紫菜中无机砷含量的分析研究

本文针对紫菜中的无机砷超标问题,采用高效液相色谱—原子荧光光度法联用技术(HPLC-AFS)对紫菜中砷的形态进行分析,并测定了紫菜中的无机砷的含量。通过分析发现,紫菜中砷主要以一甲基胂酸、二甲基胂酸及砷糖、甜菜碱、砷胆碱等多种形态的有机砷化合物的形态存在于藻体中,无机砷的含量很少,仅在0.2mg/kg左右。采用HPLC-AFS能进行无机砷的分析测定时,由于三价砷的出峰时间与第1个峰的有机砷化合物出峰时间重叠,采用过氧化氢将其氧化成五价砷,可以很好地解决这一问题。通过验证,其加标回收率可达到80.19%,方法的准确性高,回收率符合检测的要求,适用于紫菜中无机砷的测定。     

Effect of temperature on growth of the intertidal purple snail Plicopurpura pansa (Gould 1853) under laboratory conditions

To determine the optimum temperature for growth of the endangered intertidal purple snail Plicopurpura pansa (Gould 1853) (Prosobranchia, Muricidae), 731 adult specimens were reared in the laboratory on squid for 86 days at temperatures ranging from 20 to 32°C. The snails showed extremely slow growth, if at all. Males and females raised at 26°C grew the fastest, an average of 0.85 and 0.43 mm respectively. No statistically significant differences were observed at other temperatures or between males and females. The best temperature for growth for the intertidal purple snail appears to be in the range between 26 and 30°C.     

Population structure of purple sea urchin <Emphasis Type="Italic">Strongylocentrotus purpuratus</Emphasis> along the Baja California peninsula

Abstract:   Purple sea urchin Strongylocentrotus purpuratus is fished from British Columbia, Canada to Punta Baja, Mexico. The North American population has been divided into northern and southern fishery stocks at the break of Point Conception, but little is known about its southernmost distribution along the Mexican Pacific coast of the Baja California peninsula. In this study purple sea urchin populations in six sites along the Baja California peninsula were analyzed using mitochondrial deoxyribonucleic acid restriction fragment length polymorphism (mtDNA RFLP). A homogeneous distribution of three common haplotypes among all sites was observed. A significant F _ST value, however, indicated genetic structure mainly due to the haplotype array in San Miguel, Isla Todos Santos and Punta Baja sites, which were characterized by having high haplotype diversity and several unique haplotypes. Homogeneous distribution of haplotypes along the peninsula could have been influenced by the unidirectional California Current system, flowing north to south. Unique haplotypes in Punta Baja and the structure found could be the result of local oceanographic features specific to this major upwelling zone. It may be necessary to consider the Punta Baja populations individually when managing the purple sea urchin fishery in Baja California, as they show signs of being a unique stock.     

Inheritance of shell colours in the noble scallop Chlamys nobilis (Bivalve: Pectinidae)

The noble scallop Chlamys nobilis is polymorphic in its shell colour, which includes orange, orange‐purple, purple and brown. In this study, the inheritance of shell colours of this species was investigated by analysing shell colour segregation in juvenile scallops produced from cross‐fertilization. Totally, twenty two families derived from four types of parental crosses were produced. The results clearly demonstrated that shell colour in this species was consistently inherited but insensitive to the environmental factors given that each family was maintained in a common environment. The pattern of the four superficial shell colours in C. nobilis appears to be genetically controlled by the interaction of one pair of nonallelic genes, with one locus showing dominance epistasis to the other one. One‐locus‐three‐allele model was proposed to explain the genetic relationship of the orange‐purple colour dominant to the purple and brown colours, and the purple colour dominant to the brown colour. While the phenotypic ratio of 12:3:1 detected in one specific family showed that the dominant allele of the other locus controlling orange colour may mask the effect of the locus controlling orange‐purple, purple and brown colours. The present results provide a genetic basis for selective breeding in the noble scallop in terms of shell colour.     

