An assessment of chemical contaminants detected in passive water samplers deployed in the St. Thomas East End Reserves (STEER)

This report is the second in a series from a project to assess land-based sources of pollution (LBSP) and effects in the St. Thomas East End Reserves (STEER) in St. Thomas, USVI, and is the result of a collaborative effort between NOAA’s National Centers for Coastal Ocean Science, the USVI Department of Planning and Natural Resources, the University of the Virgin Islands, and The Nature Conservancy. Passive water samplers (POCIS) were deployed in the STEER in February 2012. Developed by the US Geological Survey (USGS) as a tool to detect the presence of water soluble contaminants in the environment, POCIS samplers were deployed in the STEER at five locations. In addition to the February 2012 deployment, the results from an earlier POCIS deployment in May 2010 in Turpentine Gut, a perennial freshwater stream which drains to the STEER, are also reported. A total of 26 stormwater contaminants were detected at least once during the February 2012 deployment in the STEER. Detections were high enough to estimate ambient water concentrations for nine contaminants using USGS sampling rate values. From the May 2010 deployment in Turpentine Gut, 31 stormwater contaminants were detected, and ambient water concentrations could be estimated for 17 compounds. Ambient water concentrations were estimated for a number of contaminants including the detergent/surfactant metabolite 4-tert-octylphenol, phthalate ester plasticizers DEHP and DEP, bromoform, personal care products including menthol, indole, n,n-diethyltoluamide (DEET), along with the animal/plant sterol cholesterol, and the plant sterol beta-sitosterol. Only DEHP appeared to have exceeded a water quality guideline for the protection of aquatic organisms.

Liver genomic responses to ciguatoxin: evidence for activation of Phase I and Phase II detoxification pathways following an acute hypothermic response in mice

Ciguatoxins (CTX) are polyether neurotoxins that target voltage-gated sodium channels and are responsible for ciguatera, the most common fish-borne food poisoning in humans. This study characterizes the global transcriptional response of mouse liver to a symptomatic dose (0.26 ng/g) of the highly potent Pacific ciguatoxin-1 (P-CTX-1). At 1 h post-exposure 2.4% of features on a 44K whole genome array were differentially expressed (p ≤ 0.0001), increasing to 5.2% at 4 h and decreasing to 1.4% by 24 h post-CTX exposure. Data were filtered (|fold change| ≥ 1.5 and p ≤ 0.0001 in at least one time point) and a trend set of 1550 genes were used for further analysis. Early gene expression was likely influenced prominently by an acute 4°C decline in core body temperature by 1 h, which resolved by 8 h following exposure. An initial downregulation of 32 different solute carriers, many involved in sodium transport, was observed. Differential gene expression in pathways involving eicosanoid biosynthesis and cholesterol homeostasis was also noted. Cytochrome P450s (Cyps) were of particular interest due to their role in xenobiotic metabolism. Twenty-seven genes, mostly members of Cyp2 and Cyp4 families, showed significant changes in expression. Many Cyps underwent an initial downregulation at 1 h but were quickly and strongly upregulated at 4 and 24 h post-exposure. In addition to Cyps, increases in several glutathione S-transferases were observed, an indication that both phase I and phase II metabolic reactions are involved in the hepatic response to CTX in mice.

青蒿反义鲨烯合酶基因对烟草的转化

利用反义技术研究生物代谢途径以及对其生物合成进行调控成为植物次生代谢研究领域内一个重要手段之一，并与新兴的RNAi技术一起成为本领域内重要的研究热点。在植物类异戊二烯代谢途径中存在着羟甲基戊二酰辅酶A还原酶(HMGR)、法呢基焦磷酸合酶（FPS）和鲨烯合酶（SQS）等几种关键的分支酶，他们被认为在异戊二烯类的生物合成中发挥着关键的调节作用。其中，鲨烯合酶处于HMGR和FPS的下游，并与倍半萜合酶等利用共同的前体－法呢基二磷酸（FPP），以FPP起始合成一系列的下游产物。因此，FPP成为类异戊二烯途径中的关键调节点之一。本论文基于此目的，利用反义技术研究了FPP合成鲨烯这一途径受到抑制对其他以FPP为生物合成前体的代谢支路的影响。 利用植物双元转化载体pBI121，将青蒿中鲨烯合酶基因的cDNA（约1.5kb）序列插入到pBI121中，取代原有的GUS序列，构建成植物转化载体pBIASS。以根癌农杆菌为介导，将青蒿鲨烯合酶反义基因序列导入到烟草，整合到其基因组中 ，成功获得转基因植株。对转基因烟草进行分子检测表明，外源鲨烯合酶基因的序列已经稳定整合到烟草基因组中，并对内源的烟草鲨烯合酶基因表达产生影响。转基因烟草中检测到内源鲨烯合酶基因的mRNA的水平降低。对鲨烯合酶下游产物之一的胆固醇的含量分析显示，活性减低的鲨烯合酶使胆固醇的生物合成下降约40％左右。同时，另一条以FPP为共同前体的二萜代谢途径产物之一GA3的含量得到了提高，比对照提高约30％。

