Growth inhibition and DNA damage in the earthworm (Eisenia fetida) exposed to perfluorooctane sulphonate and perfluorooctanoic acid

An artificial soil method was applied to study the effects of perfluorooctane sulphonate (PFOS) and perfluorooctanoic acid (PFOA) on earthworms (Eisenia fetida). Survival, growth inhibition and damage to DNA of earthworms were detected after 14 d acute exposure. The 14 d-LC₅₀ of PFOS and PFOA was 478.0?mg·kg^?1 dw and 759.6?mg·kg^?1 dw, respectively, indicating that they were of low toxicity. Both PFOS and PFOA could significantly inhibit the growth of earthworms after 14 d exposure, and growth inhibition rates increased with the greater concentrations of PFOS or PFOA, showing a dose–response relationship (PFOS: r?=?0.951, P r?=?0.962, P?P?50 of PFOS was lower than that of PFOA, the growth inhibition rate of earthworm exposed to PFOS was higher than that exposed to PFOA at the same concentration level, and the median values of TL, CL and OTM in PFOS treatments were also higher than those in PFOA treatments. In conclusion, both these fluorine compounds were moderately toxic to earthworms, but the PFOS effect was greater than that of PFOA.     

Cr(VI)对脊尾白虾(Exopalaemon carinicauda)幼虾暴露和恢复期肝胰脏的SOD活性、MDA及MTs含量的影响

为了探索水体中Cr(VI)对脊尾白虾(Exopalaemon carinicauda)的毒性效应,通过Cr(VI)的暴露(15 d)和清水恢复实验(15 d),研究了Cr(VI)对脊尾白虾肝胰脏的SOD活性、MDA及MTs含量的影响。结果表明,通过15 d的暴露,5 mg·L-1(高浓度)、0.5 mg·L-1(低浓度)实验组肝胰脏的SOD均显著高于对照组水平(p0.05),15 d的清水恢复后,两个浓度实验组均能够恢复到对照组水平(p0.05)。MDA在暴露阶段呈现持续上升的趋势,且在15 d后两个浓度实验组MDA的含量均显著高于对照组水平(p0.05),15 d的清水恢复后,5 mg·L-1实验组的MDA含量依然显著高于对照组水平(p0.05),0.5mg·L-1实验组的MDA含量基本恢复到对照组水平(p0.05)。通过15 d的暴露,两个浓度实验组MTs的含量均显著高于对照组水平(p0.05),15 d的清水恢复后,两个浓度实验组MTs的含量均能够恢复到对照组水平(p0.05)。     

广州市某A2/O工艺城镇污水处理厂出水对大型溞的生殖毒性作用

城镇污水处理厂废水中存在大量生物毒性物质,但该废水对动物的生殖能力影响研究还十分缺乏。利用大型溞暴露实验,检测广州市某采用A~2/O工艺的城镇污水处理厂出水的急性毒性和生殖毒性。48 h急性毒性实验表明,所有水样均未表现出急性毒性效应。慢性生殖毒性(14 d)测试结果表明:(1)从进水到缺氧池出水均能显著提高第一胎产溞数量;(2)进水和沉砂池出水显著增加受试期总产溞数量;(3)从厌氧池到出水工艺段受试溞的第一胎产溞时间均推后;(4)好氧池出水的毒性显著降低,但仍然对大型溞具有生殖毒性。研究表明,好氧池(A~2/O)工艺能够显著去除具有大型溞生殖毒性的物质,但污水处理厂废水的生殖毒性仍需要引起关注。     

Evidences showing 2,4-D ethyl ester and pencycuron-induced DNA damage in cyanobacteria and detection by PCR

