Humoral factors important in resistance of salmonid fish to bacterial disease. I. Serum antibody protection of rainbow trout (Salmo gairdneri) against vibriosis

Serum antibody is shown by passive immunization and absorption experiments to be an important component of defense mechanisms against experimentally induced vibriosis in rainbow trout (Salmo gairdneri). Antigen from heat-killed Vibrio anguillarum injected into trout with Freund's complete adjuvant induced much higher agglutinin titers than formalin-killed bacteria with or without adjuvant, or heat-killed bacteria without adjuvant. Passively transferred trout anti-Vibrio anguillarum serum provided protection against experimental vibriosis in trout for more than 2 months. Passively transferred anti-Vibrio anguillarum rabbit serum also protected trout.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.     

Epizootiology of Renibacterium salmoninarum, the causal agent of bacterial kidney disease in salmonid fish

Abstract. By use of a selective medium, Renibacterium salmoninarum was recovered from rainbow trout, Salmo gairdneri Richardson. With asymptomatic animals, small numbers of Renibacterium colonies were recovered only from the anterior part of the kidney; whereas in clinically diseased fish, dense pure culture growth was obtained from kidney, spleen, heart, blood, ascitic fluid and faeces. There was no evidence for the presence of R. salmoninarum in the water or sediment of 56 freshwater fish farms in England and Wales. Nevertheless, laboratory–based experiments showed that the pathogen was excreted in the faeces of clinically diseased fish. Moreover, there was survival of the bacterial cells for up to 21 days in sediment/faecal material.     

Stress responses and disease resistance in salmonid fish: Effects of chronic elevation of plasma cortisol

Basal levels of plasma cortisol in unstressed salmonid fish are normally in the range 0–5 ng ml⁻¹. An acute stress such as handling or 1 h confinement caused a temporary elevation of the plasma cortisol levels of both brown trout,Salmo trutta L., and rainbow trout,Salmo gairdneri Richardson, in the range 40–200 ng ml⁻¹ with a return to basal levels within 24–48 h. The extent of the cortisol elevation in response to an acute stress was dependent upon both the species and strain of trout. Chronic stresses, such as prolonged confinement or crowding, resulted in an elevation of plasma cortisol levels to approximately 10 ng ml⁻¹. Under these circumstances, blood cortisol levels remained elevated for periods of up to 4 weeks before acclimation finally occurred. It is shown, by means of intraperitoneal implantation of cortisol, that chronic elevation of plasma cortisol levels in the brown trout results in a dose-dependent increase in mortality due to common bacterial and fungal diseases. This effect is apparent at plasma cortisol levels as low as 10 ng ml⁻¹, levels below those often reported as being representative of ‘unstressed’ fish. These findings are discussed in relation to the known immunosuppressive effects of corticosteroids in teleost fish.     

魁蚶各组织溶菌酶活性对鳗弧菌侵染的响应

为了有效防治细菌及其他病原微生物对魁蚶等贝类的危害,本实验研究了魁蚶各组织中溶菌酶活性对鳗弧菌侵染的响应过程,以期探讨魁蚶体内溶菌酶的免疫功能。本实验采用注射活菌的方法侵染20月龄魁蚶个体,随机选取16只个体,在每只个体的斧足处注射1 mL(约1×10~9个)鳗弧菌菌悬液作为感染组;随机选取16只个体不注射鳗弧菌作为对照组。两组魁蚶分别于洁净海水中暂养4、12、24和48 h后,每组随机取4只魁蚶个体的血液、外套膜、鳃、斧足、肝胰腺和闭壳肌等组织,采用ELISA试剂盒测定其溶菌酶含量变化。结果显示,对于鳗弧菌的侵入,魁蚶血液中溶菌酶含量由正常低值迅速增高并一直维持较高的含量,说明血液是魁蚶机体防御病原菌的主要免疫组织之一;魁蚶外套膜在无感染的情况下,对外界水环境的干扰始终保持较高的溶菌酶含量;鳃、斧足的溶菌酶含量均在注射细菌24 h之后明显高于正常值,说明外套膜、鳃、斧足作为魁蚶机体与外界接触的第一道屏障也能应对病原菌入侵,但反应较血液延迟;肝胰腺和闭壳肌的溶菌酶含量变化不明显,推测肝胰腺和闭壳肌不是魁蚶的重要免疫组织或器官。本实验结果可为魁蚶抗病选育及免疫机理方面的研究提供相关的参数。     

