Reproductive biology,early ontogeny,and effect of temperature on development in wolffish: comparison with salmon

Peculiarities of reproduction, early ontogeny, methods of egg incubation, and influence of temperature on development are compared in wolffish (Anarhichas lupus L.) and in salmonids, mainly Atlantic salmon (Salmo salar L.). Reproductive indices of females are similar in both species, but differ in males due to the small production of sperm in wolffish, a difference explained by internal fertilization in the latter species. The rate of embryonal development until the beginning of differentiation of fin fold at the same temperature is similar in both species, but the period from activation to hatching is more than twice as long in wolffish as in Atlantic salmon. Wolffish hatch at a more advanced stage with a small remnant of yolk sac and begin to feed shortly thereafter. As in Atlantic salmon, eggs and larvae of wolffish have a wide range of temperature resistance except before and after hatching. Based on the difference of life cycles in wolffish and salmon, technologies for their artificial breeding are compared.     

大黄鱼高温适应的转录组学分析

为了探究大黄鱼高温胁迫条件下基因表达水平的变化,实验利用Illumina Hiseq 2500的125 pair-ended测序模式分别对大黄鱼高温处理组和常温对照组进行了转录组测序。对照组(3个生物学重复)和高温处理组(3个生物学重复)分别获得15.28和13.92 Gb测序数据,GC含量平均值约为51%。过滤后的高质量测序reads使用Bowtie2软件比对到大黄鱼参考转录组序列上估计基因表达量,进而进行基因表达差异统计学检验。以常温对照组为参比,高温处理组中阈值设为|log2(FC)|2和FDR0.05,共检测到1 259条显著差异表达基因。其中,821条基因为高表达,438条基因为低表达。随机选取12条差异表达基因,运用实时荧光定量PCR(qRT-PCR)进行了验证,结果证实转录组分析可靠。进一步将所有差异表达的基因进行GO功能注释和KEGG通路富集分析,结果发现大量差异表达基因的功能与氧化还原反应、蛋白质折叠和去折叠、糖和脂代谢以及某些疾病的发生相关。该研究结果为下一步深入研究大黄鱼高温适应的调控机制提供了重要的参考材料。     

Anisakid larva in the viscera of a farmed Atlantic salmon (Salmo salar)

Parasitic nematodes of the family Anisakidae occur in the visceral cavity and surrounding tissues of many marine fish species at a prevalence as high as 100% in wild salmon samples. Human consumption of fish products containing these parasites can result in the zoonotic disease anisakiasis, and Anisakis simplex is most commonly associated with human disease. Previous studies of farmed salmon have found no anisakids in viscera or muscle, presumably because formulated feed production renders parasite larvae nonviable. However, among farmed Atlantic salmon (Salmo salar) examined by histopathology as part of a provincial government auditing program in British Columbia, Canada, 1 of 894 (0.11%) had an anisakid larva partly embedded in the wall of an intestinal cecum. Skeletal muscle is not examined as part of the government's program, but other studies have correlated anisakids in the viscera and muscle. Using anisakid prevalence in viscera as an estimate of its prevalence in muscle, the risk ratio of anisakid parasites in commercial product is 570 times less in farmed than in wild Atlantic salmon.     

Effects of temperature on amoebic gill disease development: Does it play a role?

A relationship between increasing water temperature and amoebic gill disease (AGD) prevalence in Atlantic salmon (Salmo salar) has been noted at fish farms in numerous countries. In Scotland (UK), temperatures above 12°C are considered to be an important risk factor for AGD outbreaks. Thus, the purpose of this study was to test for the presence of an association between temperature and variation in the severity of AGD in Atlantic salmon at 10 and 15°C. The results showed an association between temperature and variation in AGD severity in salmon from analysis of histopathology and Paramoeba perurans load, reflecting an earlier and stronger infection post‐amoebae exposure at the higher temperature. While no significant difference between the two temperature treatment groups was found in plasma cortisol levels, both glucose and lactate levels increased when gill pathology was evident at both temperatures. Expression analysis of immune‐ and stress‐related genes showed more modulation in gills than in head kidney, revealing an organ‐specific response and an interplay between temperature and infection. In conclusion, temperature may not only affect the host response, but perhaps also favour higher attachment/growth capacity of the amoebae as seen with the earlier and stronger P. perurans infection at 15°C.     

Serum enolase: a non‐destructive biomarker of white skeletal myopathy during pancreas disease (PD) in Atlantic salmon Salmo salar L.

