Effects of induced paralysis on hemocytes and tissues of the giant lions-paw scallop by paralyzing shellfish poison

In general, bivalves are not affected by exposure to toxic dinoflagellates that produce paralyzing shellfish poisons (PSP). After injection with PSP extracted from the Gymnodinium catenatum, Nodipecten subnodosus is paralyzed, indicating that PSP provokes effects similar to what is observed in vertebrates, including paralysis and metabolic stress. To investigate the processes involved in poisoning by PSP, lions-paw scallops were injected with gonyautoxin (GTX) 2/3 epimers in the adductor muscle. Mild doses provoked adductor muscle contractions and paralysis, mantle retraction, and incapacity of shell closure, but scallops gradually recovered in a clear, dose-time recovery pattern. With high doses of GTX 2/3, scallops were permanently paralyzed, and hemocytes in hemolymph were reduced. Surprisingly, under these conditions, scallops continued normal feeding and did not show any microscopic defect in intestine or gills, but hemocytes infiltrated the adductor muscle and abnormal vitellogenesis and mantle melanization occurred. Paralysis stress was accompanied by negative scallop responses, based on visible effects, generation of nitric oxide, lipid peroxidation, and changes in antioxidant and hydrolytic enzymes in hemocytes and tissues. These data can be used to understand potential side effects of PSP in bivalves.     

Paralytic shellfish poison in Spisula solidissima: Anatomical location and ozone detoxification

The surf clam Spisula solidissima, when exposed to a northern bloom of the toxic dinoflagellate Gonyaulax tamarensis, concentrates paralytic shellfish poison (PSP) and retains it for periods of over 1 year. The purpose of this investigation was to identify those tissues in which S. solidissima concentrates PSP and to examine the efficacy of ozone gas in PSP detoxification. Various levels of the toxin were found in every untreated tissue examined: the mantle and gill containing high concentrations (>1600 g/100 g tissue); the visceral mass, siphon, and foot showing less toxicity (1100 to 200 g/100 g tissue); and the adductor muscle yielding a level of toxin considered safe for human consumption (<60 g/100 g tissue). Toxic clams exposed to ozonized seawater for 2 weeks exhibited rapid detoxification in all tissues examined.This work was supported, in part, by a grant from the Massachusetts Science and Technology Foundation, Wakefield, Massachusetts 01880, USA.     

An integrative response by Mytilus chilensis to the toxic dinoflagellate Alexandrium catenella

Physiological responses of Mytilus chilensis exposed to the toxic dinoflagellate Alexandrium catenella were measured over 21 days in the laboratory and were compared with control mussels not exposed to the dinoflagellate. Mussels were collected from culturing ropes at Yaldad Bay, southern Chile (43o08′S 73o44′W), in August 2004 and acclimated to laboratory conditions for one week prior to the experiment. After 8 days, the paralytic shellfish poisoning (PSP) toxins (i.e. saxitoxin) in the tissues of exposed mussels exceeded safe levels for human consumption. Clearance rates, ingestion of organic matter, and absorption efficiency of exposed mussels were significantly lower than those of controls on day 0, but this was followed by an increase on day 3. The exposed mussels also increased their excretion rate over time, and this increase was significantly correlated with the accumulation of PSP toxins in their tissues. Oxygen consumption was not affected by the PSP toxins. The scope for growth (SFG) on day 0 was negative in exposed mussels, but it increased during the experiment. Although feeding activity and absorption efficiency were adversely affected during the first few days of exposure to PSP toxins from A. catenella in the laboratory, the M. chilensis cultured in Yaldad Bay may have evolved mechanisms that allow them to exploit the toxic dinoflagellate as a food source.     

Accumulation of paralytic shellfish poisoning toxins in planktonic copepods during a bloom of the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis> in Hiroshima Bay,western Japan