Production of salmonid amoebic gill disease by exposure to Paramoeba sp. harvested from the gills of infected fish

Atlantic salmon, S almo salar L., were exposed to different concentrations of Paramoeba sp. The lowest concentration which induced amoebic gill disease (AGD) was 230 Paramoeba sp. L^–1 and the severity of infection increased with increasing concentration. The concentration of Paramoeba sp. positively correlated with the number of gill lesions ( R ² > 0.7). This study provides evidence that Paramoeba sp. is the causative agent of AGD and describes an experimental model that enables the severity of the induced disease to be controlled.     

Mortality related to pancreas disease in Norwegian farmed salmonid fish,Salmo salar L. and Oncorhynchus mykiss (Walbaum)

High mortality during the salmonid seawater phase is a continuous problem for the salmonid aquaculture industry, although the mortality levels show a large variation both in farms with a disease history and those without. We wanted to examine the mortality patterns in farms with pancreas disease (PD) and compare it to farms without this diagnosis. Further, we wished to investigate the factors influencing the maximum mortality in both groups. We examined data from all salmonid farms in Norway stocked after January 2003 and slaughtered before December 2007. In total, 1884 cohorts were included, and 150 of these were diagnosed with PD. We found that season accounted for more of the variation in mortality than water temperature in PD‐positive cohorts and that infection pressure influenced the mortality in non‐PD cohorts, suggesting outbreaks of disease that are not diagnosed. We also found that the mortality in PD cohorts decreased significantly from 2003 to 2007, suggesting that increased knowledge about PD and targeted actions have been effective. Our study further suggests that crude mortality figures may be of limited use when wanting to examine a particular disease and risk factors for increased mortality. We suggest farmers and legislation should turn to a more modern approach with cause‐specific mortality records.     

Growth of three salmonid species in mono and double culture (Salmo salar L., S. trutta L. and S. gairdneri Rich.)

Juvenile Atlantic salmon, sea trout and rainbow trout were used in an experiment in which two species in each tank were compared with controls of each species separately. Specific growth rate was best in the experimental groups where the two species were reared together. The theory was developed that social facilitation or moderate interaction could cause the better growht. No starvation as a result of competition was discovered in any group.     

水产养殖中使用的抗菌药物及细菌耐药性

目前，抗菌药物在水产养殖动物疾病防治中占有重要地位，然而，在水产养殖中使用抗菌药物也可产生一系列的副作用，例如细菌耐药性的产生和药物在鱼体和环境中的残留，本文就水产药物使用的历史，现状，适应症，副作用，耐药性的现状，来源，产生机制，耐药菌判定的标准，以及该领域的研究现状等问题进行综述与分析，以便为合理使用抗菌药物提供理论依据。     

牙鲆抗迟缓爱德华菌病家系建立与抗病性能评价

为筛选出抗迟缓爱德华菌的牙鲆家系,实验以已经选育出的牙鲆抗病和快速生长家系以及从韩国引进的选育群体为亲本建立了牙鲆家系56个,并对其中32个家系进行了迟缓爱德华菌人工感染实验。确定了迟缓爱德华菌对牙鲆家系的半致死浓度LD50为3.69×105CFU/mL后,从每个家系中随机抽取75尾,按照0.2 mL/10 g体质量腹腔注射半致死浓度的菌液,并设置1次重复。人工感染时水温控制在(19±1)℃。从32个家系中共选取4 800尾5月龄牙鲆幼鱼进行感染,16 d实验结束后统计各家系存活率为8.2%~66.1%,平均存活率为31.2%。不同家系对迟缓爱德华菌的抗感染能力表现出显著差异(P0.05)。筛选出6个存活率高于45%的抗病家系,发现2007年筛选出的抗鳗弧菌病家系F0768的后代家系在迟缓爱德华菌感染后的存活率普遍很高,表现出抗迟缓爱德华菌病的能力。研究为选育抗鳗弧菌病和抗迟缓爱德华菌病的牙鲆优良品系奠定了基础,对牙鲆抗细菌病的分子机理研究及抗病新品种选育具有重要意义和应用价值。     

