Survivability of Infectious Hematopoietic Necrosis Virus in Fertilized Eggs of Masu and Chum Salmon

Abstract

The possibility of vertical transmission of infectious hematopoietic necrosis virus (IHNV) was studied with the eggs of masu (cherry) salmon Oncorhynchus masou and chum salmon O. keta. The surfaces of eggs and sperm were contaminated with IHNV (10^3.8-10^4.8 50% tissue culture infective dose [TCID50]/egg) and then the eggs were fertilized. Eggs just after fertilization and embryonated eggs also were infected by injection with IHNV (10^3.8 TCID50/egg) directly into the yolk. During incubation, eggs were held in running water at 10°C. Mortality of the eggs or hatched progeny was determined and isolation of IHNV on the surface or inside of the eggs was determined during the incubation period. No mortality occurred and no virus was detected in fertile eggs from contaminated gametes. For injected eggs, IHNV was not detected on the surface of masu and chum salmon eggs after 1 d of incubation. Infectivity of IHNV inside the eggs decreased gradually and could not be detected after 1 month of incubation. This rate of IHNV reduction in the fertilized egg was similar to that found in a mixture of IHNV and homogenized yolk contents. Several individual yolk components also showed anti-IHNV activity. When eyed eggs were injected with IHNV, the embryos of both masu and chum salmon became infected, and the concentration of virus increased rapidly and reached more than 10^6.5 TCID50/fish. The cumulative mortality of eggs injected at the eyed stage for both masu and chum salmon was 90%. The susceptibilities of hatched-out larvae of masu and chum salmon to IHNV were different; cumulative mortality was more than 90% in masu salmon and 20–30% in chum salmon artificially infected with the virus. We concluded that vertical transmission of IHNV is doubtful because the virus is apparently unable to survive in eggs before the eyed stage.     

Epidermal Tumors on Rainbow Smelt and Atlantic Salmon from Nova Scotia

Abstract

Epidermal tumors on rainbow smelt Osmerus mordax are described in detail for the first time. These tumors apparently progress from small preneoplastic lesions to papillomas, and to carcinomas on some males. These tumors and similar tumors on Atlantic salmon Salmo salar have not been previously reported from Canada. Tumors were found on rainbow smelt mainly in freshwater during the spring spawning run and on Atlantic salmon grilse in freshwater in October, These tumors varied in size and number on each fish. The smaller growths were usually translucent and whitish, with a smooth surface. Some larger tumors were papillomatous and appeared reddened because of increased vascularization. Preneoplastic lesions and papillomas had a thickened epidermis with an intact basement membrane, and most of the cells were enlarged. fn papillomas of rainbow smelt, tumor cells that were not close to papillae were vacuolated, and necrotic regions were present. Carcinomas on the heads of male rainbow smelt had regions of pleomorphic cells which invaded the subepithelial tissues and penetrated the cartilage of the mandible of two specimens found in brackish and seawater. Intranuclear basophilic inclusions suggestive of a viral agent were present in the tumors of both rainbow smelt and Atlantic salmon.     

Relation of Water Temperature to Bacterial Cold-Water Disease in Coho Salmon,Chinook Salmon,and Rainbow Trout

Abstract

The effect of water temperature on the progress of experimentally induced Cytophaga psychrophila infection was investigated in juveniles of coho salmon Oncorhynchus kisutch, chinook salmon O. tshawytscha, and rainbow trout O. mykiss (formerly Salmo gairdneri). A virulent strain of C. psychrophila was administered to fish by subcutaneous injection. Infected fish were held in tanks containing pathogen-free well water at temperatures ranging from 3 to 23°C. Mean times from infection to death of the fish were shortest at 12–15°C, which were the temperatures associated with the shortest time for doubling the population of this bacterium in vitro. Juvenile steelhead (anadromous rainbow trout) injected with viable C. psychrophila cells and held in 22°C water did not become diseased.     

