Protective effects of dietary selenium on abalone Haliotis discus hannai against the toxicity of waterborne cadmium

This study was conducted to investigate protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of cadmium (Cd). A 60‐day feeding trial was conducted with abalone (initial weight: 3.17 ± 0.01 g), which were exposed to 0.34 mg/L of waterborne Cd. During a feeding trial, abalone were fed graded levels of Se at 0.10 (controls), 1.31, and 4.20 mg/kg diet respectively. Results showed that there was no significant difference in specific growth rate and survival rate of abalone among the three treatments. Compared with the controls, dietary Se significantly decreased Cd concentrations in serum, muscle, mantle, gill, mantle, and hepatopancreas of abalone. Besides, compared with the controls, dietary Se significantly increased metallothionein concentration in the hepatopancreas of abalone. Additionally, compared with the controls, dietary Se significantly decreased concentrations of malondiadehyde and protein carbonyl in hepatopancreas of abalone. Meanwhile, compared with the controls, dietary Se significantly increased activities of glutathione peroxidase, thioredoxin reductase, and thioredoxin peroxidase, and concentration of glutathione in the hepatopancreas of abalone. Based on the data above, in abalone, dietary Se showed protective effects against waterborne Cd.     

Effects of dietary α‐lipoic acid on the growth and antioxidative responses of juvenile abalone Haliotis discus hannai Ino

A feeding experiment was conducted to determine the effects of α‐lipoic acid (LA) on the growth and antioxidative responses of juvenile abalone Haliotis discus hannai Ino. Six purified diets supplemented with 0, 200, 400, 800, 1600 and 3200 mg kg^?1 of LA, respectively, were fed to abalone for 16 weeks in a flow‐through water system. The results showed that the weight gain ratio (WGR) increased with the dietary LA levels, and reached the highest value in the group with 800 mg kg^?1 dietary LA supplement. Glutathione peroxidase activity in the 800 mg kg^?1 dietary LA group was significantly higher than that in the dietary LA‐deficient (0 mg kg^?1) group. Superoxide dismutase activities in the 200, 400 and 800 mg kg^?1 groups were significantly increased. Supplements of 200, 400 and 800 mg kg^?1 dietary LA elevated the total antioxidative capacity significantly. The glutathione level in the hepatopancreas increased significantly with the dietary LA supplements in a dose‐dependent manner (except for the 200 mg kg^?1 group). Catalase and malondialdehyde in the hepatopancreas were not significantly affected by dietary LA. In conclusion, dietary LA promoted the growth and stimulated the antioxidative defence capacity of abalone. Based on the data of WGR, the optimal dietary LA supplement for juvenile abalone was found to be 709 mg kg^?1 using piece‐wise linear analysis.     

Dose‐dependent protective effects of dietary selenium on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper

This study was designed to assess the protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper (Cu). A 60‐day feeding trial was conducted in a static water system for abalone (initial weight: 3.17 ± 0.01 g) exposed to 0.02 mg L^?1 of waterborne Cu. The animals were fed one of the three experimental diets with 0.10, 1.31 and 4.20 mg kg^?1 of Se from Na₂SeO₃·5H₂O respectively. Results showed that the abalone fed 1.31 mg kg^?1 of dietary Se had the lowest Cu concentration in shell, muscle, mantle, gill, hepatopancreas and serum. Meanwhile, the significant lowest contents of malondiadehyde and protein carbonyl in hepatopancreas were also found in the treatment with 1.31 mg kg^?1 of dietary Se (P < 0.05). In addition, this treatment had significant higher glutathione content and thioredoxin reductase activity in hepatopancreas (P < 0.05). However, the activity of Se‐dependent glutathione peroxidase (Se‐GPx) was significantly decreased in the treatment with 4.20 mg kg^?1 of dietary Se (P < 0.05). In this treatment, the protein carbonyl content in hepatopancreas was significantly higher than that in the group with 1.31 mg kg^?1 of dietary Se (P < 0.05). In conclusion, in terms of anti‐oxidation and Cu accumulation, the protective effects of dietary Se on abalone against waterborne Cu were dose‐dependent. The 1.31 mg kg^?1 of dietary Se had this effect, but not 4.20 mg kg^?1 of dietary Se. Moreover, the latter increased the oxidative stress in abalone exposed to the waterborne Cu.     