Diseases which cause skeletal muscle myopathy are some of the most economically damaging diseases in Atlantic salmon, Salmo salar L., aquaculture. Despite this, there are limited means of assessing fish health non‐destructively. Previous investigation of the serum proteome of Atlantic salmon, Salmo salar L., during pancreas disease (PD) has identified proteins in serum that have potential as biomarkers of the disease. Amongst these proteins, the enzyme enolase was selected as the most viable for use as a biomarker of muscle myopathy associated with PD. Western blot and immunoassay (ELISA) validated enolase as a biomarker for PD, whilst immunohistochemistry identified white muscle as the source of enolase. Enolase was shown to be a specific marker for white muscle myopathy in salmon, rising in serum concentration significantly correlating with pathological damage to the tissue.     

Susceptibility of corkwing wrasse Symphodus melops, goldsinny wrasse Ctenolabrus rupestis, and Atlantic salmon Salmo salar smolt, to experimental challenge with Vibrio tapetis and Vibrio splendidus isolated from corkwing wrasse

The virulence of two Vibrio strains, previously isolated from diseased corkwing wrasse Symphodus melops and identified as V. tapetis and V. splendidus, to corkwing and goldsinny wrasse Ctenolabrus rupestris and to Atlantic salmon Salmo salar, was studied under laboratory conditions. Both bacteria were shown to be opportunistically pathogenic to corkwing wrasse, causing significantly higher mortality in the challenged groups than in the controls. Bacterial cultivation of kidney samples and re-isolation of V. tapetis and V. splendidus from most mortalities confirmed the two strains as the probable cause of mortality in the challenged groups. The control group also suffered relatively high mortality, but no specific pathogens that were suspected to be the main cause of death were isolated, other than a mixture of Vibrio spp. and, in the case of one individual, atypical Aeromonas salmonicida. Following injection challenge with both bacterial strains, no mortality was recorded in Atlantic salmon. In bath challenge trials with goldsinny wrasse, only slight mortality was observed in the challenged groups and the unchallenged control group. Bacterial examination showed that atypical Aeromonas salmonicida was the probable cause of death in both bath challenged and control groups of goldsinny wrasse, and no indication of infection by any Vibrio sp. was found.     

Modelling oxygen consumption rates of post-smolt Atlantic salmon in commercial-scale,land-based farms

Oxygen consumption rates of post-smolt Atlantic salmon (Salmo salar L.) were recorded during various growth trials at four land-based salmon farms in south-west Norway, during the period 1988–1992. Principle component analysis indicated that fish size, water temperature and current speed had the greatest influence on oxygen consumption of fish in all the farms. A multiple regression analysis showed that 70% of the variation in the oxygen consumption could be explained by these three parameters. The predicted effect of fish size, water temperature and current speed on oxygen consumption are consistent with those reported in studies conducted on salmonids held in tunnel respirometers. The model described in this study may be a useful tool for the estimation of oxygen and water requirements needed for the design and operation of land-based farms.     

Mycobacterium salmoniphilum infection in farmed Atlantic salmon, Salmo salar L

Multiple greyish‐white visceral nodules containing abundant rapidly growing and acid‐fast bacteria, subsequently identified as Mycobacterium salmoniphilum, were detected in moribund and newly dead market‐sized fish during a period of increased mortality in an Atlantic salmon, Salmo salar, farm in western Norway. Isolates cultured from diseased fish were phenotypically consistent with Mycobacterium sp. previously isolated from Atlantic salmon [MT 1890 (= NCIMB13533), MT1892, MT1900 and MT1901] in the Shetland Isles, Scotland. Partial sequences of 16S rDNA, ribosomal RNA internal transcribed spacer (ITS1), 65‐kDa heat‐shock protein (Hsp65) and β subunit of RNA polymerase (rpoB) revealed 97‐99% similarity with M. salmoniphilum type strain ATCC 13758^T. The source of infection was not confirmed. Koch’s postulates were fulfilled following experimental challenge of Atlantic salmon with field isolate NVI6598 ( FJ616988 ). Mortality was recorded in experimentally infected fish; however, the infection remained subclinical in the majority of affected fish over the 131‐day challenge period.     

