Molecular analysis of grass carp (<Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>) by SRAP and SCAR molecular markers

The sequence-related amplified polymorphism (SRAP) technique was used to analyze the gene differentiation between two cultured populations [Freshwater Fisheries Research Center (FFRC) and Qianzhou populations] and one wild population (Hanjian population) of grass carp (Ctenopharyngodon idella). Some loci showed quite different genetic frequencies, attributable to artificial selection, which imply that these fragments are putative markers of germplasm identification. We developed a simple and effective method to further characterize these SRAP fragments. Specific SRAP bands were cut directly from polyacrylamide gels, re-amplified, cloned, and sequenced. Twenty-one putative genetic markers were sequenced, ranging from 137 to 357 bp. The sequences were submitted to the database of the Genome Sequence Survey. A BLAST analysis showed that eight SRAP fragments were highly similar to functional genes, whereas the other 13 had no similarity, indicating that these markers are tightly linked to the germ identification trait although only eight are functional genes. Three primers were designed according to this sequence information and used for PCR amplification of the three populations. A sequence-characterized amplified region (SCAR1) was positively amplified in the artificially cultured populations but not in the wild population. The frequency of the SCAR3 marker in the cultured populations was 87% (26/174), whereas it was only 6% (6/100) in the wild population. A specific band was isolated from all individuals in the wild population with the SCAR3 primers, whereas the specific band was amplified from only seven individuals in the FFRC population and from none of the Qianzhou population. The frequency of SCAR2 in the artificially cultured populations was 96.5%. These results indicate that SCAR1 could be used as a specific molecular marker for population identification. The SCAR markers used in this study offer a powerful, easy, and rapid method for genetic analysis and the discrimination of different populations.     

The effects of tank system,water velocity and water movement on survival,somatic and gonad growth of juvenile and adult green sea urchin,Strongylocentrotus droebachiensis

The effects of water velocity and tipping frequency (water movement) on survival, somatic and gonad growth of juvenile and adult green sea urchin, Strongylocentrotus droebachiensis were investigated. Juvenile and adult urchins were held in ‘laminar’ or ‘tipper’ tanks. Both were supplied with three inlet water flows (1.5, 3.0 and 7.5 L min^?1) which converted to water velocities of 0.28, 0.57 and 1.43 cm s^?1 in the ‘laminar’ tanks and tipping frequencies of 30, 10 and 5 s in the ‘tipper’ tanks. Juvenile sea urchins had significantly lower mortality and greater somatic growth when held in ‘laminar’ flow tanks compared with ‘tipper’ tanks. The varying water velocities tested in the ‘laminar’ flow treatment had no effect on the growth of juvenile sea urchins. The juvenile sea urchins in the ‘tipping’ treatment held at high tipping frequency had significantly slower growth than those in the medium and low tipping frequency treatments. There were no differences in mortality or gonad growth in the adult sea urchins regardless of treatment. The ‘laminar’ flow tanks retained significantly more organic material compared with the ‘tipper’ tanks. The authors discuss the effects of water velocity and tipping frequencies in system design for aquaculture of sea urchins.     

Identification of RAPD‐SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp,Penaeus monodon Fabricius

White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost‐effective strategy for long‐term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10³ fold higher in disease susceptible than that in disease resistant shrimps using real‐time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease‐free shrimp aquaculture industry.     

Seasonal changes in a deep‐water fish assemblage in response to monsoon‐generated upwelling events

The coastal shelf of the Gulf of Oman experiences periodic upwelling events during the summer months that are driven by the southwest monsoon. It is unclear what role these events play in the spatial and temporal distribution of the region’s fish assemblage. We carried out trials on two different video techniques to characterize the habitat and fish assemblage along the continental shelf margin near Muscat, Oman. Exploratory surveys with a drift stereo‐video revealed three main habitat types: Sand, Reef and Megabenthos. Three areas were chosen for additional sampling using stereo‐BRUVS (‘baited’ remote underwater‐video systems). On two separate occasions (November 2005 and March 2006) replicate stereo‐BRUVS were deployed in each area stratified by the main habitat types. For each teleost and elasmobranch species encountered on the video, an estimate of total body length and the relative abundance (MaxN_i) was made. The stereo‐BRUVS recorded a wide range of demersal and pelagic teleosts including species of conservation interest such as sharks, rays and groupers. The drift stereo‐video recorded significantly fewer species than the stereo‐BRUVS (N = 15 versus N = 43). Species diversity from the stereo‐BRUVS increased by 96% in March 2006 (N = 41) compared to November 2005 (N = 23), a pattern consistent at all three areas. The structure of the overall fish assemblage (using canonical analysis of principal coordinates analysis) was highly variable both in time and space. There was ample evidence of strong habitat associations, particularly with depth and seasonal shifts in abundance and diversity. We argue the upward migration of oxygen‐depleted water into the shallow depths during the late monsoon displaces the demersal fish community along this coast.     

