贪食迈阿密虫(M.avidus)对4种海水养殖鱼类组织匀浆液的趋化性

通过对贪食迈阿密虫(M. avidus)对4种养殖鱼类组织匀浆液的趋化特性进行研究,以探讨贪食迈阿虫对养殖鱼类的侵染特点。试验结果显示,贪食迈阿密虫对大黄鱼、大菱鲆、黄姑鱼、鲈鱼的眼、脑、脑脊液匀浆液有强烈的趋化效果(P<0.01),对皮肤黏液、鳃、肝、肌肉、肾组织匀浆液的趋化性较弱(P>0.05);对不同鱼类组织匀浆液对贪食迈阿密虫的趋化诱导差异比较结果推测,贪食迈阿密虫对宿主具有一定的选择性。本研究有助于深化贪食迈阿密虫致病性的认识,并为海水养殖鱼类盾纤虫病的预防提供参考资料。     

Pharmacokinetics and Pharmacodynamics of a Depolymerized Glycosaminoglycan from Holothuria fuscopunctata,a Novel Anticoagulant Candidate,in Rats by Bioanalytical Methods

dHG-5 (Mw 5.3 kD) is a depolymerized glycosaminoglycan from sea cucumber Holothuria fuscopunctata. As a selective inhibitor of intrinsic Xase (iXase), preclinical study showed it was a promising anticoagulant candidate without obvious bleeding risk. In this work, two bioanalytical methods based on the anti-iXase and activated partial thromboplastin time (APTT) prolongation activities were established and validated to determine dHG-5 concentrations in plasma and urine samples. After single subcutaneous administration of dHG-5 at 5, 9, and 16.2 mg/kg to rats, the time to peak concentration (T_max) was at about 1 h, and the peak concentration (C_max) was 2.70, 6.50, and 10.11 μg/mL, respectively. The plasma elimination half-life(T_1/2β) was also about 1 h and dHG-5 could be almost completely absorbed after s.c. administration. Additionally, the pharmacodynamics of dHG-5 was positively correlated with its pharmacokinetics, as determined by rat plasma APTT and anti-iXase method, respectively. dHG-5 was mainly excreted by urine as the unchanged parent drug and about 60% was excreted within 48 h. The results suggested that dHG-5 could be almost completely absorbed after subcutaneous injection and the pharmacokinetics of dHG-5 are predictable. Studying pharmacokinetics of dHG-5 could provide valuable information for future clinical studies.     

土壤调理粉剂处理下茶园土壤重金属污染特征及潜在风险评价

本文以贵州安顺不同土壤调理粉剂处理下“瀑布毛峰”茶园土壤为供试样品,通过样品测试与理化分析,对CK处理(0 kg/hm²)、A处理(600 kg/hm²)、B处理(750 kg/hm²)和C处理(900 kg/hm²)下茶园土壤重金属含量特征及污染水平进行探讨。结果表明：不同土壤调理粉剂处理下,研究区域土壤重金属Cr、Hg、Pb、Cu、Zn、Ni、As和Cd的变异程度各异,且土壤中这8种重金属含量也各不相同,其中pH、Cr、Pb和Ni含量在C处理中最高,As、Hg和Cd含量在A处理中最高,Cu和Zn含量在B处理中最高;对各重金属元素进行相关性分析可知,Ni与Pb之间存在极显著相关性,表明两者之间的同源性极高,其余元素之间相关性特征不明显;由综合污染指数（P_综）得出A处理为轻度污染,CK处理、B处理和C处理均为中度污染,不同处理土壤中Cr均在国家Ⅱ级标准(GB15618—2008)规定范围内,样点达标率为100%,在所选定的污染评价因子中,Cu的贡献率最高,为土壤主要影响因子,其次是Cd和Ni;研究区土壤重金属单项潜在生态风险指数($E_{r}^{i}$)中Hg属于强生态风险,其余7种重金属处于轻微潜在生态风险等级;综合潜在生态风险指数(RI)平均值为196.79,总体处于中等潜在生态风险水平,Hg对RI的贡献率最大,是最主要的生态风险元素。     

电感耦合等离子发射光谱法测定水质中的金元素

应用电感耦合等离子体发射光谱法测定了废水中的金元素,探讨了金测定的谱线选择、离子干扰等问题,实验结果表明:在最佳仪器条件和测定条件下,该方法的检出限为0.008 mg/L,方法精密度的方法标准偏差(RSD)在4.28%~1.44%之间,回收率为96%~104%。该方法操作简便,快捷。     

四川老鹰茶醇提物的抑菌及抗氧化活性研究

以四川3个不同地区的老鹰茶及老鹰茶植物的鲜叶为原料,采用超声波辅助提取技术提取醇提物,测定多酚和黄酮含量以及抑菌和抗氧化性活性.结果表明:6种样品醇提物的提取率最高的为石棉老鹰茶鲜叶,为(26.73±0.68)％;总多酚和总黄酮含量最高的是石棉老鹰茶鲜叶,分别为(22.10±0.11)％和(16.77±0.14)％.6种样品醇提物对4种细菌均有抑制作用,且提取物浓度越高抑菌效果越强;其中石棉老鹰茶鲜叶对痢疾志贺氏菌(Shigella dysenteriae)抑制效果最好,抑菌圈直径达到(2.56±0.09)cm.6种样品醇提物的DPPH(1,1-二苯基-2-三硝基苯肼)自由基清除能力和总还原能力均高于对照品BHT(2,6-二叔丁基对甲酚).6种样品抑菌和抗氧化能力结果由强到弱为石棉老鹰茶鲜叶>叙州老鹰茶鲜叶>青川老鹰茶鲜叶>石棉老鹰茶>叙州老鹰茶>青川老鹰茶.试验表明四川3个不同地区的老鹰茶及老鹰茶植物的鲜叶均具有较强的抑菌和抗氧化活性,其中石棉老鹰茶植物的鲜叶相对更好,可优先开发利用.     

