Molecular genetic structure and relationship of Chinese and Hungarian common carp (Cyprinus carpio L.) strains based on mitochondrial sequence

China and Hungary are major providers of the common carp (Cyprinus carpio L.) in East Asia and Europe respectively. However, the genetic variation and relationship of this species in the two countries have been poorly understood. In this study, mitochondrial COII‐tRNA^Lys and D‐loop sequences were analysed to investigate the genetic structure and relationships of the representative wild and domesticated common carps distributed in China and Hungary. The results indicated that the genetic diversities of the Chinese common carps are higher than those of Hungarian common carps, and the diversities of the wild common carps are higher than those of domesticated common carps in both the countries. Analysis of molecular variance and pairwise F_ST demonstrated a significant genetic divergence between the Chinese and the Hungarian common carps, and between the wild and the domesticated common carps. Bayesian phylogenetic analysis and statistical parsimony network showed an obvious genetic differentiation between the Chinese and the Hungarian common carps, between the wild and the domesticated common carps. However, a few specimens and haplotypes from the Chinese wild common carps appeared in the Hungarian common carps, demonstrating that there was no absolutely isolated and possible genetic linker between the Chinese and the Hungarian common carps.     

2种多糖对鲤鱼离体培养免疫细胞活性的影响

采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖(Astragalus polysaccharides,APS)和香菇多糖(Lentinan,LNT)后,分别采用MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸暴发的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧暴发活性,黄芪多糖则没有显著的诱导作用;香菇多糖低浓度时对细胞的氮呼吸暴发活性无显著影响,黄芪多糖能显著诱导氮呼吸暴发活性,随着两者作用浓度的增加均表现为高浓度抑制作用;香菇多糖和黄芪多糖都能显著促进鲤鱼外周血淋巴细胞的增殖。结果表明黄芪多糖和香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是有潜力的鱼用免疫增强剂。     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

The use of immunostimulating herbs in fish. An overview of research

The use of immunostimulants as an alternative to the drugs, chemicals and antibiotics currently being used to control fish diseases in fish culture is attracting the attention of many researchers. In this context, many have focused on the use of medicinal plant products as potential therapeutic measures for modulating the immune response and, specifically, on the use of herbs to prevent and control fish diseases. Medicinal plants (plant remedies) are a deeply rooted component of the cultural heritage of many people from diverse cultures and countries and are, as such, closely linked to the maintenance of good health. The aim of this paper is to review research currently being carried out on the herbs and herbal extracts that have been shown to modulate the immune system of fish. Special attention is given to the use of Chinese and Indian herbs.     

Behaviour of thiocarbamate herbicides in soils: Adsorption and volatilization

Adsorption and volatilization of cycloate, EPTC and vernolate applied to six types of soils were studied in the laboratory. Cycloate proved to be the most and EPTC the least adsorptive molecule in all systems examined, while the volatilization order of the compounds was the reverse EPTC>vernolate>cycloate. Neither the adsorption nor the volatilization of thiocarbamates was markedly affected by formulating agents. The adsorption isotherm of several systems did not obey the classical Freundlich equation. A reciprocal relationship which was found between the adsorption coefficients and the volatilization rates of thiocarbamates permits calculation of one of these properties if data on the other are available.     

Immunostimulants added to injected Aeromonas salmonicida bacterin enhance the defense mechanisms and protection in rainbow trout (Oncorhynchus mykiss).

Immunostimulants were given to rainbow trout for assaying effects on modulating non-specific defense mechanisms, specific immune response, and protection levels against pathogen challenge with Aeromonas salmonicida. Three drugs, levamisole (an approved veterinary drug in the USA), a quaternary ammonium compound (QAC), and a short-chain polypeptide (ISK) were found to affect the non-specific defense mechanism activities, which were measured by changes in circulatory neutrophil and phagocytic activity levels, and the specific immune response factors, which were measured by numbers of plaque-forming cells, and circulatory antibody levels. When given alone, the immunostimulants elevated the non-specific factors. When injected in combination with an A. salmonicida O-antigen bacterin, the non-specific factors were further elevated, and the specific response was raised over samples taken from fish given the bacterin without the immunostimulants. Challenge tests with the virulent pathogen, A. salmonicida, showed a 5-6 day delay in the onset of mortalities in the fish given the immunostimulants alone, and a 12-14 day delay when immunostimulants given were combined with the bacterin. In the groups given the QAC or ISK with the bacterin, there was a 20% and 40% survival rate, respectively.     

Effectiveness of dietary vitamin supplementation to the performance of common carp (Cyprinus carpio L.) larvae in intensive rearing condition

Study of enrichment of different vitamins (C, B₁, B₆ and E) in common carp larvae through 4 weeks feeding in recirculation system was carried out. For this purpose, procedure for enrichment of Artemia nauplii with different vitamins was developed and investigated the effectiveness in larvae nutrition. The habituation of larvae to dry feeds was done using supplemented dry diets with the same vitamins. To study the effect of vitamins in young fish, the most common stress situation was planned for carp nursed fries, which occurs during transportation. Levels of vitamins and parameters of immune response in mucus were monitored before and after stress situation. At the end of the feeding trials using enriched Artemia nauplii, level of vitamin B₁, B₆ and C were increased in the body of fish, but remained on similar level or decreased after dry feeding period. Therefore, we concluded that Artemia is a good delivery vector of these vitamins. While handling stress, vitamin C and E concentration was decreased and sparing effect of antioxidant vitamins was confirmed in groups supplemented with both vitamins. Vitamin B₁ was consumed intensively during the stress, vigorously in groups without supplementation. Contrary to this, vitamin B₆ level increased in stress condition in group without supplementations. The levels of immunoglobulins in mucus were increased in all treated groups independently of treatment. Our results confirm that carp larvae reared in tank condition needs sufficient amount of vitamins C, B₁ and B₆ supplements to protect them from the mild handling stress.     

香菇多糖对鲤鱼（cyprinus carpio）离体培养免疫细胞的活性调节作用*

采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖（APS）和香菇多糖（LNT）后,分别采用 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸爆发的影响;实时定量PCR法测定头肾巨噬细胞细胞因子IL-1β诱导表达的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧爆发活性;其低浓度时对细胞的氮呼吸爆发活性无显著影响,随着作用浓度的增加表现为高浓度抑制作用;香菇多糖可以显著促进鲤鱼外周血白细胞的增殖,且能显著增强IL-1β的体外诱导表达。结果表明香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是一种有潜力的鱼用免疫增强剂。     

The endosomal epsilon-coatomer protein is involved in human adenovirus type 5 internalisation

The effects of bafilomycin A1 and of the reduced level of endosomal epsilon-COP (coatomer protein) on the infectivity of human adenovirus type 5 were investigated in Coxsackie adenovirus receptor- (CAR-) transfected Chinese hamster ovary (CHO) cells. The endosomal proton pump inhibitor bafilomycin A1 was able to cause only partial inhibition. Using Id1F cells (an epsilon-COP thermosensitive mutant CHO cell line) the reduction of epsilon-COP level also had partial inhibitory effect. Based on these results and comparing them to existing models of the adenovirus entry, we propose a refined model in which there are two pathways of adenoviral entry: the first one involves the epsilon-COP as the downstream effector of the acidification and can be blocked by bafilomycin A1 and the second one is a pH-independent pathway.