Evaluation of Gambierdiscus survival after exposure to ballast water

The dinoflagellate Gambierdiscus was exposed to ballast water from a trans-oceanic vessel, and maintained at a variety of temperatures in the dark to determine if viability would be maintained. Logarithmically growing Gambierdiscus inocula were admixed (1:6, vol:vol) with ballast water, maintained in the dark at 22.6 °C, 24.6 °C, 26.8 °C and 29.0 °C and assessed for numerical abundance over six days. Calculated growth rates from the biomass time series showed no indication that ballast water negatively impacted Gambierdiscus viability; accompanying microscopic inspections supported this conclusion. Filtration of large volumes of collected ballast water failed to show the presence of any Gambierdiscus cells contained therein. Recovery and microscopic examination of the experimental inocula after 10 weeks in the dark, failed to show cyst development at any temperature regime. This examination of ballast water showed no evidence of cytotoxicity to Gambierdiscus spp.     

An examination of the epiphytic nature of Gambierdiscus toxicus, a dinoflagellate involved in ciguatera fish poisoning

Twenty-four specimen of macroalgae were collected in nearshore waters of the island of Hawaii, identified, and maintained to examine how the epiphytic relationship between Gambierdiscus toxicus (isolate BIG12) varied among the macroalgal species. Gambierdiscus cells were introduced to Petri dishes containing 100 g samples of each macroalgal host, which were examined at two, 16, 24, and every 24–72 h thereafter, over a 29-day period. Gambierdiscus proliferated in the presence of some host species (e.g., Galaxaura marginata and Jania sp.), but grew little in the presence of other species (e.g., Portieria hornemannii). Gambierdiscus exhibited high survival rates (>99%) in the presence of Chaetomorpha sp., but died before the end of the experiment (after 21 days) with other host species (e.g., Dictyota and Microdictyon spp.). Gambierdiscus avoided contact with P. hornemannii, but averaged up to 30% attachment with other host species. The numbers of Gambierdiscus cells belonging to one of three classes (alive and attached; alive and unattached; and dead) were determined for each time point. The 24 algal hosts were grouped according to their commonalities relative to these three classes using a Bray-Curtis similarity index, similarity profile (SIMPROF) permutation tests, and Multi-Dimensional Scaling (MDS) analysis (PRIMER 6). The resultant six groupings were used to construct different Gambierdiscus growth profiles for the different algal hosts. Group A is characterized by a preponderance of unattached cells and high mortality rates. Groups B, C, E, and F also displayed high proportions of unattached cells, but mortality either occurred later (Groups B and C) or rates were lower (Groups E and F). Group D had the highest proportion of attached cells. Group E contained three out of the four chlorophyte species, while Group F contained the majority of the rhodophytes. Over 50% of the species in Group F are considered to be palatable, whereas Groups A, B, and C are composed of species that exhibit chemical defenses against herbivory. The results of this study coupled with previous findings indicate that Gambierdiscus is not an obligate epiphyte; it can be free-swimming and found in the plankton. The conditions that lead to changes between epiphytic and planktonic stages need to be better studied in order to determine how they affect Gambierdiscus growth and physiology, connectivity and dispersion mechanisms, and toxin movement up into the foodweb.     

Ciguatera fish poisoning: Incidence,health costs and risk perception on Moorea Island (Society archipelago,French Polynesia)

