Routes of water loss in South African abalone (Haliotis midae) during aerial exposure

South African abalone, Haliotis midae, were exposed to air at 12 °C for 36 h to simulate the extent and rate mass loss experienced by animals during long distance live exports. Animals lost 15.1 ± 0.94% of their mass during the 36 h air exposure, an approximation of the highest mass losses sustained by industry.The total mass loss was attributed to water loss, as the contribution of dry mass to the total mass remained constant under all conditions. Water content decreased from 64.8% of the body mass (M_b) under control conditions to 58.8% M_b after 36 h in air. In real terms, however, animals had lost 22% of the body water pool.Abalone exhibited a typically high water turnover rate when in water (125 μL g^− 1 h^− 1), which decreased markedly during air exposure (2.2 μL g^− 1 h^− 1). Haemolymph volume decreased from 43% M_b in water to 14% M_b in air. The concomitant decrease in haemolymph pressure probably limited the first step in urine formation (ultra-filtration through the pericardium). Thus we observed that while urine flow represented about 26% of the total water loss when the animals were in water, urine flow ceased during air exposure.The decrease in haemolymph volume in air represents a redistribution of water to the tissues and not a bulk loss of haemolymph. This is supported by the concentration of haemolymph ions by a factor of 1.2 during aerial exposure, which was predicted based on the 22% decrease in water content. Under the same conditions, evaporation from water containers with similar surface to volume dimensions as abalone, accounted for only an 8.25% mass loss. As all other water loss routes were accounted for, we measured pedal mucus production rates of abalone in water and air. During 36 h aerial exposure, the pedal mucus production represented a loss of 6.8% M_b. We conclude that water loss during 36 h air exposure is attributable to evaporation (8.25% M_b) and pedal mucus production (6.8% M_b). This paves the way for directed research into mitigating water loss during the live export process.     

Ammonia toxicity and its effect on the growth of the South African abalone Haliotis midae Linnaeus

The current study investigated acute toxicity to ammonia of the South African abalone, Haliotis midae, from three size classes relevant to mariculture operations, and the chronic impact of sub-lethal ammonia levels on growth of juvenile abalone.Results showed that tolerance to ammonia (at pH 7.8 and T_a = 15 °C) increases with body size (i.e. age) as indicated by 36 h LC₅₀ values: juvenile abalone (1-2.5 cm shell length) had the lowest LC₅₀ of 9.8 μg l^− 1 FAN, whereas LC₅₀ was 12.9 μg l^− 1 FAN in “cocktail”-size abalone (5-8 cm shell length). The highest LC₅₀ of 16.4 μg l^− 1 FAN was observed in “brood stock”-size animals (10-15 cm). When “cocktail”-size abalone were allowed to acclimatize to sub-lethal ammonia levels for 48 h, their ammonia tolerance increased compared with non-acclimatized abalone of the same size: LC₅₀ was 2.0 μg l^− 1 FAN higher at 14.8 μg l^− 1 FAN.Growth of juvenile abalone (1-2.5 cm shell length) during chronic exposure to sub-lethal FAN levels is inhibited: specific growth rate (SGR) is significantly reduced by 58.7% to 0.10 ± 0.03% d^− 1 (weight) compared with 0.24 ± 0.06% d^− 1 of abalone of a control group (no ammonia).The results demonstrate the negative effects of ammonia not only on survival but also on growth of farmed abalone, both impair profitability of the farming operation. The information from the present study will be helpful in determining water quality requirements in South African abalone farms.     

Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828)

The thermoregulatory behaviour of green abalone Haliotis fulgens and pink abalone H. corrugata was investigated. Haliotis fulgens juveniles ranging in wet weight from 3.0 to 3.3 g and from 28.7 to 30.5 mm shell length and of H. corrugata 2.0 g and 25.7 mm in shell length were exposed to 19°C for 30 days in a flow‐through water system. Temperature preference was determined in a horizontal thermal gradient and was found to be 25.4°C for green abalone and 25.0°C for pink abalone. Displacement velocity was 4.3 cm h⁻¹ for H. fulgens and 12.8 cm h⁻¹ for H. corrugata. The optimum temperature for growth calculated for both abalone species was 24.6 and 24.5°C respectively. The critical thermal maxima (CTMax) of H. fulgens and H. corrugata were determined as a measure of thermal tolerance. Abalones were subjected to increasing water temperatures at a rate of 1°C on 30 min until they detached from the substrate. The CTMax at 50% were 33.6 and 32.0°C for green and pink abalone respectively. The results are discussed in relation to site selection and commercial rearing.     

Comparative analysis of the predominant carotenoids and chemical components in the common and orange‐muscle mutant of Haliotis gigantea

The Xishi abalone (Haliotis gigantea) is an economically significant aquaculture species in southern China. We identified a novel orange‐muscle mutation present in less than 2% of the cultured population. High‐performance liquid chromatography and electrospray ionization mass spectrometry identified zeaxanthin and β‐carotene as the two main pigments present in the orange‐muscle abalone. The concentrations of zeaxanthin and β‐carotene detected in the orange‐muscle abalones were significantly higher than those detected in the common abalones (p < 0.01). Notably, the concentration of zeaxanthin was approximately 16.5 times higher in orange‐muscle abalones than in common abalones, which was also the species with highest carotenoid content reported in shellfish to date. Furthermore, the contents of total amino acids and essential amino acids in the orange‐muscle abalone were higher than that in the common abalone, though not significantly, while the content of non‐essential amino acids was significantly higher in the orange‐muscle abalone than that in the common abalone (p < 0.05). The saturated fatty acid content in the orange‐muscle abalones was slightly lower than that in the common abalones, and the unsaturated fatty acid content was higher in the orange‐muscle abalones than that in the common abalones. These results could provide a basis for characterizing the mechanism by which carotenoids accumulate in abalones and further guide the breeding of abalone with orange‐muscle mutants in the future.     

四种鲍45S rDNA在染色体上的比较定位

为了研究皱纹盘鲍、西氏鲍、绿鲍和杂色鲍等4种鲍的核型特征,实验利用荧光原位杂交(fluorescence in situ hybridization, FISH)技术比较定位了上述4种鲍的45S rDNA位点。皱纹盘鲍中约83%的中期细胞均检出2对45S rDNA位点,分别位于13号和16号染色体的长臂端部。西氏鲍中约75%的中期细胞均检出3对45S rDNA位点,分别位于6号染色体短臂端部、14号和17号染色体长臂端部。绿鲍中约85%的中期细胞均检出3对45S rDNA位点,分别位于4号、6号和8号染色体长臂的端部。杂色鲍中约65%的中期细胞均检出3对45S r DNA,位点,分别位于3号、4号和12号染色体短臂的端部。此外,4种鲍均有少数中期相的45S rDNA位点数高于众数,这提示,除了明确的45S rDNA位点外,4种鲍可能均有若干个不稳定的45S rDNA位点。实验结果丰富了鲍细胞遗传学研究资料,同时为鲍的遗传育种研究提供了基础数据。     

Effects of dietary cholesterol on growth, survival and body composition of juvenile abalone Haliotis discus hannai Ino

An experiment was conducted to determine the dietary cholesterol requirement of juvenile abalone Haliotis discus hannai. Eight isoenergetic (18.15–18.61 kJ/g) and isonitrogenous (29.00–29.78% protein) diets, supplemented with 0.00, 0.15, 0.30, 0.45, 0.60, 0.75, 0.90 and 1.05% cholesterol were evaluated. Juvenile abalone (initial weight: 0.67–0.72 g) were reared in a flow-through water system for 24 weeks. During the feeding trial, water temperature was maintained at 14–17 °C, salinity 31–33, pH 7.4–7.9. Abalone fed diet without cholesterol supplementation had the lowest weight gain ratio (WGR, 356.70%). Survival ranged from 98.52 to 100.00% and was not significantly different among treatments. There were no significant effects of dietary cholesterol on composition and cholesterol concentration in the muscle and viscera of abalone. Based on data of WGR using broken-line analysis, the optimal dietary cholesterol requirement of juvenile abalone was found to be 0.23%.     

