QTL for IHNV resistance and growth identified in a rainbow (Oncorhynchus mykiss) × Yellowstone cutthroat (Oncorhynchus clarki bouvieri) trout cross

Infectious hematopoietic necrosis virus (IHNV) is a major constraint to rainbow trout culture. Yellowstone cutthroat trout (Oncorhynchus clarki bouvieri) have greater resistance to this virus than do rainbow trout (O. mykiss), but the genetic mechanism of this resistance is not understood. We conducted a genome scan using a backcross of cutthroat trout into a rainbow trout background to estimate the number and locations of quantitative trait loci (QTL) associated with IHNV resistance and growth in trout. IHNV resistance was considered in terms of both survival (binary trait) and days to death (quantitative trait). The genetic map was scanned using interval mapping via two different approaches: one model considered survival alone and a second two-part model combined both survival and days to death. Three QTL were significantly (P ≤ 0.05) associated with virus resistance genome-wide, explaining 32.5% of the phenotypic variation. Cutthroat alleles at two of these QTL resulted in increased resistance to the pathogen, as expected. No growth QTL were detected in this cross. We suggest that these traits are genetically independent.     

Occurrence of two forms of Y chromosome in rainbow trout (Oncorhynchus mykiss) males from Rutki strain

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 Y^S 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 Y^L chromosome (longer form, like the X chromosome).     

虹鳟肠炎红嘴病病理模型的构建

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。     

Designing an in‐house ELISA to detect antibody against viral haemorrhagic septicaemia virus using recombinant N protein in Iranian farmed rainbow trout (Oncorhynchus mykiss)

Viral haemorrhagic septicaemia (VHS) is one of the most important viral diseases in rainbow trout that has caused great losses to Iranian rainbow trout aquaculture industry in the last 3 years. Therefore, rapid and reliable diagnosis of VHS virus infections is of great importance. An enzyme linked immunosorbent assay (ELISA) method was performed to study serum antibodies against viral haemorrhagic septicaemia virus (VHSV) using recombinant fragments of their N protein. For this purpose, the virus was first isolated from an infected farm. A part of the nucleocapsid (1–505 bp) gene was amplified by RT‐PCR using specific primers. The amplified fragment was ligated to pMALc2x vector and transferred to DH5α strain of Escherichia coli. Then, recombinant plasmids were tested for protein expression in E. coli Rosetta strain. SDS‐PAGE analysis indicated the production of a recombinant protein with an expected molecular weight of 61 KDa. Analysis of trout serum samples from seven previously infected farms and two VHS free farms showed that the designed ELISA method was effective in diagnosing the infected fish. The results revealed that the developed serological assay using designed ELISA based on recombinant protein (N) has the potential to be used in monitoring studies and to determine the prevalence of VHS in rainbow trout farms. The present data allow evaluating the levels of nonneutralizing antibodies without crude virus preparations.     

Effects of soybean meal based diet on growth performance, gut histopathology and intestinal microbiota of juvenile rainbow trout (Oncorhynchus mykiss)

Effects of soybean meal (SBM) based diet on growth performance, histology of the intestinal epithelium and on the gut microbiota of juvenile rainbow trout (Oncorhynchus mykiss) were investigated on two trials lasting for 8 weeks (Trial I) and 18 weeks (Trial II). The microbiological characterization was done both with conventional plating techniques, biochemical profiling and length heterogeneity analysis of PCR amplified 16S rDNA (LH-PCR).Typical histological changes were found after 18 weeks of SBM feeding (Trial II). Mean height of simple foldings were significantly higher within the group fed with fish meal (FM) based diet. These changes were not, however, reflected in the appearance, behavior and weight gain of the fish, although the feed conversion rate (FCR) was higher, compared to fish fed with the traditional FM feed.In both SBM and FM groups the counts of cultivable intestinal bacteria dropped within the first 4 weeks of feeding (Trial I). Subsequently, the bacterial numbers increased in the FM group, but not in the SBM group by the 8th week of the trial. The LH-PCR data suggested also that diet-related qualitative changes in the microbiota had occurred (Trial II). The main identified bacteria were mainly representatives of the genera Aeromonas, Sphingomonas and Chryseomonas, and among the lactic acid bacteria, Lactococcus and Lactobacillus.Future research will be directed to the identification and eventual elimination the SBM-associated factors involved in the inflammatory responses and unfavorable FCR, in particular by modifying the intestinal microbiota.     

Evidence of genotype–diet interactions in the response of rainbow trout (Oncorhynchus mykiss) clones to a diet with or without fishmeal at early growth

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.