Sequence characterized amplified regions (SCAR) reveal candidate markers linked to growth traits in the Chinese shrimp Fenneropenaeus chinensis

Growth traits in 150 Chinese shrimps Fenneropenaeus chinensis were analyzed by the sequence characterized amplified regions (SCAR) technique in two populations of sixth-generation cultured shrimps with different ranges of body lengths (CP-a and CP-b) and in the wild-type population (WP). First, 240 random primers were used to screen polymorphic fragments by random amplified polymorphic DNA (RAPD). Population analysis revealed nine RAPD markers correlated with growth traits, including seven positively and two negatively correlated ones. The sequences of the RAPD markers were then used to design longer primers for SCAR. Six primer pairs were obtained, and two of these produced polymorphic fragments among the three groups. One amplified 39, 26 and 27 polymorphic fragments, with band frequencies of 78, 52 and 54% in the CP-a, CP-b and WP groups, respectively. The distributions of polymorphic fragments in the three groups were significantly different (P < 0.05) according to a χ ² test, indicating that they may be candidate markers linked to growth traits. Another primer pair amplified three alleles, resulting in six combinations of genotypes among three groups. Since allele A₁ was only found in the population of shorter shrimps, it may be a negative growth marker.     

A census of maerl beds in Irish waters

1. Records of maerl beds in Irish waters were compiled from the following sources: Admiralty Charts, published/unpublished ecological/geological accounts, and personal communications/observations. 2. Geographic and depth distributions of maerl records in Irish waters are discussed. Two main areas, Galway Bay–Connemara and South West Ireland, were identified as harbouring the highest concentrations of confirmed records, with maximum depth of confirmed records being approximately 25–30 m. 3. Deposits shown as ‘crl’ markings on older Admiralty Charts may refer to maerl and are situated at greater depths (to 95 m) and are further offshore than confirmed deposits (to 32 m), the latter being based on more recent observations. 4. Although the precise number of maerl beds in Irish waters cannot be assessed from the available records, the minimum number of larger beds (>1 km²), based on confirmed records only, is 35–40 beds located in 18 areas. In addition, along the south‐west coastline numerous smaller spatial aggregations (as small as 1–5 m²) occur. Copyright © 1999 John Wiley & Sons, Ltd.     

Understanding fishing‐induced extinctions in the Amazon

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Could artificial structures such as fish passes facilitate the establishment and spread of the ‘killer shrimp’ Dikerogammarus villosus (Crustacea: Amphipoda) in river systems?

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Effects of feeding frequency and dietary protein levels on juvenile allogynogenetic gibel carp (Carassius auratus gibelio) var. CAS III: growth,feed utilization and serum free essential amino acids dynamics

A 8 week growth trial of three feeding frequencies (2, 4 and 6 meals day^?1) and three dietary protein levels (30%, 34% and 38%) was conducted to investigate the influence of feeding frequency and dietary protein level on growth, feed utilization, serum free essential amino acids (EAA) dynamics and nitrogen (N), phosphorus (P), carbon (C) retention in juvenile allogynogenetic gibel carp (Carassius auratus gibelio) ‘CAS III’. The results showed that feeding frequency and dietary protein level increased feed intake (FI), specific growth rate (SGR) and feed efficiency (FE). SGR increased significantly with the increase in feeding frequency and dietary protein level. But, the increase in dietary protein level from 30% to 38% in group 6 meals day^?1 did not result in better growth. Compared with the group of 4 meals day^?1 × 38%, the fish fed the diet containing 30% crude protein at 6 meals day^?1 obtained similar SGR but showed lower FIp (protein intake). Compared with 2 meals day^?1, NREs (nitrogen retention efficiency) of 4 and 6 meals day^?1 were relatively higher. PRE (phosphorus retention efficiency) and CRE (carbon retention efficiency) increased significantly with feeding frequency while TNW (total nitrogen waste output), TPW (total phosphorus waste output) and TCW (total carbon waste output) decreased significantly. Feeding frequency and dietary protein level increased trypsin activity. Serum free EAA concentrations varied more synchronously at 6 meals day^?1 than 2 meals day^?1. It could be concluded that increasing feeding frequency could improve growth by increasing feed intake and feed efficiency for juvenile gibel carp. The increase in feeding frequency showed protein‐sparing effect.     