Hybridization between Haliotis rufescens and Haliotis discus hannai: evaluation of fertilization,larval development,growth and thermal tolerance

Two introduced abalone species are currently produced in Chile, red abalone Haliotis rufescens and Japanese abalone Haliotis discus hannai. However, red abalone accounts for 99% of total production, while the Japanese abalone has not adapted well to Chilean coastal waters. This study reports the hatching, growth and thermal tolerance performance in interspecific hybrids produced between red (R) and Japanese (J) abalone. Our results show that egg age and sperm concentration were critical factors to produce hybrids. The cross R♀ × J♂ showed a fertilization rate of 55.3 ± 3.5% using 20‐min‐old eggs and sperm concentrations of 14 × 10⁶ cells mL^?1, while the reciprocal cross (J♀ × R♂) was not successful. Further, larval development stages were similar in RR, JJ and RJ hybrid abalones. Among the experimental trials, settlement rate varied from 12.3% to 18.6% and final survival from 20.1% to 31.7%, being the RJ hybrid rates intermediate between parental species. The final shell lengths were similar between RR and RJ hybrids, but significantly higher in JJ abalones. In addition, thermal tolerance was ascertained due its pivotal role for the abalone physiology. Thus, RJ hybrids showed the highest HSP70 gene expression and offers new possibilities to expand Chilean abalone production in warm waters zones.     

皱纹盘鲍染色体C带和rDNA定位

为了提高对皱纹盘鲍染色体的辨识水平,实验利用 Ba(OH)₂ 处理显示了皱纹盘鲍染色体的C带,并用荧光原位杂交分析(fluorescence in situ hybridization,FISH)研究了核糖体大亚基rDNA在皱纹盘鲍中期染色体上的数目与位置。核型结果显示,皱纹盘鲍染色体组包含7对中部着丝粒染色体和8对亚中部着丝粒染色体,另有3对染色体介于中部着丝粒染色体与亚中着丝粒染色体之间(m/sm)。C显带结果显示,8对染色体有稳定的着丝粒C带,5～7对染色体上有中期相间多态的端部C带,3对染色体上有同源染色体异态的臂间C带。FISH 分析显示,皱纹盘鲍中期染色体上分布着4个大亚基 rDNA位点,分别位于2号短臂(2S)、7号短臂(7S)、12号短臂(12S)和18号长臂(18L)的端部。研究结果为皱纹盘鲍染色体辨识提供了新的特征与标记,为进一步研究皱纹盘鲍种群的染色体多态和鲍属染色体进化提供了基础资料。     

试述皱纹盘鲍的受精作用及其精卵结合机制

皱纹盘鲍是重要的海产珍贵贝类 ,也是北方沿海鲍类人工养殖的主要品种 ,因其具有肉质细嫩、味道鲜美、营养丰富、可作为中药材等特点倍受人们重视。鲍在世界范围内的养殖面积也在不断增大 ,这就迫切需要对鲍进行人工育苗 ,以满足鲍类大批养殖的需求。因此 ,掌握鲍的受精作用及精卵结合机制将为其进行人工繁殖提供重要的理论依据 ,提高育种效率。介绍了皱纹盘鲍的精子结构、卵子结构、受精过程、精卵结合机制 ,以求为皱纹盘鲍的人工育苗技术提供理论参考。     

Parentage assignment in hybrid abalones (Haliotis rufescens × Haliotis discus hannai) based on microsatellite DNA markers