Ciguatera Fish Poisoning (CFP) is a non-bacterial seafood poisoning well characterized in the remote archipelagos of French Polynesia, yet poorly documented in the Society archipelago, most notably on Moorea, the second most populated island in French Polynesia, which counts a high proportion of fishermen fishing on a regular basis. To address this knowledge gap, a holistic study of the ciguatera issue was conducted on Moorea. First, ciguatera risk was analysed in terms of incidence rate, fish species most commonly involved and risk stratification in Moorea lagoon based on 2007–2013 epidemiological data. A mean incidence rate of 8 cases per 10,000 inhabitants for the study period and an average under-reporting rate of 54% were found. Taking into account hospitalization and medication fees, and loss of productive days, the health-related costs due to CFP were estimated to be USD $1613 and $749 for each reported and unreported case, respectively, with an overall cost of USD $241,847 for the study period. Comparison of the present status of CFP on Moorea with a risk map established in the late 1970’s showed that the spatial distribution of the risk has stayed relatively stable in time, with the north shore of the island remaining the most prone to ciguatera. Evaluation of the current knowledge on CFP among different populations groups, i.e. fishermen, residents and visitors, was also conducted through direct and indirect interviews. About half of the fishermen interviewed were actually able to identify risky fishing areas. While, overall, the CFP risk perception in the fishing community of Moorea seemed accurate, although not scientifically complete, it was sufficient for the safe practice of their fishing activities. This may be due in part to adaptive responses adopted by 36% of the fishermen interviewed, such as the avoidance of either high-risk fishing sites or toxic species. At the residents and visitors’ level, the study points out a striking lack of awareness of the CFP issue among visitors, as compared to local residents. Indeed, less than 25% of Moorea visitors vs. an average of 98% in residents were aware of CFP or of its presence on the island. Interestingly, evaluation of the fish consumption preferences showed that 70% of visitors do not consume lagoon fish during their stay, not for fear of CFP, but mainly due to the lack of availability of these species in recreational facilities or because they have nutritional preference for pelagic fish. This lack of awareness, along with the report by several CFP patients of the consumption of fish species yet banned for sale, stress the need for improved communication efforts on this critical issue among both residents and visitors on Moorea. The implementation of a public outreach strategy is proposed, based on both existing information networks and low-cost communication actions through information displays at various strategic locations, e.g. Tahiti-Faa’a international airport, the ferry boat station, recreational facilities, as well as the major trading points on Moorea Island.     

Sexual reproduction of Acropora reef corals at Moorea, French Polynesia

Little information is available on reproductive processes among corals in isolated central Pacific reef regions, including French Polynesia. This study examined the timing and mode of sexual reproduction for Acropora reef corals at Moorea. Spawning was observed and/or inferred in 110 Acropora colonies, representing 12 species, following full moon periods in September through November 2002. Gamete release was observed and inferred in four species of Acropora between 9 and 13 nights after the full moon (nAFM) in September 2002. Twelve Acropora spp. spawned gametes between 5 and 10 nAFM in October 2002, with six species spawning 7 nAFM and four species spawning 9 nAFM. In November 2002, spawning of egg and sperm bundles was observed and inferred in 27 colonies of Acropora austera, 6 nAFM. These are the first detailed records of spawning by Acropora corals in French Polynesia.     

节杆菌环境适应性的基因组学研究进展

节杆菌分布广泛,能适应多种环境条件,而且多数节杆菌具有营养多功能性,能降解多种环境污染物,因而受到人们的广泛关注。近年来,随着多株节杆菌基因组的测序完成,人们对节杆菌环境适应性的分子机制有了全面的认识。基因组学研究结果表明,节杆菌在σ因子、氧化应激、渗透应激、饥饿应激、温度应激等胁迫应激反应相关基因方面的特点使其能够在多种环境条件下生存。本文挑选部分具有代表性的节杆菌基因组学研究,对其环境适应性的基因组学基础进行综述,以期为利用节杆菌进行环境污染修复提供理论基础,并为其它细菌的环境适应性机制研究提供参考。     

Petroleum production platforms as sites for the expansion of ciguatera in the northwestern Gulf of Mexico

Ciguatera is a common human disease of tropical, coral reef ecosystems acquired by consuming finfish-containing ciguatoxins (CTX). There are few records of this disease in the northwestern Gulf of Mexico, a region characterized by soft muddy bottoms that are considered poor habitat for the CTX source dinoflagellate Gambierdiscus toxicus. However, the approximately 4000 petroleum production platforms and hundreds of state-sponsored artificial reefs located in the Gulf of Mexico provide hard substrate and often support coral and other components of the tropical benthos. In addition to their role in their resource extraction, these oil production platforms are also popular sites for recreational fishing and sport diving. We examined these platforms as potential substrate for G. toxicus and report a first record of this species in the NW Gulf of Mexico. All the platforms (n = 6) examined harbored the dinoflagellate as an epiphyte on the fouling community, with three finds of G. toxicus associated with the pelagic macroalga Sargassum. Only minor toxicity (<0.15 ppb) was noted in two of 20 great barracuda (Sphyraena barracuda) examined. Tagging data suggest trans-Gulf migrations by barracuda are common; thus, we cannot determine if the toxicity was acquired locally or transported in migrating fish.These platforms are a clear example of how human activity has altered the environment in a way that allows expansion of a HAB population. The rapid increase in production platforms since 1942 has provided novel substrate in a sandy/muddy bottom environment generally considered to be poor habitat for these benthic dinoflagellates. These platforms create a unique habitat in the upper euphotic zone and serve as intersection points for fishermen and potentially toxic fish. Many Gulf of Mexico states have active programs to turn non-producing platforms into artificial reefs. Our results suggest that the use of these platforms as fisheries enhancement structures could have unintended consequences for human health, particularly if projected rising sea-surface temperatures over the next century alter benthic distributions and fish migration patterns. These concerns also extend to mariculture operations around oil production rigs or offshore wind farms, both of which would also add substrate for epibenthic microalgae.     

