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1.
Inactivated infectious haematopoietic necrosis virus (IHNV) vaccines   总被引：1，自引：0，他引：1  
The inactivation dynamics of infectious haematopoietic necrosis virus (IHNV) by b-propiolactone (BPL), binary ethylenimine (BEI), formaldehyde or heat and the antigenic and immunogenic properties of the inactivated vaccines were evaluated. Chemical treatment of IHNV with 2.7 mm BPL, 1.5 mm BEI or 50 mm formaldehyde abolished virus infectivity within 48 h whereas heat treatment at 50 or 100 degrees C rendered the virus innocuous within 30 min. The inactivated IHNV vaccines were recognized by rainbow trout, Oncorhynchus mykiss, IHNV-specific antibodies and were differentially recognized by antigenic site I or antigenic site II IHNV glycoprotein-specific neutralizing monoclonal antibodies. The BPL inactivated whole virus vaccine was highly efficacious in vaccinated rainbow trout challenged by waterborne exposure to IHNV 7, 28, 42 or 56 days (15 degrees C) after immunization. The formaldehyde inactivated whole virus vaccine was efficacious 7 or 11 days after vaccination of rainbow trout but performed inconsistently when tested at later time points. The other vaccines tested were not efficacious.  相似文献   

2.
In this study, we reasoned that if we challenged rainbow trout with the causative agents of enteric redmouth disease (ERM), rainbow trout fry syndrome (RTFS), and viral haemorrhagic septicaemia (VHS), we would: 1) detect additive genetic variation for resistance to ERM, RTFS, and VHS; and 2) find that resistance of the trout to ERM and RTFS are favourably correlated genetically, while resistance to VHS is unfavourably correlated with resistance to ERM and RTFS. We tested these premises by challenging 63 full-sib families of rainbow trout (50 sires, 38 dams) with Yersinia ruckeri, Flavobacterium psychrophilum, and VHS virus, the causative agents of ERM, RTFS, and VHS. Resistance to each disease was assessed as both a binary trait (i.e., died/survived) and a longitudinal trait (i.e., time until death following challenge). Additive genetic variation and genetic correlations for resistance to ERM, RTFS, and VHS were estimated by fitting a threshold liability model to resistance assessed as a binary trait. As a longitudinal trait, additive genetic variation and genetic correlations were estimated by fitting a Weibull frailty model to the times until death. Our findings support the first of our premises as we detected additive genetic variation for resistance to ERM, RTFS, and VHS. The heritability for resistance to ERM, RTFS, and VHS ranged between 0.42 and 0.57 on the underlying liability scale when resistance was assessed as a binary trait. As a longitudinal trait, the heritabilities ranged between 0.07 and 0.21 for time until death on the logarithmic-time scale. We were, however, unable to support our second premise as we found that resistance to each of the diseases tended to be weakly correlated genetically. The genetic correlations between the resistances ranged between −0.11 and 0.15 when resistance was assessed as a binary trait, and between −0.23 and 0.16 when resistance was assessed as a longitudinal trait. These findings are encouraging for commercial trout production. The additive genetic variation detected for resistance demonstrates that selectively breeding trout for resistance to ERM, RTFS, and VHS will be successful, providing a complementary approach to control these diseases. The weak genetic correlations suggest that it should be relatively easy to improve resistance to each of the diseases simultaneously.  相似文献   

3.
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4.
Morphologically differentiated sex chromosomes are found in only few salmonid species. Some populations of rainbow trout, Oncorhynchus mykiss, exhibit chromosomal polymorphism related to sex. We found sex-related chromosomal polymorphism in fish from the synthetic Rutki strain, Poland, in approximately 85% (n = 22) of examined males (XY-like) whereas the remaining males (n = 4) possessed chromosomes similar to these observed in females (XX-like). To investigate whether males possessing XX-like chromosomes were genotypic males or genotypic females with altered phenotypic sex, androgenetic progeny of four males (representing both XY-like and XX-like forms, n = 2 + 2) was examined. Androgenetic progeny (F1) of all four fathers consisted of both phenotypic females and males. F1 male progeny of two fathers showed XX-like chromosomes whereas F1 male progeny of the other two fathers possessed YY-like (supermale) chromosomes. F1 were reared further until they were sexually mature. Two males from each of four F1 families were used to produce F2 androgenetic and control F2 generation. All F2 individuals, androgenetics and control, were phenotypic males at sexual maturation. The results indicate that males possessing XX-like chromosomes are genetic males and they are not sex-reversed females. Thus, the Y chromosome can exist in different morphological forms in farmed rainbow trout. The YS chromosome (shorter form, unlike the X chromosome) has a shorter or absent p-arm and lacks the pericentromeric cluster of AT-rich chromatin and the 5S rDNA sequences that are found in the YL chromosome (longer form, like the X chromosome).  相似文献   