Parentage analysis in aquaculture determines genealogical relationships between broodstock and progeny when the parents are unknown. Thus, parentage analysis is a useful tool to establish pedigree reports in molecular‐assisted selection programs. Here, we evaluated 10 heterologous microsatellite markers for parentage assignment in abalone hybrids produced from 43 abalone broodstocks of red abalone (Haliotis rufescens) and Japanese abalone (H. discus hannai). The allele frequencies, exclusion probabilities and broodstock contributions were calculated using CERVUS, PAPA and GERUD software. The polymorphic information content (PIC) values showed that most of the microsatellite loci were highly informative (>0.7) and more than 90% of parentage assignment was possible with a minimum of 5–6 microsatellite markers. Parentage assignment for hybrid and pure‐red progeny showed a better performance than pure‐Japanese progeny. This result could be due to the high level of allele loss in the parental genotypes. In addition, results indicated that only two sires contributed over 80% and 90% of red and hybrid progenies, respectively. This study gives a new molecular tool to support marker‐assisted selection in abalone hybrids produced in Chile.     

Molecular cloning,expression and functional analysis of a predicted HdiQM gene in small abalone,Haliotis diversicolor

The small abalone (Haliotis diversicolor) is a mollusc and was cultured in south of china. Problems such as the decreasing pathogen‐resistance and their mass mortality during the summer. The increased immunity of small abalone populations is a key factor in resolving these problems. Thus, the study of immunity‐related genes in small abalone has become important. In this study, three bacterial species were initially isolated from small abalone carcasses. The regression of infection was analysed, which revealed that the bacteria species could cause rapid morbidity in small abalone. A QM‐like gene (HdiQM) was found and bacterial challenge tests showed that HdiQM gene expression was induced by the bacterial isolates from small abalone carcasses. Therefore, our results implied that HdiQM was found to be an inflammatory stress‐inducible gene associated with pathogen infection, with important functions in small abalone immunity.     

紫外线消毒海水培育九孔鲍苗的研究

2000年以来自福建省南部海域至广东、海南岛沿海，陆续发生九孔鲍人工培育种苗大量死亡现象一“脱板症”，造成九孔鲍苗的严重短缺。经研究初步认为“脱板症”是一种疑似病毒性疾病。本文针对九孔鲍人工培育种苗疑似病毒性疾病，利用紫外线发生装置消毒海水培育九孔鲍苗，取得了一定的成效，比较药物处理海水培育九孔鲍苗提高出苗率达50％以上。     

The effects of density on the behaviour and growth of juvenile blacklip abalone (Haliotis rubra)

The growth of juvenile abalone in aquaculture is known to be affected by density as a result of competition for food and decreases in water quality. Our results suggest that behaviour is also affected by density and this also has a significant impact on the growth of individual abalone. 1800 juveniles of Haliotis rubra were individually tagged and reared for 5 months in 12 gravity-fed tanks at two levels of density. The experimental design allowed the differentiation of the direct from the indirect effects of density. The abalone growth and distribution was monitored monthly along with the water quality. The distribution of abalone during daytime was closely related to the availability of preferred shelter space. The percentage of abalone stacked on the top of others increased with density. Preferred shelter space was characterized by low light intensity and a corner or edge for the abalone to rest against. Hides at the ends of the tanks were not occupied as much as others. Abalone were found crawling on the side of the tank during daytime when tanks were shaded. Abalone juveniles of 15–60 mm showed fidelity to their resting shelter during daytime but this fidelity was significantly reduced at the higher density. Shading of the tanks totally changed the distribution of the abalone and their daytime behaviour. Competition for shelter space reduced growth more than water quality. This revised version was published online in August 2006 with corrections to the Cover Date.     

