DNA barcoding of fishes from Zhoushan coastal waters using mitochondrial COI and 12S rRNA genes

Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.

Scale and Associated Factors of Using Pyrethroid-Impregnated Mosquito Nets as Fishing Tools on Nokoué Lake within Sô-Ava Municipality in Benin, 2020

Background: In lacustrine communities, whether in Benin or elsewhere, populations use impregnated mosquito nets (IMNs) as fishing nets. This depletes the lake of its fishery resources, which in turn are contaminated by the pyrethroids impregnated in the nets. This study aims to determine the scale and factors associated with the use of pyrethroid-impregnated mosquito nets as fishing tools in the municipality of Sô-Ava in Benin. Methods: This is a cross-sectional and analytical study with two components. First, a quantitative component was made up of 280 volunteers who were interviewed in the seven districts of the city. Data collection was done in two phases: the first from September to October 2020, and the second in August 2022. Data were analyzed with Stata and logistic regression was used. Another qualitative component was made up of forty participants chosen by reasoned choice, of which 32 were split into four focus groups of eight members each, and the rest participated in semi-structured interviews. Triangulation of the different sources was used to analyze the data. Results: Around 67% of the population reported using impregnated mosquito nets as fishing tools and 33% exclusively for malaria. Seasonal fishermen (ORa = 2.03, CI = 1.35 - 4.97, P = 0.004) and years of professional experience (ORa = 1.53, CI = 1.00 - 2.05, P = 0.021) increase the risk of using these nets as fishing tools. The use of insecticide-treated mosquito nets against mosquitoes causes skin scratching and impairs breathing because of the impregnation products, as reported by respondents. Conclusion: A high prevalence of inappropriate use of impregnated mosquito nets in fishing practices is highlighted in this study. Interviews in the field revealed that nets are diverted for fishing purposes to increase the volume of catches. Consequently, it will be appropriate to assess the pyrethroid content in water and fishery products.

黄褐棉45S rDNA的FISH定位及核型分析

黄褐棉是棉属5个四倍体棉种之一,利用荧光原位杂交技术将45S rDNA定位在黄褐棉2、4、9号染色体,2号染色体上的45S rDNA特别大,信号位于随体并覆盖了染色体的短臂,比二倍体和四倍体棉种的45S rDNA都要大得多;另外的2对信号很小,形状与陆地棉中的弱信号类似。黄褐棉的核型公式为:2n=4x=52=50m(2SAT)+2sm,属于2B类型,第2对染色体为亚中着丝粒染色体,其余都为中部着丝粒染色体。黄褐棉的核型、随体数、45S rDNA与其他四倍体棉种区别很大,黄褐棉是一个非常特殊的四倍体棉种。

植物45S rDNA的染色体位置的CPD染色和FISH分析

采用PI和DAPI组合(CPD)染色结合45SrDNA探针的荧光原位杂交(FISH)对分属6个科的16种植物的45S rDNA的染色体位置进行了分析。在所有供试植物中,共检测到53个45S rDNA位点。大多数45S rDNA位点分布在染色体的短臂;位于染色体臂内和染色体末端的位点的比例大体相当;多数位于染色体臂内的45S rDNA位点有次缢痕形成,但rDNA重复单位簇所处的位置存在差异。根据45S rDNA所处的染色体臂的不同、距着丝粒远近的差异、形成次缢痕与否以及rDNA重复单位簇相对于次缢痕的位置等特征,将植物的45S rDNA位点划分为12种染色体分布类型。基于我们的结果和其他的报道对45S rDNA位点、核仁组织区(NOR)、次缢痕和随体相互之间的关系进行了分析。

16轮Blow-CAST-Fish的弱密钥攻击

Blow-CAST-Fish的轮函数F由作为算法子密钥的四个S盒构成。如果S盒产生碰撞,则可构造F函数的输入差分为非零、输出差分为零的差分特征,进而对算法进行差分分析以获取其他子密钥。研究单个S盒的碰撞,构造F函数的差分特征,并测试使特征成立的弱密钥比例,给出了特征概率为2-49,弱密钥比例为2-52.4的14轮差分特征,并以此为基础,攻击16轮Blow-CAST-Fish。