5.
In spring 2008, infectious hematopoietic necrosis virus (IHNV) was detected for the first time in the Netherlands. The virus was isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), from a put‐and‐take fishery with angling ponds. IHNV is the causative agent of a serious fish disease, infectious hematopoietic necrosis (IHN). From 2008 to 2011, we diagnosed eight IHNV infections in rainbow trout originating from six put‐and‐take fisheries (symptomatic and asymptomatic fish), and four IHNV infections from three rainbow trout farms (of which two were co‐infected by infectious pancreatic necrosis virus, IPNV), at water temperatures between 5 and 15 °C. At least one farm delivered trout to four of these eight IHNV‐positive farms. Mortalities related to IHNV were mostly <40%, but increased to nearly 100% in case of IHNV and IPNV co‐infection. Subsequent phylogenetic analysis revealed that these 12 isolates clustered into two different monophyletic groups within the European IHNV genogroup E. One of these two groups indicates a virus‐introduction event by a German trout import, whereas the second group indicates that IHNV was already (several years) in the Netherlands before its discovery in 2008.  相似文献   

6.
应用重复力模型估计虹鳟生长性状的遗传力和育种值   总被引：6，自引：0，他引：6  
采用单性状重复观测值动物模型（重复力模型）对中国水产科学研究院黑龙江水产研究所渤海冷水性鱼类试验站5个品系后代的4026尾虹鳟 (Onchorynchus mykiss) 的生长性状（体重、体长和肥满度）进行了遗传参数和育种值的估计,并对各个性状的表型和遗传趋势进行预测。在此模型中考虑了年份-季节固定效应、父本固定效应和母本固定效应;随机效应有个体的加性效应和个体永久环境效应。结果表明：该5个品系虹鳟及后代体重的遗传力为0.35,体长为0.10、肥满度为0.34。个体间的育种值差异较大,仅排名前20位相差近10倍;从不同评定方法的秩相关来看,用综合育种值法与单个性状方法之间存在极显著的相关,其相关度分别为0.998, 0.877, 0.85. -0.071, -0.064和-0.13。用综合育种值对虹鳟个体的选择价值的评定和名次排列, 以及与单个性状的评定名次有着一定程度上的差别。体重、体长和肥满度性状,其表型趋势与遗传趋势基本一致的品系,均可通过表型值进行选择。  相似文献   

7.
为探讨鲁氏耶尔森菌侵染虹鳟的致病机制,本实验建立了鲁氏耶尔森菌感染虹鳟引起的肠炎红嘴病的病理模型,制定相应的临床症状及组织病理学评分系统,并对该模型进行研究。将43尾平均体质量约为12 g的健康虹鳟随机分成5组:3个实验组(n=30)、对照组(n=10)和哨兵组(n=3)。3个实验组分别采用2.0×106、2.0×107和2.0×108 CFU/m L的鲁氏耶尔森菌感染浓度,通过腹腔注射方式进行人工感染试验。对感染鲁氏耶尔森菌的虹鳟肠、肝脏、脾脏和肾脏组织进行镜检及临床症状、剖检病变判断,结合细菌学检测,按制定的评分系统评价各组肠炎红嘴病造模效果,确定最佳造模方案。结果显示,各攻毒组虹鳟感染后72 h均出现不同程度死亡,临床症状表现为红嘴、肛门红肿、鳍(胸鳍、腹鳍、臀鳍)等出现不同程度充血,下颌部、腹部出现出血点等。组织病理学可见肝脏、脾脏、肾脏及肠组织均有炎性细胞浸润现象出现,肝细胞、肠上皮细胞和肾小管上皮细胞等实质细胞变性、坏死,脾脏部位淋巴细胞减少、红细胞死亡堆积。综合各组得分发现,2.0×107 CFU/m L组的鲁氏耶尔森菌感染虹鳟造模效果最佳,患病虹鳟的临床症状显著且组内差异较小,病程迁延较长,便于研究。研究表明,对体质量约12 g的虹鳟幼鱼腹腔注射0.1 m L浓度为2.0×107 CFU/m L的鲁氏耶尔森菌可成功构建肠炎红嘴病病理模型。  相似文献   