秋石斛钙依赖蛋白激酶基因的克隆及表达分析

钙依赖蛋白激酶(calcium dependent protein kinase,CDPK)在植物的生长发育及逆境胁迫方面发挥着重要作用。该研究以秋石斛品种‘水芙蓉’为试验材料,采用 RT PCR方法克隆钙依赖蛋白激酶基因,并对其进行生物信息学分析;采用qRT PCR方法对CDPKs基因在‘水芙蓉’不同组织及不同低温胁迫下的表达情况进行分析,为秋石斛CDPK基因的功能验证以及抗寒分子机制研究奠定基础。结果表明：(1)成功克隆获得了DenCDPK1(GenBank登录号为MZ322902)、DenCDPK2(GenBank登录号为MZ322903)、DenCDPK3(GenBank登录号为MZ322904)基因,序列长度依次为1 934、1 971和2 302 bp,分别编码534、541和536个氨基酸。 (2)序列一致性结果显示,DenCDPKs蛋白质与铁皮石斛相应CDPK蛋白质序列的一致性均高于97%,且DenCDPKs均存在STKc_CAMK结构域和4个EF hand结构域。(3)qRT PCR分析显示,DenCDPK1、DenCDPK2和DenCDPK3基因均在叶中高表达,分别在茎、花、茎中低表达;且‘水芙蓉’叶片中DenCDPK1和DenCDPK2的表达量高于其他3个供试品种;经8 ℃胁迫12 h后,叶片中DenCDPKs的表达量均显著高于对照组(25 ℃条件下),且DenCDPK3的表达量在0 ℃胁迫12 h时达到最高值。(4)常温(25 ℃)下‘水芙蓉’叶片中的相对电导率显著低于其他3个供试品种;经低温(8 ℃、0 ℃)胁迫12 h后,叶片中的相对电导率均显著高于对照组,且随温度的降低呈上升趋势。研究认为,DenCDPKs基因可能参与秋石斛低温胁迫的响应过程,特别是DenCDPK2可能在秋石斛抗寒过程中发挥着重要作用。     

柽柳水浸提液对猪毛菜种子萌发和幼苗生长的化感作用

以一年生草本植物猪毛菜的种子和幼苗为受体,采用室内培养皿法,研究塔里木河下游柽柳水浸提液对猪毛菜种子及幼苗的化感作用。结果表明:(1)柽柳茎、根浸提液对猪毛菜种子萌发的抑制作用较为显著;柽柳茎浸提液0.08～0.20g/mL浓度处理组的猪毛菜种子发芽率、发芽指数和活力指数与对照组之间均差异显著,其中种子发芽率分别减少了84.78%、89.12%、76.09%和67.39%;柽柳根浸提液对猪毛菜种子活力指数的化感作用较为显著,0.12g/mL和0.16g/mL浓度处理组与对照组相比差异极显著,化感效应指数分别为-0.960和-0.930。(2)柽柳茎浸提液处理中的猪毛菜幼苗苗长和根浸提液处理中的猪毛菜幼苗鲜重与对照组差异显著,两者在0.08g/mL浓度处理下分别增加了61.49%和52.63%;在其他浓度处理下则主要受抑制作用。(3)柽柳叶浸提液的综合化感效应呈现"W"型变化趋势;茎浸提液在高浓度时具有较强的综合化感效应;根浸提液的综合化感效应在0.12g/mL浓度最强(RI=-0.749),在0.08g/mL浓度时最弱(RI=-0.389)。     

