Seasonal cycle and regulation by temperature of antifreeze protein mRNA in a Long Island population of winter flounder

The seasonal cycle and regulation by temperature of antifreeze protein mRNA (AF mRNA) were investigated in a Long Island population of winter flounder (Pseudopleuronectes americanus) by Northern blot hybridization and by in vitro translation of liver RNA. AF mRNA was expressed at high levels in the fall and winter (Nov.–Feb.) and at low or undetectable levels in the summer. The time of accumulation of AF mRNA coincides with the time during which water temperature and photoperiod decrease to 4°C and 9 h of light per day, respectively. A temperature and photoperiod decrease in the laboratory during this time also resulted in high levels of AF mRNA. The levels of other mRNAs, as assayed by in vitro translation, were relatively constant during both seasonal acclimation and laboratory acclimation. The seasonal cycle of AF mRNA in Long Island winter flounder is similar to that of a more northern, Newfoundland population of winter flounder and different from that of an intermediate, New Brunswick population. These similarities and dissimilarities are discussed in light of potentially different exogenous and endogenous regulatory cues in the different populations.     

Protein content and freezing avoidance properties of the subdermal extracellular matrix and serum of the Antarctic snailfish,Paraliparis devriesi

The Antarctic snailfish, Paraliparis devriesi (Liparididae), occupies an epibenthic habitat at a depth of 500–650 m in the subzero waters of McMurdo Sound, Antarctica. This species has watery (97%) gelatinous subdermal extracellular matrix (SECM) comprising a mean of 33.8% of the body weight, the largest known proportion of any adult fish. The protein concentration of the SECM was found to be 6–7 mg ml^–1 (0.6–0.7% w/v). Separation of the polypeptides of the SECM by SDS-PAGE revealed 11 polypeptides ranging in relative molecular mass (M_r) from 67,000 to 13,000, with other unresolved polypeptides of less than 13,000. The isoelectric points of these proteins ranged from 4.85 to 8.05. Partial N-terminal amino acid sequence data were obtained for four of the major SECM polypeptides. The N-terminal amino acid sequences of three of these were not identical to or homologous with any other known sequences, whereas the N-terminal sequence of one polypeptide (M_r 51,000) was identical to partial sequence from the apolipoprotein A-I precursor of Atlantic salmon (Salmo salar). Although not isolated from either SECM or serum, melting point-freezing point behavior of body fluids suggest that Paraliparis possess modest amounts of a noncolligative antifreeze compound. Since relatively small amounts of antifreeze are present in the serum and even less in the SECM, freezing avoidance results from the combined effects of antifreeze and the elevated osmolality of body fluids. There are no special adaptations to prevent freezing in the superficially located high water content SECM.     

Comparison of flavor components in shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> cultured in sea water and low salinity water

To compare the flavor components of shrimp Litopenaeus vannamei cultured in sea water and low salinity water, the chemical composition of muscle was analyzed, muscle extracts prepared, and their sensory components and sensory properties were assessed. Shrimp cultured in sea water had a higher content of crude protein, lower moisture, and a higher flesh pH compared to the low salinity samples (P<0.05), whereas no significant differences were found in crude lipid and ash. The free amino acid composition of muscle extracts and the amino acid composition of peptides from shrimp cultured in sea water and low salinity water were similar, that is, the former had a very high content of glycine, arginine, proline and alanine, and the latter a very high content of glutamate, glycine, arginine, proline and alanine. The major components of nucleotides were adenosine monophosphate and inosine monophosphate, and the major components of organic acids were acetic acid and malic acid. Extracts from shrimp cultured in sea water had enhanced umami, sweetness and overall flavor, and less of an earthy-musty taste compared to samples cultured in low salinity water (P<0.05). Aftertaste did not differ between the two shrimp extracts (P>0.05).     