8.
The aim of this work was to explore the efficacy of a veterinary drug, Triclabendazole (5-chloro-6-(2, 3-dichlorophenoxy)-2-methylthio-1H-benzimidazole), in an inclusion complex with β-cyclodextrin as a suitable treatment for parasitic diseases caused by Ichthyophthirius multifiliis in rainbow trout. The efficacy was determined by the reduction in the infection intensity. The complexes were prepared by the kneading method and were characterized by DSC and X-ray diffractometry. The selected stoichoimetry was 1:3 because of the higher percentage of Triclabendazole complexed with cyclodextrin. Administration of Triclabendazole and complex was carried out by including appropriate doses in animal feed. Our studies suggest that the Triclabendazole–cyclodextrin complex results in a reduction in the infection degree and trophont size is decreased in the animals treated. The oral treatment of Triclabendazole in inclusion complexes may be an alternative to bath treatments in trout farming.  相似文献   

9.
A DNA vaccine against infectious haematopoietic necrosis virus (IHNV) is effective at protecting rainbow trout, Oncorhynchus mykiss, against disease, but intramuscular injection is required and makes the vaccine impractical for use in the freshwater rainbow trout farming industry. Poly (D,L-lactic-co-glycolic acid) (PLGA) is a U.S. Food and Drug Administration (FDA) approved polymer that can be used to deliver DNA vaccines. We evaluated the in vivo absorption of PLGA nanoparticles containing coumarin-6 when added to a fish food pellet. We demonstrated that rainbow trout will eat PLGA nanoparticle coated feed and that these nanoparticles can be detected in the epithelial cells of the lower intestine within 96 h after feeding. We also detected low levels of gene expression and anti-IHNV neutralizing antibodies when fish were fed or intubated with PLGA nanoparticles containing IHNV G gene plasmid. A virus challenge evaluation suggested a slight increase in survival at 6 weeks post-vaccination in fish that received a high dose of the oral vaccine, but there was no difference when additional fish were challenged at 10 weeks post-vaccination. The results of this study suggest that it is possible to induce an immune response using an orally delivered DNA vaccine, but the current system needs improvement.  相似文献   

10.
All evidence to date suggest that sperm motility is the primary determinant of fertilization success in externally fertilizing fish species. Ovarian fluid, which comprises 10–30% of the total egg volume in salmonids, enhances sperm motility with respect to swimming speed, trajectory and the duration of movement. It was recently demonstrated that there is individual variability in sperm motility enhancing potential of ovarian fluid of particular females. In the present study we examined the effect of particular ovarian fluids collected from 31 females on the sperm motility parameters of one male of rainbow trout (Oncorhynchus mykiss) using computer-assisted sperm analysis (CASA). During our experiment we also monitored the pH of ovarian fluid. We found that particular fluids differed in the ability to activate spermatozoa; sperm remained immotile in four fluids and exhibited 50–100% motility in 27 samples. The percentage of motile sperm, velocity and duration of movement positively correlated with ovarian fluid pH (r2 = 0.34–0.62). These data strongly suggest that the pH of the ovarian fluid is the primary determinant of sperm motility in rainbow trout under natural conditions of fertilization.  相似文献   

