Changes in the proximate composition of juvenile white suckers following re-feeding after a prolonged fast

Cultured juvenile white suckers, Catostomus commersoni, are commonly held over winter without the provision of exogenous feed. This study examined the ability of suckers to recover from prolonged fasting, and the energy partitioning strategies employed during the re-feeding phase. In one trial, white suckers held, without exogenous feed, in commercial lake cages for a prolonged period were fed a semi-moist salmonid diet. Although the fish consumed the feed, they exhibited negligible growth over the 12 weeks of the trial, and there was no evidence of increases in tissue lipid, glycogen or protein. In the second trial, previously well-fed juvenile white suckers were starved for 20 weeks and then re-fed. Condition factor (K), but not body weight was increased during the 8 week trial. However, hepatosomatic index (HSI), hepatic and somatic glycogen, and somatic lipid and protein reserves (expressed as a percentage of tissue) were significantly increased during the re-feeding period. Further, changes in plasma thyroid hormone levels indicated an important role of these hormones in the energy partitioning events. The poor growth of juvenile white suckers in intensive culture may be caused by deleterious changes in feed conversion ability resulting from extended fasting of the fish prior to grow-out.     

Evaluation of commercial catfish, tilapia and salmonid diets for growth promotion of white suckers

In an attempt to identify appropriate diets for use in intensive baitfish culture, the efficacy of three commercial diet formulations, a high-protein, high-lipid, low-carbohydrate diet [a semi-moist salmonid diet (SD)] and two low-protein, low-lipid, high-carbohydrate diets [a catfish diet (CD) and a tilapia diet (TD)] on growth performance of juvenile white suckers, Catostomus commersoni, was examined. All three diets permitted growth during the 16 week trial. The SD promoted highest growth and somatic protein and lipid deposition, whereas the CD and TD formulations promoted markedly higher accumulations of glycogen in the carcass and liver. The high hepatic glycogen content was associated with an elevated hepatosomatic index in the CD- and TD-fed groups. Plasma T3 levels were lower in the CD- and TD-fed groups, possibly a response to the low dietary protein or indicative of dietary insufficiency in these fish. Of the three diets evaluated, the SD was considered to be the best in terms of the growth performance parameters measured, and offers an economical and appropriate diet for intensive culture of juvenile white suckers.     

Tumours in white suckers from Lake Michigan tributaries: pathology and prevalence

The prevalence and histopathology of neoplastic lesions were assessed in white sucker Catostomus commersonii captured at two Lake Michigan Areas of Concern (AOCs), the Sheboygan River and Milwaukee Estuary. Findings were compared to those observed at two non‐AOC sites, the Root and Kewaunee rivers. At each site, approximately 200 adult suckers were collected during their spawning migration. Raised skin lesions were observed at all sites and included discrete white spots, mucoid plaques on the body surface and fins and large papillomatous lesions on lips and body. Microscopically, hyperplasia, papilloma and squamous cell carcinoma were documented. Liver neoplasms were also observed at all sites and included both hepatocellular and biliary tumours. Based on land use, the Kewaunee River was the site least impacted by human activities previously associated with fish tumours and had significantly fewer liver neoplasms when compared to the other sites. The proportion of white suckers with liver tumours followed the same patterns as the proportion of urban land use in the watershed: the Milwaukee Estuary had the highest prevalence, followed by the Root, Sheboygan and Kewaunee rivers. The overall skin neoplasm (papilloma and carcinoma) prevalence did not follow the same pattern, although the percentage of white suckers with squamous cell carcinoma exhibited a similar relationship to land use. Testicular tumours (seminoma) were observed at both AOC sites but not at the non‐AOC sites. Both skin and liver tumours were significantly and positively associated with age but not sex.     