Hyperparasitism by Mesochorus spp. (Hymenoptera: Ichneumonidae) in Peristenus sp. (Hymenoptera: Braconidae) and development of PCR primers for hyperparasitoid detection

Peristenus sp. pupae collected from Lygus spp. nymphs in 2001 and 2002 were over-wintered in the laboratory. In both years, more than 30% of adults emerging from over-wintering pupae were identified as ichneumonid hyperparasitoids, Mesochorus curvulus Thomson and Meschorus sp. (Hymenoptera: Ichneumonidae). At the end of the over-wintering period, Peristenus sp. males emerged first followed by Peristenus sp. females and finally Mesochorus spp. The male:female ratio in emerging Peristenus sp. adults was skewed towards males. The Internal Transcribed Spacer (ITS) region and the cytochrome oxidase I (COI) gene from Mesochorus spp. were sequenced. ITS sequences were used to develop PCR primers to detect Mesochorus spp. hyperparasitism in the primary host, Lygus spp. PCR analysis of field-collected Lygus spp. nymphs gave similar estimates of Mesochorus spp. hyperparasitism to the rearing protocols (25–28%). Sequence analysis of COI and ITS regions and subsequent restriction endonuclease analysis of ITS PCR products from Mesochorus spp. indicate the presence of two genotypes in the population. The possibility that these two genotypes represent separate or cyrptic species is discussed.     

蝴蝶兰几丁质酶基因的克隆及表达特性分析

利用RT-PCR及RACE技术,克隆到蝴蝶兰1个几丁质酶基因PhCHT(GenBank登录号为KT992851),该基因cDNA全长1 210bp,包含37bp的5′-UTR、933bp开放阅读框和240bp 3′-UTR,编码310个氨基酸;该蛋白为糖苷水解酶第19家族成员,兼具有溶菌酶活性;生物信息学分析显示,该蛋白具N-端信号肽和跨膜结构,为胞外分泌蛋白;该蛋白与海枣、谷子、油棕和拟南芥的几丁质酶类似蛋白相近,并且在系统进化树上与甘蔗和陆地棉的Ⅶ类几丁质酶同属一个分支。PhCHT基因的表达分析表明,PhCHT在蝴蝶兰营养器官和生殖器官中均有表达,根中表达量最高;13℃/8℃低温处理3、6、9和15d时该基因的表达被抑制,4℃低温处理1、2和4h表达量升高。研究表明,PhCHT基因能够响应短期的冷胁迫。研究结果为进一步研究蝴蝶兰几丁质酶的系统进化及抗性育种奠定了基础。     

Impact of tillage on the incidence ofFusarium spp. in soil

The influence of several long-term conventional and conservation tillage treatments on the incidence and the diversity ofFusarium spp. in soil was studied. Soil samples were randomly collected from naturally contaminated field trials and Fusarium species were isolated by using the dilution plate method. The identification of the species was done by direct microscopic observation onFusarium-specific media. The isolation frequency ofFusarium species and the total number of colony forming units was affected by the sampling year and the cultivated crop and showed significant differences between the tillage treatments. Moldboard plough-treatments resulted in a lower diversity of Fusarium species than the chisel plough and rotary tiller treatments. Besides the tillage system the tillage depth also appeared to affect theFusarium populations. The deeper the tillage the lower was the number of isolatedFusarium spp. TwentyFusarium species were identified over both years of investigation. In conservation tillage plots a higher diversity ofFusarium species was found than in the moldboard plough-based tillage plots. A correlation betweenFusarium species producing the mycotoxin deoxynivalenol (DON) isolated from soil and the DON-content of grain could not be observed. However, these investigations indicate that conservation soil tillage results in conditions which increase the incidence ofFusarium species in soil.     