11.
In previous proof-of-concept work, it was shown that the use of treated coal mine water for rainbow trout (Oncorhynchus mykiss) culture in a cage was technically feasible, though only a 50-fish bioassay was grown and no work on production-related issues was conducted. To further advance the use of treated mine water, an under-utilized water resource throughout Mid Appalachia, work was conducted to assess the effects of using treated coal mine water for the intensive production of rainbow trout in a flow-through system. During this study, comprehensive water quality data were collected to supplement fish weight and length data taken during routine monthly sampling events. The 8000 fish grew well in the raceway system over the 9 months of production, where a feed conversion ratio of 1.4 and a condition factor of 5.1 × 10−4 were measured with stocking and harvest densities of 26.4 and 50.2 kg/m3, respectively. Further, total net production was 3275 kg (7220 lb) with 98.6% survival. Throughout the study, dissolved ion concentrations (Fe, Al, Mg, Ca, and SO4) often exceeded recommended tolerance limits. Further, elevated ammonia nitrogen concentrations generated from a component of the mine water-treatment process were identified as a potential limiting factor for aquaculture development. However, when the non-ideal effects of high ionic strength and the speciation of dissolved metal–ligand complexes were taken into account, the concentrations of free metal ions were within recommended tolerance limits.  相似文献   

12.
This study examined the genetic variability and genotype × diet interactions during early growth (initial mean body weight 1.2 g) among seven heterozygous clones of rainbow trout, Oncorhynchus mykiss. The clones were hand-fed a diet containing either fishmeal or plant proteins during a 49-day trial divided into two periods (P1, 26 days, and P2, 23 days). Weight, variation of weight within clone, feed intake, feed efficiency and mortality were calculated for both periods.There was a highly significant effect of diet and of clone for all traits at both periods, except for feed efficiency and mortality at P1. Highly significant interactions between diet and clone were also recorded for all these traits, except for mortality at P1. The occurrence of genotype × diet interactions when feeding juvenile rainbow trout with an all plant-protein diet indicates that a highly performing genotype on a fishmeal diet may perform poorly when fed a plant-protein diet. Interactions were found for the two major determinants of growth, i.e. feed intake and feed efficiency, showing that the dietary response differs according to the genotype. Monitoring of the within-clone variability of weight showed that a plant-based diet is likely to enhance the overall phenotypic variance in a population, whatever its initial genetic variability.  相似文献   

13.
Abstract. Sublethal exposure of rainbow trout fry, Salmo gairdneri Richardson, to polychlorinated biphenyls (PCBs) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) did not significantly affect mortality or mean time to death following challenge with infectious haematopoietic necrosis virus (IHNV). Rainbow trout fry were fed experimental diets containing 0, 5. 50 or 500 μg/g of the technical polychlorinated biphenyl mixture, Aroclor 1254, for 30 days or were injected intraperitoneally with graded single doses of TCDD. Fish from each treatment group were challenged by immersion exposure to IHNV. At early time points following virus challenge, histopathologic lesions due to virus disease were more severe and occurred more frequently in virus-challenged fish which received either toxicant than in virus-challenged control fish. Taken together, these findings suggest that rainbow trout are less sensitive than mammalian species to the sublethal effects of these structurally related toxicants on resistance to viral disease. However, a subtle effect of these chemicals on the pathogenesis of the virus disease is suggested by the histopathologic data.  相似文献   

