有“fish”,而无“鱼”意

“fish”是“鱼”的意思，这是稍识英文的人都知道的。但并非含有“fish”的短语和句子都包含有“鱼”的意思。

IHC与FISH在乳腺癌HER2蛋白表达和基因扩增检测中的应用

目的比较免疫组织化学法(immunohistochemisity,IHC)和荧光原位杂交法(fluorescence in situ hybridization,FISH)在乳腺癌患者中检测人表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)的应用。方法2019年2月至2022年1月本院收治158例乳腺癌患者作为研究对象,均采用IHC和FISH检测HER2的蛋白表达和基因扩增情况,比较两种方法的检测结果和一致性。结果IHC结果显示158例患者中HER2(1+)、HER2(2+)、HER2(3+)分别为5例(3.16%)、97例(61.39%)、56例(35.44%),FISH结果显示HER2扩增73例(46.20%),未扩增85例(53.80%),其中,HER2(1+)患者FISH均显示未扩增,阴性符合率为100%;HER2(3+)患者FISH均显示扩增,阳性符合率为100%;97例HER2(2+)患者FISH显示扩增17例,阳性符合率为17.52%。两种方法检测HER2一致性较好且具有统计学意义(Kappa值=0.775,P<0.05)。结论HIC与FISH在乳腺癌患者HER2表达检测中一致性较好,可在临床推广使用,必要时需结合两种方法检测。

基于FISH和GISH技术的芝麻栽培种和野生种染色体组特征比较分析

为揭示胡麻属物种进化特征,探究胡麻属基因组结构演变及物种进化,促进野生资源的开发利用,以芝麻栽培种豫芝11号和2n=26类型野生种S.alatum 3651为试验材料,采用荧光原位杂交(FISH)和基因组荧光原位杂交(GISH)技术对芝麻栽培种和野生种染色体组特征进行分析。结果表明,芝麻栽培种和野生种均为2n=2x=26类型;rDNA-FISH杂交结果显示,栽培种的13对染色体中,3对染色体(第7、8、9对染色体)的短臂端部携带45S rDNA特异信号,显示为随体特异染色体;2对染色体(第5、11对染色体)的短臂携带5S rDNA特异信号,5S rDNA与45S rDNA信号位于不同染色体上。野生种的13对染色体中,有2对染色体(第4、7对染色体)携带45S rDNA杂交信号,1对染色体(第4对染色体)携带5S rDNA特异信号,5S rDNA与45S rDNA信号位于同一染色体不同位置,表明栽培种与野生种的染色体组特征差异较大。GISH杂交结果显示,分别以栽培种和野生种的基因组DNA作探针与自身染色体杂交,每条染色体上携带不同强弱的杂交信号,而与对方染色体杂交,均表现出极少的杂交信号。芝麻栽培种和野生种具有相同的染色体数目,但染色体rDNA数量、分布及其GISH信号均存在明显差异,说明栽培种和野生种亲缘关系较远。

IHC、FISH、qRT-PCR检测NSCLC ALK融合基因的对比分析

目的对比分析免疫组化(IHC)、荧光原位杂交(FISH)、实时荧光定量PCR(qRT-PCR)检测非小细胞肺癌(NSCLC)ALK融合基因情况。方法收集453例NSCLC标本。随机选取IHC检测的ALK融合基因不同表达程度标本40例[ALK(-)、(+)、(2+)、(3+)标本各10例],采用FISH和qRT-PCR法分别检测各组ALK融合基因表达水平,并进行一致性分析。结果IHC与FISH、qRT-PCR的ALK融合基因检测结果比较,IHC ALK(-)/(3+)与FISH、qRT-PCR检测结果一致率均为100%,具有高度一致性(r=0.781、r=0.740,P<0.05)。FISH与qRT-PCR的检测结果一致率为97.5%,具有高度一致性(r=0.943,P<0.05)。结论FISH、qRT-PCR法与IHC法检测在不同表达程度ALK融合基因样本中检测结果存在差别,IHC敏感性更高。FISH、qRT-PCR法在ALK融合基因样本的检测结果具有高度一致性。