牙鲆和半滑舌鳎5S rDNA基因的染色体定位及鲽形目5种鱼类的分子系统学分析

利用荧光原位杂交(FISH)技术将5S rDNA基因定位在牙鲆和半滑舌鳎染色体上,结果表明,5S rDNA基因在雌、雄性半滑舌鳎的一对同源染色体上分别存在2个杂交信号位点;在二、三倍体牙鲆的同源染色体上分别存在2个和3个杂交信号位点,杂交信号明显且特异。本研究首次将5S rDNA基因定位在半滑舌鳎和三倍体牙鲆的染色体上,为牙鲆及半滑舌鳎倍性鉴定、染色体鉴别提供了有效方法。此外,根据牙鲆5S rDNA基因编码区序列设计引物,扩增出大菱鲆和半滑舌鳎5S rDNA基因编码区序列,与鲽、塞内加尔鳎、大口黑鲈、七鳃鳗的5SrDNA基因序列进行同源性比较后发现,5种鲽形目鱼类5S rDNA基因序列同源性高达98.3%,系统发生分析结果显示5种鲽形目鱼类聚为一支;各序列中GC含量均显著高于AT含量。     

大型溞的营养成分分析及其作为渔用饲料原料的潜在利用价值

为了评估大型溞在渔用饲料中的的营养价值,测定了不同季节人工培养的大型溞和夏季污水中大型溞风干样的营养成分与重金属含量。结果显示,各季节人工培养的大型溞水分含量存在显著的季节差异,冬季最高为5.14%,夏季最低为2.51%;夏季污水中大型溞水分含量(9.12%),显著高于人工培养的大型溞。人工培养的大型溞粗蛋白含量为30.93%~50.21%,在不同的季节也存在显著性差异,冬季人工培养、夏季污水中大型溞粗蛋白含量显著高于其他组。大型溞的15种氨基酸含量在各季节中差异显著,冬季最高(36.65%),夏季最低(24.74%);夏季污水中氨基酸含量为41.79%,9种必需氨基酸占22.15%,赖氨酸含量达到3.4%,蛋氨酸的含量为1.14%,苏氨酸含量为2.45%。大型溞粗脂肪含量随季节变化差异显著,冬季最高(18.88%),秋季最低(4.19%),夏季污水与冬季人工培养的大型溞无显著性差异。秋季人工培养的大型溞与夏季污水大型溞必需脂肪酸含量最高,为17%左右,所有大型溞中均没有检测到花生四烯酸和亚麻酸。重金属铬、铜和锌、铅最高含量分别出现在夏季污水、秋季、春季、夏季人工培养大型溞;夏季污水和冬季人工培养大型溞中未检测到铅元素,所有大型溞中均没有检测到镉元素。除铬外,铜、铅、砷和锌的最低含量均出现在夏季污水大型溞中。研究表明,大型溞蛋白质、脂肪营养丰富,而夏季污水大型溞必需氨基酸和必需脂肪酸营养价值普遍高于人工培养的大型溞,且重金属潜在毒性较低,作为渔用饲料蛋白源进行资源化利用的可行性更高。     

Growth of early juvenile winter flounder (Pseudopleuronectes americanus Walbaum)

Metamorphosis is a critical developmental stage in marine fish species, but mortalities are also elevated at the settling phase. Our objective was to evaluate the effects of different rearing systems and larval diets on survival, general condition and pigmentation of newly settled winter flounder (Pseudopleuronectes americanus) juveniles. At settlement, winter flounder juveniles were reared in two different types of tanks (plankton kreisel and rectangular raceway). The early juveniles were followed for 1 month and their condition was evaluated using biochemical indices. Juveniles did not show signs of fin rot and pigmentation was well developed, but they had different RNA/DNA ratios. This study highlights the fact that rearing conditions at the onset of the juvenile stage may be critical for successful juvenile production.     

Genetic characterization, morphometrics and gonad development of induced interspecific hybrids between yellowtail flounder, Pleuronectes ferrugineus (Storer) and winter flounder, Pleuronectes americanus (Walbaum)

Viable interspecific hybrids between yellowtail flounder (Pleuronectes ferrugineus, Storer) and winter flounder (Pleuronectes americanus, Walbaum) were produced by artificial insemination of yellowtail flounder eggs with winter flounder sperm. However, mean fertilization rate, hatching success and early survival up to 3 weeks post hatch were significantly lower than those of parental pure cross controls (P < 0.01). Overall, cytogenetic traits (karyological analysis and estimation of cellular DNA contents using flow cytometry) of hybrid flounder were intermediate between the two parental species. Microsatellite assay was used to distinguish the parental genomes in the hybrids; in most cases, one allele was specific to each of the parents. Morphometrics assessed by body proportions indicated that hybrids generally displayed a morphology intermediate between the maternal and paternal species. Interspecific hybrids exhibited abnormal and retarded gonad development in both sexes based on histological analysis of gonads from adult fish. The sterility of the hybrids presents a significant advantage for their use in aquaculture, as potential escapees would not be capable of reproducing in the wild and contaminating natural stocks.     