14.
本研究将传染性造血器官坏死病病毒(infectious hematopoietic necrosis virus,IHNV)分离株SD-12糖蛋白(glycoprotein,G)基因克隆进商业化载体pc DNA3.1(+),构建了IHNV G的表达载体,即传染性造血器官坏死病(infectious hematopoietic necrosis,IHN)核酸疫苗,命名为p IHNsd-G。采用背鳍基部肌肉注射的方式,以2μg/尾的剂量免疫虹鳟(Oncorhynchus mykiss)鱼苗(5.0±0.5)g。于免疫后第4天及第7天,利用real-time PCR技术检测免疫虹鳟头肾及接种部位肌肉组织Mx-1基因表达情况;于免疫后第21天,以100倍半数组织培养感染剂量(tissue culture infective dose,TCID50)采取腹腔注射的方式进行攻毒实验,计算核酸疫苗相对保护率(relative percent survival,RPS);于免疫后第60天及150天检测免疫虹鳟血清IHNV中和抗体效价;最后,以p IHNsd-G的启动子序列和氨苄青霉素抗性基因序列为目标基因,利用PCR方法监测p IHNsd-G在免疫虹鳟接种部位的动态分布情况。结果显示:Mx-1基因在头肾和接种部位肌肉中均显著上调表达,并且在接种部位肌肉组织中明显高于同一时间点的头肾组织;攻毒实验中p IHNsd-G对虹鳟的相对保护率高达94.4%;而在免疫后第60天,所有免疫虹鳟血清中均存在中和抗体,其最高效价高达320,在免疫后第150天,最高抗体效价为80,自此,说明已成功获得有效的IHN核酸疫苗。p IHNsd-G在虹鳟接种部位的PCR监测结果显示:在免疫后的第1天即可在注射部位的肌肉中检测到全部p IHNsd-G目标片段,在第84天时已经无法从注射部位肌肉中扩增出全长氨苄青霉素抗性基因,而所有目标基因在第150天时均消失不见。本研究在成功构建IHN核酸疫苗并系统地验证了其有效性的基础上,开展了该疫苗在接种部位的动态分析研究,为IHN核酸疫苗的研发和安全性评价研究提供了基础数据。  相似文献   

15.
Infectious haematopoietic necrosis virus (IHNV) is a serious pathogenic threat to rainbow trout (Oncorhynchus mykiss) and other salmonids. Major histocompatibility complex (MHC) polymorphisms have been closely linked to disease susceptibility. In this study, polymerase chain reaction‐single‐strand conformation polymorphism, cloning and sequencing were used to examine MHC class II β chain gene (DAB) polymorphisms and analyse their association with IHNV in rainbow trout. The DAB exon 2 was amplified from 104 resistant and 91 susceptible fish. Forty‐five different alleles were identified, 30 of which were novel. Individuals possessed between one and four alleles, indicating that the genome harbours at least two closely related DAB exon 2 loci. The percentages of amino acid mutations in the peptide binding region (PBR) and non‐PBR were 66.67% and 45.28% respectively. The rate of non‐synonymous substitutions occurred at a significantly higher frequency than synonymous substitutions, suggesting the existence of balancing selection for maintenance of polymorphisms at the DAB exon 2 locus. The alleles Onmy‐DAB*0201 (P < 0.001), Onmy‐DAB*0904 (P < 0.001) and Onmy‐DAB*1102 (P < 0.01) were associated with susceptibility to IHNV, while Onmy‐DAB*0103, Onmy‐DAB*0601 and Onmy‐DAB*1402 were associated with resistance (P < 0.001). These alleles could be used as molecular markers for IHNV resistance breeding in rainbow trout.  相似文献   

16.
Co‐infection of rainbow trout with infections haematopoietic necrosis virus (IHNV) and Flavobacterium psychrophilum is known to occur, and it has been speculated that a combined infection can result in dramatic losses. Both pathogens can persist in fish in an asymptomatic carrier state, but the impact of co‐infection has not been well characterized or documented. In this study, it was hypothesized that fish co‐infected with F. psychrophilum and IHNV would exhibit greater mortality than fish infected with either pathogen alone. To test this, juvenile rainbow trout were co‐infected with low doses of either IHNV or F. psychrophilum, and at 2 days post‐initial challenge, they were given a low dose of the reciprocal pathogen. This combined infection caused high mortality (76.2%–100%), while mortality from a single pathogen infection with the same respective dose was low (5%–20%). The onset of mortality was earlier in the co‐infected group (3–4 days) when compared with fish infected with F. psychrophilum alone (6 days) or IHNV (5 days), confirming the synergistic interaction between both pathogens. Co‐infection led to a significant increase in the number of F. psychrophilum colony‐forming units and IHNV plaque‐forming units within tissues. This finding confirms that when present together in co‐infected fish, both pathogens are more efficiently recovered from tissues. Furthermore, pathogen genes were significantly increased in co‐infected groups, which parallel the findings of increased systemic pathogen load. Extensive tissue necrosis and abundant pathogen present intracellularly and extracellularly in haematopoietic tissue. This was pronounced in co‐infected fish and likely contributed to the exacerbated clinical signs and higher mortality. This study provides novel insight into host–pathogen interactions related to co‐infection by aquatic bacterial and viral pathogens and supports our hypothesis. Such findings confirm that mortality in fish exposed to both pathogens is greatly elevated compared to a single pathogen infection.  相似文献   

