Effect of feeding frequency and water salinization on early development of Pyrrhulina brevis, an Amazonian ornamental fish / Efeito da frequência alimentar e da salinização da água sobre o desenvolvimento inicial de Pyrrhulina brevis, um peixe ornamental da Amazônia

Pyrrhulina brevis is an endemic fish from the Amazon basin that is valued in the ornamental fish market. In larviculture, fish are very sensitive to stressors and have a high mortality rate. Salinized water may provide more energy to overcome stress and feeding frequency is an important strategy to improve productive performance. Therefore, the aim of this study was to evaluate the best feeding frequency and the best level of water salinization for larval development of P. brevis. A total of 360 post-larvae of P. brevis (5.26 ± 1.65 mg, 5.57 ± 0.68 mm) were randomly distributed in 36 aquariums (1 L) in a 3x4 factorial scheme, to evaluate three concentrations of salt in water (0, 1 and 2 g L-1) and four feeding frequencies (once, twice, three and four times a day). We offered 150 Artemia nauplii per day and per post-larvae during 15 days. The fish kept at 1 g L-1 salinized water, and those fed 3 and 4 times a day presented significantly greater final length and weight, weight gain, and specific growth rate for length and weight. An interaction between salinized water and feeding frequency was observed for survival rate. We conclude that, during the initial rearing phase of P. brevis, feeding frequency of three times a day and salinized water at 1 g L-1 is recommended.(AU)

Pyrrhulina brevis é um peixe endêmico da bacia amazônica que apresenta bons valores no mercado de peixes ornamentais. Na larvicultura, os peixes são muito sensíveis aos estressores e apresentam alta taxa de mortalidade. A água salinizada pode fornecer mais energia para superar o estresse e a frequência alimentar é uma estratégia importante para melhorar o desempenho produtivo. Portanto, o objetivo deste trabalho foi avaliar a melhor frequência alimentar e o melhor nível de salinização da água para o desenvolvimento larval de P. brevis. Um total de 360 pós-larvas de P. brevis (5,26 ± 1,65 mg, 5,57 ± 0,68 mm) foram distribuídos aleatoriamente em 36 aquários (1 L) em esquema fatorial 3x4, para avaliar três concentrações de sal na água (0, 1 e 2 g L-1) e quatro frequências de alimentação (uma, duas, três e quatro vezes ao dia). Foram oferecidas 150 náuplios de Artemia por dia e por pós-larva, durante 15 dias. Os peixes mantidos com 1 g L-1 de água salinizada e os alimentados 3 e 4 vezes ao dia apresentaram comprimento e peso finais, ganho de peso e taxa de crescimento específico para comprimento e peso significativamente maiores. Uma interação entre água salinizada e frequência alimentar foi observada para a taxa de sobrevivência. Concluímos que, durante a fase inicial de criação de P. brevis, recomenda-se uma frequência alimentar de três vezes ao dia e água salinizada a 1 g L-1.(AU)

The fatty acid elongation genes elovl4a and elovl4b are present and functional in the genome of tambaqui (Colossoma macropomum).

Long-chain (C20-24) polyunsaturated fatty acids (LC-PUFA) are physiologically important nutrients for vertebrates including fish. Previous studies have addressed the metabolism of LC-PUFA in the Amazonian teleost tambaqui (Colossoma macropomum), an emerging species in Brazilian aquaculture, showing that all the desaturase and elongase activities required to convert C18 polyunsaturated fatty acids (PUFA) into LC-PUFA are present in tambaqui. Yet, elongation of very long-chain fatty acid 4 (Elovl4) proteins, which participate in the biosynthesis of very long-chain (>C24) saturated fatty acids (VLC-SFA) and very long-chain polyunsaturated fatty acids (VLC-PUFA), had not been characterized in this species. Here, we investigate the repertoire and function of two Elovl4 in tambaqui. Furthermore, we present the first draft genome assembly from tambaqui, and demonstrated the usefulness of this resource in nutritional physiology studies by isolating one of the tambaqui elovl4 genes. Our results showed that, similarly to other teleost species, two elovl4 gene paralogs termed as elovl4a and elovl4b, are present in tambaqui. Tambaqui elovl4a and elovl4b have open reading frames (ORF) of 948 and 912 base pairs, encoding putative proteins of 315 and 303 amino acids, respectively. Functional characterization in yeast showed that both Elovl4 enzymes have activity toward all the PUFA substrates assayed (18:3n-3, 18:2n-6, 18:4n-3, 18:3n-6, 20:5n-3, 20:4n-6, 22:5n-3, 22:4n-6 and 22:6n-3), producing elongated products of up to C36. Moreover, both Elovl4 were able to elongate 22:5n-3 to 24:5n-3, a key elongation step required for the synthesis of docosahexaenoic acid via the Sprecher pathway.