Spawning success and early life history of longnose suckers in Great Lakes tributaries

Fish eggs and larvae are often subject to very high mortality, and variation in early life survival can be important for population dynamics. Although longnose suckers (Catostomus catostomus) are widespread in northern North America, little is known about their early life history. We examined fecundity and early larval survivorship during sucker spawning events in three small Lake Michigan tributaries. Although egg deposition varied 25% among spawning events, estimated larval export to the lake varied over 25,000‐fold from around 1000 to 26 million. Based on variation in environmental conditions across years, it appears that spring flow and temperature may be important determinants of egg survival to larval outmigration. Larval age data suggest that most individuals that survived to outmigration hatched during a 2‐day period despite adult spawning across at least 10 days. Most larvae spent <2 weeks in the stream and emigrated around the time of transition from endogenous to exogenous feeding before substantial growth occurred. In two of three cases, larvae drifted exclusively at night; however, high drift rates occurred during both day and night in the case where larvae were very abundant, suggesting density‐dependent drift behaviour. Our results indicate that survival in tributary streams from egg deposition to larval export is highly variable in longnose suckers. These large differences in early life survival may translate into variability in recruitment, thereby influencing population structure and dynamics.     

Effect of some repetitive factors on turbot stress response

The stress response of the turbot, Scopthalmus maximus (L.), to repetitive factors including netting, air exposure, blood sampling and hand-stripping, were tested using two different tank sizes as well as two blood sampling techniques. Exposure of juvenile fish to air for 1-4 min had no immediate effect on plasma cortisol concentrations or haematocrit values. Similarly, the serial netting of immature fish from tanks did not significantly modify plasma cortisol concentrations, haematocrit or osmolarity values. Hand-stripping of mature males was more disturbing than air exposure. The cumulative effect on plasma cortisol levels and osmolarity of stress factors such as netting, air exposure, blood sampling and stripping applied simultaneously to mature males in a 16 m3 tank and repeated twice daily for several days was recorded. The level of cortisol increased from 5 ng ml-1 to 300 ng ml-1 after 10 days of treatment, while an osmoregulatory imbalance and fish death were observed. Moreover, adaptation of fish to smaller tanks seemed to improve the increased plasma cortisol levels and death rate. Reduction in the number of stress factors applied greatly decreased both the range of physiological responses and the death rate. In order to avoid a cumulative stress response, handling of fish should therefore be reduced to a minimum.     

Consistency of stress response to repeated handling in the gilthead sea bream Sparus aurata Linnaeus, 1758

Gilthead sea bream Sparus aurata Linnaeus, 1758 were subjected to repeated stresses, namely confinement and handling once a month for 5 consecutive months. Plasma cortisol concentration was determined as a primary stress response, and plasma glucose and osmolality, serum agglutination and growth rate were analysed as indicators of the secondary and tertiary effects of stress. The results of the mean values after the five trials showed that most indicators followed a consistent pattern with time. Cortisol and growth rate were positively correlated, and cortisol and osmolality were negatively correlated after stress. Analysis of the individual responses throughout all 5 months showed that 6–20% of the fish displayed a consistently high or low response depending on the parameter analysed.     

Plasma growth hormone levels in fed and fasted rainbow trout (Oncorhynchus mykiss) are decreased following handling stress

Plasma growth hormone concentrations of rainbow trout,Oncorhynchus mykiss, fasted for six weeks, were significantly (p < 0.01) higher than in comparable fed animals; in the fasted fish, the levels fell progressively following acute stress (by displacing the fish within their home aquarium), with significant differences from pre-stressed fish evident between one and thirty-two hours after application of the stressor. Plasma growth hormone concentrations also fell significantly in the fed group, but differences were evident only between two and eight hours after stressor application.Plasma cortisol concentrations in pre- and post-stressed fed and fasted fish were similar. There was a bimodal response to stressor application in both groups, with significantly higher values relative to the pre-stressed sample evident one and eight hours after disturbance, but not after two, four or thirty-two hours. The changes in plasma cortisol levels between the initial (09:00h) sample and the sample taken eight hours later resembles the diet pattern seen in trout given access to self-demand feeders.Plasma glucose concentrations in pre-stressed fed animals were higher than in pre-stressed fasted fish. This relationship was also evident between one and four hours and thirty-two hours after stressor application. The post-stress rise in plasma glucose concentration was evident between one and four hours in the fed group, and between four and eight hours in fasted fish.The diel changes in plasma growth hormone and glucose concentrations could not be attributed to normal circadian patterns, and there was no apparent correlation between changes in plasma growth hormone and cortisol concentrations. There was a significant inverse correlation between plasma glucose and growth hormone concentrations when the total data set were analyzed, but these correlations were not apparent when the treatment groups were analyzed separately.     