Microbial Quality and Molecular Identification of Enterotoxigenic Staphylococcus Strains Isolated from Dried, Smoked, and Braised Fish Sold in Ouagadougou Markets

Background: The investigation of toxin genes in strains involved in staphylococcal food poisoning contributes to food safety. The aim of this study was to isolate and identify enterotoxigenic Staphylococcus strains from dried, smoked, and braised fish sold in Ouagadougou markets. Methodology: Staphylococci were isolated using standard microbiology methods. Staphylococcus strains were identified using API Staph kit (Reference # 20500, BioMerieux S.A., Marcy l'Etoile, France). The molecular identification of isolated Staphylococcus aureus strains was specifically confirmed by PCR using the Staur4 and Staur6 primers. The genes encoding enterotoxins, enterotoxin-like toxins, exfoliative toxins, and TSST-1 toxin were detected by multiplex PCR using specific primers from Inquaba Biotec West Africa Ltd, Africa's Genomics Company. Results: The results of the microbiological quality assessment indicated that most of the samples analyzed were found to be of unsatisfactory microbiological quality according to the Staphylococcus aureus microbiological criteria (m = 102). Overall, only 12.55% of samples were satisfactory, while 97.45% were unsatisfactory. The STAPH API gallery allowed the identification of the following species: Staphylococcus aureus, Staphylococcus xylosus, Staphylococcus lugdunensis, Staphylococcus hominis, Staphylococcus haemolyticus, Staphylococcus lentus, Staphylococcus sciuri and Staphylococcus capitis. Of the 108 Staphylococcus isolates, 81 (75%) showed at least one (1) toxin gene. Among the 21 toxin genes tested in this study, 20 genes were detected in all strains analyzed. The staphylococcal toxin genes detected were present in both Staphylococcus aureus and the other coagulase-negative strains isolated in this study. In addition, these genes are found individually or in association in certain strains. The most frequent genes detected in toxin gene-positive strains were: the tsst-1 gene in 45 isolated strains (41.7%), sei (16/14.8%), seg (13/12%), ser (7/6.5%) sec (6/5.5%), and sea (5/4.6%) for staphylococcal enterotoxins, seln (14/12.9%), selq (8/7.4%), for enterotoxin-like toxin gene and eta (3/2.7%) for exfoliative toxin genes. Conclusion: This study highlighted the pathogenicity of Staphylococcus strains isolated from dried, smoked, and braised fish sold in Ouagadougou markets. Monitoring toxin-producing strains of Staphylococcus is invaluable for better prevention of food poisoning.

Measurement of Malondialdehyde as a Biomarker of Lipid Oxidation in Fish

Oxidative reactions are key part of the stress response in marine organisms, which are exposed to a wide variety of environmental stressors. Lipid peroxidation (LPO) occurs in response to oxidant attack, giving rise to unsaturated aldehydes as malondialdehyde (MDA), the main biomarker for LPO assessment. Levels of MDA are measured in biological samples of different fish matrices to determine the oxidative effect of physical or chemical agents, particularly represented by metals. The most used assay is the spectrophotometric determination of thiobarbituric acid (TBA) as MDA-TBA2 adduct. Selectivity is enhanced by separating the adduct by chromatographic methods such as high-performance liquid chromatography or by using alternative derivatization reagents. Because, in biological organism, MDA is found free or protein bound, the two forms should be measured, and the extraction procedures are a crucial step in the processing of the biological sample. This review focused on MDA determination procedures used to assess the effect of experimental and environmental stress induced in fish.