17.
In this study, we wanted to evaluate genetic variation in resistance to infectious pancreatic necrosis (IPN) in a breeding population of rainbow trout. Two hundred families were challenged at a commercial test station, using methods that were previously developed for testing resistance to IPN in Atlantic salmon. Thirty‐five days after the challenge the accumulated mortality was 26%. The results show that resistance to IPN is moderately heritable in the tested population (h2=0.30). The genetic correlation between IPN resistance and body weight was found to be low and non‐significant. The significant additive genetic variation found in IPN resistance after a controlled challenge test gives promise for successful breeding for increased resistance to IPN in rainbow trout.  相似文献   

18.
Infectious haematopoietic necrosis virus (IHNV) is the causative agent of infectious haematopoietic necrosis, a disease of salmonid responsible for great economic losses. The disease occurs in most parts of the world where rainbow trout is reared but has not been previously reported in Kenya. In this study, rainbow trout fry and growers from two farms in Nyeri County were screened for IHNV. Whole fry (n = 4 from each farm) and kidney samples from growers (n = 15 and n = 6 from the two farms, respectively) were collected and preserved for cell culture examination or PCR analysis. Screening of samples was done by PCR followed by sequencing of the glycoprotein gene of the virus. Demonstration of the virus was done by propagation in EPC cells followed by the indirect fluorescence antibody test (IFAT). The results revealed the presence of IHNV at low prevalence of 0.1 and 0.4 for the two farms. The virus was confirmed both by IFAT and by partial sequencing of the G gene. Phylogenetic analysis revealed that the Kenyan isolates were identical to those of the J genogroup found mostly in Asia. The findings have implications for biosecurity measures and import regulations for the Kenyan rainbow trout industry.  相似文献   

19.
The parasitic ciliate Ichthyophthirius multifiliis has a low host specificity eliciting white spot disease (WSD) in a wide range of freshwater fishes worldwide. The parasite multiplies rapidly whereby the infection may reach problematic levels in a host population within a few days. The parasite targets both wild and cultured fish but the huge economic impact of the protozoan is associated with mortality, morbidity and treatment in aquacultural enterprises. We have investigated the potential for genetic selection of WSD-resistant strains of rainbow trout. Applying the DNA typing system Affymetrix® and characterizing the genome of the individual fish by use of 57,501 single nucleotide polymorphisms (SNP) and their location on the rainbow trout chromosomes, we have genetically characterized rainbow trout with different levels of natural resistance towards WSD. Quantitative trait loci (QTL) used for the selection of breeders with specific markers for resistance are reported. We found a significant association between resistance towards I. multifiliis infection and SNP markers located on the two specific rainbow trout chromosomes Omy 16 and Omy 17. Comparing the expression of immune-related genes in fish—with and without clinical signs—we recorded no significant difference. However, trout surviving the infection showed high expression levels of genes encoding IgT, T-cell receptor TCRβ, C3, cathelicidins 1 and 2 and SAA, suggesting these genes to be associated with protection.  相似文献   

20.
Rainbow trout from 23 families were evaluated for growth and resistance to the bacterial coldwater disease (BCWD) caused by Flavobacterium psychrophilum and infectious haematopoietic necrosis (IHN) caused by IHN virus. Average family weights were between 161 and 263 g with an average of 225 g at 213 days post‐fertilization with specific growth rates ranging from 2.37 to 2.88. Per cent survival of fish challenged with F. psychrophilum was between 18% and 100%, while for those challenged with IHNV, the range was between 12% and 93%. Significant positive correlations were found for end body weight and resistance to IHN (P < 0.05) and for early body weight and resistance to BCWD (P < 0.1). However, no significant correlations were detected between resistance to both pathogens or disease resistance and overall genetic diversity or diversity within the major histocompatibility locus.  相似文献   

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