Concentrations of paralytic shellfish poisoning (PSP) toxins in toxic dinoflagellate cells and in marine planktonic copepods were monitored during the bloom of Alexandrium tamarense in Hiroshima Bay, western Japan. Concentration of the toxins retained by copepods was a function of the ambient toxin concentration, i.e. the product of A. tamarense cell density and cellular toxicity. The toxin concentration in copepods increased with the increase of toxicants in the seawater then leveled off, but decreased significantly at higher concentrations. In the field, the maximum toxin concentration was 1.2 pmol ind^-1, whereas in the laboratory, the copepod Acartia omorii accumulated a much higher concentration of PSP toxins (24 pmol ind^-1). Feeding avoidance against Alexandrium tamarense and a shift to alternative food sources such as diatoms in the field might keep their toxin levels lower than their potentially maximum level. The copepod toxin levels in the field were not so high as to cause an instantaneous lethal effect on their predator fishes but may reach possibly lethal levels after a few days' continuous feeding. Overall toxin retention by copepods after 12 h feeding and 2 h starvation was only 2.5% of total ingested toxins, which suggested that a significant amount of toxins was released into the seawater. Measurements of toxin reduction and gut evacuation suggested that the toxins were removed through both fecal evacuation and metabolism (e.g. excretion, decomposition and transformation). The results, as a whole, imply that copepods can be a link for PSP toxin flux in both pelagic and benthic food webs and can also be a sink for toxins by metabolizing and removing them from the environment.Communicated by T. Ikeda, Hakodate     

Use of high-performance liquid chromatography to investigate the production of paralytic shellfish toxins by Protogonyaulax spp. in culture

Procedures have been developed for the extraction and high-performance liquid chromatography (HPLC) analysis of paralytic shellfish poisoning (PSP) toxins from Protogonyaulax spp. grown in batch culture. Using these procedures, the toxin content of two isolates of P. tamarensis (NEPCC 183 and 255) and one isolate of P. catenella (NEPCC 355) were examined. Total toxin and individual toxin concentrations were measured for each isolate during the exponential and stationary phases of growth in batch culture. The total toxicity of each isolate as measured by HPLC analysis was found to agree with toxicity as determined by the standard mouse bioassay. Two of the isolates (255 and 355) were found to be toxic and the third (183) was non-toxic. The toxic isolates (255 and 355) both showed higher average total PSP toxin content during the exponential phase (35 and 23 fmol toxin cell^-1, respectively) than during the stationary phase (21 and 8 fmol toxin cell^-1, respectively). These cultures differed dramatically in their toxin composition. P. tamarensis (255) contained a large proportion of the N(21) sulfo toxins (B₁, B₂, C₁, C₂) while P. catenella (355) contained primarily Gonyautoxins 1 through 4. The percent composition of individual toxins was found to be constant throughout the growth cycle for both toxic isolates, even though the total toxin concentration varied. Our results suggest that PSP toxin profiles might be useful as chemotaxonomic indicators.     

液相色谱结合串联质谱方法研究中国沿海贝类中脂溶性藻毒素的种类结构和分布规律

一些海洋微藻能分泌产生藻毒素,滤食性的双壳贝类食用有毒的浮游微藻并在体内累积藻毒素,进而引发贝类食用者或其它高等级动物中毒或死亡。脂溶性藻毒素是一些易溶于有机溶剂、因而易富集于动物脂肪组织中的微藻毒素,通常包含6大类,每种类均包括众多结构。使用高效液相色谱结合串联质谱（HPLC-MS/MS）技术方法,研究了来自中国沿海55个站位的34种贝类中脂溶性藻毒素的种类结构及分布。结果表明,在中国沿海多种双壳贝类体内,存在多种脂溶性藻毒素组份,包括软海绵酸毒素OA（Okadaic acid）、扇贝毒素PTXs（Pectenotoxin）、原多甲藻酸Azaspiracid（AZAs）毒素、虾夷扇贝毒素 YTXs（Yesssotoxin）、环亚胺类毒素（Cyclic imines）中的 Gymnodimine（GYM）和 Spiroloides（SPX）毒素;其中南部海域的广东、广西和海南,主要以环亚胺类毒素 GYM 为主;北黄海主要以虾夷扇贝毒素 YTXs 为主,且虾夷扇贝毒素YTXs只在黄海海域检出,特别是主要存在于北黄海的虾夷扇贝（Patinopecten yessoensis）和栉孔扇贝（Chlamys farreri）中;渤海沿岸,主要分布有扇贝毒素PTXs;扇贝毒素PTXs和虾夷扇贝毒素 YTXs含量和检出频率都比较高;扇贝毒素共发现三种结构,其中酸式结构PTX-2Sa的检出率最高（44%）,最高值为53.2μg·kg-1;其次是环亚胺类毒素中的GYM毒素,检出率为35.3%,最高值为39.1μg·kg-1;虾夷扇贝毒素 YTXs共发现三种结构,北黄海浮筏养殖虾夷扇贝中的虾夷扇贝毒素YTXs总量最高值出现在秋季的9月份;中国北黄海虾夷扇贝中YTXs毒素长年存在,秋季含量较高。微藻毒素的分布取决于产毒微藻的分布,目前中国贝类中的这三种主要微藻毒素的生物学来源还尚未确定。简言之,中国海产贝类中存在扇贝毒素PTXs、虾夷扇贝毒素 YTXs和环亚胺类毒素等多种脂溶性微藻毒素?     