A soft technology to improve survival and reproductive performance of Litopenaeus stylirostris by counterbalancing physiological disturbances associated with handling stress

The consequences of handling stress (fishing, transfer, eyestalk ablation) on shrimp broodstock are poorly documented. The weakness of farmed shrimp, Litopenaeus stylirostris, during winter is a major problem in New Caledonia, because of seasonal climate (tropical–sub-temperate). The transfer of broodstock in winter from earthen outdoor ponds to indoor maturation tanks in the hatchery (T = 20 °C, Salinity = 35‰, fed shrimp) usually leads, after 48 h, to high mortality (up to 70%). Eyestalk ablation to induce ovarian maturation in females leads to further mortality.

Starting from a background analysis of physiological disturbances (initial osmoregulatory imbalance) associated with handling stress (Wabete, N., Chim, L., Lemaire, P., Massabuau, J.-C., 2004. Caractérisation de problèmes de physiologie respiratoire et d'échanges ioniques associés à la manipulation chez la crevette pénéide Litopenaeus stylirostris à 20 °C. Styli 2003. Trente ans de crevetticulture en Nouvelle-Calédonie. Ed. Ifremer. Actes Colloq. 38, 75-84.), we developed a protocol using a soft technology, based on modifications of water salinity, temperature and feeding regime. The aim was to minimize problems of osmoregulatory imbalance and associated mortalities. The protocol we developed, called the LSD OT protocol (Low Salinity and Diet, Optimal Temperature), was first evaluated on sub-adult shrimp (20–25 g) and then applied to broodstock. Survival after transfer and following eyestalk ablation, as well as reproductive achievement (spawning rate, nauplii number) was considerably improved when shrimps were transferred under “physiological comfort” i.e. warmed isosmotic water (26 °C and 26‰) and unfed for 3 d. This new handling protocol, based on a better control of salinity, temperature and feeding conditions, has been transferred successfully to private hatcheries and already contributes to an increased profitability of New-Caledonian shrimp industry.     

Reduction of transport stress of ayu by obligated schooling

We investigated stress responses of ayu Plecoglossus altivelis under the hypothesis that overcrowding during transport can be a stressor and the stress predisposes fish to disease by decline in immunocompetence. The extent of stress response was determined under different fish densities. As inferred from the serum concentration of cortisol 1 day after the treatment, the density at more than 23 kg/m³, which was within the range of normal transporting procedures, acutely evoked intense stress responses. To examine a possible way to reduce the stress response of fish under crowding conditions, we compared two fish groups established in water tanks with circulating (whirling) and turbulent currents at the same density. The cortisol levels 1 day after the treatment were 2.5-fold higher in turbulent current than in whirling current. Fish in whirling current formed a school, while those in turbulent current aggregated with each other. Schooling of fish induced by water flow may reduce their stress responses during transport. A transport system with a directional water flow should keep fish in good condition without reducing the number of fish contained.     

