Production of Homozygous Transgenic Rainbow Trout with Enhanced Disease Resistance

Abstract

Previous studies conducted in our laboratory showed that transgenic medaka expressing cecropin B transgenes exhibited resistant characteristic to fish bacterial pathogens, Pseudomonas fluorescens and Vibrio anguillarum. To confirm whether antimicrobial peptide gene will also exhibit anti-bacterial and anti-viral characteristics in aquaculture important fish species, we produced transgenic rainbow trout expressing cecropin P1 or a synthetic cecropin B analog, CF-17, transgene by sperm-mediated gene transfer method. About 30 % of fish recovered from electroporation were shown to carry the transgene as determined by polymerase chain reaction (PCR) amplification assay. Positive P1 transgenic fish were crossed to non-transgenic fish to establish F1 transgenic founder families, and subsequently generating F2, and F3 progeny. Expression of cecropin P1 and CF-17 transgenes was detected in transgenic fish by reverse transcription (RT)-PCR analysis. The distribution of body sizes among F1 transgenic fish were not significantly different from those of non-transgenic fish. Results of challenge studies revealed that many families of F2 and F3 transgenic fish exhibited resistance to infection by Aeromonas salmonicida and infectious hematopoietic necrosis virus (IHNV). All-male homozygous cecropin P1 transgenic families were produced by androgenesis from sperm of F3 heterozygous transgenic fish in one generation. The resistant characteristic to A. salmonicida was confirmed in progeny derived from the outcross of all-male fish to non-transgenic females. Results of our current studies confirmed the possibility of producing disease-resistant homozygous rainbow trout strains by transgenesis of cecropin P1 or CF-17 gene and followed by androgenesis.

Effects of Cecropin Transgenesis and Interspecific Hybridization on the Resistance to Ichthyophthirius multifiliis in Channel Catfish and Female Channel Catfish × Male Blue Catfish Hybrids

“…Recently, cecropin P1 and CF‐17 (which contains a synthetic cecropin B analog) were found to enhance the resistance to Aeromonas salmonicida and IHNV in both F 2 and F 3 transgenic Rainbow Trout (Chiou et al. ).…”

“…Homozygous cecropin P1 and CF‐17 transgenic Rainbow Trout Oncorhynchus mykiss exhibited improved resistance to Aeromonas salmonicida and infectious hematopoietic necrosis virus (IHNV; Chiou et al. ). Cecropins inhibited the replication of human immunodeficiency virus 1 by suppressing viral gene expression (Wachinger et al.…”

Transcriptomic responses to yam ( Dioscorea oppositifolia L .) extract dietary supplementation in rainbow trout ( Oncorhynchus mykiss ) liver

“…Rainbow trout is an important economical cold‐water fish. In the global aquaculture industry, parasitic, bacterial and viral diseases cause huge economic losses every year (Chiou et al, ). Yam was determined to exert immune response‐modifying effects (Amat et al, ; Jeon et al, ; Kong et al, ).…”

Disease reduction in aquaculture with genetic and genomic technology: current and future approaches

“…In fish, resistance to infection by A. salmonicida and infectious haematopoietic necrosis virus (IHNV) has been improved in homozygous cecropin P1‐ and CF‐17‐transgenic rainbow trout (Chiou et al . ). Cecropins also exhibit strong antifungal activity (Ekengren & Hultmark ; Ji et al .…”

“…For example, cecropins inhibited the replication of human immunodeficiency virus 1 (HIV-1) by suppressing viral gene expression (Wachinger et al 1998). In fish, resistance to infection by A. salmonicida and infectious haematopoietic necrosis virus (IHNV) has been improved in homozygous cecropin P1-and CF-17-transgenic rainbow trout (Chiou et al 2014). Cecropins also exhibit strong antifungal activity (Ekengren & Hultmark 1999;Ji et al 2014) against fungi including Aspergillus, Fusarium (De Lucca et al 1998 and Candida species (Park et al 1997) in vitro.…”