Effects of the toxic dinoflagellate <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis> on uptake and fate of paralytic shellfish poisons in the Pacific giant lions-paw scallop <Emphasis Type="Italic">Nodipecten subnodosus</Emphasis>

Juvenile Pacific giant lions-paw scallops Nodipecten subnodosus were fed the toxic dinoflagellate Gymnodinium catenatum, a producer of paralytic shellfish poison (PSP), supplied with Isochrysis galbana (a nontoxic microalgae). Short-term (<24 h) experiments were performed to determine clearance and ingestion rates of G. catenatum. Kinetics of PSP was examined in longer-term experiments (>2 days). At high food concentrations, juvenile scallops showed production of pseudofeces, partial shell valve closure, and reduction in feeding. According to HPLC analysis, the only toxins present in the dinoflagellate G. catenatum and in the scallops were the gonyautoxins (GTXs), except in the labial palps and digestive gland, where trace amounts of saxitoxin (STX) were present in scallops. These tissues could play an important role in toxin biotransformation. The ranking of toxin concentration in tissues was: digestive gland > labial palps > intestine > gills > mantle > adductor muscle, where the total contribution of viscera was more than 80% of the total toxin body burden. Juvenile scallops exhibited no apparent detrimental physiological responses during the long-term feeding experiment. The dinoflagellate may contribute nutrients to the scallop, in addition to the microalgae I. galbana. The dinoflagellate may enhance cell uptake and byssus production. Once PSP accumulated during the first 12 days, it was slowly eliminated. The limited capacity for accumulating toxins in the adductor muscle favors domestic marketing of scallops.     

Toxic dinoflagellates (Alexandrium fundyense and A. catenella) have minimal apparent effects on oyster hemocytes

The possible effect of Alexandrium spp. containing paralytic shellfish poisoning (PSP) toxins on the hemocytes of oysters was tested experimentally. In one trial, eastern oysters, Crassostrea virginica Gmelin, were exposed to bloom concentrations of the sympatric dinoflagellate, Alexandrium fundyense Balech, alone and in a mixture with a non-toxic diatom, Thalassiosira weissflogii (Grun) Fryxell et Hasle. Subsequently, another experiment exposed Pacific oysters, Crassostrea gigas Thunberg, to a mixed suspension of the sympatric, toxic species Alexandrium catenella (Whedon et Kofoid) Balech, with T. weissflogii. Measurements of numbers of oyster hemocytes, percentages of different cell types, and functions (phagocytosis, reactive oxygen species (ROS) production, and mortality) were made using flow-cytometry. During and after exposure, almost no significant effects of Alexandrium spp. upon hemocyte numbers, morphology, or functions were detected, despite observations of adductor-muscle paralysis in C. virginica and measured toxin accumulation in C. gigas. The only significant correlation found was between toxin accumulation at one temperature and higher numbers of circulating live and dead hemocytes in C. gigas. The PSP toxins are known to interfere specifically with sodium-channel function; therefore, the finding that the toxins had no effect on measured hemocyte functions suggests that sodium-channel physiology is not important in these hemocyte functions. Finally, because oysters were exposed to the living algae, not purified toxins, there was no evidence of bioactive compounds other than PSP toxins affecting hemocytes in the two species of Alexandrium studied.     