Development of a Piscirickettsia salmonis immersion challenge model to investigate the comparative susceptibility of three salmon species

Piscirickettsia salmonis, the aetiological agent of salmonid rickettsial septicaemia (SRS), is a global pathogen of wild and cultured marine salmonids. Here, we describe the development and application of a reproducible, standardized immersion challenge model to induce clinical SRS in juvenile pink (Oncorhynchus gorbuscha), Atlantic (Salmo salar) and sockeye salmon (O. nerka). Following a 1‐hr immersion in 10⁵ colony‐forming units/ml, cumulative mortality in Atlantic salmon was 63.2% while mortality in sockeye salmon was 10%. Prevalence and levels of the bacterium in kidney prior to onset of mortality were lower in sockeye compared with Atlantic or pink salmon. The timing and magnitude of bacterial shedding were estimated from water samples collected during the exposure trials. Shedding was estimated to be 82‐fold higher in Atlantic salmon as compared to sockeye salmon and peaked in the Atlantic salmon trial at 36 d post‐immersion. These data suggest sockeye salmon are less susceptible to P. salmonis than Atlantic or pink salmon. Finally, skin lesions were observed on infected fish during all trials, often in the absence of detectable infection in kidney. As a result, we hypothesize that skin is the primary point of entry for P. salmonis during the immersion challenge.     

Early morphological predictors of maturity in one-sea-winter Atlantic salmon

Early separation of maturing from immature fish would be very valuable in Atlantic salmon (Salmo salar) culture. Possible morphometric predictors of maturation in one sea-winter Atlantic salmon were investigated from measurements of body weight, length and depth and adipose fin length undertaken at 14 day intervals from May until August in a population of 80 individually tagged fish, 61 (male and female) of which were subsequently found to be maturing (i.e. grilse). Stepwise discriminant analysis revealed that a combination of forklength and condition factor proved to be the strongest and most consistent predictor of maturation (over 88% accurate) until August. In August when maturing fish could be easily distinguished by shape and colour, forklength and condition factor no longer had any discriminatory power, but body depth at the anal and dorsal fins were good predictors. A second trial was performed on a separate stock of fish for which the same predictors proved robust. The discriminant functions derived from one stock of fish did not, however, produce the same discriminatory power when applied to the other stock. Hence there does not appear to be a simple mathematical means for the general discrimination of maturing and immature salmon. However the morphological predictors described above could be used for visual discrimination when ‘grilse grading’.     

Investigating the threat of non‐native North American signal crayfish (Pacifastacus leniusculus) to salmon redds

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Size heterogeneity can reduce aggression and promote growth in Atlantic salmon parr

Aggression in groups of 0+ Atlantic salmon (Salmo salar) was monitoredat weekly intervals in two tanks containing 100 fish each. Three 1 + salmonparr were added to one of these. After 5 weeks, fish weights were measuredin both tanks and the conditions reversed. At ten weeks, weights of fish inboth tanks were measured again. In both populations, levels of aggressionamong the smaller fish were significantly lower and growth ratessignificantly higher when the large fish were present. Although the largefish attacked the small ones, the rate at which they did so was an order ofmagnitude lower that the rate at which small fish attacked each other in theabsence of larger conspecifics. This raises the possibility that levels ofaggression among farmed salmon might be reduced by the addition of a fewlarge conspecifics.     

Optimisation of gene expression analysis in Atlantic salmon lenses by refining sampling strategy and tissue storage

Analysis of gene expression in the lens is one of the analytical tools employed to investigate cataract formation in Atlantic salmon (Salmo salar L.). High quality RNA preparations are an essential prerequisite for gene expression analysis. The first aim of the present study was to investigate the possible effects of two methods of tissue preservation on the quality of RNA extracted from Atlantic salmon lenses. RNA was extracted from lenses either stored in RNAlater or flash-frozen in liquid nitrogen. Both tissue preservation methods yielded RNA of similarly high quality. The second aim was to examine if stress related to fish handling and the choice of anaesthesia during the sampling procedure affected gene expression in the lens. Six sampling procedures were tested on groups of sea water adapted Atlantic salmon smolt. Fish were either killed instantaneously (control group) or sampled after 30 min anaesthetised with isoeugenol, after 30 min without anaesthesia, after 120 min anaesthetised with isoeugenol, after 120 min with 15 min anaesthesia with metacaine or after 120 min without anaesthesia. The expression levels of specific genes, of special interest in the study of molecular mechanisms of cataractogenesis, were analysed in lenses by real-time RT–PCR. Fish not anaesthetised had significantly increased levels of heat shock protein 70 (HSP70) mRNA after 30 min compared to the control group. Glutathione reductase (GR) and manganese superoxide dismutase (Mn-SOD) were expressed at significantly lower levels in groups of Atlantic salmon sampled after 120 min anaesthetised with isoeugenol or metacaine, and anaesthetised with isoeugenol, metacaine or without anaesthesia, respectively. The same expression patterns were found in corresponding gill tissues for these two antioxidant genes. In conclusion, preservation in liquid nitrogen instead of RNAlater is recommended due to practical conditions in RNA extraction. A quick sampling protocol with the use of anaesthetics and not exceeding 30 min should be preferred to avoid effects of the sampling procedure on lens gene expression in Atlantic salmon.     