The impact of 2,4-D ethyl ester and pencycuron in inducing DNA damage in three species of cyanobacteria-Anabaena fertilissima, Aulosira fertilissima, and Westiellopsis prolifica as evidenced by PCR-based assays: RAPD and 16S rRNA amplification was examined. Exposure of genomic DNA (in vitro) to pencycuron for 4 days did not produce severe damage in DNA fragments of all three cyanobacterial species whereas exposure to 2,4-D ethyl ester markedly inhibited the template activity of genomic DNA compared to untreated cultures of A. fertilissima. In A. fertilissima a single band of approximately 1000?bp was observed even after 16 days of exposure to 60?ppm pencycuron which suggests that certain segments of DNA are resistant to pencycuron DNA damaging effects. However, a significant effect was observed in the case of W. prolifica for 2,4-D ethyl ester and pencycuron where complete disappearance of fragments was not recorded even after 16 days of incubation and interestingly some new DNA bands were induced. Similar to the effects with RAPD profile, amplification of rRNA was significantly inhibited following exposure of genomic DNA to 2,4-D ethyl ester and pencycuron. Our findings clearly demonstrate that pesticide concentrations affected cyanobacterial DNA and lethality of these microbes might be due to irreversible DNA damage. Thus, it is postulated that PCR assays may be conveniently used for screening DNA damage produced by 2,4-D ethyl ester and pencycuron in all three cyanobacteria examined in this study.     

磺胺甲恶唑暴露对小麦叶片蛋白和叶绿素含量及SOD酶活性的影响

为了探明磺胺甲恶唑(sulfamethoxazole,SMZ)的生态毒理效应,通过实验室人工控制毒理实验,研究了低浓度SMZ暴露对小麦体内超氧化物歧化酶(SOD)活性、叶绿素(CHL)和蛋白质(SP)含量的影响。结果显示:染毒7d后,各浓度组小麦叶片的SOD活性均被显著诱导(P<0.01),并且染毒浓度的升高增强了SOD的活性,表明SMZ暴露胁迫下,小麦可启动自身的保护机制以最大限度地减少自由基损伤。而小麦叶片的CHL含量,随着SMZ染毒浓度的增加而逐渐降低。并且当SMZ暴露浓度较高时,小麦叶片的CHL含量被显著抑制(P<0.05))。当SMZ染毒浓度为0.05～0.50mg·L-1时,SP的含量被显著诱导(P<0.01));1.00mg·L-1SMZ对SP的含量产生显著(P<0.01)抑制,这说明SMZ染毒剂量将对蛋白质的合成产生严重影响。综上,SOD的活性变化可反映出SMZ暴露对小麦的污染效应及其生态毒性作用,但将其作为评估SMZ污染暴露的生物标志物有待于进一步深入研究。     

汞和砷单一及联合暴露对日本沼虾的氧化应激效应

为深入探讨汞(Hg)和砷(As)对水生生物的慢性毒性效应,以日本沼虾(Macrobrachium nipponense)为受试生物,以超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、金属硫蛋白(MT)和丙二醛(MDA)为测试分子标志物,研究上覆水中不同浓度Hg和As单一及联合暴露对日本沼虾肝胰腺的氧化损伤及联合作用机制。结果表明:Hg和As单一及联合暴露对4种分子标志物均可产生不同程度的抑制或激活效应。其中,SOD活性均受到抑制,且高浓度As(1000μg·L-1)暴露10 d的抑制率最大,高达59.13%;而CAT活性整体被激活,Hg单独暴露下的激活效应要高于As,且在联合暴露组中随着暴露时间的增加呈现出显著的剂量效应关系(P0.05);MT和MDA含量在Hg和As的暴露下均显著增加(P0.05)。其中MT对Hg的螯合能力大于As,As单独暴露3 d后MDA变化较小,但10 d后其含量显著增加(P0.05)。析因方差分析发现,Hg-As对机体的联合作用机制主要表现为协同作用。综合生物标志物响应指数(IBR)评价证实,联合毒性要高于单独毒性,Hg与As在高浓度下联合暴露10 d的IBR值最大,毒性最强,高浓度As单独暴露处理组的毒性次之;同时,机体自身随暴露时间延长对Hg也表现出一定的解毒功能。     

Influence of habitat on the reproductive biology of the deep-sea hydrothermal vent limpet <Emphasis Type="Italic">Lepetodrilus fucensis</Emphasis> (Vetigastropoda: Mollusca) from the Northeast Pacific