Molecular Characterization of the Von Willebrand Factor Type D Domain of Vitellogenin from Takifugu flavidus

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val¹¹⁵, ASP¹¹⁶, Val¹¹⁷, and Lys¹²²) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.     

Intestinal Microbiota in Large Yellow Croaker,Larimichthys crocea,at Different Ages

The polymerase chain reaction–denaturing gradient gel electrophoresis (PCR‐DGGE) of 16S ribosomal RNA gene was used to investigate bacterial communities in the intestines of large yellow croaker at six different ages (12 d, 18 d, 26 d, 40 d, 3 mo, and 1 yr old) as well as within the corresponding feed and culture water. In addition, Illumina Miseq sequencing was utilized to compare intestinal microbiota between 12‐d‐old and 1‐yr‐old individuals. PCR‐DGGE results revealed that the culture water had the highest bacterial diversity, followed by the feed, while the intestines had the lowest diversity. The intestinal microbiota at six ages changed severely; however, the change did not follow any trend. The large yellow croaker intestines harbored specific bacterial communities that differed from those in both feed and water. Illumina Miseq sequencing results revealed that the diversity of intestinal bacteria in 12‐d‐old fish was higher than that in 1‐yr‐old fish, and the bacterial composition differed significantly between them. γ‐Proteobacteria and Pseudoalteromonas supplied the most abundant phylum and genus in the 12‐d‐old fish intestine. However, in the 1‐yr‐old fish intestine, Firmicutes and Clostridium were the most dominant, respectively. The study may contribute to a better understanding of gut microbiota and dynamics of the large yellow croaker and the relationship with their surrounding environment.     

Molecular cloning and functional characterization of the fatty acid delta 6 desaturase (FAD6) gene in the sea cucumber Apostichopus japonicus

The sea cucumber (Apostichopus japonicus), an important echinodermata, had high value in nutrition and medicine for its rich collagen, sulphated polysaccharide, glycosides and polyunsaturated fatty acids (PUFA). The cDNA of the fatty acid desaturase gene in A. japonicus (AJFAD6) was cloned and was found to encode a desaturase with delta 6 FAD activity. Sequence analysis indicated that AJFAD6 included an open reading frame of 1392 bp, encoding 463 amino acids. AJFAD6 has all the conserved motifs found in other members of the FAD6 family, including an N‐terminal cytochrome b5 domain and three histidine‐rich regions. qRT‐PCR showed that AJFAD6 was expressed in all tissues tested during juvenile development and was mainly expressed in the respiratory tree at 150 days after adherence (150 days) and in the intestine at 100 days. Furthermore, AJFAD6 mRNA was also detected in the analysed adult tissues, with higher expression in the intestine and testis. Functional characterization of AJFAD6 in a recombinant yeast, Pichia pastoris, showed that AJFAD6 could catalyse exogenous linoleic acid (LA) and α‐linolenic acid (ALA) to produce γ‐linoleic acid (GLA) and stearidonic acid (STA), respectively, at conversion rates of 11.1% for LA to GLA and 3.4% for ALA to STA. Our results suggested that the biosynthetic pathway of PUFA existed in the sea cucumber, but endogenous production of eicosapentaenoic acid, arachidonic acid and docosahexaenoic acid from either LA or ALA precursor appeared to be limited.     

Apparent digestibility coefficients of animal and plant feed ingredients for juvenile Pseudobagrus ussuriensis

A study was conducted to determine apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein (CP), lipid, gross energy, phosphorus and amino acids (AAs) from Peruvian fish meal (PFM), native fish meal (NFM), meat bone meal (MBM), dried daphnia meal (DDM), soybean meal (SBM), full‐fat soybean (FSBM), cottonseed meals (CSM), rapeseed meal (RSM) and corn gluten meal (CGM) for Pseudobagrus ussuriensis (7.8 ± 0.06 g). Test diets were formulated to contain a 70:30 mixture of reference diet to test ingredient with chromic oxide (5 g kg^–1) serving as the inert marker. ADCs of DM ranged from 49.94% (CGM) to 87.11% (PFM), the highest ADCs of DM were observed in PFM and the lowest was found in CGM. ADCs of CP in the test ingredients ranged from 74.92% (CGM) to 93.41% (SBM). ADCs of crude lipid ranged from 51.80% (CGM) to 94.07% (PFM) for fish. ADCs of energy ranged from 56.42% (CGM) to 90.09% (PFM). ADCs of phosphorus ranged from 21.81% (CSM) to 65.70% (PFM). AA availability for the test ingredients followed a similar pattern to ADCs of CP, in which values for PFM, NFM and MBM were generally higher (p < .05) than those of other protein ingredients. Among all plant meals, the AA availability values in SBM were higher (p < .05) than those in CGM, CSM and RSM. The availability of most AAs in CGM was the lowest (p < .05) among all tested ingredients. In conclusion, PFM and NFM are good sources of available protein and AAs. When fishmeal supply is limited, MBM and DDM are substitutes for fishmeal reduction in P. ussuriensis diets; at the same time, SBM and FSBM are also very acceptable protein feedstuffs. However, CSM, RSM and CGM could not be used at high levels in P. ussuriensis diets because of their inferior digestibilities.