皱纹盘鲍脯氨酰内肽酶的分离纯化及性质

脯氨酰内肽酶(Prolyl Endopeptidase,PEP)广泛分布在动植物和微生物体内,参与细胞生长和代谢调控。PEP在哺乳动物生殖器官的生长发育过程中起着重要作用,但贝类PEP与性腺发育是否有关尚未有相关报道。本研究以皱纹盘鲍为对象,采用荧光定量PCR技术(qRT-PCR)在性腺检测到有最高的基因表达量和较高的酶活性。通过硫酸铵分级沉淀和连续柱层析,从皱纹盘鲍性腺中分离纯化得到分子量约为82 ku,等电点约为5.5的PEP,其最适温度和最适pH分别为25 °C和6.0,在15~25 °C和pH 5~8能够保持较高的酶活性。利用LC/MS/MS分析纯化蛋白,得到182个氨基酸残基,与皱纹盘鲍脯氨酰内肽酶一致。圆二色谱分析显示,温度对PEP二级结构有较大的影响,拟合热变性温度为51.4±0.2 °C。为了进一步探究PEP在鲍鱼性腺发育过程中的作用,对不同发育时期(生长初期、前期、中期和后期)的雌雄鲍鱼,采用免疫印迹(Western Blot)和qRT-PCR分析性腺中PEP的蛋白和基因表达水平。结果表明,PEP在雄性和雌性性腺的各个时期均能被检测到,在雄性性腺的成熟中期和雌性性腺的成熟后期表达量最高。PEP在鲍鱼性腺不同发育时期时表达量的差异表明其可能参与到性腺发育的过程。     

“东优1号”杂色鲍及其亲本群体的形态特征和养殖性能比较

以杂交种东优1号杂色鲍及其亲本群体为研究对象,对三者的外形特征和养殖性能进行了比较。结果表明,在壳形和密度相关的参数中,壳宽/壳长指标差异最为显著,日本群体的壳宽/壳长指标显著地大于台湾群体(P<0.05),而东优1号的值则介于双亲群体之间。采用海区延绳式吊养和陆地工厂化养殖两种养成模式,分别在福建、广东和海南对东优1号杂色鲍及其亲本群体进行为期一年的养殖对比试验。结果表明,杂色鲍台湾群体受养成期暴发性死亡症的严重影响,存活率极低;日本群体虽然具有较好的抗病能力,但生长速度却显著慢于东优1号和台湾群体;东优1号杂色鲍的生长速度和存活率在3个实验点都表现出较为显著的优势。研究结果表明东优1号杂色鲍是适合在亚热带和热带地区进行养殖的优良鲍新品种。     

杂色鲍一种编码CRAD功能域基因的克隆及其在发育和应激中的表达

为探讨Caspase募集功能域(CRAD)在鲍发育和应激中的作用,实验通过比对杂色鲍转录组序列和RACE扩增的方法,获得了杂色鲍一条含CRAD的基因全长c DNA,命名为Hd CRAD;采用实时定量PCR的方法获得了该基因在发育和应激中的表达谱。结果显示,Hd CRAD的c DNA全长1 371 bp,开放阅读框735 bp,编码244个氨基酸。编码蛋白具有保守的CRAD功能域。该基因在各检测组织中均可表达,在黏液腺中表达量最高;在发育各阶段也均有表达,面盘幼虫晚期表达量最高。在细菌感染、高温或缺氧应激处理后,血细胞的Hd CRAD表达水平均有显著性变化。在担轮幼虫时期,RNA干扰该基因会显著提高幼虫畸形率;幼虫的caspase-8和DAD1的表达水平会显著上升,而caspase-3的表达水平会下降。研究表明,该基因参与了鲍幼虫发育过程;在成体免疫、耐高温和抗低氧中发挥了一定作用。     