水稻GA3应答基因表达谱微阵列分析与OsGAS1功能初步研究

利用反义技术研究生物代谢途径以及对其生物合成进行调控成为植物次生代谢研究领域内一个重要手段之一，并与新兴的RNAi技术一起成为本领域内重要的研究热点。在植物类异戊二烯代谢途径中存在着羟甲基戊二酰辅酶A还原酶(HMGR)、法呢基焦磷酸合酶（FPS）和鲨烯合酶（SQS）等几种关键的分支酶，他们被认为在异戊二烯类的生物合成中发挥着关键的调节作用。其中，鲨烯合酶处于HMGR和FPS的下游，并与倍半萜合酶等利用共同的前体－法呢基二磷酸（FPP），以FPP起始合成一系列的下游产物。因此，FPP成为类异戊二烯途径中的关键调节点之一。本论文基于此目的，利用反义技术研究了FPP合成鲨烯这一途径受到抑制对其他以FPP为生物合成前体的代谢支路的影响。 利用植物双元转化载体pBI121，将青蒿中鲨烯合酶基因的cDNA（约1.5kb）序列插入到pBI121中，取代原有的GUS序列，构建成植物转化载体pBIASS。以根癌农杆菌为介导，将青蒿鲨烯合酶反义基因序列导入到烟草，整合到其基因组中 ，成功获得转基因植株。对转基因烟草进行分子检测表明，外源鲨烯合酶基因的序列已经稳定整合到烟草基因组中，并对内源的烟草鲨烯合酶基因表达产生影响。转基因烟草中检测到内源鲨烯合酶基因的mRNA的水平降低。对鲨烯合酶下游产物之一的胆固醇的含量分析显示，活性减低的鲨烯合酶使胆固醇的生物合成下降约40％左右。同时，另一条以FPP为共同前体的二萜代谢途径产物之一GA3的含量得到了提高，比对照提高约30％。

Physiological ecology of juvenile chinook salmon (Oncorhynchus tshawytscha) at the southern end of their distribution, the San Francisco Estuary and Gulf of the Farallones, California

Juvenile chinook salmon, Oncorhynchus tshawytscha, from natal streams in California’s Central Valley demonstrated little estuarine dependency but grew rapidly once in coastal waters. We collected juvenile chinook salmon at locations spanning the San Francisco Estuary from the western side of the freshwater delta—at the confluence of the Sacramento and San Joaquin Rivers—to the estuary exit at the Golden Gate and in the coastal waters of the Gulf of the Farallones. Juveniles spent about 40 d migrating through the estuary at an estimated rate of 1.6 km/d or faster during their migration season (May and June 1997) toward the ocean. Mean growth in length (0.18 mm/d) and weight (0.02 g/d) was insignificant in young chinook salmon while in the estuary, but estimated daily growth of 0.6 mm/d and 0.5 g/d in the ocean was rapid (P≤0.001). Condition (K factor) declined in the estuary, but improved markedly in ocean fish. Total body protein, total lipid, triacylglycerols (TAG), polar lipids, cholesterol, and nonesterified fatty acids concentrations did not change in juveniles in the estuary, but total lipid and TAG were depleted in ocean juveniles. As young chinook migrated from freshwater to the ocean, their prey changed progressively in importance from invertebrates to fish larvae. Once in coastal waters, juvenile salmon appear to employ a strategy of rapid growth at the expense of energy reserves to increase survival potential. In 1997, environmental conditions did not impede development: freshwater discharge was above average and water temperatures were only slightly elevated, within the species’ tolerance. Data suggest that chinook salmon from California’s Central Valley have evolved a strong ecological propensity for a ocean-type life history. But unlike populations in the Pacific Northwest, they show little estuarine dependency and proceed to the ocean to benefit from the upwelling-driven, biologically productive coastal waters.