Habitat selection by the hydrothermal vent limpet, Lepetodrilus fucensis, in Northeast Pacific hydrothermal vent ecosystems, may influence its reproductive output, as it occupies habitats with varying physico-chemical conditions that reflect the availability of nutritional resources. Histological techniques were used to determine size at first reproduction, gametogenesis, reproductive output, and fecundity in relation to shell length (SL), through examination of the gonads of male and female L. fucensis, collected from five different hydrothermal vent habitat types with different temperature anomalies and hydrothermal fluid flow vigour: vigorous (VIG), diffuse (DIF), tubeworm bushes (TWB), peripheral (PER), and senescent areas (SEN). Both male and female L. fucensis exhibited early maturity, with the first reproductive event occurring at 3.8 and 3.9 mm shell length, respectively. All stages of gamete development were present in the gonads of males and females, suggesting continuous gametogenesis and asynchronous reproduction in this species. Gametogenic maturity of limpets did not vary among actively venting habitats (VIG, DIF, TWB, and PER), but was significantly lower in males and females from SEN habitats. Mean oocyte diameter was largest in females from VIG habitats, and smallest in females from SEN habitats, than in those from the other habitats (DIF, TWB, and PER). Females from actively venting habitats also had greater actual fecundity than those from senescent habitats. While the gametogenic pattern of L. fucensis appears phylogenetically constrained, selection of actively venting habitats by L. fucensis maximizes its reproductive output. The multiple feeding strategies of L. fucensis may allow for a constant supply of energy to be allocated to reproduction in any habitat except senescent vents. Early maturity, high fecundity, and continuous production of gametes suggest a reproductive strategy characteristic of an opportunistic species, and may be contributing to the extremely abundant populations of L. fucensis observed in the Northeast Pacific vent ecosystem.     

Release of photoassimilated carbon as dissolved organic matter by marine phytoplankton

The release of photoassimilated carbon as dissolved organic matter was studied in situ in oligotrophic and eutrophic marine waters and in axenic laboratory cultures. Percentage extracellular release (PER), integrated for the trophogenic zone, ranged from 6 to 12% in eutrophic waters and from 17 to 27% in oligotrophic seas. Most of the algal cultures released low amounts of dissolved organic matter (5%) during exponential growth. The highest levels of PER were observed in surface and deep samples, possibly as a result of elevated photorespiration and senescent cells. The generally lower values for extracellular release reported in this work as compared to other studies may be partly due to improved experimental techniques which minimized previously encountered artifacts.     

Effects of UVB radiation in a microbenthic community of a marine shallow-water sandy sediment

The role of ambient and enhanced ultraviolet-B radiation (UVB; 280 to 315 nm) in a natural sand-associated microbenthic community was studied in a 3-week experiment by incubating intact sediment cores from a shallow bay in an outdoor flow-through system with 27 aquaria. After sampling of initial cores, the remaining cores (one per aquarium) were given one of three treatments: no, ambient, and moderately enhanced UVB, and sampled, nine at a time, after 5, 12, and 19 d. The response of the community was studied by analysing algal and meiofaunal composition and biomass, chlorophyll a content, composition of pigments and fatty acids, and content of UV-absorbing compounds (state variables), as well as carbon fixation and allocation, and bacterial productivity (rate variables). Among rate variables, significant effects of UVB-treatments were found for carbon fixation and allocation, while bacterial productivity was not affected. For state variables, a significant response was observed for the composition of microalgae and fatty acids, and for chlorophyll a content. The effect of treatments was mainly observed as differences in development with time (two-way analysis of variance, treatment × time interaction). Towards the end of the experiment, the no-UVB treatment most often differed from one or both of the two treatments with UVB exposure, the latter showing lower values. There were marked successional changes in the community, irrespective of treatment. The microalgal community changed from being dominated by coccoid cyanobacteria and epipsammic diatoms to a dominance of epipelic diatoms and filamentous cyanobacteria. The pattern of carbon allocation, as well as an increased C/N ratio of the sediment, suggested limitation of growth, perhaps by nutrients, at the end of the experiment. This may possibly have acted synergistically with UVB exposure to create the treatment effects. The new knowledge gained from our experiment is that ambient UVB can exert a stress on the function of sand-associated microbenthic communities in shallow waters and that this effect coincides with structural differences in the community. More experiments in natural or semi-natural systems are needed to allow better prediction of microbenthic community-level responses to UVB. Received: 11 November 1997 / Accepted: 12 June 1998     