Distribution and Contamination of Arsenic in Fish, Gastropods and Bivalves in the Aby and Tendo Lagoons in East of Ivory Coast

Lagoons are ecosystems for biodiversity and the livelihoods of coastal communities. The main objective of the study was to analyze the variability of arsenic concentrations in gastropods and bivalves in the Aby and Tendo lagoons, taking into account spatial, seasonal and hydrological variations. The study was carried out in four stages spread over two successive hydroclimatic cycles, including two seasons during the rainy season and two more during the dry season. The samples were taken in two areas of the Aby and Tendo lagoons. Arsenic levels were measured by ICP-MS. The results showed that mean arsenic concentrations in the muscles of organisms in Aby Lagoon ranged from 0.01 to 1.26 μg As/g, with a mean and median of 0.17 and 0.06 μg As/g, respectively. Fish had the highest levels of arsenic, followed by crustaceans, while molluscs and plants had lower and comparable concentrations of As. Arsenic concentrations in tilapia and jawbones varied significantly between sites and seasons, with higher concentrations at Tendo and during the rainy season. Arsenic concentrations in gastropods and bivalves were significantly higher than those of other species, with averages of 0.74 and 1.03 mg As/kg, respectively.

Evaluation of Agricultural Contamination Level by Heavy Metals and Pesticides in Sediments of Rivers and Water Bodies for Shrimp Fishing in the Lower Ouémé: Case of the Aguigadji, Ahlan and Sele Stations in Benin

This study assessed sediment contamination by heavy metals and pesticide active ingredients linked to chemical inputs used in agricultural activities in the lower Ouémé. Pesticide residues from the organochlorine, pyrethroid and organophosphorus families were investigated by gas chromatography, and heavy metals (Cd, Pb, As, Ni, Zn, Fe, Mg, Cr and Hg) by atomic absorption spectrophotometry. The metallic pollution indices, the contamination factor (CF) and the ecological risk index were calculated. The results revealed 8 active ingredients in the rainy season and 9 in the dry season. Glyphosate was the active ingredient with the highest concentration at all stations, 9.65 ± 0.84 mg/kg recorded in the dry season at the Aguigadji station. All glyphosate values were above the EQS. DDT, Atrazine and Endosulfan also showed high concentrations in the dry and rainy seasons. Emamectin, Abamectin and Lambda Cyhalothrin also showed high concentrations in the dry season at Aguigadji, Ahlan and Sele. Only glyphosate was recorded at the control station (Toho), but in very low concentrations. Lead showed the highest concentrations at all the stations, 265.96 ± 21.02 mg/Kg in the rainy season and 255.38 ± 79.09 mg/Kg in the dry season, all detected at the Aguigadji station and above the EQS. Zn, Ni, Fe, Cu and Cr were all representative in both the dry and rainy seasons at the contaminated stations. Manganese showed high concentrations in the rainy season. Pb showed very high contamination (FC ≥ 6) at the Aguigadji and Ahlan stations and significant contamination (3 ≤ FC 6) at the Sele station in both the rainy and dry seasons. Ni, Hg and Cd, showed either moderate or significant contamination at the contaminated stations. The risk values showed a considerable ecological Ri (190 ≤ Ri < 380) in the rainy season and a moderate ecological Ri (95 ≤ Ri < 190) in the dry season at these contaminated stations.

Effect of the Dietary Substitution of Fish Meal with Achatina fulica Meat Meal on the Growth Performance and Production Cost of African Catfish (Clarias gariepinus) Fingerlings