异硫氰酸荧光素标记鳗弧菌条件优化及应用条件对荧光信号的影响

为了实现对目标细菌的有效示踪,实验以异硫氰酸荧光素(FITC)为标记物,分别设置FITC标记浓度0.2、2、20、50、100、200μg/mL,标记时间1、15、30、60、120、240 min,研究了FITC标记浓度与标记时间对鳗弧菌标记效果的影响,同时对在不同温度(25、4、-20、-70℃)及不同时间(24、48、96、192 h)存储条件下鳗弧菌标记荧光的稳定性进行了比较,对标记鳗弧菌经甲醛灭活后的荧光变化进行了观察,研究了不同光照条件(0、150、1 500、25 000 lx)对标记鳗弧菌的荧光衰减影响,以及在-20和-70℃条件下反复冻融对标记鳗弧菌荧光强度的影响。结果表明,FITC的最适标记浓度范围为20~100μg/mL,最适标记时间范围60~120 min,储藏在-20和-70℃条件下,标记后的鳗弧菌荧光信号比储藏在4和25℃条件下稳定,标记后的鳗弧菌经甲醛灭活后的荧光信号稳定性优于对照组,不同强度自然光照对标记鳗弧菌的荧光衰减效果不明显,标记细菌经3次冻融,发现在-70℃条件下冻存的细菌荧光信号稳定性保持良好。本研究结果为FITC用于标记海洋细菌提供了实用的技术参数。     

Metabolism of fatty acids in fish. II. Biosynthesis of fatty acids in relation to diet in the carp, Cyprinus carpio Linnaeus 1758

Carp, weighing 15–25 g, were maintained in the laboratory on a control diet, or on diets prepared from wheat or corn, and the incorporation of sodium (1?¹⁴C)-acetate into their liver total fatty acids as well as into liver triglycerides and phospholipids was followed. The control diet was characterized by a high concentration of total lipids and linolenic acid, and the other two feeds by a low concentration of total lipids including linolenic acid and high concentration of carbohydrates. The highest rate of fatty acid biosynthesis was observed in fish given the corn diet, and the lowest rate was found in the carp on the control diet. Levels of radioactivity became very high in triglycerides of corn-fed fish. The major products of fatty acid biosynthesis were palmitic and oleic acids, in the corn-fed fish, while in the control fish almost 50% of the total radioactivity was in the long chain polyunsaturated fatty acids. The degree of labeling of oleic and palmitic acids appeared to depend on the level of linolenic acid in the diet. The results suggest that about 1% of linolenic acid in the diet is required to keep lipogenesis at a low level as well as to prevent hyperproduction of oleic acid. Linoleic acid seems to play a subordinate role in regulating lipogenesis in carp liver.     

Water quality affects SDH activity, protein content and RNA:DNA ratios in fish (Catla catla, Labeo rohita and Oreochromis mossambicus) raised in ponds of a sewage-fed fish farm