A pilot study on the effect of Cu,Zn, and Cd on the spectral curves and chlorophyll of wheat canopy at tiller stage

The aim of this study was to investigate canopy spectral reflectance responses to different levels of heavy metals copper (Cu), zinc (Zn), and cadmium (Cd) induced stress. Random blocks design experiment was conducted to simulate Cu, Zn, and Cd at five concentration levels. Chlorophyll and visible and near infrared canopy reflectance were measured for each treatment 56 days after seeding using chlorophyll meter SPAD-502 and spectroradiometer, respectively. New vegetative indices termed ratio of inclination angles (RIA), MERIS (the medium resolution imaging spectrometer) terrestrial chlorophyll index (MTCI), and normalized difference vegetation index (NDVI) were used to assess the response of canopy spectral reflectance to different heavy metal levels. Significant spectral variability especially in blue, red, and near infrared reflectance was observed for different heavy metal treatments. One-sample test showed significant difference for NDVI, MTCI, and RIA among five-level treatments at the 0.01 level. Correlation analysis demonstrated that the two vegetative indices (RIA and MTCI) were significantly correlated with chlorophyll meter values for Cd, Cu, and Zn treatments. However, NDVI was only significantly correlated with chlorophyll meter values for Cu and Zn treatments. Linear regression analysis also illustrated that RIA and MTCI were potential indices for predicting chlorophyll concentration with significant F test under Cu-, Zn-, and Cd-mediated stress.     

全氟辛烷磺酸盐(PFOS)对半滑舌鳎(Cynoglossus semilaevis)免疫相关基因表达的影响

为了解全氟辛烷磺酸盐(perfluorooctane sulfonate, PFOS)暴露对半滑舌鳎(Cynoglossus semilaevis)免疫功能的影响,在实验室条件下,运用RT-PCR方法分析了PFOS暴露对半滑舌鳎热休克蛋白hsp70、hsp90、C型凝集素(c-type lectin)和细胞色素c氧化酶(cytochrome c oxidase, cox)等4种免疫相关基因表达水平的影响。实验测定了上述4种基因在半滑舌鳎肝、鳃、肠及肌肉4种不同组织中随时间(0、24 h、48 h、96 h和7 d)的表达变化情况。结果表明,在4种组织中,hsp70基因的表达与对照相比为上调,其中,肝组织hsp70基因的表达量显著高于其他各组织,且表达高峰值的出现也早于其他各组织;hsp90基因在肝和鳃组织中表达量随时间不同而波动,在肠组织中表达上调,在肌肉中表达显著下调;c-type lectin基因表达量与对照组相比表达显著下调或无明显差异;cox基因在肝组织和肠组织中表达下调,在鳃和肌肉中表达上调。上述研究结果表明,PFOS能引起免疫相关基因的表达变化,对半滑舌鳎具有潜在的免疫毒性。肝组织中各免疫基因对PFOS胁迫的响应高于其他组织。本研究可为阐明全氟辛烷磺酸盐对半滑舌鳎的免疫毒性提供基础数据。     

Performance evaluation of isoproturon-degrading indigenous bacterial isolates in soil microcosm