Differences in stable isotope ratios of Dall’s porpoises (Phocoenoides dalli) between coastal and oceanic areas of the North Pacific

Carbon and nitrogen stable isotope ratios in the muscle of Dall’s porpoises were measured. Samples were collected from the catches of the hand harpoon fishery, incidental catches of drift net, and scientific research on the use of drift nets. Samples were from the North Pacific, Sea of Japan, Sea of Okhotsk, and Bering Sea. Although no variation in δ¹⁵N was observed, δ¹³C was significantly different between population groups near Japan and the oceanic North Pacific and Bering Sea. The difference may be due not only to local variation in prey species, but also to an overall difference in carbon stable isotope ratios that originate from coastal benthic or oceanic pelagic‐based food webs. We differentiated Dall’s porpoise population groups from both areas using carbon stable isotope ratios with an error rate of <5%. Although further study is needed, our results suggest that carbon stable isotope ratios could possibly be an indicator of whether a Dall’s porpoise belongs to a coastal benthic or oceanic pelagic food web.     

Tissue and organ distribution of 14C‐activity in dextrin‐adapted Atlantic salmon after oral administration of radiolabelled 14C1‐glucose

Accumulation of ¹⁴C in various tissues and organs was studied in three different groups of 0.8‐kg Atlantic salmon Salmo salar force‐fed with ¹⁴C₁‐glucose in order to evaluate if metabolism of glucose depended on adaptation to dietary carbohydrate level. The salmon had been fed diets supplemented with 0, 100 and 200 g maize dextrin kg^?1 for 10 months before the experiment. The fish were force‐fed 6.65 × 10⁴ Bq of ¹⁴C₁ glucose kg^?1 BW, in gelatin capsules. Fish for analysis were obtained 16 h later. ¹⁴C was measured in blood plasma, gill, kidney, liver and white muscle, and in lipid extract of liver. The liver contained most ¹⁴C, followed by heart, blood plasma, gill and liver lipid extract, while kidney and muscle contained the least ¹⁴C per gram or millilitre tissue. The muscle contained most radioactivity, on an estimated total tissue basis, followed by liver, blood plasma, gill, liver lipid extract, kidney and heart tissue. Thirty‐eight per cent of the orally administered ¹⁴C was recovered in the salmon adapted to the diet without dextrin after 16 h. This was significantly (P < 0.05) higher than the 30% and 32% recovered in the salmon adapted to diets with 10% and 20% dextrin. This effect on adaptation to dietary dextrin level in glucose uptake or metabolism was supported by a trend (P < 0.10) toward higher radioactivity per gram or millilitre of each individual tissue in the fish adapted to the diet without dextrin, when compared with the other two adaptation regimes.     

Evidence for genetic distinction among sympatric ecotypes of Arctic char (Salvelinus alpinus) in south‐western Alaskan lakes

Resource polymorphism may play an important role in the process of speciation. The Arctic char (Salvelinus alpinus) exhibits great phenotypic and genetic diversity across its range, making it an ideal species for studies of resource polymorphism and divergence. Here, we investigated genetic variation at 11 microsatellite loci among 287 Arctic char from five isolated yet proximate postglacial lakes in south‐western Alaska that were previously examined for resource polymorphism. Significant differences in pairwise F_ST were detected among all lakes (range from 0.05 to 0.28, all P < 0.02). In one lake (Lower Tazimina Lake), we found evidence for two genetic groups of char and for significant differences in the distribution of microsatellite variability among at least two of the three previously described body size morphotypes (‘large’‐, ‘medium’‐, and ‘small’‐bodied char; maximum F_ST = 0.09; differences in admixture proportions). We also found a significant association between genetic admixture proportions and gill raker counts among body size morphs (r = ?0.73, P < 0.001). Our data represent the first record of genetically distinct sympatric morphs of Arctic char in Alaska and provide further evidence that differences in morphology associated with feeding (gill rakers) and growth trajectories reflect niche diversification and promote genetic divergence in Holarctic populations of Arctic char.     