The spatial and seasonal variability of the respiratory enzyme succinate dehydrogenase and the protein content were examined in different tissues of fish cultured in three ponds along the effluent gradient of a sewage-fed fish farm. Indian major carp Catla catla (150-230 g) and Labeo rohita (60-190 g) cultured in both the middle and last points of the sewage effluent (stocking pond 1) and (stocking pond 4) and Oreochromis mossambicus (50-160 g), a naturally growing fish of the inlet (facultative pond) and the out let of the sewage effluent (stocking pond 4) were procured every month during the period of January-December, 2005 and were subjected to determination of succinate dehydrogenase activity, total protein, DNA and RNA contents from gill, liver and muscle tissue respectively. The SDH activity of all three test fishes (Catla catla, Labeo rohita and Oreochromis mossambicus) was reduced significantly (ANOVA; P < 0.05) when cultured in SP-4 compared to SP-1 in the case of Catla catla and Labeo rohita and in facultative pond in the case of Oreochromis mossambicus.Conspicuous differences in the SDH activity of fish between the last and first stocking pond or the facultative pond were clearly due to the result of the differences in water quality. There was a direct relationship between SDH activity in gill tissue of any of the fish investigated and ammonia-N concentration of water or water pH. This shows that the respiratory activity of these fishes was strongly affected by the ammonia and pH of water. In other words, this suggests that as the distance from the point source increases, there was a substantial improvement of water quality in the ponds located along the sewage effluent gradient. Evidently, there is a progressive pattern of growth, survival and physiological health of fish and abundance of favorable diversity of food organisms with rich biodiversity.     

Effects of silver carp and the small indigenous fish mola Amblypharyngodon mola and punti Puntius sophore on fish polyculture production

A sustainable semi-intensive pond aquaculture technology including major carp species as cash-crop and small indigenous fish species (SIS) as food for the farmers' families is being optimized in Bangladesh. The inclusion of silver carp (Hypophthalmichthys molitrix), a cheap large species affordable by poor farmers, is now being considered. As part of a study on the effects of this filter feeder on polycultures including the large carps rohu (Labeo rohita), catla (Catla catla) and common carp (Cyprinus carpio) and the SIS punti (Puntius sophore) and mola (Amblypharyngodon mola), an experiment was carried out under farm conditions to test the effects of silver carp and of each SIS species on the growth, survival and yield of the large and small species and on pond ecology.The experiment was performed in 38 farmers' fishponds of different sizes, from 220 m² to 1200 m². The results show that the larger the fish pond the better rohu performance, the larger punti fry weight and the lower punti fry harvested biomass. Pond size did not affect other fish species. The addition of 250 mola and/or punti per 100 m² fishponds affected rohu and catla and did not affect common and silver carps. The addition of mola alone reduced rohu's parameters by 15%. The addition of SIS in the three combinations tested (250 mola, 250 punti, 125 of each species) reduced catla's parameters by 20-24%. Punti fry were larger when both SIS were stocked and punti fry biomass was larger when only punti were present. Total mola harvested biomass and yield were larger when the entire SIS stocked were only mola.The addition of 10 silver carp over the 99 large carps stocked per 100 m² fishponds negatively affected rohu and catla growth and yield by about 15-21% and 45-50% respectively but not their survival, did not affect common carp performance, did not affect punti and mola reproduction in the ponds, reduced punti yields by 25%, reduced mola performance by about 35%, and silver carp own biomass increased total yield and total income in about 12% each. These effects are explained and discussed considering fish interactions through the food web. The decreased income from selling the more expensive large carps is more than compensated by that obtained from silver carp, which allows the option to the farmer to sell part of the silver carp to complete the cash income that would have been obtained from large carps only if silver carp would not be stocked, and consume the rest with the family.     

吉富罗非鱼与奥利亚罗非鱼自繁与杂交 F1遗传特性与抗病力分析

以吉富罗非鱼与奥利亚罗非鱼为繁育亲本,采用完全双列杂交繁育4组F1,将初始规格基本一致的4组罗非鱼饲养100 d后,运用“加性-显性”遗传分析模型,分析了4组F1罗非鱼8个生长相关性状杂种优势、遗传效应以及与性状间的相关性.结果表明:(1)F1群体平均优势为0.129 4～0.368 4.除尾柄长超亲优势较大外,其他性状的群体超亲优势较小或表现出负向超亲优势.(2)8个性状的广义遗传率(HB)为0.714 2～0.995 3,表明加性效应和显性效应对性状的遗传变异影响极显著(P＜0.01).除尾柄长外,其他性状的狭义遗传率(HN)介于0.469 4～0.737 9,表明加性遗传方差在表型方差中所占比率较高.(3)体质量、全长、体长、体高、体宽、头长、尾柄长、尾柄高性状之间表型相关在0.776 6～0.999 7范围内,而遗传相关在0.994 1～1.000 0之间,表明这些性状间都存在极显著的正相关.取样结束后,采用3.95×106 CFU/mL的海豚链球菌菌液进行腹腔感染,吉富罗非鱼自繁组F1代12 h后出现死亡,而奥利亚罗非鱼自繁组F1192 h后才出现死亡.384 h后,吉富罗非鱼自繁组F1死亡率为40％,正反交组F1分别为20％和23.3％,奥利亚罗非鱼自繁组F1死亡率最低,为6.67％.研究结果表明,除尾柄长外,杂交F1的其他性状不具备超亲优势,然而杂交可以提高选育后代的抗病力.     