Stress hormones and the cellular stress response in salmonids

The relationship between stress protein (SP) levels and the hormonal stress response in salmonids was examined through the measurement of gill SP70 and SP30 levels together with plasma cortisol, glucose and ion concentrations, in response to handling stress (45 s holding in a net), intraperitoneal cortisol implants, and heat shock (+10 °C). Handling and cortisol implants resulted in increased plasma cortisol and glucose levels. Heat shock following handling reduced plasma [Na⁺] below that observed in response to the handling stress alone, and heat shock following cortisol implant significantly lowered both plasma [Cl^–] and [Na⁺] below that of fish receiving the cortisol implant alone. Increased SP70 levels occurred 1 h following the 2 h +10 °C heat shock. Handling the fish prior to the application of heat shock suppressed the increase of SP70 levels in the gills. However, increased plasma cortisol concentrations alone did not attenuate gill tissue SP70 increase caused by heat shock. Physiological (10^–7 M) and pharmacological (10^–5 M) concentrations of adrenaline caused increased levels of SP70 in hepatocytes. Addition of the -blocker propanolol blocked this response to adrenaline. The results indicate that handling procedures do not result in an increase of hsp30 or hsp70 and may suppress hsp synthesis under certain circumstances.     

Noninvasive assessment of nitrate‐induced stress in koi Cyprinus carpio L. by faecal cortisol measurement

This study determined effects of nitrate (NO₃^?)‐induced stress on faecal cortisol levels in koi (Cyprinus carpio). NO₃^? was presented in two formats: (1) bulk increase followed by dilution to assess cortisol response and recovery and (2) bulk increase followed by incremental increases to assess cortisol response limit. Fish were maintained group‐wise in treatment and control aquaria (n=6 per group), and 0.5 g L^?1 NO₃^? (as NaNO₃) was added to the water. Faecal samples were collected daily and blood samples were taken pretreatment and 72‐h posttreatment and were assayed for cortisol via ELISA. NO₃^? increased plasma and faecal cortisol 4.4‐fold and 3.9‐fold respectively. Plasma cortisol was not measured further. After a 74% NO₃^? decrease by dilution, faecal cortisol decreased to baseline within 24 h, and restimulation by NO₃^? (1.0 g L^?1) elevated faecal cortisol to maximal study levels. In a separate experiment, exposure to 0.4 g L^?1 NO₃^? increased faecal cortisol 6.9‐fold. However, three additional 0.4 g L^?1 NO₃^? increments across 9 days did not further increase cortisol. This study demonstrates that faecal cortisol measurement in fish via an ELISA can be useful as an indicator of NO₃^?‐induced stress in the aquatic environment.     

拥挤胁迫后鲫鱼血液皮质醇和溶菌酶水平的变化及对病原的敏感性

分别在拥挤胁迫后第3、10、20、30天对鲫鱼(Carassiusauratus)血液皮质醇水平和血液溶菌酶水平进行了检测,并在每次采集血样后用嗜水气单胞菌(Aeromonashydrophila)进行人工感染以评估鱼体抵抗力。结果显示,血液皮质醇水平在胁迫后几天内显著升高(P<0.005),随后虽有所下降,但至第30天高密度组仍保持较高水平(216.32±11.91)ng/mL。实验初期,血液溶菌酶水平小幅升高,但经过长时间的拥挤后,高密度组血液溶菌酶活性受到极大抑制,显著低于同期对照组(P<0.01)。攻毒后的死亡率与胁迫时间成正相关,在最后一次采样期,高密度组死亡率高达58.3%,而同期对照组仅为16.7%,两者具统计学差异(P<0.01)。实验表明,经过拥挤胁迫后,鲫鱼血液皮质醇水平和溶菌酶水平均发生了变化,短期内,两者成正相关,而长期胁迫则表现为负相关。另外,拥挤胁迫也引起了鱼体抵抗力的下降。     

克氏原螯虾自噬相关基因PcAtg2的克隆及其在白斑综合征病毒胁迫下的表达分析

为了解自噬相关基因Atg2在克氏原螯虾(Procambarus clarkia)先天免疫中的作用,本研究克隆了克氏原螯虾Atg2 (PcAtg2)基因全长序列。生物信息学分析显示,PcAtg2蛋白编码序列全长为9 966 bp,推测其编码2 189个氨基酸。组织定量表达分布显示,PcAtg2在克氏原螯虾的各个组织中均有表达,其中在肝胰腺中表达最高,在眼柄中表达最低。在白斑综合征病毒(WSSV)感染实验中,PcAtg2基因表达量在不同组织中均呈现显著上调趋势。RNA干扰(RNAi)实验显示,PcAtg2基因沉默后,WSSV在克氏原螯虾体内的增殖明显被抑制,同时,自噬相关基因的表达量上调。透射电镜分析结果显示,在WSSV感染后,PcAtg2基因沉默组中克氏原螯虾肝胰腺组织中的自噬小体多于对照组。本研究结果可为了解克氏原螯虾应对WSSV胁迫下的调控机制提供理论参考。     