皱纹盘鲍4个养殖群体杂交子代生长与存活状况比较

为利用杂种优势培育皱纹盘鲍(Haliotis discus hannai)优良新品种,本研究以4个不同养殖群体[黄岛(HD)、荣成(RC)、日本(JP)、大连(DL)]的皱纹盘鲍为亲本,设计4×4完全双列杂交,建立了4个自交家系和12个正反杂交家系,在利用微卫星标记进行亲子鉴定的基础上,对各家系的F1在1、5、13、17月龄时的生长性状、杂种优势率、生长速度和存活情况进行比较,分析杂交效应。结果表明,在各个生长阶段均有部分杂交家系与自交家系相比表现出显著的生长优势;HDRC、HDDL和JPDL家系的生长速度较高;HDDL、HDJP、RCDL、JPRC及RCHD家系有着较高的存活率;在杂种优势方面,HDRC、HDDL与DLHD家系在各生长参数与生长速度上有明显的杂种优势,HDDL、RCDL、DLHD家系在存活率上表现出明显的杂种优势。本研究筛选出的具有优势的交配组合,可作为皱纹盘鲍生产上种苗来源的参考,也为利用杂种优势培育皱纹盘鲍新品种提供了基础资料。     

Temperature and size range for the transport of juvenile donkey's ear abalone Haliotis asinina Linne

Live transport of hatchery‐produced juvenile donkey's ear abalone Haliotis asinina Linne was examined to evaluate the effect of transportation on the survival of juvenile abalone. Simulated transport experiments were conducted to determine the appropriate temperature using 5, 10 and 20 g L^?1 of ice to air volume for 8 h and the appropriate size using two size groups (Size A, 15–20 mm, 0.5–1.3 g, and Size B, 30–35 mm, 5.3–8.5 g) up to 24‐h out‐of‐water live transport. Survival was significantly higher (P<0.001) when 10 g L^?1 of ice was used to decrease the temperature to the range of 17–23 °C. At this temperature, both size groups subjected to simulated transport for 8 and 10 h had 100% survival after 48 h, while mortality occurred in abalones subjected to 16 and 24 h of simulated transport. The Size B abalone subjected to 24 h of transport had significantly higher survival (64.4 ± 2.9%) (P<0.001) than the Size A abalone (5.5 ± 1.6%) after 48 h. Live juvenile abalone were successfully transported to the field applying the protocols developed in the lab experiment. This study serves as a guide for handling and shipping live juvenile abalone.     

The growth of disk abalone, Haliotis discus hannai at different culture densities in a pilot-scale recirculating aquaculture system with a baffled culture tank

The growth rate of disk abalone, Haliotis discus hannai, energy consumption and changes in water quality were monitored in a pilot-scale recirculating aquaculture system (RAS) for 155 days. Baffles were installed in the RAS culture tanks to enlarge the attachment area and clean out solid waste materials automatically by hydraulic force only. The experimental disk abalones, of shell length 24.5 ± 0.5 mm, were cultured at three stocking densities, 700, 1300 and 1910 individuals/m² bottom area, in triplicate. The abalones were fed with sea mustard, Undaria pinnatifida, once a week. The abalone feed conversion rates and daily growth rates ranged from 24.5 to 25.9 and 0.32 to 0.36%, respectively. Their daily shell increments and survival rates ranged from 67.7 to 78.6 μm/day and 87.6–92.2%, respectively. The growth in weight tended to decrease at a culture density of 1300 individuals/m² bottom area, while shell increments and survival rates were acceptable at this density. The total power consumption for heating was 1185.4 kW, comprising 30.2% of the total power consumption, while the average water exchange rate was only 2.9% per day. The total ammonia nitrogen stabilized below 0.07 mg/L, after conditioning of the biofilter. The NO₂⁻–N, NO₃⁻–N and total suspended solid concentrations were also maintained within acceptable ranges for the normal growth of disk abalone. The use of the RAS with these newly designed culture tanks for disk abalone culture produced 1300 individuals/m² bottom area with a water exchange rate of only 2.9% per day and used about one-tenth of the heat energy of a conventional flow-through system.