青蒿素生物合成相关基因对烟草遗传转化的研究

　　青蒿素是存在于中药青蒿(Artemisia annua L.)中的一种含有过氧桥的倍半萜内酯化合物，是中国科学家研发出的当今最有潜力的抗疟药剂，较传统抗疟药很少或无毒副作用，因此青蒿素的生产备受人们关注。目前，青蒿素的生产主要以植物提取为主，但由于青蒿植株中青蒿素的含量很低（约占干重的0.01%～0.8%），从而导致青蒿素价格昂贵，使许多贫困地区的疟疾患者无法得到医治，故提高青蒿植株中青蒿素的含量或扩大青蒿素的来源，降低生产青蒿素的成本具有重要的意义。　　 　　本论文基于扩大青蒿素的来源和提高青蒿植株中青蒿素含量的目的，开展了以下两方面的工作： 一、紫穗槐二烯在烟草中组合生物合成的研究 　　紫穗槐二烯合酶（amorpha-4,11-diene synthase，ADS）是青蒿素生物合成的关键酶之一，为了能在烟草中合成青蒿素的前体，本研究将青蒿的紫穗槐二烯合酶基因置于CaMV 35S启动子控制下，通过根癌农杆菌介导转入烟草（Nicotiana tobacum L.），并获得了转ADS基因烟草植株。经PCR及Southern杂交分析表明，ADS基因已经整合到转基因烟草基因组中；RT-PCR及对转基因烟草中ADS酶活性和产物中紫穗槐二烯和植物甾醇的测定分析，进一步证明整合的ADS基因在转录、翻译水平上均已经表达。上述结果表明，利用基因工程将青蒿素生物合成途径的关键酶基因导入植物，转基因植物中能够合成青蒿素的前体，这一研究结果为利用转基因植物生产青蒿素或其前体奠定了基础。 二、青蒿鲨烯合酶双链干涉基因对烟草的遗传转化研究 　　鲨烯合酶（squalene synthase, SQS）是甾醇类生物合成分支途径的关键酶之一，利用RNA干扰技术(RNA interference，RNAi)抑制目标基因表达的技术已日趋成熟。本文根据植物中hpRNA(hairpin RNA)的原理，在与烟草SQS同源性高达80％的青蒿ASQS序列的5/端保守区选择622 bp作为构建RNAi的序列，借助中间克隆载体，经过三次亚克隆，最后形成含ASQS－RNAi表达盒的双元表达载体pART27－ASQS，并转入农杆菌EHA105。采用农杆菌介导的烟草叶盘转化法，共获得了12棵转基因植株。转基因植株经过PCR和PCR-Southern blotting 检测，证实外源ASQS基因已经导入烟草中，并已经成功整合到烟草基因组中；通过RT-PCR分析说明，转基因烟草中SQS基因的表达已被成功抑制，部分转基因植株中内源SQS的干扰效果高达90％以上。对SQS的直接产物鲨烯和最终产物植物甾醇的检测显示，转基因烟草的植物甾醇和鲨烯的含量明显低于对照。本实验的结果为下一步将此RNA干扰载体导入青蒿，抑制青蒿中ASQS基因的表达，从而提高青蒿素的含量提供了可能。

Biochemical variation during ovarian vitellogenic growth in a hill stream teleost Garra mullya (Sykes) due to cadmium toxicity

Some biochemical variations during ovarian vitellogenic growth in hill-stream teleost Garra mullya due to sublethal concentration of cadmium has been discussed. Total protein, cholesterol and glycogen in ovary and liver along with gonadosomatic index (GSI) and hepatosomatic index (HSI) in Cd-treated fish exhibited significant decrease while liver glycogen remained unaltered.

Sterol composition of marine algae from Karachi coast of Arabian Sea

During the course of chemical investigation of marine algae collected from Karachi coast of Arabian Sea, five sterols named as sarangosterol(1), 23-methyl cholesta-5, 25-dien-3ß-ol(2) from Endarachne binghamiae (brown alga), sargasterol(3) from Dictyota indica (brown alga), cholesterol(4) from Laurencia obtusa (red alga) and clerosterol(5) from Codium iyengarii (green alga) have been isolated. Their structures were elucidated with the help of spectroscopic means.