Inter- and intraspecific variation in infectivity ofSteinernema spp. to larvae of the mushroom flyLycoriella solani

Laboratory bioassays were done to assess the susceptibility of larvae of the mushroom sciarid flyLycoriella solani Winnertz (Diptera; Sciaridae) to 16 isolates comprising five species of the entomopathogenic nematode,Steinernema (Nematoda; Steinernematidae). Each isolate was tested by challenging sciarid larvae with single nematodes. Six isolates were also tested in dose-response experiments. Variations in nematode infectivity occurred (p<0.001) at both inter- and intraspecific levels.Steinernema feltiae (Filipjev) was the most virulent species: two isolates, Nemasys (LD₅₀=1.5–3.9) and Sus 94 (1.8–3.9) were superior to the rest.Steinernema kraussei (Steiner) was the least infectious nematode tested as its infection probability never exceeded 0.10. In addition, lethal dose₅₀ values for this species ranged from 9.0–62.6 and two isolates failed consistently to infect any hosts.     

Predicting the distribution of the crested tinamous, Eudromia spp. (Aves, Tinamiformes)

A bioclimatic analysis of the crested tinamous was conducted to explore climatic factors underpinning the distribution of both Eudromia elegans and E. formosa and to evaluate its potential application in paleontological studies. The study utilized records throughout the entire known range of Eudromia spp. in southern South America. Relationships between 20 environmental parameters and the presence of Eudromia species were established, mapping and characterizing their spatial distribution in a geographic information system using BIOCLIM and MAXENT algorithms. The MAXENT prediction map shows a more homogeneous pattern while BIOCLIM showed a patchier pattern. The models applied here generated maps that adjust to the well-known previous distributions of both species. Nevertheless, for Eudromia elegans, the distribution predicted by MAXENT includes areas where it is actually considered absent, and the BIOCLIM prediction does not include some areas where it is presumed present. Eudromia formosa were found in warmer and wetter sites than E. elegans. Low precipitation areas were identified as suitable for Eudromia elegans. Strong differences between the climatic profiles for both Eudromia spp distributions occurred, with the precipitation the most important influence. E. formosa tolerates the highest maximum temperatures, whereas E. elegans supports the lowest temperatures.     

石斛兰转ACS反义基因抗性原球茎及转化植株的筛选与鉴定

该试验就石斛兰转化ACS(1-氨基环丙烷-1-羧酸合成酶)反义基因的不同筛选方法和筛选处理对抗性原球茎筛选的影响,以及石斛兰转基因植株的再生与鉴定进行研究.结果表明:(1)石斛兰原球茎经带有gus报告基因和ACS反义基因的农杆菌LBA4404侵染共培养5d后除菌,采用逐渐提高选择压浓度的延迟筛选,并在低选择压浓度下切割而高选择压浓度下不切割的处理方式为抗性原球茎的最佳筛选途径,抗性原球茎获得率可达14.97％.(2)抗性原球茎繁殖时应逐渐降低选择压浓度,且在低选择压浓度下进行切割处理,繁殖倍数达到1.15倍,且原球茎生长势好.(3)抗性原球茎在1/2 MS+0.5 mg/L 6-BA培养基中的分化率达到73.85％;107株无根小苗培养于1/2 MS+1.0 mg/L NAA+50.0 mg/L Km(卡那霉素)+100.0 mg/L Cef(头孢霉素)培养基中进行生根培养,共获得了13株具有卡那霉素抗性的转化植株,转化效率达到12.15％.(4)转化植株经报告基因产物GUS组织化学检测和gus的PCR检测,证实带ACS反义基因的T-DNA已整合进石斛兰基因组中,且转基因植株在形态上与未转基因植株无明显差别,3株转基因植株移栽2个月后均已成活.     

Occurrence ofDinophysis spp. and toxic shellfish in the Northern Adriatic

The dynamics of the toxicity of the musselMytilus galloprovincialis was compared between two different shellfish farms, 5 km apart, but using the same cultivation technique. The main differences concerned the freshwater influx and the open aspect to the Gulf of Trieste. It is suggested that a deep closed bay and abundant fresh water inflow are the two main conditions for the low toxicity levels in mussels and for shorter periods of danger. A detailed study of the phytoplankton samples revealed the presence of eight species ofDinophysis in the area of both shellfish farms. During the period of the DSP outbreak in Slovenia (autumn and winter 1989).D. fortii andD. acuminata were the most frequentDinophysis species. There was a high positive correlation between the onset of mussel toxicity and the appearance ofDinophysis spp.     