Determining an optimal release site for juvenile winter flounder Pseudopleuronectes americanus (Walbaum) in the Great Bay Estuary, NH, USA

One of the main elements in developing an optimal release strategy for an enhancement effort is to evaluate and select release sites that will support growth and survival of newly released, cultured fish. Three potential release sites (New Castle (NC), Broad Cove (BC) and Oyster River (OR)) in the Great Bay Estuary, NH, USA were evaluated for pilot‐scale releases of winter flounder (Pseudopleuronectes americanus Walbaum). Cultured juvenile flounder were placed in cages at each of the three sites. Sites were evaluated based on growth and survival of the fish in relation to water temperature, prey availability and sediment composition. Fish grew faster in the two upper estuarine sites BC (0.54 mm day^?1) and OR (0.56 mm day^?1) than at the site at the mouth of the estuary (NC=0.37 mm day^?1). Fish survival (44–53%) and water temperature (17.8–19.7°C) did not vary between sites. Benthic samples showed that prey was available to, and eaten by, the flounder. Sediment composition was the main difference between the three sites, with one site (NC) characterized by gravel whereas the other two sites were sandy. These results corroborate other studies showing the importance of sediment quality for the distribution of flatfish populations. From these results, we can confidently eliminate NC as a potential release area and recommend that sandy sites are better for stocking cultured juvenile winter flounder than gravely sites.     

Is the protected Atlantic cod (Gadus morhua) population in Gilbert Bay,Labrador, nearing a tipping point?

1. The Gilbert Bay Marine Protected Area (MPA), Labrador, was created in 2005 to protect a resident, locally adapted population of Atlantic cod and its habitat. Twenty years of monitoring has shown a 90% decline in this bay-cod population since the creation of the MPA.
2. The decline of large commercial-sized cod in the bay was accompanied with increases in the abundance of other fish species, including rock cod (Gadus macrocephalus ogac), sculpin (Myoxocephlalus scorpius), and winter flounder (Pseudopleuronectes americanus), suggesting loss of strong top-down community control.
3. These changes suggest a positive feedback mechanism leading to a fish community tipping point, which could severely impact the recovery potential of the protected Atlantic cod population in the MPA.
4. We suggest that managers should be including this possibility in their management decisions for the Gilbert Bay MPA.

     

鲫肠道乳酸菌的分离及益生特性

为获得鱼源益生乳酸菌,本研究从健康鲫肠道内分离鉴定得到38株乳酸菌,并选择其中8株乳酸菌进行体外益生特性分析。结果显示,8株乳酸菌均能在p H=4.5和10%鲫胆汁环境中存活,对嗜水气单胞菌、豚鼠气单胞菌、无乳链球菌和副溶血弧菌均有较强的抑菌能力,但菌体表面疏水性和自凝集能力具有菌株特异性。选择5株疏水性较高(63%~89%)、自凝集能力强(37%~45%)的乳酸乳球菌进行体外黏附能力测定,结果显示,黏附能力在菌株间具有差异性(4.5%~7.9%),但均能显著降低嗜水气单胞菌NJ-35的体外黏附率,黏附抑制率达30%~35%;最后对筛选出的3株高黏附力的乳酸乳球菌进行安全性评价,发现3株菌对鲫均无致病力。鱼源乳酸菌的筛选,为鲫养殖生产中益生菌的实际应用提供了理论依据。     

尼罗罗非鱼制作传统上海熏鱼过程中的风味变化

为探究上海熏鱼制作过程中不同加工阶段风味物质的变化,本研究以尼罗罗非鱼肉为对象,通过高效液相色谱法、氨基酸自动分析法和固相微萃取—气相色谱—质谱联用技术对上海熏鱼4个加工阶段(生鲜罗非鱼、一次浸渍、一次浸渍后油爆和上海熏鱼成品)的风味物质进行分析和鉴定。结果显示,各阶段IMP含量呈逐渐上升趋势,是主要的鲜味核苷酸。游离氨基酸总量和呈味氨基酸含量也逐渐升高,并呈显著性差异,其中天冬氨酸和谷氨酸对上海熏鱼风味影响最大。生鲜罗非鱼、一次浸渍、一次浸渍后油爆和上海熏鱼成品中挥发性物质依次为50、84、78和82种,主要由醛类、酮类、醇类和烃类构成。因此,浸渍和油爆是上海熏鱼风味形成的主要原因,使得罗非鱼鱼体的腥味得以有效改善。     