急性操作胁迫对四指马鲅幼鱼肝脏组织结构和氧化应激的影响

为探究急性操作胁迫对四指马鲅(Eleutheronema tetradactylum)幼鱼肝脏组织结构以及氧化应激的影响,该研究在离水操作胁迫后不同时间取样,观察肝脏显微结构的变化,并检测相关抗氧化酶活性。结果显示,随着离水胁迫时间的延长,肝脏组织损伤程度呈现先上升后下降的趋势,胁迫第24小时肝脏基本结构与对照组相似。超氧化物歧化酶(SOD)和过氧化氢酶(CAT)有相同的变化趋势,胁迫后第2小时显著增加(P0.05)并达到最高值,然后开始下降,最后其值显著低于对照组(P0.05);丙二醛(MDA)含量在胁迫第2~第12小时呈现先升高后下降的变化趋势,第12小时后又有所上升;还原型谷胱甘肽(GSH)含量胁迫2 h没有显著变化(P0.05),之后呈先下降后上升的趋势,但其含量仍显著低于对照组(P0.05);总抗氧化能力(T-AOC)胁迫第2小时显著上升(P0.05),之后下降,实验结束时显著低于对照组(P0.05)。结果表明离水操作胁迫对四指马鲅幼鱼肝脏组织结构以及抗氧化酶产生一定的影响,但随着胁迫时间的延长,其组织结构以及抗氧化能力都有一定的恢复趋势。     

PCR检测对虾白斑综合征病毒(WSSV)中使用UNG防遗留污染

设计了一对引物用于PCR检测对虾白斑综合征病毒，PCR扩增过程中用尿嘧啶-DNA糖基酶(UNG)防遗留污染。使用UNG时，该对引物的适宜dUTP和Mg^2 浓度分别为0．4mmol／L和2．0mmol／L；UNG存在时，PCR检测WSSV DNA的最低量为1pg，UNG的使用使PCR的检测灵敏度降低了1个数量级；进行正常PCR扩增前，0．5U UNG可消除至少10ng含dU的WSSV DNA的PCR扩增产物。     

Reduced stress and increased immune responses in Nile tilapia kept under self-feeding conditions

ABSTRACT: Stress and immune responses were studied in Nile tilapia Oreochromis niloticus under both conditions of self-feeding and scheduled feeding. Self-feeding fish had a significantly lower cortisol level in their blood plasma than that of scheduled feeding fish. Furthermore, skin color was found to be paler in the self-feeding fish compared with that of the scheduled feeding fish. Thus, the self-feeding fish had a less stressful physiological status than the other experimental fish. Immunological analyses revealed that the self-feeding fish significantly exceeded the scheduled feeding fish in the phagocytic activity of their macrophages, antibody production, and the number of blood-circulating lymphocytes. The higher immune responses of self-feeding fish seem to be attained through a less stressful environment. This self-feeding behavior could be exploited to create reduced stress for disease control in fish farming.     