Isoproturon (IPU)-degrading soil bacteria were isolated from herbicide-applied wheat fields. These isolates were identified using cultural, morphological, biochemical and 16S rRNA sequencing methods. 16S rRNA sequences of both the bacterial isolates were compared with NCBI GenBank data base and identified as Bacillus pumilus and Pseudoxanthomonas sp. A soil microcosm study was carried out for 40 days in six different treatments. Experimental results revealed maximum 95.98% IPU degradation in treatment 6 where bacterial consortia were augmented in natural soil, followed by 91.53% in treatment 5 enriched with organic manure as an additional carbon source. However, only 14.03% IPU was degraded in treatment 1 (control) after 40 days. In treatments (2–4), 75.59%, 70.92% and 77.32% IPU degradation was recorded, respectively. IPU degradation in all the treatments varied significantly over the control. 4-Isopropylaniline was detected as IPU degradation by-product in the medium. The study confirmed that B. pumilus and Pseudoxanthomonas sp. performed effectively in soil microcosms and could be employed profitably for field-scale bioremediation experiments.     

Effects of ultraviolet radiation on different life cycle stages of the south Pacific kelps, Lessonia nigrescens and Lessonia trabeculata (Laminariales, Phaeophyceae)

The effects of exposure to ultraviolet radiation (UVR), 280–400 nm, in different life histories and development stages of the kelps, Lessonia nigrescens and L. trabeculata, collected in the south-east Pacific coast (30°S) were evaluated in the laboratory. Germination and viability (motile zoospores, settled spores), diameter of the primary cell of the gametophytes, percentage of female gametophytes, fertility and sporophytes production were measured after exposure to three radiation treatments (PAR; PAR + UVA; PAR + UVA + UVB). The effects of UVR in young sporophytes (diploid stage) were evaluated as changes in maximal quantum yield of chlorophyll fluorescence of photosystem II (PSII) (F _v/F _m). A significant decrease in all variables was observed for the treatment that included UVB (PAR + UVA + UVB) after 2 and 4 h of exposure, in relation to the control. The motile spores were more sensitive to UVR exposure compared to settled spores and gametophytes, suggesting that along with an increase in ontogenetic development; there is an increase in the tolerance to UVR. In addition, it was observed that early stages of the intertidal L. nigrescens were more tolerant to UVR compared to the subtidal L. trabeculata. These results allow initially to infer that UVR may be regarded as an important environmental factor influencing the upper limit of distribution of these species, mainly through its detrimental effects on the early stages of the life cycle.     

Mercuric chloride induced proteotoxicity and structural destabilization in marine fish (Therapon jarbua)

The reaction byproducts derived from lipid peroxidation (LPO), as well as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) enzyme activities were measured in tissues of marine fish (Therapon jarbua) exposed to mercuric chloride (HgCl) in water dispersions of 0.125 or 0.25 ppm. LPO was significantly increased in various tissues relative to control values after 96-h exposure. SOD and GPx activities significantly decreased after exposure to first two doses but significantly elevated CAT in Dose II kidney and liver tissues. The elevated levels of LPO, decreased activities of SOD and GPx, and increased CAT activities in all tissues examined in T. jarbua are an index of oxidative stress in fish. Structural analysis by scanning electron microscopy studies revealed the structural deformation in HgCl₂-exposed animals. These observations suggest that HgCl acts as a mediator in free radical generation. The increase in CAT and decrease in SOD and GPx activities in these tissues may be an adaptive response.     

镉对单环刺螠非特异性免疫及组织蓄积的影响

为探究镉(Cd)对海洋动物的毒性效应,考察了不同质量浓度(0.005、0.05和0.5 mg·L~(-1)) Cd处理对单环刺螠相关免疫生理指标及主要组织中Cd蓄积量的影响。结果表明:Cd胁迫下,血细胞密度(density of hemocyte, DHC)先升后降再升,酚氧化酶(phenoloxidase, PO)活力先升后降,溶菌酶(lysozyme, LSZ)活力先降后升再降,至处理96 h,处理组DHC均显著高于对照组(P0.05),总蛋白含量变化不显著;除低浓度胁迫组(0.005 mg·L~(-1))PO活力与对照组相近外,其他处理组的PO和LSZ活力均显著低于对照组(P0.05)。超氧化物歧化酶(superoxide dismutase, SOD)活力和丙二醛(malondialdehyde, MDA)含量总体呈上升趋势,过氧化氢(H_2O_2)含量先升后降再升,至胁迫96 h,各处理组SOD活力、MDA和H_2O_2含量均显著高于对照组(P0.05)。总之,Cd胁迫对单环刺螠非特异性免疫影响显著,Cd的短期胁迫可刺激DHC、PO和LSZ升高,免疫力增强,而高浓度Cd的胁迫时间较长会造成机体解毒能力下降,持续刺激机体产生过氧化反应,MDA含量升高,可作为单环刺螠在Cd长期胁迫下的免疫检测指标。不同胁迫浓度组单环刺螠肌肉和消化道中的Cd含量随胁迫时间均呈上升趋势,且存在显著的时间-剂量-效应关系(P0.05),其蓄积量水平为:消化道肌肉,具有组织特异性。     

