidence (13-33%) is well within the range reported for other waters. Because of their enhanced growth rate, it is likely that some of the precocious males in the River Leven were 0+ fish. A very high incidence of hybridization (>18%) between Atlantic salmon and brown/sea trout was found in Troutbeck but not in the other rivers. Mitochondrial DNA analysis of these hybrids revealed them to be the product of several, independent cross-fertilizations involving both sexes of both species. The implications of this finding are discussed in relation to the availability of suitable spawning sites in Troutbeck.

Genetic variation between outer-coastal and fjord populations of Pacific herring (Clupea pallasii) in the eastern Gulf of Alaska

Pacific herring (Clupea pallasii) from the Gulf of Alaska were screened for temporal and spatial genetic variation with 15 microsatellite loci. Thirteen collections were examined in this study: 11 from Southeast Alaska and 2 from Prince William Sound, Alaska. Although FST values were low, a neighbor-joining tree based on genetic distance, homogeneity, and FST values revealed that collectively, the Berners Bay and Lynn Canal (interior) collections were genetically distinct from Sitka Sound and Prince of Wales Island (outer-coastal) collections. Temporal genetic variation within regions (among three years of Berners Bay spawners and between the two Sitka Sound spawners) was zero, whereas 0.05% was attributable to genetic variation between Berners Bay and Sitka Sound. This divergence may be attributable to environmental differences between interior archipelago waters and outer-coast habitats, such as differences in temperature and salinity. Early spring collections of nonspawning Lynn Canal herring were nearly genetically identical to collections of spawning herring in Berners Bay two months later—an indication that Berners Bay spawners over-winter in Lynn Canal. Southeast Alaskan herring (collectively) were significantly different from those in Prince William Sound. This study illustrates that adequate sample size is needed to detect variation in pelagic fish species with a large effective population size, and microsatellite markers may be useful in detecting low-level genetic divergence in Pacific herring in the Gulf of Alaska.

Genetic and morphological differences between Sebastes vulpes and S. zonatus (Teleostei: Scorpaeniformes: Scorpaenidae)

The taxonomic status of Sebastes vulpes and S. zonatus were clarified by comprehensive genetic (amplif ied fragment length polymorphisms [AFLP] and mitochondrial DNA [mtDNA] variation) and morphological analyses on a total of 65 specimens collected from a single locality. A principal coordinate analysis based on 364 AFLP loci separated the specimens completely into two genetically distinct groups that corresponded to S. vulpes and S. zonatus according to body coloration and that indicated that they are reproductively isolated species. Significant morphological differences were also evident between the two groups; 1) separation by principal component analysis based on 31 measurements, and 2)separation according to differences in counts of gill rakers and dorsal-fin spines without basal scales, and in the frequencies of specimens with small scales on the lower jaw. Restriction of gene flow between the two groups was also indicated by the pairwise ΦST values estimated from variations in partial sequences from the mtDNA control region, although the minimum spanning network did not result in separation into distinct clades. The latter was likely due to incomplete lineage sorting between S. vulpes and S. zonatus owing to their recent speciation.