Nutritional value and production of three species of purple non‐sulphur bacteria grown in palm oil mill effluent and their application in rotifer culture

Three species of purple non‐sulphur bacteria (PB), Rhodopseudomonas palustris, Rhodobacter sphaeroides and Rhodovulum sulfidophilum, grown in palm oil mill effluent (POME) were successfully used for the first time as feed for rotifers (Brachionus rotundiformis). Rp. palustris cultured in both POME and synthetic medium gave the highest rotifer density (332–395 individuals mL⁻¹) from 3 to 5 days at 10 g L⁻¹ salinity. Other PB cultured in synthetic medium generally support higher rotifer density than PB cultured in POME. Rb. sphaeroides had the highest biomass (1.91–3.34 g L⁻¹) and growth rate (0.64–1.11 g day⁻¹) in both types of culture medium. Nevertheless, only Rv. sulfidophilum grown in POME contained both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), indicating its ability to biosynthesize them from POME nutrients. Rotifers fed Rv. sulfidophilum grown in POME had significantly higher amounts of protein, arachidonic acid, EPA and DHA than rotifers fed Rv. sulfidophilum grown in synthetic medium. The nutritional profile of lipid‐deficient PB can be improved by growing them in POME, and these enriched PB produced at an estimated cost of USD 8.71–35.35 kg⁻¹ dry biomass, depending on species, can support rotifer production in a batch culture system.     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

3种不同体色刺参体壁营养成分的比较研究

本研究分别对青色、紫色和白色刺参(Apostichopus japonicus)体壁的营养成分,包括蛋白、脂肪、粗多糖、氨基酸组成、脂肪酸组成及微量元素进行了比较分析。结果显示,青色刺参蛋白含量显著低于紫色和白色刺参(P<0.05),脂肪含量三者无显著差异(P>0.05),青色刺参和紫色刺参的粗多糖含量高于白色刺参(P<0.05)。3种体色刺参检测出17种氨基酸,白色和紫色刺参氨基酸含量相对较高,为48%左右,而青色刺参体壁氨基酸含量为44%左右。紫色和白色刺参的赖氨酸、苯丙氨酸、苏氨酸、缬氨酸等氨基酸的含量均显著高于青色刺参(P<0.05)。亚油酸(LA)、花生四烯酸(AA)、二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)在3种刺参占较高比重,其中,LA作为重要必需脂肪酸在紫色和白色刺参中均显著高于青色刺参(P<0.05),为青色刺参的4.6倍左右;紫色和白色刺参的AA含量也显著高于青色刺参(P<0.05);而青色刺参中的EPA和DHA均显著高于紫色和白色刺参(P<0.05)。在检测的矿物质中,Fe在3种刺参中含量最高,紫色刺参Fe的含量显著低于青色和白色刺参(P<0.05);青色刺参Mn的含量显著高于紫色和白色刺参(P<0.05);白色刺参Cr的含量显著高于青色和紫色刺参(P<0.05)。研究表明,3种体色刺参的营养成分存在较大差异,均具有进一步开发利用价值。     

6-DMAP诱导栉孔扇贝异源雌核发育二倍体

采用紫外线遗传灭活的长牡蛎精子激活栉孔扇贝卵子,运用L9（3^4）设计不同6-DMAP的处理条件来诱导染色体加倍,获得第二极体抑制型栉孔扇贝雌核发育二倍体早期胚胎。以雌核发育担轮幼虫为材料,滴片法制备染色体并采用计数法统计倍性,分析不同6-DMAP浓度、诱导时机和诱导持续时间对雌核发育二倍体诱导率的影响,并统计早期胚胎畸形率。结果表明,不同6-DMAP浓度、诱导时机和诱导持续时间对异源精子诱导栉孔扇贝雌核发育二倍体的诱导率均有显著影响;早期胚胎畸形率与二倍体的诱导率呈负相关,y=46.632-0.891x（r=-0.813,P〈0.01）。6-DMAP诱导栉孔扇贝雌核发育二倍体的最佳处理条件为：20%～25%受精卵排放出第1极体时,60μg/ml的6-DMAP持续处理25min可得到最高的诱导率为29.5±5.36%。研究结果为6-DMAP诱导栉孔扇贝雌核发育染色体加倍提供了一定的细胞学依据和技术参数。     