Analysis of lipid and lipid-fractions of some freshwater fishes and their inter-relationship

Two fish species each from carnivorous (Clarias batrachus, Channa punctatus), omnivorous (Cyprinus carpio, Cirrhinus reba), and plankton feeder (Catla catla, Labeo rohita) were collected from freshwater sources under natural habitat to study their total lipid (TL) and lipid-fractions. Significant relationship between these parameters was also worked out. The variation of total lipid and lipid-fractions in tissues of freshwater fishes were not significantly different (P>0.05). But a higher trend of total lipid and glyceride (TGL) contents were found in carnivores followed by omnivores and least in plankton feeders. The trend was reverse for total phospholipid, cholesterol and free fatty acids. TGL content in all class of fishes was significantly related with TL (P<0.01), phospholipid (PL) (P<0.001), cholesterol (P<0.05), free fatty acids (P<0.05) and monoglycerides (P<0.001). Similarly total lipid was linearly related with total glycerides (TL=-3.02 + 0.10 TGL) and phospholipid (TL=7.13-0.12 PL). From this study it is concluded that almost all lipid-fractions of freshwater fishes can be predicted easily from total lipid content of the tissue.

Changes in lipids of mussel during storage

Proximate composition, lipid and fatty acid components of dried mussel and changes in lipids during 1 year storage were studied. Male mussel contained lower fat contents and higher contents of polyunsaturated fatty acids of C20:5n-3, and C22:6n-3. High percentages of Cl6:1, Cl7:1, Cl8:3n-3, C20:3n-8 existed in NL and C!6:0, C18:0, Cl8:1n-9, C20:2n-6, C20:5n- 3, C22:6n-3 were very rich in PL. Triglycerides phosphatidylcholine, cholesterol were major components of mussel lipids. Free fatty acids (FFA) increased greatly and phospholipids decreased during storage, saturated fatty acids showed an increase trend and polyunsaturated fatty adds decreased differently. Dried mussels were vacuum packed and air packed and packaging methods had a great influence on the oxidation of mussellip,ids, indicating preference of vacuum packaging.

Phycochemical studies on sterols from three brown seaweeds of northern Arabian Sea

Three brown algae i.e. Dictyota hauckiana Nizamuddin, Levringia boergesenii Kylin and Spatoglossum variabile Figari et De Notaris, collected from the coastal areas of Karachi, have been investigated for their sterol composition. Four sterols with cholesta skeleton including a new one, 17α-hydroxy-24β-vinyl cholesterol have been isolated from them. Their structures were elucidated with the help of spectroscopic analysis. The new sterol has been named as hauckiosterol after its first source of isolation.

Some biochemical constituents and the calorific values of different regions of the body musculature of pond murrel Channa punctatus Bloch

A study of the distribution of some chemical constituents in the musculature of Channa punctatus Bloch showed it to be fairly heterogeneous. The increase in the levels of protein, fat, ash, total carbohydrates, glycogen, RNA, DNA and cholesterol towards the posterior region of the fillet was the result of increase in the number of cells per unit weight of the sample and in the concentration of myocommata. The distribution of water in the musculature was inversely related to that of the fat. The concentration of protein appeared to be associated with the RNA level. The posterior region of the fillet exhibited more calorific value than the anterior region.

Chemical changes in skin mucin as an index of early stages of spoilage in fish

Results of a preliminary investigation on the overall chemical nature of fish skin mucin in lung fish, Clarias batrachus, with special reference to water soluble low molecular weight compounds, are presented. Changes observed during room temperature spoilage have been studied with a view to present a new approach towards the assessment of freshness in fish inspection. pH of the mucin was distinctly alkaline (8.2) and remained unchanged during spoilage. Much of the nitrogen was found to be present in the glycoprotein fraction. Free amino acids and purine bases were present in appreciable quantities in the aqueous extracts which registered a significant increase after 10 hrs. Post-mortem increase in total solids was accompanied by a slight rise in protein nitrogen which may indicate tissue breakdown. Increase in TVN was also observed to occur earlier in the outside mucin as compared to the inside muscle. Presence of free sugars or sialic acid could not be confirmed nor was there any indication of cholesterol and lipoid material as stated in earlier literature.

Seasonal changes in the biochemical composition of ovary in Heteropneustes fossilis (Bloch)

Seasonal changes in the biochemical composition of ovary in H. fossilis are reported. An inverse relationship was noted in fat and water contents. Maximum fat was observed in June and lowest in December. Protein and ash were generally low during winter and high during summer or monsoon months. Variations in the cholesterol content were more or less identical to those of the fat.

Growth and survival of Penaeus monodon postlarvae fed shrimp head meal and fish meal as primary animal source of protein

Although shrimp head meal alone does not provide for good growth and survival, fish meal can provide high survival rate. The addition of shrimp head improves this diet. It is suggested that cholesterol present in shrimp could have caused this difference. Composition of the test diets is tabulated, as are proximate chemical analysis of the diets, and the mean initial weights, final weights, weight gains, survival rate, feed consumed, protein consumed, of Penaeus monodon postlarvae, feed conversion and protein efficiency ratio.