养殖博氏鱼芒肉营养成分的分析及评价

采用国家标准方法测定湖北英山博氏(鱼芒)肉一般营养成分、氨基酸、脂肪酸和矿物质元素含量.结果表明,博氏(鱼芒)肉中水分、粗蛋白、粗脂肪和粗灰分的含量分别为75.36％±1.07％、17.90％±0.43％、5.13％±0.18％和0.98％±0.09％;18种氨基酸总量为(69.01±0.07) g/100 g(干重),其中5种鲜味氨基酸占氨基酸总量的41.70％,8种人体必需氨基酸占氨基酸总量的48.68％,必需氨基酸与非必需氨基酸比值为95.06％;粗脂肪中含24种脂肪酸,油酸含量38.41％±0.29％最高,单不饱和脂肪酸含量占脂肪酸总量的42.68％±0.10％,n-6多不饱和脂肪酸与n-3多不饱和脂肪酸的比值为1.48;矿物质元素中钾含量最高,为(1 118.51±30.67) mg/100 g(干重),微量元素中,铁、锌含量最高,分别为(1.19±0.10)和(1.18±0.05)mg/100 g(干重).与其它鱼类相比,博氏(鱼芒)肉蛋白质质量较高,氨基酸组成比例均衡,富含n-3多不饱和脂肪酸,矿物质元素含量丰富,有较高的营养价值.     

Bacterial colonization of winter flounder Pseudopleuronectes americanus fed live feed enriched with three different commercial diets

The proliferation of bacteria in intensive aquaculture systems may be responsible for poor growth and mass mortality of marine fish larvae. Essential fatty acids provided in the diet could protect larvae by modulation of the immune response via arachidonic acid (AA) and eicosapentaenoic acid (EPA). Winter flounder Pseudopleuronectes americanus larvae were fed rotifers Brachionus plicatilis enriched with three commercial diets containing different fatty acid profiles. Bacterial colonization on the gills and skin and in the intestinal lumen was evaluated at the end of the rotifer feeding period (day 26), and growth was surveyed until metamorphosis. At 26 days post hatching, larvae fed rotifers containing the higher AA content and with a higher docosahexaenoic acid (DHA) to EPA ratio showed better growth and the lowest bacterial colonization of the intestinal lumen compared to larvae fed rotifers with the lowest AA and DHA : EPA levels. AA had been selectively incorporated into the polar lipids of larvae fed the rotifers enriched with the three diets. This is the first study in winter flounder larvae to report a link between different commercial rotifer enrichments and bacterial density in intestinal lumen.     

The Effects of Live and Artificial Diets on Feeding Performance of Cultured Winter Flounder,Pseudopleuronectes americanus,in the Wild: Survival,Feeding, Growth,and Nucleic Acid Analyses

This research, which is part of a larger study designed to assess the feasibility of winter flounder, Pseudopleuronectes americanus, stock enhancement in New Hampshire, identifies hatchery feeds that optimize feeding‐related performance of fish once released in the wild. Fish reared on post‐nauplii of brine shrimp, Artemia sp., white worms, Enchytraeus albidus, common burrower amphipods, Leptocheirus plumulosus, and formulated pellets were evaluated post‐release from in situ cages using survival, growth rate, feeding onset and incidence, stomach fullness, diet composition, and nucleic acid‐based condition as indicators of hatchery diet suitability. Amphipod‐reared fish had the highest mean stomach content index of all feed types, including wild fish. Wild and worm‐reared fish exhibited the most similar survival, overall stomach fullness, and diet composition profiles over time. Amphipod‐reared fish ranked highest in overall performance; however, if wild fish performance is viewed as the ideal for a stocked fish, worm‐reared fish performed optimally. This study describes hatchery feeding strategies that may ease the transition of flatfish released into the wild for stock enhancement.     