Fishmeal is the most preferred source of protein in aquafeeds, but it is expensive and scarce. Hence, Achatina fulica meat meal (AFM), which is much less preferred for human consumption out of three species of African giant land snails, was tested as a fishmeal substitute for Clarias gariepinus growth. Five iso-nitrogenous and iso-calorific diets were formulated, in which AFM substituted fish meal at 0% (control or Diet A), 25% (Diet B), 50% (Diet C), 75% (Diet D) and 100% (Diet E). These dietary treatments were each replicated thrice in a completely randomized design experiment, using 36-L plastic tanks in which the fish were fed daily rations corresponding to 5% of their body weight, for 8 weeks. Water quality parameters in the tanks were monitored. Proximate analyses were conducted on the fish meal, snail meal and experimental diets before the feeding trials. Cost-benefit analysis of the different diets was performed. The crude protein content of AFM (69.18%) was significantly higher than that of fish meal (55.81%). There was no significant difference (P > 0.05) in the mean weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio and survival rate in fish fed Diet A and Diet B. The best protein efficiency ratio (0.77) was recorded in fish fed Diet B. Furthermore, the survival rate of fish increased with increased levels of AFM substitution. Water quality parameters were within a suitable range for tropical fish culture, indicating that the AFM did not pollute the water. The fish fed 25% AFM diet significantly (P Clarias gariepinus diets at a 25% substitution level. The aquaculture industry can thus exploit the availability of this feed resource.

Study of the Biological Quality of the Water in the Manantali Dam Reservoir: Analysis of the Fish Fauna and Plankton Communities

The aim of this study of the Manantali dam reservoir is to investigate the biological quality of the water through inventories and analysis of the ichthyofauna and planktonic communities (zooplankton and phytoplankton). Inventories of ichthyofauna and plankton communities were carried out quarterly from 2003 to 2022. The fish populations studied on the basis of bioindicators (species richness, diversity index, equitability index, diversification index) identified 29 species belonging to 10 families. The Cichlidae family is the most represented, while the Schilbeidae and Tetraodontioae families are less represented in terms of number of species. The species Hydrocynus forskalii and Brycinus nurse with relative abundances of 40.12% and 21.4% respectively, both from the family Alestidae, are the most abundant. The species H. forskalii has a frequency of occurrence of 100%. The specific diversity index of 2.9 and the equitability index of 0.6 show an average diversity of fish species. The diversification index of 2.9 indicates a theoretical number of habitats of 2 to 3. The planktonic community is made up of zooplankton and phytoplankton. The inventory of zooplankton shows the presence of 28 zooplankton species belonging to 9 families of the three zooplankton groups Rotifera, Cladocera and Copepoda. Analysis of the frequencies of occurrence of the various taxa showed that the characteristic species of Lake Manantali are the omnipresent rotifers Keratella cochlearis, Keratella quadrata and Trichocerca chattoni. For phytoplankton, the inventories carried out show the presence of 65 species belonging to 5 families (Chlorophycees, Diatomees, Dinophycees, Chrysophycees and Cyanophycees). An analysis of the frequency of occurrence of the various taxa shows that the characteristic species of Lake Manantali are Chlorophycees, with 7 species: Desmidium baileyi, Micrasterias alata, Sphaerocystis schroeteri, Spondylosium sp., Staurastrum heimii, Staurastrum subanchora, Staurodesmus wandae;Diatoms, with one species: Navicula sp. and Cyanophycees, with one species: Microcystis aeruginosa.

鱼类性别可塑性的分子机制

鱼类性别具有高度的可塑性,具体表现为天然性逆转、原发性逆转和次发性逆转。近年来,一系列研究都证明鱼类性别可塑性与雌激素密切相关。一旦阻断性腺雌激素的合成,无论是未分化还是已分化卵巢都将性逆转为精巢。鱼类的性别决定通路基因缺失诱导的性逆转也与雌激素相关。重要的是,发现生殖细胞的可塑性依赖于foxl3和dmrt1的同时存在,缺失其中一个都不能通过改变雌激素水平从而诱导性逆转。因此,foxl3和dmrt1是生殖细胞响应雌激素的关键基因。另外,表观遗传调控基因kdm6bb通过选择性剪接介导温度诱导的性逆转。这些研究增进了我们对鱼类性别可塑性分子机制的认识。

棉花体细胞染色体rDNA-FISH技术

介绍了棉花体细胞染色体为靶、r DNA为探针的荧光原位杂交实验技术 ,并着重分析和讨论了棉花 r DNA- FISH技术的关键因素 ,包括染色体制片、探针与染色体共变性等。作为靶染色体的棉种包括陆地棉、海岛棉、草棉、亚洲棉、克劳茨基棉和比克氏棉等 ,作为封阻的是鲑鱼精 DNA。