Feeding and absorption of the toxic dinoflagellate Alexandrium tamarense by two marine bivalves from the South China Sea

Paralytic shellfish poisoning (PSP) toxins can be accumulated by bivalves through the feeding process; therefore, knowledge on feeding and the assimilation of PSP-toxin-containing algae is critical to understand the kinetics of PSP toxins in these bivalves. In the South China Sea, it has been documented that the scallop Chlamys nobilis has a much higher PSP toxin burden than the clam Ruditapes philippinarum. Experiments were therefore carried out to assess whether the difference in toxin burden between these two species of bivalves was due to differences in feeding and absorption. In a mixed diet of Alexandrium tamarense (a PSP-toxin-producing dinoflagellate) and Thalassiosira pseudonana (a non-toxic diatom), the maximum clearance and filtration rates were about two times higher in the scallop C. nobilis than in the clam R. philippinarum. Furthermore, the clams produced pseudofeces at a lower cell density than the scallops. However, we found that the clams were unable to selectively exclude the toxic dinoflagellates by pseudofeces production. The scallop C. nobilis also possessed a greater ability to assimilate A. tamarense with a comparable carbon absorption efficiency to the diatom T. pseudonana. In contrast, the carbon absorption in the clam R. philippinarum was lower when feeding on A. tamarense than on the diatom. In general, the absorption efficiency decreased with increasing concentration of A. tamarense. Thus, it is likely that the higher PSP toxin levels in the scallops compared with clams can be partly explained by differences in their feeding and absorption behavior. Other processes, especially the biotransformation and biokinetics of PSP toxins, may also play a significant role in defining the inter-species differences in PSP body burden in marine bivalves.     

环境激素烯菌酮引发小鼠前列腺癌细胞的毒性机制

为探讨环境激素烯菌酮引发小鼠前列腺癌(RM-1)细胞毒性机制,设计了3组实验,分别为对照组、烯菌酮染毒组(染毒组)和烯菌酮染毒后添加维生素E组(实验组),各组给药培养48h后,观察细胞形态变化。检测细胞增殖活性、丙二醛(MDA)含量、抑制羟自由基能力、乳酸脱氢酶(LDH)活性、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性。结果表明,与对照组相比,染毒组RM-1细胞圆缩脱落明显,贴壁细胞减少;细胞增殖率、抑制羟自由基能力、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性明显降低(p<0.05),LDH活性和MDA含量显著升高(p<0.05)。与染毒组相比,染毒后添加维生素E,圆缩脱落细胞减少,而梭形贴壁细胞明显增加;细胞增殖率、抑制羟自由基能力、Na+-K+-ATP酶活性和Ca2+-Mg2+-ATP酶活性显著升高(p<0.05),LDH活性和MDA含量明显下降(p<0.05)。这表明维生素E可能通过其抗氧化性拮抗了烯菌酮引发的氧化应激,缓解了细胞膜的损伤,维持了细胞膜的完整性和膜蛋白的功能,防止了细胞内LDH外逸、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性的下降,从而逆转了烯菌酮引发的细胞死亡和增殖率的下降。     

腹泻性贝毒素软海绵酸的昆明系小鼠生物学检测法的建立

为完善腹泻性贝毒素软海绵酸(Okadaic acid,OA)的小鼠生物学检测技术,以昆明系小鼠为实验材料,应用小鼠生物学检测法检测了OA的毒性.用不同浓度OA标准溶液通过腹腔注入19～21g昆明系小鼠体内,观察其中毒症状,确定1个鼠单位(MU)所代表的毒素剂量,并且建立剂量-死亡时间曲线.结果表明,昆明系小鼠经OA腹腔注射后中毒症状和死亡时间具有明显剂量效应关系,其剂量-死亡时间曲线方程为y=600.57x-2.257,其中y代表致死时间(h),x代表剂量(μg);将死亡时间t变为时间倒数1/t,对OA的毒性MU取对数(即logMU),得直线回归方程为y=0.5363x+0.1295,R2=0.9488,p<0.01,其中y代表OA毒素剂量单位的对数(1ogMU),x代表小鼠死亡时间倒数(1/t),R2为可决系数.通过该方程计算得出,1个鼠单位(MU)所对应的OA剂量值约为4.015μg,其95%置信限为3.826～4.204μg.     