Ability of salmon growth hormone to accelerate somatic growth of juvenil abalone Haliotis discus hannai

ABSTRACT:   The ability of salmon growth hormone (sGH) to accelerate the somatic growth of juvenile abalone Haliotis discus hannai by immersion was examined. After immersion for one hour into a sGH-rich solution at a concentration of 30 mg/L, the sGH immunoreactivity in the body fluid of abalone was maximal after one day, and levels were still detectable at two days. No immunoreactivity was observed in the control group. Following immersion for one hour at one or two-week intervals for 120 weeks into the sGH-rich solution at 30 mg/L, the sGH-immersed abalone exhibited a significant increase in shell length and body weight. On the other hand, abalone immersed into a salmon prolactin (sPRL)-rich solution at 30 mg/L for one hour showed a lower increase in both shell length and body weight than the sGH-immersed abalone. No increases in shell length and body weight were observed in either the bovine serum albumin-immersed abalone or untreated controls. These results provide evidence that sGH can be transported from ambient water into the circulatory system of abalone, and can subsequently improve the somatic growth of juveniles.     

Effects of Cu,Zn and Cd on larval development and respiratory metabolism of pacific abalone (Haliotis discus hannai)

Trochophore and veliger are vulnerable pelagic larvae and susceptible to environmental toxicants, which are extremely crucial for the population renewal. Cu, Zn and Cd pollution are common elements in metal pollution in coastal areas. Though Pacific abalone is the main economic mollusc in China, its larval response to metals remains unclear. To understand this, the effect of Cu, Zn and Cd toxicity on malformation classification and respiratory metabolism of trochophore and veliger are analysed and measured in this paper. The 48 hr EC₅₀ for abalone trochophore was (18.9 ± 1.0) μg/L for Cu, (65.1 ± 0.3) μg/L for Zn and (1,377.8 ± 68.9) μg/L for Cd respectively. The LC₅₀ of veliger was (45.8 ± 0.9) μg/L for Cu, (50.0 ± 0.9) μg/L for Zn and (1,049.3 ± 37.6) μg/L for Cd respectively. Trochophore died with no shell at 128 μg/L Cu, yet remained alive with multiple broken‐shell bodies at 128 μg/L Zn and failed to form well‐developed shells at 20,480 μg/L Cd. All veliger died with complete shells at the mentioned concentrations. A self‐made device was used to study the effect on respiratory metabolism. The respiratory metabolism level was significantly fluctuating under Cu and Zn exposure and declined under Cd. The results indicated a decreasing order of the metal toxicity to trochophore and veliger as follows: Cu > Zn ? Cd. Cu and Zn were the main causes of shell abnormality, and Cd was less toxic. Moreover, the affected respiratory has a negative impact on further larval development.     

Inbreeding depression traits in pacific abalone Haliotis discus hannai by factorial mating experiments

ABSTRACT:   'Inbreeding depression' may be an avoidable phenomenon for abalone culture. However, only a few studies have been carried out on inbreeding depression. In the present study, using six families produced in 1994, a factorial mating system including inbreeding and outbreeding was constructed in order to demonstrate inbreeding depression traits of the Pacific abalone. In total, 24 inbreeding and 21 outbreeding crosses were produced during three years (1999–2001) and these offspring were reared for approximately one year. Significant differences in fertilization rate and growth were not observed between inbreeding and outbreeding crosses. However, the deformity rate of veliger larvae was always higher in inbreeding crosses than that of outbreeding crosses in all experiments. Moreover, a significantly high deformity rate was observed in some full-sib families of inbreeding. Alternatively, the survival rates of inbreeding crosses were much lower than for outbreeding crosses after about 4 months and one year in two rearing localities. These results indicate that inbreeding depression is observed in the traits of deformity rate and survival, but not in fertilization rate nor growth in the first generation of a full-sib family of the Pacific abalone.     

Mining expressed sequences for single nucleotide polymorphisms in Pacific abalone Haliotis discus hannai

Although single nucleotide polymorphisms (SNPs) are important resources for population genetics, pedigree analysis and genomic mapping, such loci have not been reported in Pacific abalone so far. In this study, a bioinformatics strategy was adopted to discover SNPs within the expressed sequences (ESTs) of Pacific abalone, Haliotis discus hannai , and furthermore, polymerase chain reaction direct sequencing (PCR-DS) and allele-specific PCR (AS-PCR) were used for SNPs detection and genotype scoring respectively. A total of 5893 ESTs were assembled and 302 putative SNPs were identified. The average density of SNPs in ESTs was 1%. Fifty-two sets of sequencing primers were designed from SNPs flanking ESTs to amplify the genomic DNA, and 13 could generate products of expected size. Polymerase chain reaction direct sequencing of the amplification products from pooled DNA samples revealed 40 polymorphic SNP loci. Using a modified tetra-primer AS-PCR, seven mitochondrial and six nuclear SNPs were typed and characterized among 37 wild abalones. In conclusion, it is feasible to discover SNPs from number limited ESTs and the AS-PCR as a simple, robust and reliable assay could be a primary method for small- and medium-scale SNPs detection in abalones as well as other non-model organisms.     