Effect of Temperature on Growth,Energy Budget,and Physiological Performance of Green,White, and Purple Color Morphs of Sea Cucumber,Apostichopus japonicus

As an intertidal species, Apostichopus japonicus is subject to diel and seasonal temperature fluctuations. Color variation is a distinct characteristic of A. japonicus, and a new color morph, purple A. japonicus, was recently found on the coast of Rushan, Weihai City. This study was conducted to compare the performance of green, white, and purple color morphs of A. japonicus at different water temperatures to help improving aquaculture technology and management. In this study, green, white, and purple color morphs of juvenile A. japonicus (1.5 ± 0.1 g) were cultured at 10, 14, 18, 22, and 26 C for 60 d, according to the suitable temperature range for sea cucumber growth. Temperature was shown to have a significant effect on growth, energy budget, and physiology of sea cucumbers (P < 0.05). Specific growth rates, feed conversion efficiencies, and the proportion of energy allocated to growth of all three color morphs were optimal at 18 C. Activities of the enzymes related with oxidative stress, immune response, and digestion were determined. Superoxide dismutase (SOD) and trypsin activities generally reached a maximum in all three color morphs at 18 C. The purple color morph of A. japonicus was higher in growth rate, food conversion efficiency, and proportion of energy allocated to growth than green and white color morphs at 18 and 22 C, consistent with its higher trypsin and SOD activity, indicating the mutual effect of growth and physiology. At 18, 22, and 26 C, the green color morph had the highest level of lysozyme activity and purple color morph had the lowest, indicating possible infection by external pathogens of green color morph, which might be caused by high temperature. These results suggested that the optimum temperature for culture of the three color morphs was approximately 18 C, the purple had a better adaption to high temperature, and the green and white color morphs may be more susceptible to thermal stress than the purple color morph.     

Significant improvement of shrimp growth performance by growth hormone‐releasing peptide‐6 immersion treatments

Growth hormone‐releasing peptide‐6 (GHRP‐6) is one of the earliest developed synthetic peptidyl growth hormone secretagogue receptor agonists. These compounds mimic the effect of the endogenous ligand ghrelin. In vertebrates, ghrelin is a potent circulating orexigenic hormone with functional roles in controlling food intake, energy expenditure, adiposity, growth hormone secretion and immunity. Ghrelin has been studied mainly in vertebrates; thus, little is known about its role in invertebrates, including crustaceans. We first evaluated the effect of GHRP‐6 injection over feed intake in shrimp and its effects on shrimp growth when the peptide was administrated by successive immersion baths. GHRP‐6 increased feed intake, body weight and size, the number of rostral spines and gill branches, protein concentration and haemocyte number in treated shrimps. We also evaluated the peptide uptake and clearance in a pharmacokinetics, using [H³]GHRP‐6 administered to postlarvae. Given a limited exposure and efficient clearance of the peptide‐associated radioactivity from larvae, our findings suggested that GHRP‐6‐treated Litopenaeus vannamei can be consumed safely by humans after aquaculture applications. These results propose that GHRP‐6 could be an additional tool to study growth physiology in crustaceans and also a promising candidate for development into a new biotechnology product for improving shrimp growth and quality.     

Color effects of lures on the hooking rates of mackerel longline fishing

SUMMARY: This study attempts to understand the color effects of artificial lures on the hooking rate of mackerel longline fisheries in Taiwan waters. From the results obtained from field work, the order of hooking rates from the different colored lures was red > black > yellow-green > yellow-orange = reddish orange = pink > white > blue > purple > transparent. Transparent, purple and blue lures had the smallest hooking rates and showed significant differences (least significant difference test) from the other colored lures. However, no significant differences could be found among the hooking rates of the other colored lures. In addition, the lumiunous reflectance of nine different gray lures did not show any significant difference in the hooking rates. Therefore, it is likely that the 'spectral' (colored) effects are the major influence causing the differences in hooking rates and the luminous effects play only a minor role in artificial lures.     