利用FISH和DGGE对产甲烷颗粒污泥中微生物种群的研究

利用FISH和DGGE技术对厌氧反应器内处于不同运行阶段的4个产甲烷颗粒污泥进行研究,考察其中真细菌和古细菌的种群结构,并对其中的优势古细菌进行系统发育分析.FISH结果表明,颗粒污泥中真细菌含量明显高于古细菌,真细菌主要分布在颗粒污泥外层,古细菌则主要分布在内层;DGGE结果表明,随着反应器COD负荷的增加以及运行时间的延长,真细菌种群结构相对较稳定,而古细菌种群结构则发生了较明显变化,其中占优势的古细菌种类逐渐减少;将有代表性的7个古细菌条带切胶回收并测序,结果显示,反应器运行后期占优势的菌种主要包括甲烷微粒菌(Methanocorpusculum)、甲烷杆菌(Methanobacterium)和甲烷髦毛菌(Methanosaeta)等.

大麦45S和5SrDNA定位及5SrDNA伸展纤维的FISH分析

用荧光原位杂交技术对45S和5SrDNA在大麦(HordeumvulgareL.)有丝分裂中期染色体进行了确定分析,较强的45SrDNA信号共有2对,分别分布在大麦的第1染色体的短臂和第2染色体的长臂。而5SrDNA则只有1对杂交信号,位于第3染色体的长臂,但信号较弱。用伸展DNA纤维的荧光原位杂交(Fiber-FISH)技术测定了5SrDNA在大麦的基因组中的拷贝数,计算出5SrDNA的拷贝数约为408～416。对大麦品种中rDNA位点数目的可变性进行了讨论。

棉花gDNA体细胞染色体FISH技术

介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ，简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程，并着重分析和讨论了影响试验结果的关键因素，包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉；探针和封阻均采用ｇＤＮＡ，材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等），分别交互使用。试验结果比较理想，获得良好的ＦＩＳＨ片子，而且重复性好。

鳜鱼生殖生长调控与良种培育技术研究进展

鳜鱼分布在东亚国家,是我国主要养殖淡水鱼类,具有重要的经济价值。近年来,随着全基因组测序的完成,鳜鱼生殖生长遗传基础被逐步揭示。研究人员在鳜鱼中开发了性别染色体分子标记,实现了性别控制育种;开展了细胞工程育种,获得多倍体鱼;进行生长选育和饲料驯化,培育出了多个鳜鱼优良新品种。本文结合自己研究团队的工作,对鳜鱼生殖生长调控机制和良种培育技术进行了总结整理,并讨论了遗传育种方法的应用现状,以期为鳜鱼乃至经济鱼类产业的发展提供方向和目标性思考。

荧光原位杂交(FISH)技术研究窖泥微生物群落

为了探索荧光原位杂交(FISH)技术,研究窖泥微生物群落结构特征多样性,进行了纯培养微生物及窖泥微生物的FISH检测。以Escherichia coli、Bacillus subtilis、Lactobacillus planetarium、Acetobacter rancens、Clostridiumacetobytylicum 5株不同种属的细菌为对象,研究了影响荧光原位杂交(FISH)技术对其定量表征的因素,并将该技术应用于窖泥微生物群落的研究。实验结果表明,E.coli及L.planetarium的定量表征受到生长期的影响,在对数和稳定期的菌体检出率>82%,衰退期的L.planetarium则明显减少;死菌体比活菌体的检出信号显著减弱;经硫酸铝处理,除去腐植酸的窖泥样品明显地提高了可辨力,加入窖泥中的E.coli,FISH检出量为光学显微镜计数量的46.89%。采用FISH技术可视化定量表征了窖泥中细菌和古菌的特征,有益于从细胞水平研究其微生物群落的关系。

