The effect of feeding frequency on growth performance of juvenile cobia,Rachycentron canadum (Linnaeus, 1766)

The present study was carried out to investigate the number of daily feeding sessions that results in maximum growth of juvenile cobia under laboratory conditions. Groups of eight fish (110 g) were randomly distributed in twenty 500‐L tanks and hand‐fed a commercial diet for 60 days. The same amount of feed was offered daily, divided in 1, 2, 3, four or six meals. None of the parameters associated with growth performance (survival, weight gain, specific growth rate, feed intake, condition factor or size variation) showed any significant differences among treatments. Although under the present conditions feeding frequency had no effect on the growth performance of cobia larger than 110 g, in commercial farming operations where large numbers of fish are kept within a single rearing structure, fish may have aggressive interactions during feeding. Under these conditions, it is difficult to ensure that all cobia are fed to satiation and thus it is usual to provide two or more meals per day. The present results indicate that for an individual cobia the provision of more than one daily meal has no significant effect on growth performance.     

Gonad maturation and spawning of cobia,Rachycentron canadum (Linnaeus, 1766) off the Dungun coast,Malaysia

The reproductive biology of cobia, Rachycentron canadum, from the coastal waters of Dungun, Malaysia was studied from June 2014 to May 2015. From commercial trawls, a total of 201 samples (combined sexes) were collected (fork lengths [FL] 37.5–124.0 cm; body weights 0.5–20.4 g). The overall sex ratio of females to males was 1:0.9, which was not significantly different (χ² = 2.12, df = 1; p < .05). Estimations of length at 50% maturity (L₅₀) showed that both sexes matured at approximately 75 cm FL; estimated spawning frequency was 6 days. Mean batch fecundity (BF) ranged from 0.55 to 4.32 million eggs. The average number of eggs per gram of ovary was from 2,100 to 5,400 eggs, and relative fecundity 147 eggs/g. There was a weak positive correlation (r² = .48) between BF and female FL as well as BF with an ovary‐free body weight (r² = .56), possibly due to females being in a continuous spawning condition and some possibly half‐spent, based on the histological examination of the female gonads. Despite cobia being asynchronous spawners, the gonadosomatic index in both males and females showed peaks in June, November, and particularly March. Based on histological examination, spawning‐capable males were encountered throughout the study period, whereas spawning‐capable females in the late developing subphase were found mostly in March and April. This is the first study on the reproductive aspects of cobia in Malaysian waters.     

Microsatellite DNA markers for population genetic studies and parentage assignment in cobia,Rachycentron canadum

Twenty nuclear‐encoded microsatellites from a genomic DNA library of cobia, Rachycentron canadum, were isolated and characterized. The microsatellites include two tetranucleotide, one trinucleotide, three combination tetranucleotide/dinucleotide, nine dinucleotide, and five imperfect (dinucleotide) repeat motifs. Gene diversity ranged between zero to 0.910; the number of alleles among a sample of 24 fish ranged from one to 15. Cobia support an important recreational fishery in the southeastern United States and recently have become of interest to aquaculture. The microsatellites developed will be useful tools for studying both population genetics (e.g. stock structure, effective population size) and inheritance of traits important to aquaculture.     

Ontogeny of the gastrointestinal tract and selected digestive enzymes in cobia Rachycentron canadum (L.)