盐胁迫对油菜幼苗生长和光合特征的影响

采用盆栽砂培试验,研究了不同浓度(0、50、100、200、300mmol·L-1)NaCl胁迫10和30d对油菜(Brassica napus)幼苗干质量、叶绿素(Chl)含量、净光合速率(Pn)、气孔导度(Gs)、细胞间CO2浓度(xi)、蒸腾速率(Rt)、水分利用效率(Ew,u)和气孔限制值(Ls)等的影响.结果表明,在NaCl胁迫下,油菜幼苗植株干质量显著降低,长期高盐胁迫下油菜干质量降低更显著;随NaCl浓度的增加,Chl含量、Chl a/Chl b比值均呈先升高后降低的变化趋势,处理10d,Chl含量、Chl a/Chl b比值在NaCl浓度为200mmol·L-1条件下达最大值,处理30d,在NaCl浓度为100mmol·L-1条件下达最大值.在50～100mmol·L-1NaCl胁迫下,油菜叶片的Pn、xi和Ls所受影响均很小;高盐胁迫下,其Pn、Gs、xi和Rt均显著下降,而Ew,u和Ls则显著上升.相关分析显示,植株干质量与Chl含量、Chl a/Chl b比值间无相关性,与Na+、Cl-含量,Ew,u和Ls间呈显著负相关(P＜0.01),与根冠比,K+、Ca2+含量,K+/Na+、Ca2+/Na+比值,K+与Na+的选择性比率[S(K+,Na+)],Ca2+与Na+的选择性比率[S(Ca2+,Na+)],Pn,Gs,xi和Rt间呈显著正相关(P＜0.01).上述结果表明,200mmol·L-1 NaCl胁迫10和30d、300mmol·L-1 NaCl胁迫10d,油菜幼苗光合抑制主要来自气孔限制,而300mmol·L-1 NaCl胁迫30d,气孔限制和非气孔限制在油菜幼苗光合抑制中均具有重要作用.Na+、Cl-、K+、Ca2+含量,Ew,u,Ls,根冠比,K+/Na+、Ca2+/Na+比值,S(K+,Na+),S(Ca2+,Na+),Pn,Gs,xi和Rt均可作为油菜生长盐适应性的评价指标.     

Toxin accumulation and feeding behaviour of the planktonic copepod Calanus finmarchicus exposed to the red-tide dinoflagellate Alexandrium excavatum

The planktonic copepod Calanus finmarchicus is a dominant member of the zooplankton community in the lower St. Lawrence Estuary in eastern Canada. Blooms of the toxic marine dinoflagellate Alexandrium excavatum which produces high cellular levels of paralytic shellfish poisoning (PSP) toxins, occur during the period of high C. finmarchicus production in summer in this region. To study the feeding behaviour of C. finmarchicus in the presence of Alexandrium spp., experiments were conducted in which female adult copepods collected from the St. Lawrence Estuary between May and September 1991 were exposed under controlled conditions to two toxic isolates of A. excavatum (Pr18b and Pr11f) from the estuary and to a non-toxic control (PLY 173) of a closely related species, A. tamarense isolated from the Tamar Estuary, Plymouth, U.K. Clearance rates on non-toxic A. tamarense cells averaged 5.5 ml ind^-1 h^-1 but were nearzero with either toxic isolate. When presented with a mixture of A. excavatum and the non-toxic diatom Thalassiosira weissflogii in varying proportions, C. finmarchicus fed upon the diatom but avoided the toxic dinoflagellate. Although feeding rates on A. excavatum were very low, toxin analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD) revealed that the PSP toxins were accumulated in copepods exposed to toxigenic dinoflagellates.The toxin composition in copepods was similar to that of the toxic dinoflagellate, but not necessarily identical, particularly after short-term (2-h) exposure, when relatively elevated levels of N-sulfocarbamoyl toxins were detected. The evidence suggests that C. finmarchicus ingests toxic dinoflagellate cells, either mistakenly or during exploratory bouts of feeding, and accumulates PSP toxins in its gut system and perhaps in other tissues.     

皮质醇影响青鳉鳃内钠钾ATP酶基因表达的研究

目前河流中存在较高浓度的天然和人造糖皮质激素(如皮质醇等)污染,其可能对鱼类的渗透压调节和海河洄游产生影响.鱼类鳃内钠钾ATP酶对调节渗透压平衡、适应海河洄游等起关键性作用,有研究表明鱼类鳃内钠钾ATP酶基因表达受皮质醇调控.为此,论文克隆了青鳉钠钾ATP酶α和钠钾ATP酶β基因的序列片段,建立了实时定量RT-PCR测定方法,研究了盐度的增加对其基因表达的影响,并重点探讨了皮质醇对鱼类适应盐度时鳃内钠钾ATP酶基因表达变化的影响.结果表明,适应15‰盐水的青鳉鳃内钠钾ATP酶α和钠钾ATP酶β基因表达显著升高;在转入淡水48h后,钠钾ATP酶α和钠钾ATP酶β基因表达基本恢复到正常水平,但是转入含有100ng·L-1皮质醇暴露水平的淡水中,48h后钠钾ATP酶α基因表达仍处于较高水平,表明100ng·L-1浓度的皮质醇能够显著诱导鳃内钠钾ATP酶α的基因表达,可能干扰渗透压调节,对一些洄游鱼类的溯河过程可能构成潜在影响.     