低轮Blow-CAST-Fish算法的差分攻击

对作为Blow-CAST-Fish算法子密钥的4个S盒的碰撞性进行分析,构造输入差分为非零、输出差分为零的轮函数F的差分特征,通过对算法进行差分分析,获取相关子密钥,并测试使特征成立的弱密钥概率。在此基础上,成功利用特征概率为2-61、弱密钥概率为2-12的6轮差分特征攻击8轮Blow-CAST-Fish算法。

白缘(鱼央)5S rDNA的扩增分析及其在染色体上的FISH定位

本文分别对雌雄白缘(鱼央)(鱼央)的5SrDNA进行了PCR扩增和序列分析,并采用双色荧光原位杂交(FISH)技术,以白缘(鱼央)的5SrDNA序列和小麦的45SrDNA为探针,对其在白缘(鱼央)雌雄中期染色体上进行了FISH定位研究。结果表明,白缘(鱼央)5SrDNA序列无雌雄差异;5SrDNA的保守区序列为117bp;5SrDNA和45SrDNA分别被定位于白缘(鱼央)的性染色体和第5号染色体上。同时从GenBank中获得了22种鱼的5SrDNA,运用DNAman软件构建了23种鱼的系统发育树,对白缘(鱼央)的进化地位进行了初步研究。本研究为阐明白缘(鱼央)在鱼类系统进化中的位置、重复序列在脊椎动物性染色体上的分布状况以及与性别决定与分化的关系,提供了资料积累和分析依据。

海岛棉基于双色FISH的核型分析

以45S r DNA作探针对海岛棉体细胞中期染色体进行荧光原位杂交(Fluorescence in situ Hybridization,FISH),检测出6个强的荧光信号(4大2小),结合DAPI图像观察发现信号分布在各自染色体随体位置,称为NOR(Nucleolar Organizer Region)信号。以二倍体A基因组棉种亚洲棉基因组DNA(g DNA)为探针对海岛棉进行FISH,结果发现52条染色体中有无杂交信号的各占一半,荧光信号分布在较长的A亚组染色体上,直接区分开海岛棉A、D亚组染色体,直观证实了海岛棉的异源双二倍体起源。以45S r DNA和A组棉种亚洲棉g DNA作探针对海岛棉进行双色FISH时,结果发现海岛棉染色体上既检测到了6个强的NOR信号又检测出了g DNA杂交信号,且清楚观察到2个NOR信号分布在A亚组上,另4个在D亚组上。基于该双色FISH图像的核型分析表明,海岛棉的核型公式为:2n=4x=52=40m(2 SAT)+12sm(4 SAT),属于2A类型,且有3对随体,其中1对定位于A9号染色体,属于中部着丝点(m)类型;另2对分别定位于D8和D12号染色体上,属于近中部着丝点(sm)类型;且发现海岛棉A亚组染色体的相对长度的并非全部大于D亚组,两亚组染色体间在长度上存在交叉,说明基于FISH的核型分析比传统方法更为准确。

野生黄瓜5S rDNA-FISH技术体系建立

目的通过改进黄瓜中期染色体制片和rDNA-FISH技术的关键因素,对野生种(Cucumis sativus var.hardwickii)PI 183967进行核型分析为继续研究黄瓜各条染色体的准确鉴定和进一步开展细胞学研究奠定基础。方法使用8-羟基喹啉0.002mol·L-1预处理2h,可以得到形态清晰的中期染色体,分裂相最高比例为17.2%。染色体制片以改良酶解空气干燥制片法较好,流程简单,可以获得大量清晰、分散的有丝分裂中期染色体,且背景干净。荧光原位杂交过程中染色体制片采用胃蛋白酶37℃处理30min,70℃变性3min时,探针使用缺口平移法地高辛进行标记,75~100℃变性10min,能获得杂交信号清晰的染色体图像。结果在黄瓜野生种PI 183967中只有一对5SrDNA信号,位于5号染色体短臂上,相对远离着丝点位置。结论野生黄瓜PI 183967的5号染色体可以用5SrDNA鉴定。

A Qualitative Appraisal of China’s Efforts in Fishing Capacity Management

This paper provides a critical review of some major efforts made by China to manage the fishing capacity of its marine capture fisheries. It begins by reviewing the key concepts and issues closely related to fishing capacity management. Following on this, a brief introduction to China’s marine capture fisheries is offered, especially the trends of its marine landings and marine fishing fleets and the status of fish stocks in waters under its jurisdiction. Next comes the main concern of the paper, i.e., some major actions recently initiated by China to deal with the overcapacity problem confronting it. The paper ends with conclusions and suggestions.