The ontogeny of the digestive system of cobia Rachycentron canadum from hatching to 22 days post-hatch (dph) (20·1 mm standard length) was examined with light microscopy. The activities of selected pancreatic enzymes were also determined during this period in order to optimize current rearing methods for this species. At hatching (3·6 mm), the digestive tract consisted of a relatively undifferentiated, straight tube positioned dorsally to the yolk sac. The major morphological changes in the digestive tract primarily occurred over the first 1–4 dph (3·6–4·4 mm). During this time, larvae began exogenous feeding (3 dph) and the digestive tract differentiated into five histologically distinct regions: buccopharynx, oesophagus, stomach anlage, anterior intestine and posterior intestine. Yolk reserves were exhausted by 5 dph (4·5 mm) and the oil globule began rapidly decreasing in size disappearing entirely by 9–10 dph (6·3–6·8 mm). Gastric glands differentiated at this time, and by 12 dph (8·1 mm) surface mucous cells of the stomach anlage stained positive for neutral mucosubstances. By 16 dph (11·6 mm), the blind sac (fundic region) of the stomach formed as did the pyloric caecae which initially appeared as a single protrusion of the anterior intestine just ventral to the pyloric sphincter. Generally, enzyme activities (U larva⁻¹) for amylase (0·0–1·8), chymotrypsin (0·0–7902·4), trypsin (0·2–16·6) and lipase (9·3–1319·0) were measurable at or soon after hatching and increased steadily from c. 8–22 dph (5·7–20·1 mm). The results of this study are discussed in terms of current and future weaning practices of this species.     

Identification and expression analysis of cobia (Rachycentron canadum) Toll-like receptor 9 gene

Cobia culture is hindered by bacterial infection (Photobacterium damselae subsp. piscicida) and in order to study the effect of P. damselae subsp. piscicida challenge and CpG ODN stimulation on cobia Toll like receptor 9 (RCTLR9), we used PCR to clone RCTLR9 gene and qRT-PCR to quantify gene expression. The results indicated that RCTLR9 cDNA contains 3141 bp. It encodes 1047 amino acids containing 16 typical structures of leucine-rich repeats (LRRs) including an LRRTYP, LRRCT and a motif involved in PAMP binding was identified at position 240–253 amino acid. Broad expression of RCTLR9 was found in larval, juvenile and adult stages irrespective of the tissues. In larval stage, RCTLR9 mRNA expression decreased at 5 d and then increased at 10 dph. At juvenile stage cobia, the expression was significantly high (p < 0.05) in spleen and intestine compared to gill, kidney, liver and skin. However, at adult stage, the significant high expression was found in gill and intestine. Cobia challenged with P. damselae subsp. piscicida showed significant increase in RCTLR9 expression at 24 h post challenge in intestine, spleen and liver, while in kidney the expression was peak at 12 h and later it decreased at 24 h. The highest expression was 40 fold increase in spleen and the lowest expression was ∼3.6 fold increase in liver. Cobia stimulated with CpG oligonucleotides showed that the induction of these genes was CpG ODN type and time dependent. In spleen and liver, CpG ODNs 1668 and 2006 injected group showed high expression of RCTLR9, IL-1β, chemokine CC compared to other groups. Meanwhile, CpG ODN 2006 has induced high expression of IgM. The CpG ODNs 2395 have induced significant high expression of Mx in spleen and liver. These results demonstrates the potential of using CpG ODN to enhance cobia resistance to P. damselae subsp. piscicida infection and use as an adjuvant in vaccine development.     

盐度对军曹鱼胚胎和仔鱼发育的影响

观察比较了不同盐度梯度(20、23、26、29、32、35、38、41和44)下军曹鱼受精卵的沉浮性、孵化率和畸形率以及测定了不同盐度梯度(10、14、18、22、26、30、34、38、42和44)下仔鱼不投饵存活系数SAI值。结果表明,受精卵在海水盐度32以上为完全浮性,26以下为完全沉性。军曹鱼受精卵孵化的最适盐度范围为29～38,适宜盐度范围为26～41,其中23～26和41～44分别视为受精卵孵化在低盐区和高盐区的两个临界区域,在此盐度上下,军曹鱼受精卵孵化率大幅度下降而仔鱼畸形率大幅度上升。试验盐度范围内军曹鱼仔鱼SAI值为2.32～16.24,仔鱼生长和存活的最适盐度范围为26～34,适宜盐度范围为22～38,而盐度18～22及38～42可分别视为军曹鱼仔鱼存活率在低盐区和高盐区的两个临界区域。     

Elevated oxygen uptake and high rates of nitrogen excretion in early life stages of the cobia Rachycentron canadum (L.), a fast-growing subtropical fish