390份小麦-黑麦种质材料主要农艺性状分析及优异材料的GISH与FISH鉴定

将小麦近缘属植物黑麦中的优良基因导入小麦可以拓宽小麦的遗传基础,丰富小麦的遗传变异。本研究调查并分析了390份小麦-黑麦种质材料。在这390份种质材料中,6个主要农艺性状值均有较大的极差,说明其遗传多样性丰富。与10份小麦主栽品种相比,90%以上的材料具有穗长和分蘖数的显著优势,60%以上的材料具有小穗数优势,约30%的材料穗粒数和千粒重显著高于主栽品种。利用基因组原位杂交(genomic in situ hybridization,GISH)和多色荧光原位杂交(multicolor fluorescent in situ hybridization,mc-FISH)技术,对8份农艺性状优良的代表性材料进行染色体组成分析,发现3份为六倍体小黑麦(AABBRR),2份为八倍体小黑麦(AABBDDRR),1份为1RS?1BL易位系,其余2份不具有可见的黑麦染色体或染色体片段。值得指出的是,3份六倍体小黑麦与2份八倍体小黑麦所含的黑麦染色体不完全相同。八倍体小黑麦中有1对来源于黑麦的小染色体,而六倍体小黑麦中没有类似小染色体;并且,不同材料中黑麦4R染色体端部的GISH杂交带有明显差异。本研究结果为这些小麦-黑麦种质材料进一步应用于小麦育种提供了依据。

采用FISH、DGGE和Cloning对短程脱氮系统中硝化菌群的比较分析

针对4种不同的实际污水短程生物脱氮系统（SBR大型中试反应器、UASB-A/O小型反应器、A/O中试反应器和SBR小型反应器）,采用Fish、PCR-DGGE和PCR-Cloning-Sequencing分子生物学方法对系统中硝化菌群AOB和NOB进行定性与定量化分析.Fish结果表明,在4种短程脱氮系统中,AOB相比于NOB已成为明显的优势菌群,占总菌群的3%～12%;在SBR中试和小试反应器中没有检测出NOB;A/O中试反应器中存在极少量的Nitrospira（＜0.2%）,而UASB-A/O小型反应器中存在极少量的Nitrobacteria（＜0.2%）.PCR-DGGE结果表明SBR中试、A/O和UASB-A/O 3种短程脱氮系统中的AOB均以Nitrosomonas-like为主.SBR大型中试反应器中污泥样品的PCR-Cloning-Sequencing结果表明,所有的克隆相似于Nitrosomonas,其中60%以上的克隆相似于Nitrosomonas europaea.

性别特异分子标记在斑鳢全雄育种上的应用

为了快速筛选培育出全雄斑鳢,本研究结合性别特异分子标记和三系配套育种技术,开发了全雄斑鳢的育种方法。将健康的七日龄斑鳢幼鱼随机分成3组进行雌化处理,在饲料中分别添加雌二醇(E2)100、300、600 mg/kg,饲养60 d。利用性别特异分子标记引物M12、P2鉴定筛选出决定型为XY型斑鳢,将XY型正常雄鱼与XY型伪雌鱼交配,获得的子代分为两组,一组为投喂正常饲料,另一组进行雌激素投喂,利用MX1和MX3引物筛选出YY超雄鱼,最后将YY超雄鱼和正常雌鱼作为亲本交配生产出全雄斑鳢子代。结果显示,600 mg/kg激素浓度组的性逆转率最高,达75%,从508尾经雌激素E2投喂的家系中筛选获得235尾具有XY基因型斑鳢。XY伪雌鱼与正常雄鱼交配获得的子代在2月龄时检测到22尾YY超雄鱼,7月龄时检测到14尾YY超雄鱼,筛选获得YY超雄鱼个体用于生产全雄子代。本研究方法显著提高了全雄化斑鳢育种效率,缩短了育种周期,展现出巨大的经济潜力和应用价值,为其他鱼类开展性别特别分子标记辅助育种提供借鉴或参考。