Relationship between dietary lipid source, oxidative stress, and the physiological response to stress in sub-yearling chinook salmon (Oncorhynchus tshawytscha)

Relationships between dietary lipid source, stress, and oxidative stress were examined in juvenile chinook salmon (Oncorhynchus tshawytscha). Four different experimental diets were used: menhaden oil (MHO; elevated 20:5n-3 and 22:6n-3), soybean oil (SBO; elevated 18:2n-6), linseed oil (LSO; elevated 18:3n-3), and a mixture of 55% linseed oil and 45% soybean oil (MIX; approximately equal levels of 18:2n-6 and 18:3n-3). Juvenile salmon (initial body weight of 16.0 g) were fed experimental diets for 12 weeks (early March to early June). At the end of feeding, fish subjected to a low-water stressor for 96 h had greater liver and brain lipid peroxidation compared to unstressed controls; peroxidation was not influenced by diet. Diet and stress affected plasma cortisol levels. Stressed fish fed SBO had the greatest cortisol concentrations, followed by MIX, MHO, and LSO (mean concentrations for the SBO and LSO diets differed significantly). The cortisol response to stress may have been influenced by the ratio of prostaglandin 1- and 2-series to prostaglandin 3-series precursor fatty acids provided by the different diets. The results of this study suggest a connection between the physiological response to stress, dietary lipid quality, and oxidative stress. This is the first evidence of such a relationship in fish. Abbreviations: AA - arachidonic acid; ACTH - adrenocorticotropin; BHT - butylated hydroxytoluene; BLPO - brain lipid peroxidation; dGLA - dihomo-γ-linolenic acid; DHA - docosahexanoic acid; EPA - eicosapentanoic acid; FER - feed efficiency ratio; FOX - ferrous oxidation-xylenol orange; GLA -γ-linolenic acid; LA - linoleic acid; LCO3 - long-chain n-3 polyunsaturated fatty acids; LLPO - liver lipid peroxidation; LN - linolenic acid; LPO - lipid peroxidation; LSO - linseed oil; MHO - menhaden oil; MIX - 55% linseed oil + 45% soybean oil; PC - plasma cortisol; PG - prostaglandin(s); PGE₂- prostaglandin E₂; PUFA - polyunsaturated fatty acid; SBO - soybean oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

白斑综合症病毒（WSSV）对中国对虾卵及各期幼体人工感染的试验研究

用感染白斑综合症毒病的中国对虾头胸甲，对健康的仔虾进行了人工投喂感染实验，同时从病虾中分离出病毒悬液作为毒种，无节幼体，蚤体 幼体和糠虾体进行不同温度条件下的人工浸浴实验，结果表明，卵和无节幼体，蚤状幼体，糠虾幼对病毒悬液不敏感,闰理切片观察未见病毒粒子，光镜病理切片可观空到鳃上皮，前肠上皮有包涵体样病变，同时在肝胰组织中发现了肝胰腺细小病毒（HPV）包涵体。     

Whole-body corticosteroid and plasma cortisol concentrations in larval and juvenile Atlantic cod Gadus morhua L. following acute stress

Methods were developed to assess whole‐body immunoreactive corticosteroid concentrations (IRC) and plasma levels of cortisol in Atlantic cod subjected to several common, acute stressors. A measurable corticosteroid stress response was observed at the first sampling in whole bodies of 8‐day post‐hatch (dph) larvae. Two groups of juveniles (～5 and 30 g) were subjected to a 30 s net stressor and whole‐body IRC and plasma cortisol levels were determined. Post‐stressor IRC in smaller fish rose approximately 14‐fold, peaked at 1 h, were sustained for 3–6 h and returned to pre‐stressor levels within 24 h. Post‐stressor plasma cortisol levels in larger fish rose approximately 18‐fold, peaked at 0.5–1 h, were sustained for 1–3 h and then returned to near pre‐stressor levels after 24 h. Immunoreactive corticosteroid concentrations appeared to remain elevated longer than plasma cortisol levels suggesting that steroids other than cortisol were contributing to total immunoreactivity in homogenates. Juveniles exposed to either a grading procedure or high density transport had maximal IRC and plasma cortisol levels within 90 min which returned to pre‐stressor levels within 24–72 h. A reduction in water salinity (20 g L^?1) did not moderate the corticosteroid response in juveniles. The results show that Atlantic cod respond to common, acute stressors in a manner similar to other teleosts. Whole‐body homogenates can be used to identify changes in IRC in response to acute stress in cod with the caveat that recovery IRC may differ from plasma cortisol concentrations.