An outbreak of acute disease and mortality in Atlantic salmon (Salmo salar) post‐smolts in Norway caused by Tenacibaculum dicentrarchi

An outbreak of disease characterized by skin ulcers, fin rot and mortality was observed a few days after the transfer of Atlantic salmon (Salmo salar) from a freshwater smolt production facility to a land‐based seawater post‐smolt site. Dead and moribund fish had severe skin and muscle ulcers, often 2–6 cm wide, particularly caudal to the pectoral fins. Microscopic examination of smears from ulcers and head kidney identified long, slender Gram‐negative rods. Histopathological analysis revealed abundance of long, slender Tenacibaculum‐like bacteria in ulcers and affected fins. Genetic characterization using multilocus sequence analysis (MLSA) of seven housekeeping genes, including atpA, dnaK, glyA, gyrB, infB, rlmN and tgt, revealed that the isolates obtained during the outbreak were all clustered with the Tenacibaculum dicentrarchi‐type strain (USC39/09^T) from Spain. Two bath challenge experiments with Atlantic salmon and an isolate of T. dicentrarchi from the outbreak were performed. No disease or mortality was observed in the first trial. In the second trial with a higher challenge dose of T. dicentrarchi and longer challenge time, we got 100% mortality within 48 hr. This is the first reported outbreak of disease caused by T. dicentrarchi in Norwegian farmed Atlantic salmon.     

Threshold values of river discharge and temperature for anglers' catch of Atlantic salmon,Salmo salar L.

Abstract The catch by anglers of adult Atlantic salmon, Salmo salar L., was studied over a 5-year period in the River Gaula, Norway. Atlantic salmon were caught over a wide range (23–570 m³ s^?1) of the observed extent of river discharge (13–950 m³ s^?1) and throughout the range of temperature (4–23 °C), but both factors strongly affected catch rate. Significant correlations between the number of Atlantic salmon caught daily, and water temperature (r = 0.33) and river discharge (r = ?0.42) were found in 1987 and 1989, respectively. The highest daily catch occurred between 50 and 150 m³ s^?1, and at temperatures between 13 and 16 °C. Threshold values for water discharge and temperature were found to exist at 250 m³ s^?1 and 8 °C, with the highest catches below and above these values, respectively.     

大西洋鲑、三倍体虹鳟和金鳟的肌肉营养成分与品质特性

为了解大西洋鲑(Salmo salar)、三倍体虹鳟(Oncorhynchus mykiss)、金鳟(Oncorhynchus mykiss)3种鱼肌肉营养成分和品质特性,利用生化分析、物性分析方法分析3种鱼肌肉的营养成分、氨基酸和脂肪酸组成、肉色、系水力和质构特性。结果表明,大西洋鲑、三倍体虹鳟、金鳟肌肉的水分质量分数分别为62.91%、67.15%、73.02%,粗蛋白质量分数分别为22.39%、21.03%、22.11%,粗脂肪质量分数分别为14.64%、17.16%、5.11%。3种鱼肌肉的滴水损失、黄色值(b~*)、羟脯氨酸含量、内聚性均显著不差异(P0.05)。3种鱼肌肉的硬度和咀嚼性由低到高依次为大西洋鲑、三倍体虹鳟、金鳟,而pH值的结果则与之相反(P0.05)。大西洋鲑和三倍体虹鳟肌肉的灰分、解冻损失、蒸煮损失、回复性、弹性和红色值(a~*)差异不显著(P0.05),但灰分、蒸煮损失、回复性均小于金鳟肌肉的对应指标(P0.05),弹性和红色值(a~*)则均大于金鳟肌肉的对应指标(P0.05)。大西洋鲑、三倍体虹鳟、金鳟肌肉中必需氨基酸含量占氨基酸总量分别为42.28%、41.84%、41.63%(质量分数),必需氨基酸/非必需氨基酸(EAA/NEAA)比值分别为73.25%、71.94%、71.32%,均符合联合国粮农组织/世界卫生组织(FAO/WHO)对优质蛋白质的评价标准;3种鱼肌肉中均检测到22种脂肪酸,组成丰富,其中不饱和脂肪酸含量较高。综上所述,3种鱼的肌肉都是符合人体营养需求的优质水产品,其中大西洋鲑和三倍体虹鳟肉质接近,且都优于金鳟的肉质。     

Reproductive success of Atlantic salmon (Salmo salar) mature male parr in a small river,the Nivelle: influence of shelters