Alongshore transport of a toxic phytoplankton bloom in a buoyancy current: Alexandrium tamarense in the Gulf of Maine

We examined the mechanisms controlling blooms of the toxic dinoflagellate Alexandrium tamarense Lebour and the concomintant patterns of shellfish toxicity in the southwestern Gulf of Maine, USA. During a series of cruises from 1987 to 1989, hydrographic parameters were measured to elucidate the physical factors affecting the distribution and abundance of dinoflagellates along this coast. In 1988 and 1989 when toxicity was detected in the southern part of this region, A. tamarense cells were apparently transported into the area between Portsmouth, New Hampshire, and Cape Ann, Massachusetts, in a coastally trapped buoyant plume. This plume appears to have been formed by the outflow from the Androscoggin and Kennebec Rivers. Flow rates of these rivers, hydrographic sections, and satellite images led us to conclude that the plume persisted for about a month, and extended alongshore for several hundred kilometers. The distribution of cells followed the position of the plume as it was influenced by wind and topography. When winds were downwelling-favorable (to the southwest), cells were moved alongshore to the south, and were held to the coast; when winds were upwelling-favorable (to the northeast),the plume sometimes separated from the coast, advecting the cells offshore. In 1987 when no plume was present, A. tamarense cells were scarce, and no toxicity was recorded at the southern stations. The alongshore advection of toxic cells within a coastally trapped buoyant plume can explain the details of the temporal and spatial patterns of shellfish toxicity along the coast. We hypothesize that (1) the source of the A. tamarense populations is in the north, possibly associated with the Androscoggin and Kennebec estuaries, that (2) toxicity patterns follow a predictable relationship with river flow volume and timing of flow peaks and that (3) wind stresses directly influence the distribution of low salinity water and the dinoflagellate cells. Local, in situ growth of dinoflagellates can be an important factor initiating toxic dinoflagellate blooms. However, these data demonstrate the significant role of alongshore transport of established populations of A. tamarense in controlling the location and timing of paralytic shellfish poisoning (PSP) outbreaks in May and June along the southwestern coast of the Gulf of Maine.     

乙烯消减盐抑制苜蓿种子萌发的机理研究

0．25mol/L的NaCl胁迫下的苜蓿种子转变1－氨基环丙烷－1－羧酸（ACC）和乙烯利为乙烯的能力明显降低。加入外源乙烯能引发受NaCl胁迫48h的苜蓿种子迅速萌发,但对种子内的Na^ 含量无明显影响。在种子吸涨过程中,外源乙烯对受NaCl胁迫的种子的水分吸收也无影响。外源乙烯能显著地提高NaCl胁迫下种子的呼吸速率和ATP含量。对乙烯消减盐抑制苜蓿种子萌发的作用机理进行了讨论。图6表1参21     

Epidemiologic Analysis of Factors Associated with Local Disappearances of Native Ranid Frogs in Arizona

Abstract:  We examined factors that may independently or synergistically contribute to amphibian population declines. We used epidemiologic case–control methodology to sample and analyze a large database developed and maintained by the Arizona Game and Fish Department that describes historical and currently known ranid frog localities in Arizona, U.S.A. Sites with historical documentation of target ranid species ( n = 324) were evaluated to identify locations where frogs had disappeared during the study period (case sites) and locations where frog populations persisted (control sites). Between 1986 and 2003, 117 (36%) of the 324 sites became case sites, of which 105 were used in the analyses. An equal number of control sites were sampled to control for the effects of time. Risk factors, or predictor variables, were defined from environmental data summarized during site surveys and geographic information system data layers. We evaluated risk factors with univariate and multifactorial logistic-regression analyses to derive odds ratios (OR). Odds for local population disappearance were significantly related to 4 factors in the multifactorial model. Disappearance of frog populations increased with increasing elevation (OR = 2.7 for every 500 m, p < 0.01). Sites where disappearances occurred were 4.3 times more likely to have other nearby sites that also experienced disappearances (OR = 4.3, p < 0.01), whereas the odds of disappearance were 6.7 times less (OR = 0.15, p < 0.01) when there was a source population nearby. Sites with disappearances were 2.6 times more likely to have introduced crayfish than were control sites (OR = 2.6, p = 0.04). The identification of factors associated with frog disappearances increases understanding of declines occurring in natural populations and aids in conservation efforts to reestablish and protect native ranids by identifying and prioritizing implicated threats.     

