Epizootics of Erythrocytic Inclusion Body Syndrome in Coho Salmon Cultured in Seawater in Japan

Abstract

Epizootics attributable to erythrocyytc inclusion body syndrome (EIBS) occurred among populations of coho salmon Oncorhynchus kisutch cultured in seawater in Japan. Onset of the disease was correlated with water temperatures declining to below 10°C. Symptoms of EIBS were severe anemia with hematocrits of less than 20% and corresponding changes in erythrocyte counts, hemoglobin concentration, and mean corpuscular hemoglobin concentration. Erythrocytes had characteristic inclusion bodies that contained enveloped viral particles with a diameter of approximately 77 nm. The disease was reproduced in artificially induced infections.     

Kinetics of Viral Load and Erythrocytic Inclusion Body Formation in Pacific Herring Artificially Infected with Erythrocytic Necrosis Virus

Abstract

Viral erythrocytic necrosis (VEN) is a condition that affects marine and anadromous fish species, including herrings and salmonids, in the Atlantic and Pacific oceans. Infection is frequently associated with severe anemia and causes episodic mortality among wild and hatchery fish when accompanied by additional stressors; VEN can be presumptively diagnosed by (1) light microscopic identification of a single characteristic—a round, magenta-colored, 0.8-μm-diameter inclusion body (IB) within the cytoplasm of erythrocytes and their precursors on Giemsa-stained blood films; or (2) observation (via transmission electron microscopy [TEM]) of the causative iridovirus, erythrocytic necrosis virus (ENV), within erythrocytes or their precursors. To better understand the kinetics of VEN, specific-pathogen-free Pacific herring Clupea pallasii were infected with ENV by intraperitoneal injection. At 1, 4, 7, 10, 14, 21, and 28 d postexposure, samples of blood, spleen, and kidney were collected and assessed (1) via light microscopy for the number of intracytoplasmic IBs in blood smears and (2) via TEM for the number of virions within erythrocytes. The mean prevalence of intracytoplasmic IBs in the blood cells increased from 0% at 0–4 d postexposure to 94% at 28 d postexposure. Viral load within circulating red blood cells peaked at 7 d postexposure, fell slightly, and then reached a plateau. However, blood cells observed within the kidney and spleen tissues demonstrated high levels of ENV between 14 and 28 d postexposure. The results indicate that the viral load within erythrocytes does not correlate well with IB prevalence and that the virus can persist in infected fish for more than 28 d.

Received September 7, 2011; accepted March 8, 2012     

Lack of Protection following Re-Exposure of Chinook Salmon to Ceratonova shasta (Myxozoa)

The recent identification of multiple genotypes of the salmonid parasite Ceratonova shasta with different virulence levels in Chinook Salmon Oncorhynchus tshawytscha suggests that it is possible to immunize fish against subsequent infection and disease. We hypothesized that exposure of Chinook Salmon to the less-virulent parasite genotype (II) prior to the more virulent parasite genotype (I) would decrease disease and/or result in fewer mature parasites compared with fish only infected with the more virulent genotype. To test this hypothesis, fish were challenged in a combination of field and laboratory exposures, and we measured infection prevalence, percent morbidity, and mature parasite production. Neither mortality nor mature parasite production were reduced when fish were exposed to genotype II prior to genotype I compared with fish exposed only to genotype I, suggesting that protection against C. shasta using a less-virulent genotype of the parasite does not occur.

Received June 9, 2014; accepted September 19, 2014     

鸡包涵体肝炎-心包积水综合征的研究进展

禽腺病毒是一群高度多样的病原体,可引起多种疾病,近年来新发现的禽病中多数与不同型的禽腺病毒感染有关[1]。各种血清型的禽腺病毒都可引起肉鸡的包涵体肝炎,但仅禽腺病毒-4型是引起仔鸡包涵体肝炎-心包积水综合征(IBH-HPS)的特异病原体。IBH-HPS最早报道于1988年。Jaffery和K     

包涵体肝炎对蛋鸡免疫及生殖影响的内分泌机制

为探明包涵体肝炎引起蛋鸡免疫抑制和生殖系统发育障碍的内分泌机制 ,给 60日龄青年蛋鸡口服感染包涵体肝炎 型腺病毒建立鸡包涵体肝炎模型 ,定期跖静脉采血 ,测定血清中甲状腺激素 ( T3、T4)、皮质醇和雌二醇水平。结果显示 ,感染鸡血清 T4水平极显著降低 ,T3呈升高趋势 ,血清皮质醇水平极显著升高 ,雌二醇水平于感染后 1周呈一过性升高 ,之后极显著降低     