Seasonal Variations of Picoeukaryote Community Structure in Zhoushan Fishing Ground,East China Sea

Picoeukaryotes(<2-3μm)are major oceanic primary producers and play a crucial role for the functioning of marine ecosystems.However,the community structure of such communities remains poorly understood,especially in the East China Sea(ECS).We investigated the seasonal variations of abundance and diversity of picoeukaryotes,and recorded environmental variables,along a transect in Zhoushan Fishing Ground.High-throughput sequencing was used for sequencing the V4-V5 variable region within the 18S rRNA gene to analyze genetic diversity and relative abundance of picoeukaryotes.A total of 40 phyla,68 classes,99 orders,126 families,and 140 genera were observed.The estimated richness and diversity indices were both higher in each season.The composition and dominant picoeukaryotes changed significantly at the class level with the seasons alternating.Basidiomycota,Ciliophora,Ascomycota and Cryptomonadales were observed throughout the year.The Linear Discriminant Analysis(LDA)revealed the predominant discriminant taxa of four seasonal groups.Redundancy analysis(RDA)showed that NO2^−and salinity played very important roles to picoeukaryotes for all the spring samples and DS5 sample in winter,PO4^3−and pH affected mostly picoeukaryotes for winter samples and autumn samples.

Chromosomal mapping of 5S and 18S-5.8S-25S rRNA genes in Saccharina japonica(Phaeophyceae)as visualized by dual-color fluorescence in situ hybridization

It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5S rDNA was unlinked to 25S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH)technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18S rDNA and 5S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18S-5.8S-25S rDNA was localized at the interstitial region of Chromosome 23,whereas 5S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5S rDNA to 18S-5.8S-25S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica.

Is there a role for fish oil in inflammatory bowel disease?

A number of animal and human studies suggest omega3-fatty acids are anti-inflammatory. As a result they may have a therapeutic role in inflammatory bowel disease(IBD). The aim of this review is to briefly assess the literature about the utility of poly-unsaturated fatty acids(PUFAs) in the management of IBD. Taken together, almost all studies suggest some beneficial effects of n-3 PUFAs in IBD but the mechanism remains controversial. In addition, clinical benefit seems to be largely confined to ulcerative colitis. However all studies have concluded that these compounds have no potential for a steroid/aminosalicylic acid sparing effect or to maintain remission. Now the question arises as to whether this treatment is of real value to IBD patients?Clearly they have some therapeutic potential but further work is needed.

Eukaryotic microbial distribution pattern and its potential effects on fisheries in the fish reserves of Qiantang River in breeding season

To examine the eukaryotic biodiversity of aquatic ecosystems in the Qiantang River,China,eukaryotic microbes in the river were investigated using 18S rRNA gene sequencing during the breeding season(July to August 2018).Four distinct distribution patterns(1.Jiande;2.Tonglu and Fuyang;3.Jiubao;4.Yanguan)of the microbial community and their potential effects on fishery activities were observed.Results show lower abundances of Dinophyta and Fungi and higher abundances of Cryptophyta and Chlorophyta in Tonglu and Fuyang than those in the other three sections.In addition,the reserves(Tonglu and Fuyang)destabilized the original eukaryotic microbial co-occurrence network.Among all the environmental factors measured,nitrogen(nitrite,nitrate,ammonium),water temperature and total chlorophyll a acted as major driving factors that controlled the eukaryotic microbial distribution.Furthermore,the existence of some algae(e.g.,Chrysophyceae,Cryptophytes,and Chlorophyceae)and fungi(e.g.,Rhizophydium)in Tonglu and Fuyang was beneficial to juvenile fish growth and water quality,although some detrimental species(e.g.,Aphanomyces)needed attention.This study provides further insights into the sustainable protection and utilization of rivers.