A complete enzymatic capacity for long-chain polyunsaturated fatty acid biosynthesis is present in the Amazonian teleost tambaqui, Colossoma macropomum.

In vertebrates, the essential fatty acids (FA) that satisfy the dietary requirements for a given species depend upon its desaturation and elongation capabilities to convert the C18 polyunsaturated fatty acids (PUFA), namely linoleic acid (LA, 18:2n-6) and α-linolenic acid (ALA, 18:3n-3), into the biologically active long-chain (C20-24) polyunsaturated fatty acids (LC-PUFA), including arachidonic acid (ARA, 20:4n-6), eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3). Recent studies have established that tambaqui (Colossoma macropomum), an important aquaculture-produced species in Brazil, is a herbivorous fish that can fulfil its essential FA requirements with dietary provision C18 PUFA LA and ALA, although the molecular mechanisms underpinning such ability remained unclear. The present study aimed at cloning and functionally characterizing genes encoding key desaturase and elongase enzymes, namely fads2, elovl5 and elovl2, involved in the LC-PUFA biosynthetic pathways in tambaqui. First, a fads2-like desaturase was isolated from tambaqui. When expressed in yeast, the tambaqui Fads2 showed Δ6, Δ5 and Δ8 desaturase capacities within the same enzyme, enabling all desaturation reactions required for ARA, EPA and DHA biosynthesis. Moreover, tambaqui possesses two elongases that are bona fide orthologs of elovl5 and elovl2. Their functional characterization confirmed that they can operate towards a variety of PUFA substrates with chain lengths ranging from 18 to 22 carbons. Overall our results provide compelling evidence that demonstrates that all the desaturase and elongase activities required to convert LA and ALA into ARA, EPA and DHA are present in tambaqui within the three genes studied herein, i.e. fads2, elovl5 and elovl2.

Bti-based insecticide enhances the predatory abilities of the backswimmer Buenoa tarsalis (Hemiptera: Notonectidae).

The backswimmer Buenoa tarsalis (Hemiptera: Notonectidae) is a naturally occurring predator of immature stages of mosquitoes. These aquatic predators can suffer from non-targeted exposure to insecticides that are commonly used in aquatic environments to control mosquitoes. Here, we evaluated whether insecticide formulations containing the bacterium Bacillus thuringiensis var. israelensis (Bti) or the organophosphate pirimiphos-methyl would affect the survival and the predatory abilities of B. tarsalis. First, we conducted survival bioassays to estimate the median survival time (LT50) of B. tarsalis when exposed to Bti-based insecticide (at 0.25 and 25 mg a.i./L) and pirimiphos-methyl (at 1, 10 and 1000 mg a.i./L). The highest concentrations of the insecticides were equivalent to the label-recommended field rates. Second, the predatory abilities of B. tarsalis exposed to insecticides were evaluated at three prey densities (3, 6 and 9 mosquito larvae/100 mL water) just after insecticide exposure or after a 24 h recovery time. While the survival of B. tarsalis was significantly reduced with pirimiphos-methyl concentrations ≥10 mg a.i./L, the Bti-exposed predators exhibited similar survival as unexposed predators. Interestingly, after a recovery time of 24 h, B. tarsalis sublethally exposed to pirimiphos-methyl or Bti-based insecticide consistently killed more A. aegypti larvae (at the intermediate density) than unexposed predators. However, for the without-recovery bioassays, the pirimiphos-methyl-exposed predators exhibited reduced predatory abilities at the lowest prey density. Because they do not reduce the survival or the predatory abilities of B. tarsalis, Bti-based insecticides can be considered a safe insecticide to use in the presence of backswimmers.