The Hydropericardium Syndrome and Inclusion Body Hepatitis in Domestic Fowl

Hydropericardium syndrome, an emerging disease of poultry, has recently been detected in some countries of Asia and America, particularly in broiler birds aged 3–6 weeks. The disease is characterized by its sudden occurrence with high mortality of up to 80% in broilers and low mortality of under 10% in layers, associated with hydropericardium. Its course is of 7–15 days under natural conditions. The causative agent is probably fowl adenovirus serotype 4, belonging to group I aviadenovirus genus of the family adenoviridae, which can be cultivated in primary cell cultures of chicken kidney and embryo liver cells. The transmission of disease occurs laterally by the oral–faecal route. The livers of affected birds show necrotic foci, and basophilic intranuclear inclusion bodies fill the entire enlarged nucleus of some of the hepatocytes. The disease can be diagnosed from its gross lesions, histopathological changes in the liver and by serological tests, such as agar gel diffusion, counter immunoelectrophoresis, indirect haemagglutination and ELISA. It has been brought under control by inactivated liver organ vaccines (0.25 ml/bird) or inactivated cell culture vaccines (10^3.5 LD₅₀/bird) given by the subcutaneous route at 10–15 days of age. The vaccine is effective in the face of an outbreak and significantly reduces the mortality.     

Influence of Infection with Renibacterium salmoninarum on Susceptibility of Juvenile Spring Chinook Salmon to Gas Bubble Trauma

Abstract

During experiments in our laboratory to assess the progression and severity of gas bubble trauma (GBT) in juvenile spring chinook salmon Oncorhynchus tshawytscha, we had the opportunity to assess the influence of Renibacterium salmoninarum (Rs), the causative agent of bacterial kidney disease, on the susceptibility of salmon to GBT. We exposed fish with an established infection of Rs to 120% total dissolved gas (TDG) for 96 h and monitored severity of GBT signs in the fins and gills, Rs infection level in kidneys by using an enzyme-linked immunosorbent assay (ELISA), and mortality. Mortality occurred rapidly after exposure to 120% TDG, with a LT20 (time necessary to kill 20% of the population) of about 37 h, which is at a minimum about 16% earlier than other bioassays we have conducted using fish that had no apparent signs of disease. Fish that died early (from 31 to 36 h and from 49 to 52 h) had significantly higher infection levels (mean ± SE ELISA absorbance = 1.532 ± 0.108) than fish that survived for 96 h (mean ± SE ELISA absorbance = 0.828 ± 0.137). Fish that died early also had a significantly greater number of gill filaments occluded with bubbles than those that survived 96 h. Conversely, fish that survived for 96 h had a significantly higher median fin severity ranking than those that died early. Our results indicate that fish with moderate to high levels of Rs infection are more vulnerable to the effects of dissolved gas supersaturation (DGS) and die sooner than fish with lower levels of Rs infection. However, there is a substantial amount of individual variation in susceptibility to the apparent cumulative effects of DGS and Rs infection. Collectively, our findings have important implications to programs designed to monitor the prevalence and severity of GBT in juvenile salmonids in areas like the Columbia River basin, and perhaps elsewhere.     

A New Eye Disease in Pen-Reared Chinook Salmon Caused by Metacestodes of Gilquinia squali (Trypanorhyncha)

Abstract

Eye infections by metacestodes of Gilquinia squali (Cestoda: Trypanorhyncha) were associated with mortality in chinook salmon Oncorhynchus tshawytscha reared in net-pens in seawater at two separate locations in British Columbia. Mortality associated with the disease was about 10% at one site and undetermined at the second site. The metacestode was found in the vitreous humor of moribund fish with eye lesions, whereas healthy fish from the same pen exhibited neither eye lesions nor the infection. The lesions were characterized by retinal folding and apparent retinal duplication, and cataractous changes; in severe cases, the cornea ruptured, and the lens was extruded. The spiny dogfish Squalus acanthias i s the definitive host for the cestode. Spiny dogfish were prevalent in and around the affected net-pen sites, thus providing coracidia to infect the first intermediate host, which is most likely a crustacean. Although the metacestode is common in the eyes of European whiting Merlangius merlangus in the northeast Atlantic region, this is the first report of the infection in a salmonid.     