Effects of artificial selection practices on loss of genetic diversity in the Pacific abalone,Haliotis discus hannai

The Pacific abalone, Haliotis discus hannai Ino, is one of most important aquaculture species in China. Artificial selection has been the common and inevitable approach in breeding. In present study, the effect of artificial selection on the genetic structure of two abalone lines (JJ selection strain, three successive selection lines based on the fast‐growing trait, which was named JJF0, JJF1, JJF2, and JJF3; and R selection strain, the offspring of the red shell colored variants) were evaluated using 10 microsatellites. Loci showed from low to high polymorphism, with the number of alleles (A) ranging from 2 to 18 in each population. The mean observed (H_o) and expected heterozygosities (H_e) were 0.650 ± 0.022 and 0.711 ± 0.018, respectively. In selection strain JJ, the values for most diversity genetic indexes (A_e, the number of effective alleles, H_o and H_e) decreased from JJF0 to JJF3. Meanwhile, compared to JJ line, the genetic diversity estimates of R were close to those of JJF3. Nei's genetic distance ranged from 0.20357 to 0.51346. The unweighted pair group method with arithmetic mean tree based on Nei's genetic distance also showed that the control Japan population and JJF0 formed to a cluster firstly, which were subsequently grouped together with JJF1, JJF2, JJF3, whereas the R was isolated from the rest of the populations. Analysis of genetic information indicated that genetic diversity was lost with artificial selection practices. Mechanisms underlying the maintenance of an acceptable level of genetic diversity while pursuing economic interests should be conducted in future research studies.     

Suitability of cage culture for Pacific abalone Haliotis discus hannai Ino production in China

Pacific abalone (Haliotis discus hannai Ino) aquaculture is a thriving industry in China. This study describes a novel submerged cage culture system for abalone rearing in Fujian, South China. The cage consisted of five vertical slots that were oriented perpendicular to the flow of water. The slots were separated by six vertically connected plastic plates for abalone attachment and shelter at the bottom of the cage. Experiment 1 was designed to determine the appropriate stocking density at the start of the abalone sea‐based production cycle. Eight‐month‐old hatchery reared and size‐graded juveniles were transferred to the sea‐based culture system. For different stocking densities, shell length of juveniles obtained in this novel culture system on 2, 3.5 and 5 months, respectively, was compared with shell lengths obtained in a traditionally multi‐tier basket culture system. In Experiment 2, daily growth rates (DGRs) in shell length and biomass in terms of wet weight of 2‐year‐old abalones reared in cage and tiered basket culture systems were compared over a 6‐month period. Results of Experiment 1 showed that growth of abalone in the cage culture system is density‐dependent; the mean final shell length of juveniles obtained was 6.7–15.9% higher than in tiered baskets system even at the same initial stocking density. In Experiment 2, DGRs in shell length of 53.83–78.38 μm day^?1 obtained in cage system were significantly higher than that in tiered baskets (P < 0.01). And in terms of wet weight biomass, it was 1.48–3.01 times higher in the cage system compared with the traditional system. Abalone survival was more than 87.5% in both culture systems in both experiments. Advantages of the newly established cage culture system included better growth performance of the animals reared and potential improvement of rearing conditions, such as improved water flow velocity and dissolved oxygen.     

Growth performance of disk abalone Haliotis discus hannai in pilot- and commercial-scale recirculating aquaculture systems

The study investigated the growth performance of abalone from juvenile to marketable size in a commercial-scale recirculating aquaculture system. The rearing system consisted of 12 raceways (4.0 × 0.8 × 0.6 m) with a protein skimmer and a submerged biofilter for juveniles and 10 raceways (6.6 × 1.3 × 0.6 m) with a protein skimmer and a trickling biofilter for on-growing. Sea mustard (Undaria pinnatifida) and kelp (Laminaria japonica) were fed to the abalone. The total weight of abalone in the recirculating aquaculture system at the juvenile stage increased from 22.0 kg (average shell length 24.5 mm) to 75.5 kg (average shell length 42.5 mm) after 180 days. Feed conversion ratios increased slightly from 13.7 for the first 90 days to 16.3 thereafter. The shell growth rate of juvenile abalone between 24.5 mm and 34.8 mm was 3.4 mm month⁻¹, while for juveniles between 34.8 mm and 42.5 mm it was 2.6 mm month⁻¹. The total weight of abalone in the recirculating aquaculture system for the on-growing stage increased from 100.0 kg (average shell length 44.0 mm) to 433.3 kg (average shell length 72.7 mm) after 570 days. The feed conversion ratios for the first 173 days, the next 320 days, and the last 570 days were 19.6, 22.1, and 24.8, respectively. The growth rate of the average shell length during the on-growing period was 1.5 mm month⁻¹. Total ammonia nitrogen (TAN) concentrations were stabilized below 0.12 mg l⁻¹ in the juvenile recirculating system and 0.14 mg l⁻¹ in the on-growing recirculating system after conditioning of the biofilters.     

