Tilapias are very important to the world's aquaculture. As befitting fish of their tropical origin, their distribution, and culture practices are highly affected by low temperatures. In this study, we used genetic and genomic methodologies to reveal pathways involved in the response and tolerance of blue tilapia ( Oreochromis aureus ) to low temperature stress. Cold tolerance was characterized in 66 families of blue tilapia. Fish from cold-tolerant and cold-sensitive families were sampled at 24 and 12°C, and the transcriptional responses to low-temperature exposure were measured in the gills and liver by high-throughput mRNA sequencing. Four hundred and ninety four genes displayed a similar temperature-dependent expression in both tolerant and sensitive fish and in the two tissues, representing the core molecular response to low temperature exposure. KEGG pathway analysis of these genes revealed down-regulation of focal-adhesion and other cell-extracellular matrix (ECM) interactions, and up-regulation of proteasome and various intra-cellular proteolytic activities. Differential responses between cold-tolerant and cold-sensitive fish were found with genes and pathways that were up-regulated in one group and down-regulated in the other. This reverse response was characterized by genes involved in metabolic pathways such as glycolysis/gluconeogenesis in the gills and biosynthesis of amino-acids in the liver, with low temperature down-regulation in tolerant fish and up-regulation in sensitive fish.

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In recent years, there has been increasing demand for red tilapia, which are commercial strains of hybrids of different tilapiine species or red variants of highly inbred Nile tilapia. However, red tilapia phenotypes are genetically unstable and affected by environmental factors, resulting in nonuniform coloration with black or dark-red color blotches that reduce their market value. Solute carrier family 45 member 2 (SLC45A2) is a membrane transporter that mediates melanin biosynthesis and is evolutionarily conserved from fish to humans. In the present study, we describe the generation of a stable and heritable red tilapia phenotype by inducing loss-of-function mutations in the slc45a2 gene. For this purpose, we identified the slc45a2 gene in Nile tilapia and designed highly specific guide RNAs (gRNA) for its genomic sequence. Multiplex microinjection of slc45a2-specific ribonucleoproteins to Nile tilapia zygotes induced up to 97-99% albinism, including loss of melanin in the eye. Next-generation sequencing of the injected zygotes demonstrated that all injected fish carried mutant alleles with variable mutagenesis efficiencies. Sanger sequencing of the genomic target region in the slc45a2 gene from fin clips, sperm, and F 1 offspring of a highly mutant male identified various genomic indels and germline transmission of the sperm-identified indels. Overall, this work demonstrates the generation of somatic and germline slc45a2 mutant alleles, which leads to complete albinism in Nile tilapia.

The process of learning to associate a visual cue with food was studied in groups of common carp Cyprinus carpio and Nile tilapia Oreochromis niloticus by means of a classical conditioning paradigm. In the first phase of the study, fishes were conditioned in either single-species or mixed-species groups to associate a blinking blue light with food. In the second phase, group composition was altered so that only single species were reconditioned, composed of individuals previously paired with the same or the other species. In the first phase, in the single-species groups, Nile tilapia rapidly formed an association between the visual cue and food, whereas common carp failed to do so. In mixed-species groups, both Nile tilapia and common carp associated a visual cue with food. In the second phase of the study, Nile tilapia retained that ability irrespective of whether they had previously been trained in single-species or mixedspecies groups, common carp previously trained with Nile tilapia, were not repelled by the blinking blue light and gradually approached it from a distance, whereas common carp previously trained in homogenous groups continued to be repelled by the blinking light. Fish arrivals and departures from the feeding zones as well as fish position in the testing apparatus were analysed and contrasted in single-species and mixed-species groups outside the training sessions, revealing mutual impacts of one species on the other in mixed-species groups. The findings of this study demonstrated interspecific social facilitation of learning to associate a visual cue with food.