养殖虹鳟与大西洋鲑肉质和营养成分比较研究

试验选取10尾贵州都匀养殖场的虹鳟(1562±32)g和10尾市场出售的挪威大西洋鲑(1856±45 g),采集其背部肌肉,分别进行质构特性及营养成分分析。结果表明:质构特性上,虹鳟肌肉的失水率显著低于大西洋鲑(P<0.05),剪切力显著高于大西洋鲑(P<0.05);常规成分中,虹鳟肌肉的粗脂肪含量显著低于大西洋鲑(P<0.05),二者的粗蛋白含量无显著差异(P>0.05);脂肪酸组成上,虹鳟肌肉的n-6多不饱和脂肪酸含量显著低于大西洋鲑(P<0.05),但基于脂肪酸组成获得的饱和度(S/P)、动脉粥样硬化指数(AI)、凝血指数(TI)和降胆固醇/致高血脂比率(HH)等指标无显著性差异(P>0.05);氨基酸组成方面,虹鳟肌肉内的人体必需氨基酸比例显著低于大西洋鲑(P<0.05),鲜味氨基酸含量显著高于大西洋鲑(P<0.05)。结果表明,养殖虹鳟在肉质质构上相比大西洋鲑较“干硬”,且虹鳟肌肉多不饱和脂肪酸和必需氨基酸含量均低于大西洋鲑。     

The Impact of Egg Ozonation on Hatching Success,Larval Growth,and Survival of Atlantic Cod,Atlantic Salmon,and Rainbow Trout

The direct exposure of fish eggs to ozonated water has generated interest as a means of ensuring pathogen-free eggs without the use of harsh chemicals. However, there are numerous knowledge gaps, including safe contact times, exposure levels, and potential long-term effects on aquaculture species in both freshwater and seawater. The effect of different ozone (O₃) doses (0.5–1.0, 1.5–2.0, and 2.5–3.0 mg of O₃/L for 90 s) on recently fertilized eggs of Atlantic Cod Gadus morhua and eyed eggs of Atlantic Salmon Salmo salar and Rainbow Trout Oncorhynchus mykiss was evaluated in comparison with the effects of two commercial disinfectants: Perosan (0.004 mg/L) and Ovadine (100 mg/L). The impact of ozone application was evaluated based on hatching success, larval nucleic acid concentration, larval growth, and survival. Overall, results indicated that ozonation of Atlantic Cod eggs at a dose less than 3.0 mg/L for 90 s produced no negative effect on the larvae up to 30 d posthatch. Furthermore, ozonation of Atlantic Salmon and Rainbow Trout eggs generated no negative effect on the larvae, based on monitoring until 85% yolk sac re-absorption (16 d posthatch).

Received May 6, 2014; accepted October 24, 2014     

蛋源摩氏摩根菌分离、鉴定及遗传进化分析

本研究利用传统细菌鉴定方法、革兰氏阴性细菌生化鉴定系统及16S rDNA 序列分析对分离自市售新鲜鸡蛋中的菌株进行形态学、培养特性及生化特性等生物学特性鉴定及遗传进化分析,结果表明分离株为摩氏摩根菌。药物敏感性试验结果表明,该菌株对痢特灵、利福平、四环素、磷霉素、多黏菌素B和阿奇霉素有耐药性,属于多重耐药菌株。序列分析结果显示,该分离菌株与目前已分离的其他食源性摩氏摩根菌菌株高度同源。     

Effect of Low Dietary Magnesium on Immune Response and Osmoregulation of Atlantic Salmon

Abstract

The influences of dietary magnesium on immune response and on osmoregulation in parr of Atlantic salmon Salmo salar were determined. Groups of fish were fed a casein–gelatin diet unsupplemented (containing about 200 mg Mg/kg) or supplemented with either 300 or 500 mg Mg/kg dry diet (as MgSO₄) for 12 weeks before vaccination to produce fish with different Mg levels, and the feeding regime was continued throughout the study. No differences were observed between the treatment groups in serum-specific antibody levels every second week for 8 weeks after vaccination against Vibrio anguillarum serotypes O1 and O2. Both lysozyme levels and spontaneous hemolytic activities in serum were elevated in vaccinated fish compared with unvaccinated fish. Neither lysozyme activity, complement hemolytic activity, total protein in serum nor blood hemoglobin were affected by dietary Mg. The spontaneous hemolytic activity in serum was lower in fish fed the unsupplemented diet and highest in fish fed the diet supplemented with 500 mg Mg/kg. After 28 weeks on the diets supplemented with graded levels of Mg, a salinity tolerance test (32.5 g/L, 24 h) was performed. High mortality and elevated serum chloride concentrations in all groups after 24 h reflected a general salinity intolerance, but the highest serum chloride level was observed in fish fed the unsupplemented diet. This indicates that low dietary Mg affects the osmoregulation of Atlantic salmon.     