Effect of Fish Meal Replacement by Unconventional Meal in the Post-Larval Diet of Clarias Gariepinus in Benin (West Africa)

To reduce the pressure on aquatic resources due to the use of fishmeal in fish feed, the mastery and use of alternative sources of fishmeal in all stages of fish development are essential. Five diets including 4 experimental and one control diet were considered. Experimental diets of post-larvae were developed from unconventional ingredients in which fishmeal was completely replaced by a mixture of brewer’s yeast, chicken viscera and maggots. These foods were tested on Clarias gariepinus post-larvae with an initial average weight of 80 ± 1 mg. 1500 fish individuals were equally distributed in 15 concrete basins with a volume of 500 L forming five treatments in triplicate. Fish individuals were fed, manually at a ration rate of 20%, four times per day. The specific growth rates were 8.38%/day in imported food and 6.35%/day in the experimental food receiving 45.5% of protein from unconventional meal. Similar results were obtained with the feed consumption index. These encouraging results show that it is possible to partially or completely replace fishmeal in catfish feed and obtain an economically profitable production. The recovery of waste such as chicken viscera, brewer’s yeast and soybean meal in fish food formulation as tested in this study is a good contribution to environmental sanitation.

"Teaching to Fish"Rather Than"Giving a Fish"Is China Contributing to the Global Sustainable Futures?

"Give a man a fish and you feed him for a day;teach a man to fish and you feed him for a lifetime".This proverb now becomes the slogan of China's international aid.This paper is based on my previous empirical studies on China's"donor logic"of aid.Rather than looking at the motives of Chinese aid and how it is challenging the Western ideas,this paper focuses on policy discourse and tries to seek the recent trends and knowledge transformations of China's international cooperation of education.

Metazoan Parasites of <i>Geophagus proximus</i>, a Cichlidae Fish from the Eastern Amazon (Brazil)

The present study investigated the fauna of metazoan parasites of a Geophagus proximus population from the lower Tapajós River, in the state of Pará, northern Brazil. A total of 137 monogeneans were collected from the gills of G. proximus, including Sciadicleithrum kritskyi, Sciadicleithrum paranaensis and Sciadicleithrum geophagi, while 119 Raphidascaris (Sprentascaris) lanfrediae nematodes and 28 metacercariaes of digenea undetermined were collected from the intestine. Hosts harboring four species of parasites were predominant. The parasites had an aggregated dispersion pattern. The present study represents the first record of S. geophagi parasitizing G. proximus, increasing the geographic distribution of these parasite species to the Tapajós River basin.

Identification of Bacterial Fish Pathogens in Brazil by Direct Colony PCR and 16S rRNA Gene Sequencing

Intensive fish farming systems in Brazil have increased the disease incidence, mainly of bacterial origin, due to higher stocking density, high organic matter levels and poor quality of the aquatic environment that causes high mortality rates during outbreaks. The identification of pathogenic species using a fast and reliable method of diagnosis is essential for successful epidemiological studies and disease control. The present study evaluated the use of direct colony PCR in combination with 16S rRNA gene sequencing to diagnose fish bacterial diseases, with the goal of reducing the costs and time necessary for bacterial identification. The method was successful for all 178 isolates tested and produced bands with the same intensity as the standard PCR performed using pure DNA. In conclusion, the genetics methods allowed detecting the most common and important pathogens in Aquaculture, including 12 species of occurrence in Brazilian fish farms. The results of the present study constitute an advance in the available diagnostic methods for bacterial pathogens in fish farms.

5S rDNA在百合染色体上的定位及其联会特点研究

利用FISH技术追踪不同倍性百合中5S rDNA在染色体上的位置,发现5S rDNA位点稳定位于3号染色体长臂近端部。通过观察铁炮百合小孢子母细胞减数分裂过程中染色体的联会配对特点,发现不同染色体联会特点不同,以5S rDNA重复序列为探针的FISH结果可发现铁炮百合3号染色体上5S rDNA区域在减数分裂过程中不发生联会,这是5S rDNA能稳定在3号染色体遗传的主要原因。

45S rDNA在7种竹子植物染色体上的定位

编码18-5.8-25S核糖体RNA的45S rDNA基因,是1个简单的多基因家族基因,一般以串联方式相连,对应于核仁组织区(NOR)。首次利用荧光原位杂交技术(FISH),研究45S rDNA在散生竹类的毛竹和斑竹,混生竹类的茶秆竹、日本矮竹、菲白竹和铺地竹,以及丛生竹类的白绿竹染色体上的分布。结果表明:本研究所涉及的散生竹和混生竹的几个竹种,只观察到1对随体染色体的次缢痕区域有45S rDNA位点,而丛生竹类的白绿竹除随体染色体具有45S rDNA位点外,某些非随体染色体上也有不同拷贝数的45SrDNA位点存在。