Grimardias D, Merchermek N, Manicki A, Garnier J, Gaudin P, Jarry M, Beall E. Reproductive success of Atlantic salmon (Salmo salar) mature male parr in a small river, the Nivelle: influence of shelters.
Ecology of Freshwater Fish 2010: 19: 510–519. © 2010 John Wiley & Sons A/S Abstract – The breeding activity of a small population of Atlantic salmon (Salmo salar L.) was monitored on the Nivelle River in southwest France to estimate the overall contribution of mature male parr and the numbers involved in egg fertilisation. In the twelve redds sampled, 563 eggs were collected for parental assignment and the physical habitat was characterised to investigate the possible relationship between parr reproductive success and habitat complexity. The overall contribution of mature male parr was particularly high (87%, one of the highest estimates ever reported). Concerning habitat complexity, granulometry and shelter diversity were correlated with the number of parr breeders contributing to egg fertilisation in each redd. A complex habitat increased multiple paternity, which could affect the effective size and genetic variability of small salmon populations.     

花鲈ncc、nkcc基因在海、淡水适应中鳃组织的表达与定位分析

为探究ncc、nkcc基因在花鲈渗透调节中发挥的作用,实验通过全基因组鉴定、多重序列比对、系统进化树构建以及蛋白结构预测对花鲈ncc进行了鉴定及序列分析,利用实时荧光定量PCR (qRT-PCR)检测ncc和nkcc在海水、淡水花鲈鳃组织中的表达水平,利用原位杂交技术确定ncc2和nkcc1a在海水及淡水花鲈鳃中的表达位置。结果显示,从花鲈中鉴定出2个ncc基因,即ncc1和ncc2,其编码序列(CDS)长度分别为2 691和3 120bp,编码896和1 039个氨基酸,在进化上具有保守性。ncc2在淡水花鲈鳃组织中的表达量显著高于海水,而nkcc1a在海水花鲈鳃组织中的表达量显著高于淡水,ncc1、nkcc1b、nkcc2在海淡水中的表达量则无显著差异。淡水适应过程中花鲈鳃组织中的ncc2的表达量逐渐上调,而nkcc1a的表达量逐渐下调;海水适应过程则呈现相反的表达趋势。此外,原位杂交结果显示,ncc2和nkcc1a基因分别位于淡水与海水中鳃组织的相邻鳃小片间的鳃丝上皮。以上结果表明,ncc2和nkcc1a基因分别编码淡水及海水花鲈鳃中重要的Na⁺及Cl     

Aeromonas salmonicida infects Atlantic salmon (Salmo salar) erythrocytes

Aeromonas salmonicida subsp. salmonicida (hereafter A. salmonicida) is the aetiological agent of furunculosis in marine and freshwater fish. Once A. salmonicida invade the fish host through skin, gut or gills, it spreads and colonizes the head kidney, liver, spleen and brain. A. salmonicida infects leucocytes and exhibits an extracellular phase in the blood of the host; however, it is unknown whether A. salmonicida have an intraerythrocytic phase. Here, we evaluate whether A. salmonicida infects Atlantic salmon (Salmo salar) erythrocytes in vitro and in vivo. A. salmonicida did not kill primary S. salar erythrocytes, even in the presence of high bacterial loads, but A. salmonicida invaded the S. salar erythrocytes in the absence of evident haemolysis. Naïve Atlantic salmon smolts intraperitoneally infected with A. salmonicida showed bacteraemia 5 days post‐infection and the presence of intraerythrocytic A. salmonicida. Our results reveal a novel intraerythrocytic phase during A. salmonicida infection.     

Identification of differential broodstock contribution affecting genetic variability in hatchery stocks of Atlantic salmon (Salmo salar)

Supportive breeding of Atlantic salmon (Salmo salar) is commonly employed to maintain numbers of fish where the species has become locally endangered. Increasingly, one of the main aims of population management is the preservation of natural genetic diversity. If the stocks employed in supportive breeding exhibit reduced variation they can alter the natural pattern of genetic variation observed in wild populations. In northern Spain, wild adult salmon are caught every year from local rivers and artificially crossed in order to create supportive stocks. The offspring are hatchery reared until the juvenile stage, then released into the same river where their parents were caught. In the current study, our findings demonstrate that although adult broodstock exhibit a pattern of variation similar to the wild populations, variability at microsatellite loci was drastically reduced in the juveniles released into one of three rivers analyzed. The contribution of broodstock to this juvenile stock was examined by pedigree analysis. A restricted number of females contributing to the hatchery stock was identified as the main cause of loss in genetic variation, possibly due to overmaturity of some multi-sea-winter females. We suggest that better monitoring and control of parental contribution will help in solving the problem of loss of genetic diversity in hatchery populations.