GO通过氧化损伤激活秀丽线虫未折叠蛋白反应

以秀丽线虫为受试生物,探讨氧化石墨烯(GO)对秀丽线虫未折叠蛋白应答的激活及其与氧化应激的关系。将L4幼虫暴露于GO中24 h,通过检测活性氧(ROS)水平、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等指标评估GO对秀丽线虫所致氧化损伤;以hsp-4和hsp-6分别作为秀丽线虫内质网和线粒体未折叠蛋白反应(UPR)的报告基因评价GO对秀丽线虫UPR应答的激活;同时以谷胱甘肽(GSH)作为抗氧化剂探讨GO所致UPR应答与氧化应激的关系。研究结果显示,GO暴露后秀丽线虫体内ROS、LDH和MDA水平升高,抗氧化酶活性上升;内质网和线粒体UPR反应增强,GSH预处理可以降低内质网和线粒体UPR应答。研究表明,GO短期暴露可以诱导秀丽线虫机体氧化应激进而激活UPR应答,抗氧化保护可以降低UPR应答反应。     

Effect of nutrient enrichment on the complementary (secondary) metabolite composition of the soft coral Sarcophyton ehrenbergi (Cnidaria: Octocorallia: Alcyonaceae) of the Great Barrier Reef

 A long-term study of the effects of nutrient enrichment on coral reefs (ENCORE Experiment) was carried out at One Tree Island (23°30′S; 152°96′E), Great Barrier Reef, between 1992 and 1996. The experiment involved the addition of water-soluble nutrients to 12 microatolls which contained a range of organisms and were situated within the larger lagoon. Three replicates of each of three nutrient treatments (nitrogen, phosphorus, nitrogen plus phosphorus) and an untreated set of three control atolls were prepared using the 12 selected microatolls. As part of the larger ENCORE experiment, changes in the chemical composition of the alcyonacean soft coral Sarcophyton ehrenbergi Marenzeller (Octocorallia: Alcyonacea), placed in the treatment microatolls, were monitored for a 1 yr period in an attempt to detect any responses attributable to nutrient enrichment. Multivariate analyses were performed to determine whether there were any patterns of response in the different nutrient treatments. At the level of individual metabolites, there were no clear treatment effects. However, the ratio of bioactive or stress metabolites (terpenes) to energy storage metabolites (lipids), referred to as the “physiological-change indicator”, revealed effects of nutrient enrichment. Nitrogen enrichment resulted in a trend towards higher physiological-change indicators than control or phosphorus treatments in the majority of cases, while phosphorus enrichment significantly decreased the ratio relative to controls. In most cases, the physiological-change indicator increased in soft soft corals relocated into contact with the scleractinian Pocillopora damicornis. The potential of soft corals to serve as indicators of a changed nutrient regime is discussed. Received: 23 November 1998 / Accepted: 23 September 1999     

