Growth hormone transgenesis and feed composition influence growth and protein and amino acid content in transgenic G3 mutiara catfish (Clarias gariepinus)

Abstract

Clarias gariepinus growth hormone (CgGH) transgenesis was previously used to develop a population of second-generation (G2) transgenic mutiara catfish (C. gariepinus). The third generation of these fish (1-month old fingerlings) had 2–3 times improvement in growth compared to non-transgenic fish in the commercial feed test for 6 weeks of rearing. We assessed the impact of CgGH transgene expression on growth and protein and amino acid content of the G3 generation of these transgenic mutiara catfish relative to non-transgenic catfish. Since variation in composition of feed mixes can affect protein and amino acid content of fish, we tested three mixtures of commercial feed and boiled tuna (Euthynnus affinis): feed A (50:50 feed to tuna), B (65:35), and C (80:20) to transgenic catfish. Feed A* (50:50) was fed as a control to non-transgenic catfish. Feed efficiency, including feed conversion ratio and protein use efficiency (i.e., protein retention and protein productive value), was assessed. Feed efficiency, protein content, and essential amino acid content in G3 transgenic catfish (feed A and B) were higher than in non-transgenic fish (feed A*). The latter were deficient in lysine and methionine. Transgenic catfish fed with feed C (80:20) showed lysine deficiency and lower growth than fish fed feeds A and B. Feed B (65:35) was the optimal feed mixture utilized; it increased growth, protein levels, and feed conversion efficiency in G3 catfish. The growth of transgenic fish was higher than non-transgenic fish when supported by feeding with balanced nutrients.

The rate of embryonic development and hatching in transgenic G3 mutiara catfish (Clarias gariepinus) eggs at room temperature

“…This phenomenon is similar to the growth rate of oocytes from G 2 transgenic Mutiara catfish eggs [3] , and in egg embryogenesis from female A indicates the rapid development of cleavage, morula, blastula, gastrula, organogenesis phases compared to B and C (Table 2). On the other hand, the development of G 3 embryos from female B and C was slower than A, which suspected to be related to the low level of CgGH expression in these eggs causing late oocyte growth [8] .…”

“…PCR programme settings as follows: 48°C for 20 min; 40 cycles of 95°C for 1 min, 95ºC for 10 s, 60°C for 30 s and 72°C 30 s; and 72°C for 5 min. Transgenic positive broodfish were indicated by the presence of a 600 bp band parallel to the CgGH gene band in the pCMV-CgGH plasmid (positive control) as the primer amplification product of GH-F (5'-ATGGCTCGAGTTTTGGTGCTGCT-3') and GH-R (5'-CTACAGAGTGCAGTTGGAATCCAGGG-3') [7,8] .…”

CgGH insertion functional domain analysis in transgenic G1 and G2 and G3 mutiara catfish (Clarias gariepinus) broodstock

“…Transgenesis of GH causes excessive fish growth that is several times higher than that of non-transgenic fish, having the potential to increase fish culture yields. The growth of transgenic mutiara catfish (inserted CgGH sequence, GenBank accession no.MN249238.1) reported by Buwono et al (2016;2019b;2021) was high (2-3 times that of non-transgenic fish) because of exogenous GH (CgGH) insertions at G 0 , G 1 , G 2 , and G 3 . This inserted transgene was successfully inherited in three generations through the reproduction of transgenic broodstock with CgGH transmission rates of 42.85% in G 1 , 50% in G 2 and 70% in G 3 (Buwono et al, 2021).…”

“…The growth of transgenic mutiara catfish (inserted CgGH sequence, GenBank accession no.MN249238.1) reported by Buwono et al (2016;2019b;2021) was high (2-3 times that of non-transgenic fish) because of exogenous GH (CgGH) insertions at G 0 , G 1 , G 2 , and G 3 . This inserted transgene was successfully inherited in three generations through the reproduction of transgenic broodstock with CgGH transmission rates of 42.85% in G 1 , 50% in G 2 and 70% in G 3 (Buwono et al, 2021). Transgene inserts integrated into the fish genome have been shown to be inheritance on to offspring through broodstock reproduction.…”

Sosialisasi Budidaya Ikan Lele Mutiara Padjadjaran Di Kelompok Peternak Lele Cileunyi

“…10, No. 4, Desember 20212021. Stabilitas pewarisan karakter pertumbuhan G0 sampai G3 pada ikan lele mutiara Padjadjaran (G4) ini menunjukkan bahwa GH eksogen (CgGH) diwariskan secara permanen pada setiap generasi.…”