Physiological energetics of cobia Rachycentron canadum were quantified for 18 to 82 days post-hatch (dph) hatchery-reared juveniles to better understand energy transformation and its implications in growth and survival. Mean oxygen consumption rates ( ; mg O₂ h⁻¹) of fish fed ad libitum and fish that were starved significantly increased with increasing wet mass (M; g), = 1·4291 M ^0·8119 and = 1·1784 M ^0·7833, respectively, with a significant reduction in mean metabolic rates of starved fish (19 to 27% specific dynamic action; SDA). Total ammonia nitrogen excretion rates ( A _MM, μmol h⁻¹) also scaled with M and significantly decreased after starvation. Mean mass-specific A _MM and urea excretion rates are the highest reported in the literature, with urea accounting for approximately half the total nitrogen excretion measured in both fed and starved fish. Relatively high energetic rates may allow cobia to develop rapidly into pre-juveniles and be less susceptible to predation and starvation at a comparatively early age.     

Whole-genome sequence diversity and association analysis of 198 soybean accessions in mini-core collections

We performed whole-genome Illumina resequencing of 198 accessions to examine the genetic diversity and facilitate the use of soybean genetic resources and identified 10 million single nucleotide polymorphisms and 2.8 million small indels. Furthermore, PacBio resequencing of 10 accessions was performed, and a total of 2,033 structure variants were identified. Genetic diversity and structure analysis congregated the 198 accessions into three subgroups (Primitive, World, and Japan) and showed the possibility of a long and relatively isolated history of cultivated soybean in Japan. Additionally, the skewed regional distribution of variants in the genome, such as higher structural variations on the R gene clusters in the Japan group, suggested the possibility of selective sweeps during domestication or breeding. A genome-wide association study identified both known and novel causal variants on the genes controlling the flowering period. Novel candidate causal variants were also found on genes related to the seed coat colour by aligning together with Illumina and PacBio reads. The genomic sequences and variants obtained in this study have immense potential to provide information for soybean breeding and genetic studies that may uncover novel alleles or genes involved in agronomically important traits.     

Chromosome characterization and biogeographic relations among three populations of the driftwood catfish Parauchenipterus galeatus (Linnaeus, 1766) (Siluriformes: Auchenipteridae) in Brazil

The present study employed basic and molecular cytogenetic methods to characterize three populations of Parauchenipterus galeatus from the basins of the Paraná and São Francisco Rivers, and a region of connection between the two basins. Although the diploid number was equal to 58 chromosomes, variations in karyotype formula were detected among the populations. B chromosomes were detected only in the population from the São Francisco River. Heterochromatin was located in the terminal position in almost all the chromosomes and in the pericentromeric position in some acrocentric chromosomes in the three populations. A single nucleolus organizer region was detected by silver nitrate and 18S rDNA‐fluorescent in situ hybridization in the short arm of one subtelocentric pair in the three populations, varying only in the chromosome pair bearing this site. The 5S rDNA sites were located in two submetacentric chromosome pairs in the three populations, varying only in the chromosome pairs bearing these sequences. Classic and molecular chromosome markers, along with the context of the natural history of the formation of hydrographic basins, ecological aspects, and the geographic isolation of populations between hydrographic basins and within the same basin, were important contributions to the discussion on possible biogeographic relations among the populations of Parauchenipterus galeatus. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 648–656.     

Comparative diversity analysis of halophiles at two polar saltern systems in Indramayu,West Java,Indonesia

Successive microbes in solar salt ponds are essential since it is well correlated with the quality of salt produced. This research aimed to analyse the microbial diversity of the solar salt ponds in Indonesia, which use high-density polyethylene in the ponds. There are two systems, that is, an integrated open system (In-system) and a closed system (Tt-system). The In-system uses seawater while the Tt-system uses seawater from the saline artesian well. Results showed that the In-system had richer microbes than the Tt-system. Both systems shared similar halophilic microbes profile. Ponds with low salinity (3–4 Be) had very low archaea, that is, 0·2 and 0·7% for the In-system and Tt-system respectively and were dominantly inhabited by phylum Proteobacteria. In the pond with high salinity, that is, 25 Be, both systems were dominated by the phylum Euryarchaeota, family Halobacteriaceae, and genera Halorubrum was dominantly found in In25 ponds and Tt25 ponds. Even though the two systems use the same parent seawater, that is, the Java Sea and share similar microbial composition at the phylum level, we found the dominance identified microbes in both systems were different.     