Identification of quantitative trait loci for growth-related traits in the Pacific abalone Haliotis discus hannai Ino

The locations and effects of quantitative trait loci (QTL) were estimated for nine characters for growth‐related traits in the Pacific abalone (Haliotis discus hannai Ino) using a randomly amplified polymorphic DNA (RAPD), amplification fragment length polymorphism (AFLP) and SSR genetic linkage map. Twenty‐eight putatively significant QTLs (LOD>2.4) were detected for nine traits (shell length, shell width, total weight, shell weight, weight of soft part, muscle weight, gonad and digestive gland weight, mantle weight and gill weight). The percentage of phenotypic variation explained by a single QTL ranged from 8.0% to 35.9%. The significant correlations (P<0.001) were found among all the growth‐related traits, and Pearson's correlation coefficients were more than 0.81. For the female map, the QTL for growth were concentrated on groups 1 and 4 linkage maps. On the male map, the QTL that influenced growth‐related traits gathered on the groups 1 and 9 linkage maps. Genetic linkage map construction and QTL analysis for growth‐related traits are the basis for the marker‐assisted selection and will eventually improve production and quality of the Pacific abalone.     

Influences of dietary antimicrobial peptide APSH‐07 on the growth performance,immune response and vibriosis resistance of abalone Haliotis discus hannai Ino

A 115‐day feeding trial and subsequently a 10‐day challenge test with Vibrio parahaemolyticus were conducted to investigate the effects of dietary antimicrobial peptide APSH‐07 on growth performance, immune response, antioxidative status and vibriosis resistance of abalone Haliotis discus hannai (initial body weight: 2.06 ± 0.01 g; initial shell length: 25.42 ± 0.18 mm). Four artificial diets were designed with 0 (artificial diet control), 7.5, 15.0 and 22.5 mg/kg of APSH‐07, respectively. The brown alga Laminaria japonica was used as the live food control. Results showed that the specific growth rates of abalone in the groups with 7.5 and 15.0 mg/kg dietary APSH‐07 were significantly higher than those in the controls (p < .05). The total haemocyte counts and respiratory burst activity in haemolymph of abalone in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the groups with 0 and 22.5 mg/kg of dietary APSH‐07 (p < .05). The gene expression levels of Mn‐superoxide dismutase and thioredoxin peroxidase 2, and the activities of glutathione peroxidase in the group with 7.5 mg/kg of dietary APSH‐07 were significantly higher than those in the other groups (p < .05). Cumulative mortality of abalone after the challenge test was significantly decreased in the group with 7.5 mg/kg of dietary APSH‐07 supplementation. Supplementation of 22.5 mg/kg dietary APSH‐07 significantly increased the cumulative mortality. In conclusion, 7.5 mg/kg of dietary APSH‐07 supplementation had the better growth performance, higher antioxidation, immune and disease resistance capacity of abalone. Excessive supplementation of dietary antimicrobial peptide APSH‐07 (22.5 mg/kg) had significantly negative effects. Further studies are needed to determine the optimal level of dietary APSH‐07 supplementation for abalone.     

Genetic variability revealed with microsatellite markers in an introduced population of the abalone Haliotis discus hannai Ino

One of the challenges for the culture of any species is to control the loss of genetic variability, which may result in a decrease in the quality of commercially important traits. The goal of this study is to assess the genetic diversity of a hatchery population of the Pacific abalone ( Haliotis discus hannai ) from the Center for Abalone Production of the Universidad Católica del Norte (CAP-UCN) that is maintained under a breeding programme. We used six polymorphic microsatellite markers within the cultivated population. The loci Awb033 and Awb079 had the highest number of alleles (11 and 10 respectively) and the loci Awb022 and Awb026 the lowest (two and four respectively). The mean number of alleles per locus was 6.83. The average observed and expected heterozygosities were 0.71 and 0.70, respectively, and the average F _IS ( f ) index was −0.023. We compared the population genetic parameters of the CAP-UCN population with previously published data of wild and hatchery populations of the same species. Results indicate lower genetic diversity estimated as allelic richness in the introduced population with a loss of 11–58% alleles per locus. Despite the high allelic loss, the estimated inbreeding coefficient suggests that the breeding programme carried out in the CAP-UCN has controlled and maintained heterozygosity levels successfully. A temporal study is necessary to determine whether the genetic diversity loss detected was caused during the initial introduction of breeders or to the breeding programme actually implemented.     