七彩神仙鱼皮肤色素细胞观察及类胡萝卜素组分分析

七彩神仙鱼 (Symphysodon aequifasciatus) 具独特的亲代抚育行为,前期研究发现2种C型凝集素在育幼亲本皮肤中高表达,但表达模式不同,预示着潜在的功能分化。为探究两者潜在免疫功能的差异性,克隆获得了七彩神仙鱼C型凝集素基因 SaCD302 和 SaMBL,并分析了病原刺激下的免疫应答模式。结果显示,SaCD302 和 SaMBL 的开放阅读框 (ORF) 长度分别为741和795 bp,分别编码246和264个氨基酸。SaCD302的氨基酸序列包含一个信号肽 (aa 1—29),一个CTL结构域 (aa 32—169)和一个跨膜结构域 (aa 184—206)。SaMBL的氨基酸序列包含两个低复杂度区域 (aa 32—92和 aa 104—121) 和一个典型的CTL结构域 (aa 137—263)。SaCD302 和 SaMBL 的氨基酸序列与尼加拉瓜湖始丽鱼 (Archocentrus centrarchus) 对应氨基酸序列的同源性最高。SaCD302 在各组织中均有表达,头肾中表达量最高,其次为肝脏和鳃;SaMBL 在皮肤中高表达,在肠道、肝脏、脾脏和性腺中低表达。嗜水气单胞菌 (Aeromonas hydrophila) 感染后,SaCD302 在脾脏、头肾、肠道和皮肤中的表达量均显著上调,SaMBL 仅在头肾、肠道和皮肤中表达上调,且时序性不同,表明二者可能在七彩神仙鱼免疫防御中发挥不同作用。     

In vitro antibacterial activity of the combination of sulphadiazine and trimethoprim on bacterial fish pathogens

Abstract. The in vitro bacteriostatic activity of a combination of sulphadiazine (SDZ) and trimethoprim (TMP) against representative bacterial fish pathogens was studied. In general, the minimum inhibitory concentrations (MIC) of TMP were much smaller than comparable values for SDZ. When SDZ and TMP were combined, the bacteriostatic effect was enhanced in a synergistic manner for all bacteria tested except Pseudomonas fluorescens . The synergism was most remarkable in the case of bacterial strains that were sensitive to the action of SDZ alone. This degree of synergism was observed when the ratios of SDZ and TMP in combination were in the range of 1:1 io 5:1. The MIC of SDZ plus TMP combined was less than 3.13 μg/ml for SDZ resistant strains and as low as 0.2 to 0.78 μ/ml for SDZ sensitive strains. The possible usefulness of a combination of these two compounds for therapy or control of bacterial fish disease is discussed.     

Physiological and social constraints on growth of fish with special reference to Arctic charr, Salvelinus alpinus L.

Physiological studies of growth in animals predict that growth rates should decrease with increasing size, but when Arctic charr, Salvelinus alpinus, were reared together in large groups there was often a positive correlation between initial body size and the growth rate of an individual fish. This suggested that social interactions were important determinants of growth rates and, in the absence of the establishment of direct linear hierarchies, it is suggested that growth suppression is the result of short-term bouts of aggression associated with feeding periods leading to reduced food intake by certain fish. Evidence is presented to show that growth suppression can be reduced by increasing the frequency of feeding.