Molecular cloning and characterization of sea bass (Dicentrarchus labrax L.) CD8alpha

In this work, the gene and cDNA of the sea bass CD8alpha have been isolated and characterized. The coding sequence has an ORF of 666 bp. It retains the Ig motif that interacts with MHC and the two cysteines responsible for an intra-chain disulfide bridge. The hinge region contains the two essential cysteines involved in dimerization. The transmembrane region is well conserved in all analysed sequences. Similar to other teleosts, the cytoplasmic region lacks the consensus p56(lck) motif common in higher vertebrates. Analysis of the expression pattern using RT-PCR shows the highest expression in the thymus. Like in the human gene, the sea bass CD8alpha genomic structure is organized into six exons, which roughly correspond to separate functional domains of the protein. Southern blotting shows that CD8alpha exists as a single copy gene. Together, these results support the concept that the basic structure of CD8alpha has been maintained through evolution.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Selection for weaning weight and postweaning gain in Hereford cattle. II. Response to selection

Single trait selection was practiced in three lines of Hereford cattle at two locations. Bulls were selected within sire families for increased weaning weight (WW) in the WW line (WWL), for postweaning gain (PG) in the PG line (PGL) and at random in the control line (CTL). Data include the performance of 2,467 calves produced from 1967 to 1981. Environmental effects were estimated from CTL (method I) and from multiple regression procedures (method II). Phenotypic and environmental time trends were negative for WW and generally were positive for PG. Estimated genetic gains for WW in WWL were 1.07 +/- .51 kg/yr in bulls and .62 +/- .36 kg/yr in heifers using method I and .50 +/- .31 kg/yr in bulls and .10 +/- .17 kg/yr in heifers using method II. Corresponding values for PG in PGL were .85 +/- .40 and 1.03 +/- .24 kg/yr in bulls and .30 +/- .28 and .37 +/- .12 kg in heifers. Correlated genetic gains for WW in PGL were larger than direct WW gains, whereas genetic gains for PG in WWL were smaller than direct PG gains. From method I, estimates of realized heritability (h2R) for WW were .31 +/- .18 in bulls and .22 +/- .13 in heifers. For PG, h2R was .31 +/- .13 in bulls and .06 +/- .12 in heifers. Using method II, h2R for WW was .09 +/- .08 in bulls and .02 +/- .07 in heifers. Corresponding values for PG were .29 +/- .10 and .11 +/- .08. Joint estimates of the realized genetic correlation between WW and PG were .69 +/- .18 and .46 +/- .31 for methods I and II, respectively. Variation in selection response was evaluated using quasi-replicates. Results of this study indicate that selection for PG improved both WW and PG faster than selection for WW.     

First detection of spring viraemia of carp virus in common carp (Cyprinus carpio L.) affected by a septicaemic disease in Mexico

Spring viraemia of carp (SVC) is an infectious disease responsible for severe economic losses for various cyprinid species, particularly common carp (Cyprinus carpio carpio). The causative agent is the SVC virus (SVCV), a member of the Sprivivirus genus, Rhabdoviridae family, and a List 1 pathogen notifiable by the World Organization for Animal Health. This study describes the diagnosis of an SVCV pathogen isolated in October 2015 from wild common carp inhabiting a natural lagoon in central Mexico. While neither an epidemic nor fish mortalities were reported, the collected killed specimens exhibited clinical signs of disease (e.g., exopthalmia, moderate abdominal distension and haemorrhaging, as well as internal haemorrhages and adhesions). Histological results of injuries were consistent with the pathology caused by SVCV. This finding was supported by the isolation of a virus in EPC and BF‐2 cells and subsequent RT‐PCR confirmation of SVCV. The phylogenetic analyses of partial SVCV glycoprotein gene sequences classified the isolates into the Ia genogroup. These findings make this the first report of SVCV detection in Mexico, extending the southern geographical range of SVCV within North America. However, since this pathogen was detected in fish inhabiting a natural body of water without tributaries or effluents, it is difficult to estimate the risk of SVCV for other wild/feral cohabitating cyprinid species in the lagoon. The status of this virus is also unknown for other bodies of water within this region.     

Yeast life-span extension by calorie restriction is independent of NAD fluctuation

Calorie restriction (CR) slows aging in numerous species. In the yeast Saccharomyces cerevisiae, this effect requires Sir2, a conserved NAD+-dependent deacetylase. We report that CR reduces nuclear NAD+ levels in vivo. Moreover, the activity of Sir2 and its human homologue SIRT1 are not affected by physiological alterations in the NAD+:NADH ratio. These data implicate alternate mechanisms of Sir2 regulation by CR.     

somatostatin1基因突变斑马鱼仔鱼转录组分析

生长抑素（Somatostatin,SST）是一类神经激素多肽,对脊椎动物的生长和代谢起着关键的调控作用。在斑马鱼（Danio rerio）和其它鱼类中,生长抑素由6个基因编码（sst1-sst6）,sst1存在于所有脊椎动物中,并且在进化上最为保守。本研究利用CRISPR/Cas9基因编辑技术在斑马鱼中构建了可稳定遗传的生长抑素基因sst1突变体鱼系。通过比较6 dpf（days post fertilization,dpf）的sst1-/-突变体和野生型仔鱼转录组,发现sst1-/-突变体相关生化过程和信号通路发生了显著变化。相比野生型,sst1-/-突变体仔鱼中有354个基因显著上调,504个基因显著下调。GO（Gene ontology）富集分析表明,大部分差异基因与氨基酸和蛋白质生物合成相关。KEGG（Kyoto encyclopedia of genes and genomes）通路富集显示代谢通路、氨基酸合成、氨基酰基- tRNA合成、核糖体合成和rRNA加工相关通路表达增强。结果表明,sst1-/-突变体蛋白质生物合成和代谢过程更加活跃。RT-qPCR验证结果显示,随机选取的10个基因mRNA表达变化趋势与转录组测序结果一致,说明转录组测序结果真实可靠。6月龄突变体体重体长和野生型相比并无显著差异。本研究结果为进一步挖掘生长抑素基因家族的潜在功能奠定基础。     

Kinematic Characterization of the Menorca Horse at the Walk and the Trot: Influence of Hind Limb Pastern Angle

This paper describes the handled walking and trotting kinematics (linear, temporal, and angular traits) of 35 Menorca Purebred (MEN) stallions, and the relationships among these variables is presented for the first time, along with a discussion of the influence of the hind limb pastern angle on kinematic variables at both gaits. For data collection, all animals, aged between 3 and 10 years old and belonging to 28 different studs, were recorded under the same experimental and environmental conditions, using a three-dimensional (3D) semiautomatic movement analysis system. A total of 24 kinematic variables (temporal, linear, and angular) at the walk and the trot and a morphometric variable measured at the mid stance position of walking (hind pastern angle) were included in this analysis. Angle-time diagrams of the hind pastern angle while walking and trotting normalized to stride duration were also obtained. Generally the MEN stallions' forelimb movements closely resembled the movement characteristics of other European dressage performance breeds, while the hind limb locomotion showed a greater likeness to Iberian dressage Purebreds. Despite this, their ability in collection and propulsion at the walk and the trot was relatively low. The hind limb pastern conformation was partially connected to the hind limb movements for both gaits, with an apparently negative effect of excessively upright pasterns on the amplitude at the trot, which indirectly reduced collection ability.