The stimulatory effect of LED light spectra on genes related to photoreceptors and skin pigmentation in goldfish (Carassius auratus)

This study aimed to assess differences in genes related to skin color of goldfish (Carassius auratus) exposed to light-emitting diodes (LEDs): red, green, and purple. We investigated differences in the expression of mammalian-like melanopsin (Opn4m), rhodopsin (RH), melanin-concentrating hormone (MCH), melanin-concentrating hormone receptor (MCH-R), and proopiomelanocortin (POMC) in goldfish exposed to different LED light spectra. Opn4m, RH, MCH, and MCH-R mRNA levels were significantly higher in the green and purple LED groups than in the white fluorescent bulb (control) and red LED groups. Furthermore, skin cells were isolated to measure the MCH-R mRNA expression levels. The results show that the mRNA expression levels were significantly higher in the green and purple LED groups than in the control and red LED groups. In addition, body weights in the green and purple LED groups were significantly higher than those in the control and red LED groups. However, POMC mRNA expression levels in the green and purple LED groups were significantly lower than those in the control and red LED groups. These results suggest that specific wavelengths regulate fish skin color through neuropeptide hormones and photoreceptors, and POMC, which is related to stress hormones and melatonin, is associated with stress levels as well as skin color.     

Effect of temperature on growth,digestion and immunity of green,white and purple sea cucumbers,Apostichopus japonicus (Selenka)

Three color morphs (white, green and purple strains) of Apostichopus japonicus (Selenka) were cultured in artificial seawater, for approximately 90 days, in three temperature ranges: 27–22°C (high), 22–17°C (mid) and 17–12°C (low). All strains grew in all temperature ranges. Temperature significantly affected growth rate, digestibility, digestive enzymes and immune‐related enzymes. Highest specific growth rates were exhibited in 4‐month‐old sea cucumbers at mid and high temperatures, and in 16‐month‐old sea cucumbers at mid and low temperatures. Specific growth rates of green and purple strains were not significantly different, but were significantly higher than that of the white strain at mid temperatures. The digestibility of each strain was significantly higher at 27°C, 22°C and 17°C than at 12°C. Green‐strain digestibility was higher than that of purple and white strains at specific temperatures. Protease and amylase activities of all strains followed bell‐shaped temperature curves with maximum digestive enzyme activity at 17°C. The activities of alkaline and acid phosphatases were higher in the guts of the green strain than in the white or purple strains at the same temperature. Superoxide dismutase activity was higher in the purple strain than in white and green strains.     

黄斑篮子鱼消化道组织学和组织化学研究

采用HE和AB—PAS染色方法分别从组织学和组织化学方面对黄斑篮子鱼（Siganus oramin）消化道进行了研究。结果显示,黄斑篮子鱼口小,口咽腔不宽阔。上颌具细长、尖锐齿,下颌齿短,排列紧密。犁骨、腭骨和舌上无齿。鳃耙齿分叉,食道细长,胃部稍膨大,“V”型,幽门盲囊5～6条,肠长比为2．73。胃腺发达,为管状腺体,开口于胃小凹。幽门部粘膜褶高度降低,褶皱上开始出现类似肠绒毛结构。前肠均分布有许多长条形粘膜褶,空泡状杯状细胞丰富。依据AB—PAS的染色结果将该鱼}肖化道的粘液细胞分为4种类型：I型呈红色,Ⅱ型呈蓝色,Ⅲ型呈紫红色,Ⅳ型呈蓝紫色。食道以Ⅱ型和Ⅲ型为主,胃、肠道和幽门盲囊主要以Ⅱ型和Ⅳ型为主,后肠中Ⅱ型〉III型〉IV型,其中中肠和后肠Ⅱ型占总粘液细胞的比率最大。     