Isolation and characterization of polymorphic microsatellites in European chestnut (Castanea sativa Mill.)

Thirteen polymorphic microsatellite loci were isolated from Castanea sativa (Mill.). Six contained dinucleotide repeats, six contained trinucleotide repeats, and one contained a compound microsatellite of a trinucleotide and a tetranucleotide repeat. The loci were characterized using C. sativa trees from three populations in the UK and the parents and six seedlings from a Turkish mapping population. The number of alleles revealed varied from two to 14 (mean = 5.15) per loci. Eight loci were found to be useful in the mapping family.     

Identification and characterization of 14 polymorphic EST‐derived microsatellites in eelgrass (Zostera marina)

Zostera marina, the dominant seagrass on the Northern Hemisphere, forms the basis of important but threatened marine ecosystems. Here, we report 14 microsatellite DNA markers derived from an expressed sequence tag library corresponding to a wide range of genes. All loci were moderately to highly polymorphic, with allele numbers ranging from three to eight in a single Wadden Sea population of 48 individuals. Observed heterozygosities ranged from 0.082 to 0.837. Reaction conditions for five pooled polymerase chain reactions are given. The markers will advance the population genetics of seagrasses because they allow indirect tests of selection on closely linked genes.     

Development, characterization and inheritance of new microsatellites in olive (Olea europaea L.) and evaluation of their usefulness in cultivar identification and genetic relationship studies

Twelve new microsatellites have been developed in olive. For that purpose, a genomic library of the olive cultivar ‘Arbequina’ was enriched for GA, GT and ACT repeats. Two methods of screening yielded 27 sequences containing microsatellites out of the 119 clones sequenced. The GA repeat seems to be the most abundant motif. Among sequences containing microsatellites, 4 (14.8%) were redundant, 1 (3.7%) was previously described in the literature and 12 (44.4%) could not be used for primers design because the repeat motifs were incomplete. Suitable primer pairs were obtained for the remaining 10 (37.0%) sequences plus an additional 14 recovered from a formerly developed library. For the 24 primer pairs designed, 4 failed to amplify, 8 produced a complex bands pattern and 12 succeeded in giving amplification products. Considering these 12 primer pairs, 10 showed single locus amplification, whereas the other 2 revealed two loci each. This was demonstrated by studying allele segregation in two olive progenies. Sixty-eight alleles were detected for the 12 microsatellites when 51 olive cultivars were analysed. The number of alleles per locus ranged from 1 to 13. The expected heterozygosity varied between 0 and 0.83. All pairs of cultivars could be distinguished using only three microsatellites due to their great discrimination power value. The data coming from genotyping the 51 olive cultivars for 7 out of the 12 new microsatellites were used for constructing a dendrogram by unweighted pair group method with arithmetic mean cluster analysis using the Dice similarity coefficient. Cultivar association according to their geographical origin was observed.     

Carbonic anhydrase isoenzymes in nine troops of Kenya baboons, Papio cynocephalus (Linnaeus 1766)

The distributions of alleles at the carbonic anhydrase I (CA I = CA B) and carbonic anhydrase II (CA II = CA C) loci in nine troops of Papio cynocephalus were determined. Two alleles were found at the CA I locus, and three at the CA II locus; the frequencies were: CA I^a = 0.856; CA I^b = 0.144; CA II^a = 0.784; CA II^b = 0.209; CA II^c = 0.007. Results of tests for Hardy-Weinberg equilibrium, homogeneity tests, and calculations of migration rates were used in support of the interpretation that migration and genetic drift may affect the distribution of alleles at the CA I locus and that selection is the process responsible for the distribution of alleles at the CA II locus.