Effects of dietary guaiacol on shell biomineralization of juvenile abalone Haliotis discus hannai, Ino

A study was conducted to investigate the effects of guaiacol on shell biomineralization of abalone Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol (0, 5, 10, 50, 100, 500 and 1000 mg kg⁻¹). Abalone (initial weight: 0.29±0.01 g; initial shell length: 8.55±0.27 mm) were fed with these diets in a re-circulated water system for 152 days. The results showed that the contents of calcite in the shell increased significantly with dietary guaiacol levels, but the aragonite content decreased significantly. Dietary guaiacol significantly decreased the contents of calcium and strontium, but significantly increased the magnesium content in the shell. The effects of dietary guaiacol on the contents of phosphorus and zinc in shell were not significant. It was obvious that the ratio of potential acidic/basic amino acids [(Asx+Glx)/(Lys+Arg+His)] in the shell was elevated by dietary guaiacol. In conclusion, dietary guaiacol has effects on both the mineralogy and the amino acid composition in the shell. Furthermore, changes in mineralogy are related to the ratio of (Asx+Glx)/(Lys+Arg+His) in the shell.     

Radula development in abalone Haliotis discus hannai from larva to adult in relation to feeding transitions

ABSTRACT: The radula morphology of Haliotis discus hannai was examined by scanning electron microscope from the larval to the adult stage. The radula of competent larvae contained 11–13 transverse rows of teeth after 6–7 days at 20°C. The number of rows increased to 25–30 during the first several days after settlement, but then remained approximately constant throughout the post-larval period, increasing again in abalone larger than 4 mm in shell length (SL). In post-larvae < ~1 mm SL, only two pairs of lateral teeth (L1, L2) were present in the larval radula. An additional three pairs of lateral teeth (L3–L5) were added progressively as post-larvae grew from 0.9 mm to 1.9 mm SL. Marginal teeth were added steadily from one pair in larvae to 30–40 pairs at 3–4 mm SL, 70–80 pairs in 30–40 mm juveniles, and 70–90 pairs in 90–100 mm adults. The serrations on the working edges of the rachidian (R) and lateral teeth became less pronounced as the abalone grew. Nearly all serrations disappeared from the rachidian (R) and inner lateral teeth (L1, L2) by ~2 mm SL, and from the outer lateral teeth (L3–L5) by 20 mm SL. For abalone larger than 1.5 mm SL, the L3–L5 teeth became longer and more pointed, which increased the space between adjacent rows of teeth. Post-larvae < 1 mm SL had highly curved teeth with clearance angles of approximately or less than zero, whereas larger abalone had positive clearance angles. These radula developments appear to be related to transitions in feeding habits from microbial to macroalgal diets.     

The effects of protein kinase A catalytic subunit on sperm motility regulation in Pacific abalone Haliotis discus hannai

Protein kinase A plays a central role in the regulation of sperm motility from echinoderms to mammals, but the information about its regulatory role in molluscs is very limited. In this study, a protein kinase A catalytic subunit (designated as HdPKA‐C) was identified from Pacific abalone Haliotis discus hannai. The open reading frame of HdPKA‐C was of 1,077 bp, encoding a peptide of 358 amino acids with a typical protein kinase domain. HdPKA‐C shared 82%–87% sequence similarities with other PKA‐Cs, and it was clustered first with gastropod PKA‐Cs in the phylogenetic tree. The mRNA of HdPKA‐C was constitutively expressed in examined tissues, with the highest level detected in hepatopancreas. The phosphorylated form of HdPKA‐C (p‐HdPKA‐C) was localized at the acrosome, connecting piece and flagellum of spermatozoa with variable intensity. Its phosphorylated substrates were also detected in these regions with much lower intensity at the connecting piece. The inhibition of HdPKA‐C activity with H‐89 led to a significant reduction in the percentage of motile sperm and sperm velocities. p‐HdPKA‐C was detected by Western blot in strip‐spawned sperm, naturally spawned sperm and H‐89‐treated sperm with almost the same intensity. The intensity of p‐HdPKA‐C substrates in naturally spawned sperm was higher than that in strip‐spawned sperm, and it was roughly the same as that in H‐89‐treated sperm except for two bands at 50 and 60 kDa. These results collectively indicated that HdPKA‐C played an important role in the regulation of abalone sperm motility by altering its substrates phosphorylation.