Modeled transport of winter flounder larvae spawned in coastal waters of the Gulf of Maine

Winter flounder, Pseudopleuronectes americanus, from southern New England and the Gulf of Maine were historically considered to be obligate estuarine spawners. However, recent experiments and observations document that winter flounder in the Gulf of Maine also utilize coastal waters for spawning. An individual‐based modeling approach was used to investigate the transport of winter flounder larvae from three hypothesized coastal spawning grounds in the Gulf of Maine. Transport success rates were greatest for larvae released from Ipswich Bay, intermediate for Stellwagen Bank and least successful for those released from Jeffreys Ledge. There was substantial interannual variability in larval transport and geographic patterns of potential connectivity. Furthermore, the date of spawning had an important influence on transport success. Model results suggest that certain coastal spawning grounds used by winter flounder may serve as an important source of larvae to estuaries and nearshore nursery areas. The potential influx of coastal spawned larvae could have implications for the resilience, productivity and gene flow in local populations. Model results provide further support for the conclusion that winter flounder in the Gulf of Maine may not be solely dependent upon estuaries for spawning. Results also suggest that coastal spawning groups should be considered explicitly in the management of winter flounder, and protected under Essential Fish Habitat regulations.     

Gonadal Development and Differentiation in Cultured Juvenile Winter Flounder, Pseudopleuronectes americanus

Winter flounder, Pseudopleuronectes americanus, is currently being evaluated as a stock enhancement candidate in New Hampshire, USA; however, little is known about the gonadal development or the sex ratio of cultured juveniles. To determine the size at gonadal differentiation, 327 cultured fish ranging from <20 to 110 mm total length (TL), in 10‐mm‐TL size classes, were examined histologically. Gonads had differentiated into testes and ovaries in fish ≥41 mm TL (98%), whereas the majority of fish (81%) smaller than 40 mm TL possessed undifferentiated gonads. A total of 313 cultured fish >40 mm TL were analyzed for sex ratio. In 2003, 67 females and 164 males were identified, yielding a sex ratio that was significantly skewed toward male (χ²= 40.7, df = 1, P < 0.001). This trend held true when cultured fish were sorted by age and length, with the exception of those fish 61–70 mm TL. This aberration probably was because of a small sample size in this length category. However, in both the 2004 and the 2005 cultured populations, flounder sex did not deviate from a 1:1 ratio (2004 χ²= 0.12, df = 1, P= 0.724 and 2005 χ²= 0.02, df = 1, P= 0.881). The 2003 data suggest that environmental or genetic factors may affect winter flounder sex determination; rearing manipulation studies in the hatchery are needed to confirm this hypothesis.     

Assessment of sex and evaluation of ovarian maturation of fish using ultrasonography

Ultrasonography is a non‐invasive method that can be used to assess sex and maturity of fish. This study was conducted to examine the feasibility of using ultrasound images to sex juvenile and mature halibut Hippoglossus hippoglossus (Linnaeus), mature winter Pleuronectes americanus (Walbaum) and yellowtail flounder Pleuronectes ferruginea (Storer) and mature haddock Melanogrammus aeglefinus (Linnaeus). In females, both immature and mature ovaries were relatively easy to distinguish, but in males only mature testes were consistently distinguishable. Ovarian maturation of haddock and ovulatory cycles of halibut may also be assessed using ultrasound.     

Improved method of protoplast fusion between marine luminescent bacterium and <Emphasis Type="Italic">Escherichia coli</Emphasis> cultivated with fatty acid

An attempt was made to improve the stability toward centrifugation of protoplast fusion between Shewanella sp. and Escherichia coli. Stability of the cell membrane is an important factor in protoplast fusion. In order to change the fatty acid composition of the cell membrane phospholipids, eight fatty acids [caprylic acid, capric acid, palmitic acid, oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid] were added to each nutrient medium of Shewanella sp. and E. coli. The protoplasts were treated with lysozyme, and fusion occurred in the presence of a polyethylene glycol solution. The stability of the protoplast of Shewanella sp. decreased after EPA was added to the culture medium, and the stability of the protoplast of E. coli increased after the addition of linoleic acid or linolenic acid. Some fusant colonies that developed on the regenerated medium selected for E. coli with antibiotic tolerance. The efficiency of this fusion was higher than that of initial condition using protoplasts from Shewanella sp. and E. coli incubated without fatty acids. Protoplasts improved the fatty acid composition of the phospholipids. Cell membrane stability can change in order for the weak cells to be taken in by strong cells. These results suggested that the fatty acid composition of cell membrane phospholipids affected the fusant yield of the fusion of these bacteria.