Cytopathology and Coagulopathy Associated with Viral Erythrocytic Necrosis in Chum Salmon

Abstract

The 8-month cytopathologic progression of viral erythrocytic necrosis (VEN) disease in chum salmon Oncorhynchus keta is described. Single to multiple acidophilic, cytoplasmic viral inclusion bodies developed first in mature erythrocytes and then, within 1–2 months, all morphologically identifiable hemopoietic cell types contained VEN inclusions. Cytologic analysis indicated that multinucleate giant erythroblasts, ineffective erythropoiesis, and abnormal erythroid cell maturation occurred. A significant increase in blood coagulation time occurred concomitantly. This severe and chronic blood dyscrasia accounts for some of the pathophysiologic sequelae previously observed.     

Detection and Identification of a Strain of Renibacterium salmoninarum Sourced from Cultured Chinook Salmon (Oncorhynchus tshawytscha)

A chronic and infective disease occurred on the farmed Chinook salmon (Oncorhynchus tshaivytscha) in a farm in Huairou district of Beijing and it caused high accumulative mortality.The typical clinical symptoms indicated that it might be bacterial kidney disease (BKD).The purpose of this study was to confirm its pathogen.The typical clinical symptoms of sick fish,morphology observation of bacteria in kidney tissue smears and sequence analysis of bacteria DNA were conducted.A number of gram positive brevi bacteria were observed in kidney tissue print.16S rDNA sequence of K1 strain was more than 99% homology with that of Renibacterium salmoninarum and they were in the same cluster.383 and 320 bp fragment were amplified by PCR and nested PCR.The fragments were sequenced and analyzed.The results showed the fragment was shared 100% nucleotide identity with P57 surface protein gene of Renibacterium salmoninarum. It was indicated that these diseased fish were suffering from BKD caused by Renibacterium salmoninarum.This was the first report about Renibacterium salmoninarum detected in farmed fish in China.     

一株大鳞大麻哈鱼源鲑肾杆菌的检测与鉴定

北京怀柔某养殖场的大鳞大麻哈鱼发生一种慢性,但累计死亡率很高的流行病,为研究大鳞大麻哈鱼感染病原菌的种类,本试验根据病鱼临床症状观察判断疑似感染鲑肾杆菌.试验对病鱼肾脏组织进行印片染色观察细菌形态特征,抽提病鱼的肝脏、肾脏组织DNA,扩增16S rDNA和鲑肾杆菌P57表面蛋白基因片段,测序并运用BLAST进行同源性比对.结果显示,病鱼肾脏组织印片在显微镜下可见大量形态一致的革兰氏阳性短杆菌;采用病鱼肝脏、肾脏组织扩增的16S rDNA基因构建的系统发育树显示与鲑肾杆菌聚为一支;通过PCR扩增出鲑肾杆菌P57表面蛋白基因DNA片段,测序结果与鲑肾杆菌标准株ATCC 33209完全一致,符合率为100%.结果表明病鱼感染鲑肾杆菌并引发细菌性肾病(BKD).这是中国首次在养殖鱼体内检出鲑肾杆菌.     

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.     

Ichthyophoniasis: An Emerging Disease of Chinook Salmon in the Yukon River

Abstract

Before 1985, Ichthyophonus was unreported among Pacific salmon Oncorhynchus spp. from the Yukon River; now it infects more than 40% of returning adult Chinook salmon O. tshawytscha. Overall infection prevalence reached about 45% in the Yukon River and about 30% in the Tanana River between 1999 and 2003. Mean infection prevalence was greater in females than males in the main-stem Yukon River during each of the 5 years of the study, but the infection prevalence in males increased each year until the difference was no longer significant. Clinical signs of ichthyophoniasis (presence of visible punctate white lesions in internal organs) were least at the mouth of the Yukon River (～10%) but increased to 29% when fish reached the middle Yukon River and was 22% at the upper Tanana River. However, clinical signs increased each year from 7% in 1999 to 27% in 2003 at the mouth of the river. As fish approached the upper reaches of the Yukon River (Canada) and the spawning areas of the Chena and Salcha rivers (Alaska), infection prevalence dropped significantly to less than 15% in females on the Yukon River and less than 10% for both sexes in the Chena and Salcha rivers, presumably because of mortality among infected prespawn fish. Age was not a factor in infection prevalence, nor was the position of fish within the run. The source of infection was not determined, but Ichthyophonus was not found in 400 Pacific herring Clupea pallasi from the Bering Sea or in 120 outmigrating juvenile Chinook salmon from two drainages in Alaska and Canada. Freshwater burbot Lota lota from the middle Yukon River were subclinically infected with Ichthyophonus, but the origin and relationship of this agent to the Chinook salmon isolate is unknown.     