氯代阻燃剂得克隆对孔石莼(Ulva pertusa)繁殖及早期发育的影响

得克隆(Dechlorane Plus,DP)是一种在全世界范围内广泛使用的氯代阻燃剂,具有潜在的毒性效应。但目前已有的生态毒理学数据还十分有限。本文选择分布广泛的大型绿藻孔石莼(Ulva pertusa)作为研究对象,探讨不同浓度DP对孔石莼繁殖细胞转化及早期发育的影响。结果表明,低浓度(10-6~10-8mol·L-1)DP暴露不同程度地影响孔石莼繁殖细胞的形成、释放及附着,在试验浓度范围内呈现一定的剂量效应关系。与空白对照组相比,DP处理(10-6mol·L-1)分别使繁殖细胞的形成率、释放率和附着率显著降低了76.76%、46.26%和85.64%。在丝状幼体阶段,DP处理组(10-8和10-7mol·L-1)幼体体长分别比对照组显著减少了32.74%和38.98%。结果表明,DP暴露抑制了孔石莼的繁殖及早期发育。基于绿藻繁殖特性及早期发育的生物学指标对DP暴露较为敏感,能够为海洋环境的DP生物学效应研究提供数据。     

Effects of petroleum products on bull kelp (Nereocystis luetkeana)

Although research has been conducted on the effects of oil on the giant kelp Macrocystis pyrifera, no similar studies have been completed on bull kelp, Nereocystis leutkeana, the dominant kelp in Washington State, British Columbia, and Alaska. The effects of three petroleum products [diesel fuel, intermediate fuel oil (IFO), and crude oil] were tested before and after weathering on N. luetkeana. Whole plants were exposed to petroleum product for 4 or 24 h and then transferred to the field; observations on the condition of the plants were made daily for 7 d. In addition, controlled bioassays were performed to measure the effects of petroleum exposure on net photosynthetic rate (NP) and respiration rate (R), using light-and dark-bottle techniques. These experiments verified the susceptibility of N. luetkeana tissue to the damaging effects of direct exposure to several oil types. The 4 h exposures to weathered diesel and unweathered IFO, and 24 h exposures to unweathered and weathered diesel and IFO resulted in moderate to severe damage to kelp tissue (i.e., clearly delineated bleached line accompanied by tissue necrosis). Weathered diesel was more toxic than unweathered diesel. The most severe damage to bull kelp was concentrated at the meristematic zone (junction of stipe and bulb) where new tissue growth occurs. Petroleum type significantly affected stipe and blade NP, R, and NP:R ratios. Diesel treatments had a greater negative effect on NP than did the IFO treatments. Based on these experiments, the relative ranking of the damaging effects of petroleum treatment on bull kelp are weathered diesel>unweathered IFO>unweathered diesel>weathered IFO>unweathered crude>weathered crude.     

Larval settlement and metamorphosis of the mussel <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> in response to biofilms

Biofilms were allowed to develop on glass slips immersed 1.0–1.5 m below the sea surface in Tachibana Bay, Nagasaki, Japan, for different periods of time from November 2003 to January 2005. The effects of age, immersion month, dry weight, bacterial and diatom densities of these biofilms on the settlement and metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis were investigated in the laboratory. Furthermore, biofilms were subjected to various treatments to investigate the nature of the settlement and metamorphosis cue in the biofilm. Pediveliger larvae of the mussel settled and metamorphosed in response to biofilms. Settlement and metamorphosis to the post-larval stage significantly increased with the biofilm age. In addition, the biofilm activity varied depending on the immersion month (season), e.g., for biofilms with the same age, those immersed between June and August had higher activities than those immersed between November and March. The activity of the biofilm also positively correlated with the dry weight, bacterial and diatom densities. These three quantitative parameters of the biofilm were significantly affected by the film age but were not affected by the immersion month, suggesting that other parameters (e.g., community structures, extracellular products) also affected the inductive activity of the biofilm. The fixative agents (formalin and glutaraldehyde), heat, ethanol, ultraviolet irradiation and antibiotics treatments of the biofilm resulted in significant reduction or loss of its inductive activity. The survival of bacterial cells in the treated films where activities were either reduced or lost also decreased significantly. No settlement and metamorphosis were obtained when larvae were exposed to the conditioned water of the biofilm. Thus, larvae of M. galloprovincialis settled and metamorphosed in response to a cue produced by living bacteria in the biofilm. The cue may be a bacterial extracellular product which was susceptible to the above treatments.