Survey of Pathogens in Hatchery Chinook Salmon with Different Out-Migration Histories through the Snake and Columbia Rivers

The operation of the Federal Columbia River Power System (FCRPS) has negatively affected threatened and endangered salmonid populations in the Pacific Northwest. Barging Snake River spring Chinook salmon Oncorhynchus tshawytscha through the FCRPS is one effort to mitigate the effect of the hydrosystem on juvenile salmon out-migration. However, little is known about the occurrence and transmission of infectious agents in barged juvenile salmon relative to juvenile salmon that remain in-river to navigate to the ocean. We conducted a survey of hatchery-reared spring Chinook salmon at various points along their out-migration path as they left their natal hatcheries and either migrated in-river or were barged through the FCRPS. Salmon kidneys were screened by polymerase chain reaction for nine pathogens and one family of water molds. Eight pathogens were detected; the most prevalent were Renibacterium salmoninarum and infectious hematopoietic necrosis virus. Species in the family Saprolegniaceae were also commonly detected. Pathogen prevalence was significantly greater in fish that were barged through the FCRPS than in fish left to out-migrate in-river. These results suggest that the transmission of infectious agents to susceptible juvenile salmon occurs during the barging process. Therefore, management activities that reduce pathogen exposure during barging may increase the survival of juvenile Chinook salmon after they are released.

Received May 27, 2010; accepted January 17, 2011     

Investigation of the Link between Broodstock Infection,Vertical Transmission,and Prevalence of Flavobacterium psychrophilum in Eggs and Progeny of Rainbow Trout and Coho Salmon

Abstract

The etiological agent of bacterial coldwater disease (BCWD), Flavobacterium psychrophilum, can be transmitted both vertically and horizontally. Outbreaks of BCWD can result in significant losses in salmonid aquaculture. Reduction of outbreaks in fry may be possible through implementation of a management strategy in which progeny of heavily infected broodstock are culled from the general population. Diagnostic assays to quantify F. psychrophilum concentrations in tissue samples and confirm presence of the bacterium in ovarian fluid have been previously validated. In the current study, these assays were used to screen 60 female Rainbow Trout Oncorhynchus mykiss and 60 female Coho Salmon O. kisutch broodstock at two aquaculture facilities. Eyed eggs from 10 female broodstock (five fish from each facility) exhibiting graded levels of infection were transferred to the University of Idaho and monitored through early life stages for the presence of F. psychrophilum. Female Rainbow Trout broodstock were not positive for F. psychrophilum by enzyme-linked immunosorbent assay (ELISA) and prevalence was low in these progeny. However, ELISA optical density values for kidney correlated to F. psychrophilum prevalence in progeny (r = 0.938, P < 0.05) of Coho Salmon. Nested PCR on ovarian fluid was not a reliable indicator of vertical transmission in either species as broodstock ovarian fluid results did not correlate to F. psychrophilum prevalence in eyed eggs. Further research with these assays is necessary; however, results from this study indicate that broodstock screening may be a potential tool for evaluating F. psychrophilum infection levels, which could become an important component for disease management.

Received December 19, 2012; accepted December 8, 2013     

Comparative Uptake of Orally Administered Difloxacin in Atlantic Salmon in Freshwater and Seawater

Abstract

Fluoroquinolones are synthetic antibacterials with potential use for treatment of fish diseases. This study examined the comparative uptake of difloxacin, administered as a single oral gavage, by juvenile Atlantic salmon Salmo salar in freshwater and in seawater. In separate freshwater and seawater experiments with fish of the same spawning cohort, the concentration of difloxacin in plasma and fillets was determined at 1, 2, 4, 8, 12, 18, 24, and 48 h after the fish received the dose. Although plasma difloxacin concentration began to plateau in the freshwater fish at about 24 h, the highest average, 1,195 ng/mL, was in fish sampled at 48 h; thus, the plasma levels continued to rise through the end of the test. A similar pattern was observed in fillets from fish held in freshwater, but the peak average fillet concentration at 48 h was 4,737 ng/g. Contrastingly, in the seawater study, the average peak plasma concentration of 678 ng/mL was observed at 8 h after gavage and the peak average fillet concentration of 924 ng/g was observed at 18 h. Although difloxacin concentrations were higher in the freshwater-held fish than in the seawateradapted fish, the results suggest that difloxacin could be efficacious in treating bacterial diseases of Atlantic salmon in both freshwater and seawater.     