草鱼呼肠孤病毒VP6蛋白在毕赤酵母中表达的初步研究

根据GeneBank中草鱼呼肠孤病毒(grass carp reovirus,GCRV)104株VP6蛋白的全基因序列(HM234682)设计一对特异引物,以提取的GCRV-104核酸为模板,采用RT-PCR技术扩增VP6蛋白编码基因,并将其克隆到含强启动子AOXⅠ的毕赤酵母(Pichia pastoris)表达载体pPICZB上。重组酵母质粒pPICZB-VP6经SacⅠ线性化,电击转化到毕赤酵母KM71菌株中,Zeocin抗性平板上筛选高拷贝转化子。阳性转化子在30℃,0.5%(V/V)的甲醇添加量的条件下,连续诱导3 d,表达产物经SDS-PAGE和Western-Blotting检测,结果表明成功实现了GCRV-104 VP6蛋白在毕赤酵母中的胞内表达,重组VP6蛋白相对分子质量约46 000,与天然VP6蛋白相对分子质量接近,并且可与鼠抗草鱼GCRV-104 VP6蛋白的多克隆抗体特异性结合。     

Exotic hard clam in Tokyo Bay identified as Mercenaria mercenaria by genetic analysis

ABSTRACT:   An exotic hard clam has been observed in Tokyo Bay since the mid-1990s, which has been morphologically identified as Mercenaria mercenaria . The authors have determined 444-bp partial nucleotide sequences of mitochondrial DNA (mtDNA) 16S rRNA gene of 31 hard clam individuals collected in three locations of Tokyo Bay. Phylogenetic analysis indicated all individuals to be M. mercenaria . Two shell interior colorations (white or purple) were observed, but no relationships between the colorations and mtDNA haplotypes were detected. Frequency of clam individuals having white or purple shell significantly differed among locations, which was accompanied by heterogeneous amounts of acid-volatile sulfides and organic matter and mud ratio.     

6-甲氨基嘌呤诱导栉孔扇贝三倍体的细胞学机理

以６０ｍｇ／Ｌ的６－甲氨基嘌呤（简称６－ＤＭＡＰ）诱发栉孔扇贝三倍体时,在荧光显微镜下观察受精卵的染色体行为以及核相组成,对照三倍体率和孵化率,探讨了不同减数分裂期６－ＤＭＡＰ处理对染色体倍数性的影响。结果表明,处理５￣１５ｍｉｎ可见３种类型图像,第１种图像可见第一极体和一个雌性原核及雄性原核;第２种图像可见第一极体和两雌性原核及一个雄性原核;第３种图像可见两个雌性原核和一个雄性原枋,随着处理时间     

三角帆蚌HcCUBDC基因cDNA的全长克隆与表达分析

利用cDNA末端快速扩增（Rapid Amplification of cDNA Ends,RACE）方法,获得了三角帆蚌HcCUBDC基因的全长cDNA序列,共5158bp,开放阅读框（ORF）1920bp,编码639个氨基酸,5′端非编码区576bp,3′端非编码区2662bp,GeneBank登陆号为KP067952。生物信息学分析表明,三角帆蚌HcCUBDC蛋白含有一个由19个氨基酸组成的信号肽和4个CUB结构域,但未能比对上已知的蛋白。经荧光定量PCR检测,HcCUBDC基因在紫色和白色蚌前端缘膜、后端缘膜、中央膜、鳃、斧足、肝胰腺、肠和肾8个组织中均有表达,并且都是肝胰腺表达量最低。在紫色蚌中,后端缘膜表达量极显著高于前端缘膜;在白色蚌中,前端和后端缘膜之间表达差异不显著。HcCUBDC基因在紫色蚌后端缘膜中的表达量极显著高于白色蚌,在紫色和白色蚌前端缘膜中的表达量差异不显著。外套膜原位杂交结果显示,HcCUBDC基因主要在缘膜外上皮细胞中表达。研究表明,HcCUBDC基因在三角帆蚌内壳色形成中发挥作用,可为进一步深入研究该基因在珍珠颜色形成过程中的功能及其调控机理提供基础资料。