东海近岸海域不同贝类体内挥发性酚残留的时空变化及其污染分析

2006年6月和2007年6月分别在东海近岸海域采集了4种贝类,对不同贝类体内挥发性酚残留量进行了检测。结果表明：2006年6月调查海域4种贝类体内挥发性酚残留量范围为0.25-0.59mg·kg^-1,褶牡蛎（Crassostrea plicatula）体内残留量最高;2007年6月残留量范围为0.80-1.50mg·kg^-1,紫贻贝（Mytilus edulis）体内残留量最高。在2次采样间4种贝类体内挥发性酚残留量的平均值表现为四角蛤蜊（Mactra veneriformis）〈文蛤（Meretrix meretrix）〈褶牡蛎〈紫贻贝。2006年6月调查海域各站点贝类体内挥发性酚残留量范围为ND-0.78mg·kg^-1,平均值为0.31mg·kg^-1;2007年6月残留量范围为0.20-2.22mg·kg^-1,平均值为1.19mg·kg^-1。2次采样间贝类体内挥发性酚在调查海域的分布格局为东海中部〉东海南部〉东海北部。贝类体内挥发性酚残留量主要受自身与外界环境因素的综合作用。     

Effects of a phycotoxin,okadaic acid,on oyster heart cell survival

Okadaic acid (OA) is a dinoflagellate toxin which accumulates in shellfish producing diarrhetic shellfish poisoning (DSP) in humans. It was found that OA is a highly selective inhibitor of protein phosphatase types 1 (PP1) and 2A (PP2A) which produces a marked increase in phosphorylation of several proteins, including p38 mitogen-activated protein (MAP) kinase. The cytotoxicity attributed to OA and the effects on p38 MAP kinase and calcium current were examined in the oyster Crassostrea gigas in this study. Data showed that p38 MAP kinase is strongly expressed in oyster heart and that OA bioaccumulated in cultured heart cells. Hence the effects of OA was tested in vitro and in vivo on oysters. OA was found to (i) exert a positive chronotropic effect on cultured atrial cardiomyocytes which is related to an increase in calcium current via PKC as shown by patch clamp measurements, (ii) produce an activation/phosphorylation of MAP kinase as shown by Westernblot while the non-phosphorylated p38 remained constant during treatment, (iii) did not induce a pro-apoptotic effect. Data suggest that OA may also stimulate the anti-apoptotic pathway by phosphatase inhibition.     

A study on major inorganic ion composition of atmospheric aerosols

Atmospheric aerosol samples were collected from Akola and Buldana region covering around 40 sqkm area during October-November 2002 and were analyzed for ten major inorganic ions namely F-, Cl-, NO3-, SO4(2-), PO4(2-), Na+, K+, Ca2+, Mg2+ and NH4+ using ion chromatographic technique. The average mass of aerosols was found to be 225.81 microg/m3 with standard deviation of 31.29 and average total water soluble load of total cations and anions was found to be 4.32 microg/m3. The concentration of ions in samples showed a general pattern as SO4(2-) > NO3- > Cl- > PO4(2-) > F- for anions and Na+ > Ca2+ > NH4+ > Mg2+ > K+ for cations. The overall composition of the aerosols was taken into account to identify the sources. The trend showed higher concentration of sodium followed by calcium, sulfate, nitrate, phosphate and ammoinum and found to be influenced by terrestrial sources. The presence of SO4(2-) and NO3- in aerosols may be due to re-suspension of soil particles. Ca2+, Mg2+ and Cl- are to be derived from soil materials. The presence of NH4+ may be attributed to the reaction of NH3 vapors with acidic gases may react or condense on an acidic particle surface of anthropogenic origin. The atmospheric aerosol is slightly acidic due to neutralization of basicity by SO2 and NO(x).