Chinook Salmon Epizootics in Lake Michigan: Possible Contributing Factors and Management Implications

Abstract

Stability of the Lake Michigan fishery for chinook salmon Oncorhynchus tshawytscha at high levels became questionable after stocks declined dramatically following spring epizootics in which bacterial kidney disease (BKD) was a major factor. Initially stocked in 1967, favorable survival and growth of chinook salmon through the 1970s led to increases in abundance and in popularity with anglers. Returns of chinook salmon improved annually until the late–1980s, when, with little warning, spring epizootics reduced the abundance of adult salmon by 50% or more. Reduced abundance of alewives (Alosa pseudoharengus), coupled with an increase in chinook salmon density and heavy parasite infection rates were hypothesized to have reduced chinook salmon growth and fitness and to have increased their susceptibility to BKD. Evidence of slower growth exists and low food availability may be the stressor that triggered the epizootics. Chinook salmon were a major component of the economic development and subsequent hardship of the sportfishing industry on Lake Michigan. Sustaining the chinook salmon fishery at previous levels may require managing for high abundance of alewives, which may be inconsistent with overall fish community management goals. The future sustainability and role of chinook salmon needs to be reevaluated in the context of the entire Lake Michigan fish community.     

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     

Monoclonal Antibody-Based Analysis of the Renibacterium salmoninarum p57 Protein in Spawning Chinook and Coho Salmon

Abstract

Renibacterium salmoninarum, causative agent of bacterial kidney disease of salmonid fish, produces large amounts of soluble proteins during infection and broth culture. An enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies was developed for the precise quantification of p57, a major component of these proteins. Kidney, spleen, blood, and reproductive fluids of adult Pacific salmon Oncorhynchus spp. were examined by means of the assay. Kidney and spleen harbored the highest concentrations of this antigen. In the populations of returning salmon tested, 5–25% of the fish had p57 concentrations above a baseline level of 3 ng antigenig tissue, and antigen concentrations as high as 200 μg/g tissue were detected in kidneys of individual fish. The ELISA was compared to direct fluorescent antibody analysis, in which rabbit anti-R. salmoninarum antiserum was used to identify infected fish. There was 99% agreement (199 of 201 examined fish) between the two methods. Western blot analysis was used to demonstrate that antigenic proteins present in infected fish were similar to those seen from antigen prepared from broth culture of R. salmoninarum, although less degradation of p57 appears to occur in vivo.     

An Uncommon Presentation of Fungal Infection in Atlantic Salmon Fry

Abstract

Moribund fry from several families of Atlantic salmon Salmo salar had a conspicuous cranial defect that corresponded with 18.3% (SD, 4.5%) mortality of fry. The lesion was characterized by a domed, epithelium-covered swelling centered over the optic tectum. The swelling was an edematous reaction to the extension of a fulminating necrotizing mycotic stomatitis and bronchitis into the cranial tissues including the ventral meninges. An oomycete is believed to be the pathogen. The percentage mortality of fry in each separately reared family ranged from 1.0 to 63.3%. An analysis of variance revealed that the differences in mortality among families were significant (P < 0.0001), and that the difference between replicates of each family, reared apart, were not significant. A possible genetic basis for resistance to fungal infection is discussed.     

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     

Protection of Chinook Salmon Smolts with Oral Doses of Erythromycin against Acute Challenges of Renibacterium salmoninarum

Abstract

We challenged duplicate groups of yearling smolts of chinook salmon Oncorhynchus tshawytscha held in seawater with an intraperitoneal inoculation of the kidney disease bacterium Renibacterium salmoninarum 1 d before and at intervals of 1, 11, and 29 d after a 21-d oral administration of erythromycin thiocyanate at 0.1 g/kg body weight per day. Most mortality attributable to bacterial kidney disease (BKD) in fish challenged with R. salmoninarum but not administered erythromycin occurred 2–3 weeks after challenge; the average survival 35 d after challenge was only 9%. Nearly 70% of the fish challenged 1 d before the erythromycin feeding were alive 35 d after the 21-d treatment, and more than 98% of the fish challenged the day after the 21-d erythromycin treatment survived a further 35 d. Fish challenged 29 d after the erythromycin treatment were not significantly protected against BKD. Chinook salmon that were not challenged with the bacterium but were injected with sterile phosphate-buffered saline and then fed a ration with erythromycin survived at a significantly higher rate than unchallenged fish injected with saline but not fed erythromycin. Fish that were not injected with saline or R. salmoninarum survived at a significantly higher rate than fish handled and injected with saline or pathogen. Because erythromycin protected fish challenged just before and immediately after treatment, the antibiotic should be useful early in an outbreak of BKD and as a prophylactic when stresses are expected.