牙鲆抗淋巴囊肿病相关SSR标记的筛选与鉴定

为了筛选出中国牙鲆群体的抗淋巴囊肿病相关SSR标记,实验采用分群分析法对102个牙鲆个体(感病56个,抗病46个)进行了抗淋巴囊肿病相关SSR标记的筛选。首先构建了抗、感基因池(抗病个体和感病个体各15个),并利用178对微卫星引物对其进行了扫描;对于在抗、感池间扩增出差异条带的引物,用构建基因池的30个个体对其进行了第一次单个体验证;对于在第一次单个体验证中差异显著的引物用102个个体进行了第二次验证。结果显示,有4对引物(scaffold440_22585、scaffold826_5003、scaffold703_4284和scaffold185_597)在抗、感基因池间扩增出了差异条带;经第一次单个体验证,表明scaffold826_5003(P=0.023)和scaffold185_597(P178bp=0.028,P173bp=0.009)的差异条带在抗、感个体中呈显著性差异;经第二次单个体验证,发现scaffold185_597的差异条带在抗、感个体中出现的频率分别为60.9%和14.3%(P=0.001),差异显著;对scaffold185_597在10个抗病个体中扩增出的差异条带进行了克隆并测序,经BLAST比对,证实其为黄海水产研究所水产基因组与细胞工程研究室公布的牙鲆微卫星标记scaffold185_597中的一段,同源性达到96%。研究表明,微卫星标记scaffold185_597可能与牙鲆抗淋巴囊肿病相关。     

Assigning origins in a potentially mixed‐stock recreational sea trout ( Salmo trutta) fishery

The anadromous, or sea‐going, life history form of brown trout, or sea trout ( Salmo trutta), may lead to potential mixing of populations while foraging at sea. In this article, we assess the potential that multiple populations are using common semi‐enclosed estuaries and quantify the potential levels of straying (i.e. dispersal) of foreign‐produced individuals into populations by using otolith chemical signatures as natural ‘tags’. To do so, we created a database of juvenile fish otolith chemistry (a marker of freshwater production) from four rivers and compared the chemistry of harvested fish in two estuaries important to anglers, the Renews River and Chance Cove Brook, to the database. A discriminant function analysis revealed significant differences in the otolith chemistry of juvenile fish inhabiting the four rivers with a 97% cross‐validated accuracy when classifying individual juveniles to their natal river, indicating our baseline was robust. When assigning adults caught over 3 years (2007–2009) in the recreational fishery in the Renews River estuary, it was determined that over 95% of the fish caught each year originated from Renews River. In contrast, harvested fish in Chance Cove during 2009 were disproportionately comprised of fish produced in Renews River, suggesting the potential for source‐sink population dynamics in Newfoundland. Taken as a whole, these results indicate limited population mixing in nearshore estuaries of this region, but also highlight the potential for some populations to subsidise the harvest by anglers in different areas.     

Managing European cormorant‐fisheries conflicts: problems,practicalities and policy

This article focuses on transboundary conflicts between great cormorants, Phalacrocorax carbo L., and European fisheries interests. Two races of this species commonly occur in Europe; the ‘Atlantic’ Phalacrocorax carbo carbo and the ‘Continental’ P. c. sinensis. This migratory fish‐eating bird has steadily expanded its European range and many consider this a nature conservation success. However, fishery representatives see the birds as harmful to their business and way of life, considering European Union (EU) legislation to be blocking the most intuitive and effective solution to the cormorant problem, namely co‐ordinated, pan‐European population reduction. This article argues that cormorant‐fisheries conflicts and their resolution and management cannot be characterised as, or reduced to, a simplistic ‘pan‐European’ vs ‘local’ argument. The nature of cormorant‐fisheries conflicts is explored, key issues of cormorant ecology examined, and consideration is given to how impacts on fisheries might be measured. Discussions then focus on whether current policy prevents balanced and constructive solutions or whether better use of available instruments at local‐national levels could actually be a vital step to a sustainable management solution.     

Pigmentation,carotenoids, lipid peroxides and lipid composition of red porgy (Pagrus pagrus) skin reared under open‐cage conditions

Pigmentation capability of red porgy (Pagrus pagrus) skin reared under open sea‐cage conditions and fed an astaxanthin‐enriched diet was studied. Skin lipid peroxide levels and lipid composition were also evaluated to establish the antioxidant role of astaxanthin under these sunlight‐exposure conditions. Fish placed either in an offshore sea cage system (SC) or in an inland tank facility (T) housed inside a ‘shade‐house’ enclosure were fed a commercial diet supplemented with 22 mg kg^?1 astaxanthin. No differences in growth or survival were found. Both groups displayed a red skin, but SC fish presented a darker pigmentation, which externally reflected the higher deposition of melanin, astaxanthin and tunaxanthin found in its skin. The lower level of lipid peroxides found in SC fish might be related with the higher level of astaxanthin mentioned above. Nevertheless, lipid and fatty acid profiles did not show significant differences between groups. Our results indicate that sustainable production of red porgy with a natural red hue is possible on the basis of proper adjustment of two factors illumination and dietary astaxanthin.     