Differing Resistance of Atlantic Salmon Strains and Rainbow Trout to Gyrodactylus salaris Infection

Abstract

Laboratory studies were conducted on the susceptibility of different strains of Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss to infection with the monogenean Gyrodactylus salaris. This parasite, probably originating from the Baltic Sea region, is known to minimally affect Neva River (Baltic Sea) Atlantic salmon. However, following its introduction into Norway, G. salaris has caused severe mortality and morbidity among Norwegian Atlantic salmon, which are considered a highly susceptible strain. The cohabitation experiment included one stock of rainbow trout and four different strains of Atlantic salmon from the Baltic Sea region (Mörrum River, Sweden), Europe (Skjern River, Denmark; Conon River, Scotland), and North America (Bristol Cove River, Canada). Fish were exposed to a Norwegian strain of G. salaris, and parasite population development and distribution were monitored for 7 weeks. Rainbow trout exhibited low susceptibility to G. salaris infection, whereas Conon River and Skjern River Atlantic salmon were highly susceptible and exhibited high mortality rates. Mörrum River Atlantic salmon exhibited intermediate susceptibility and low mortality. Bristol Cove River Atlantic salmon harbored relatively low parasite numbers, but fish mortality was high. Our experiment showed that the Danish Skjern River strain of Atlantic salmon is highly susceptible to G. salaris infection, further supporting the hypothesis that Atlantic Ocean strains are more susceptible to G. salaris infection than are Baltic strains.     

Epizootiological Study of Bacterial Cold-Water Disease in Pacific Salmon and Further Characterization of the Etiologic Agent,Flexibacter psychrophila

Abstract

Isolates of Flexibacter psychrophila were obtained from chinook salmon Oncorhynchus tshawytscha and coho salmon O. kisutch that had previously sustained epizootics of coldwater disease. The pathogen was readily isolated from kidney and mucus of convalescent fish. The organisms were relatively inert in most standard microbiological media but were structurally and serologically homogenous by examination of whole cell protein lysates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In contrast to the homogeneity observed in phenotypic and serologic assays, the isolates studied elaborated varied ribotypes. All isolates produced a single rDNA spacer amplification product of about 240 base pairs.     

Effect of Nanophyetus salmincola and Bacterial Co-Infection on Mortality of Juvenile Chinook Salmon

The freshwater trematode Nanophyetus salmincola has been demonstrated to impair salmonid immune function and resistance to the marine pathogen Vibrio anguillarum, potentially resulting in ocean mortality. We examined whether infection by the parasite N. salmincola similarly increases mortality of juvenile Chinook Salmon Oncorhynchus tshawytscha when they are exposed to the freshwater pathogens Flavobacterium columnare or Aeromonas salmonicida, two bacteria that juvenile salmonids might encounter during their migration to the marine environment. We used a two-part experimental design where juvenile Chinook Salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., and then challenged with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare infection was higher in N. salmincola-parasitized fish than in nonparasitized fish. In contrast, cumulative percent mortality from A. salmonicida infection did not differ between N. salmincola-parasitized and nonparasitized groups. No mortalities were observed in the N. salmincola-parasitized-only and control groups from either challenge. Our study demonstrates that a relatively high mean intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae can alter the outcomes of bacterial infection in juvenile Chinook Salmon, which might have implications for disease in wild fish populations.