Predicting population status of freshwater pearl mussel (Margaritifera margaritifera,L.) in central Sweden using instream and riparian zone land‐use data

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Effect of dietary carbohydrate‐to‐lipid ratios on growth performance,body composition,nutrient utilization and hepatic enzymes activities of herbivorous grass carp (Ctenopharyngodon idella)

Six isonitrogenous (390 g kg^?1) and isoenergetic (16.2 kJ g^?1) diets with varying carbohydrate : lipid (CHO : L) ratios (202.5–1.74), were fed to triplicate groups of 25 fish in indoor recirculation system. Over 8‐week‐growth trial, best weight gain (WG), specific growth rate, feed conversion ratio, protein efficiency ratio and protein production value (P < 0.05) were observed in fish‐fed diets with CHO : L ratio of 7.5. Fish fed either the lowest (1.7) or highest (202.5) CHO : L ratio tended to produce lower (P < 0.05) growth and feed conversion efficiencies. The values of viscerosomatic index, hepatosomatic index and intraperitoneal fat ratio increased as dietary CHO : L ratios decreased. There were no significant differences in whole body and liver crude protein among dietary treatments. Whole body and liver lipid increased as CHO : L ratios decreased. Plasma cholesterol and triacylglyceride levels increased linearly as dietary CHO : L ratios decreased. Activities of glucokinase and pyruvate kinase were stimulated by elevated levels of dietary carbohydrate; however, activities of lipase (LPS) and alkaline phosphatase were stimulated by elevated levels of dietary lipid. Based on a second‐order polynomial regression analysis of WG against dietary carbohydrate and lipid levels, 275 g kg^?1 of carbohydrate and 59 g kg^?1 of lipid, corresponding to a CHO : L ratio of 4.7, in a diet holding 390 g kg^?1 of crude protein and 16.3 kJ g^?1 of gross energy, proved to be optimal for grass carp. These results indicated that utilization of dietary lipid and carbohydrate was moderate in grass carp, but the fish were a little more capable of utilizing lipid compared with carbohydrate.     

Low levels of Aflatoxin B1 could cause mortalities in juvenile hybrid sturgeon,Acipenser ruthenus ♂×A. baeri♀

Toxicity of aflatoxin B₁ (AFB₁) was investigated in juvenile hybrid sturgeon Acipenser ruthenus ♂ × A. baeri♀, an important coldwater finfish farmed in China and other countries. Seven experimental diets (Diet A–G) containing different levels of AFB₁ (0, 1, 5, 10, 20, 40 and 80 μg kg^?1 diet) were fed to juvenile sturgeon weighing 10.53 ± 0.17 g kg^?1 to determine its effect on survival, growth, feed consumption, hematocrit, liver histology as well as muscular and hepatic toxin accumulation. The experiment lasted for 35 days and was conducted in two periods of 25 and 10 days each. No external changes or unusual behaviour was observed in the fish fed diets with AFB₁. Mortality was observed in fish fed with highest levels of AFB₁ (80 μg kg^?1– Diet G) from day 12 onwards. After 25 days, fish fed the diet of 80 μg AFB₁ kg^?1 showed significant lower survival (50 ± 5.77%) followed by those fed 40 μg AFB₁ kg^?1 diet (80 ± 5.77%) and 20 μg AFB₁ kg^?1 diet (86.66 ± 3.33%). No significant difference was observed in specific growth rate (SGR) or hepatosomatic index (HSI) between groups. Hematocrit was significantly higher in the fish fed the diet of highest AFB₁. The fish were weighed at day 25 in some treatments (Diets F and G) because of high mortality. However, feeding was continued for another 10 days to observe mortality or behavioural changes if any in the other groups. After 35 days, survival in the fish fed Diet F (40 μg AFB₁ kg^?1) was 40% and those fed Diet E (20 μg AFB₁ kg^?1) was 36.2%. Significant histopathological changes including nuclear hypertrophy, hyperchromasia, extensive biliary hyperplasia, focal hepatocyte necrosis and presence of inflammatory cells were observed in the liver of fish fed high levels of aflatoxin (40 and 80 μg kg^?1). AFB₁ accumulation in fish muscle and liver increased with increased dietary AFB₁ levels. It could be confirmed that 10 μg AFB₁ kg^?1 diet was the maximum allowable level in hybrid sturgeon diet.