Received February 24, 2015; accepted September 7, 2015     

从台湾地区进口甲鱼蛋传入疫病的风险评估

近年来,随着海峡两岸贸易的增长,来自台湾地区的甲鱼蛋输入量猛增。为了在促进两岸贸易的同时,防止有害生物传入,保护大陆地区养殖业安全和人民健康,根据世界贸易组织SPS协定的有关原则,对从台湾地区输入的甲鱼蛋携带的霍乱弧菌等10种细菌和甲鱼虹彩病毒等8种病毒进行了风险分析。评估结果表明:从台湾地区输入甲鱼蛋的主要风险是蛋表面在自然条件下可能被病原性微生物,如细菌和病毒等污染。这些病原可能会随着甲鱼蛋传播到大陆地区,造成疾病流行,或者危害人类健康。虽然其风险不高,但一旦传入,会造成一定的负面影响,其中有些病原也会引起严重后果,因而必须进行适当监管。监管重点是控制蛋表面污染的微生物。基于风险评估,本研究提出了3个风险管理备选方案。     

Primary Isolation and Characterization of Tenacibaculum maritimum from Chilean Atlantic Salmon Mortalities Associated with a Pseudochattonella spp. Algal Bloom

Abstract

This study presents the first isolation of Tenacibaculum maritimum from farmed Atlantic Salmon Salmo salar in Chile. The isolate, designated T. maritimum Ch-2402, was isolated from gills of Atlantic Salmon at a farm located in region X, Los Lagos, Chile, during the harmful algal bloom caused by Pseudochattonella spp. in February 2016. The algal bloom is reported to have caused 40,000 metric tons of mortality in this salmon farming area. The bacterium T. maritimum, which causes tenacibaculosis, is recognized as an important pathogen of marine fish worldwide. Genetic, phylogenetic, and phenotypic characterizations were used to describe the T. maritimum Ch-2402 isolate. The isolate was similar to the type strain of T. maritimum but was genetically unique. Tenacibaculum dicentrarchi isolates were also recovered during sampling from the same farm. Based on the fact that T. maritimum has been shown to cause disease in Atlantic Salmon in other regions, the presence of this bacterium poses a potential risk of disease to fish in the Chilean aquaculture industry.

Received November 6, 2016; accepted May 29, 2017 Published online July 26, 2017     

Nutritive Value of Four Soybean Products in Diets for Atlantic Salmon (Salmo salar,L.)

Abstract

Four processed soybean products were evaluated as protein sources for Atlantic salmon: solvent-extracted soybean meal (SBM44), dehulled and solvent-extracted soybean meal (SBM50), dehulled full-fat soybean meal (FFSBM) and soybean concentrate (SBC). The soybean products replaced high-quality fish meal at levels corresponding to 0, 14, 28, 42 and 56% soybean protein of total protein.

Dietary inclusion of SBC, at the expense of fish meal, did not affect weight gain, carcass lipids, fecal excretion of nutrients or fecal dry matter content significantly. The nutritive value of the SBC protein appeared comparable to that of the fish meal. The three other soybean products impaired performance increasingly with increasing levels of inclusion, indicating lower nutritive value than for the fish meal. The favourable results seen with SBC indicated a great potential of soybean to become an important protein source for Atlantic salmon through improved processing.     

春伐桑园氮磷钾肥比例对种茧育桑叶质量的影响

试验在其它桑园管理措施一致的前提下,改变N、P2O5、K2O肥的施肥比例,并对桑叶质量进行了调查。结果是N:P2O5:K2O=10:5:5的施肥比例,可适当提高4龄起蚕结茧率、虫蛹统一生命率及全茧量、茧层量、茧层率,显著提高产卵量、良卵率和实用孵化率。综合考虑桑叶的产质量,生产中可推荐采用本施肥比例,种茧育桑园尤为如此,以提高蚕种质量。     

激光灼刻在受精鸡蛋上的应用及其影响分析

为探究激光灼刻在受精鸡蛋表面灼刻后对受精鸡蛋的影响,试验比较了激光灼刻后受精鸡蛋的孵化率、健雏率及雏鸡初生重。在试验前期通过改变激光灼刻机的速度、功率寻找出在鸡蛋表面最适的灼刻参数,将试验种蛋按确定参数灼刻后与对照组种蛋一同放入孵化箱内。经过21 d孵化后,分别计算两组种蛋孵化率、健雏率及雏鸡初生重,以此判断激光灼刻对受精鸡蛋的影响。结果显示:参数设定为2 000 mm/s、8 W灼刻效果最好。一定参数下,激光灼刻的受精鸡蛋孵化率、健雏率及雏鸡初生重与未经激光灼刻的受精鸡蛋无显著差异(P>0.05),说明激光灼刻对受精鸡蛋的孵化率、健雏率及雏鸡初生重不会产生影响。