Distribution, host specificity, and overwintering of Celatoria bosqi Blanchard (Diptera: Tachinidae), a South American parasitoid of Diabrotica spp. (Coleoptera: Chrysomelidae: Galerucinae)

The genus Diabrotica (Coleoptera: Chrysomelidae) includes a great number of pest species, including some of the most important crops pests of the Americas. However, only five parasitoid species have been recorded for it. The parasitoid Celatoria bosqi Blanchard was the first parasitoid described from Diabrotica spp. in South America, where substantial parasitism has been observed. C. bosqi has been collected almost throughout the South American distribution of its main host, Diabrotica speciosa (Germar), in an area that includes temperate and tropical lowlands, and semiarid to humid highlands. Three Diabrotica species were found to host the parasitoid, D. speciosa (Germar), Hystiopsis sp., and Diabrotica viridula (F.), with a total parasitism of 2.60, 5.55, and <0.02%, respectively. Laboratory experiments with field beetles and puparia, reared in the laboratory, indicate that C. bosqi overwinters obligatorily in overwintering adult host beetles, remaining quiescent in its live host below developmental temperatures. Based on the known climatic range of C. bosqi, and its requirement of adult overwintering hosts, a potential distribution in North America is projected.     

A comparative study on the antimutagenic properties of aqueous extracts of Aspalathus linearis (rooibos), different Cyclopia spp. (honeybush) and Camellia sinensis teas

Antimutagenic activity of aqueous extracts of the South African herbal teas, Aspalathus linearis (rooibos) and Cyclopia spp. (honeybush) was compared with that of Camellia sinensis (black, oolong and green) teas in the Salmonella mutagenicity assay using aflatoxin B₁ (AFB₁) and 2-acetylaminofluorene (2-AAF) as mutagens. The present study presents the first investigation on antimutagenic properties of C. subternata, C. genistoides and C. sessiliflora. The herbal teas demonstrated protection against both mutagens in the presence of metabolic activation, with the exception of “unfermented” (green/unoxidised) C. genistoides against 2-AAF, which either protected or enhanced mutagenesis depending on the concentration. Antimutagenic activity of “fermented” (oxidised) rooibos was significantly (P < 0.05) less than that of Camellia sinensis teas against AFB₁, while for 2-AAF it was less (P < 0.05) than that of black tea and similar (P > 0.05) to that of oolong and green teas. Antimutagenic activity of unfermented C. intermedia and C. subternata exhibited a similar protection as fermented rooibos against AFB₁. Against 2-AAF, fermented rooibos exhibited similar protective properties than unfermented C. intermedia and C. sessiliflora. Unfermented rooibos was less effective than the C. sinensis teas and fermented rooibos, but had similar (P > 0.05) antimutagenicity to that of fermented C. sessiliflora against AFB₁ and fermented C. subternata against 2-AAF. Fermented C. intermedia and C. genistoides exhibited the lowest protective effect against 2-AAF, while fermented C. intermedia exhibited the lowest protection when utilising AFB₁ as mutagen. Aspalathin and mangiferin, major polyphenols in rooibos and Cyclopia spp., respectively, exhibited weak to moderate protective effects when compared to the major green tea catechin, (−)epigallocatechin gallate (EGCG). Antimutagenic activity of selected herbal tea phenolic compounds indicated that they contribute towards (i) observed antimutagenic activity of the aqueous extracts against both mutagens and (ii) enhancement of the mutagenicity of 2-AAF by unfermented C. genistoides. Antimutagenic activity of the South African herbal teas was mutagen-specific, affected by fermentation and plant material, presumably due to changes and variation in phenolic composition.     

A new species of Leucothoid Amphipod, Anamixis bananarama, sp. n., from Shallow Coral Reefs in French Polynesia (Crustacea, Amphipoda, Leucothoidae)

Both leucomorph and anamorph developmental stages of Anamixis bananaramasp. n., are illustrated and described from shallow back reef environments of Moorea, French Polynesia. Distinguished by vestigial first gnathopods that persist in post-transformational adult males, this is the second species in the genus to exhibit this unusual character. In other features such as coxae and second gnathopods Anamixis bananaramasp. n. resembles other Pacific Plate endemics of Anamixis known from the region. Specific host association is not documented but suspected to be small calcareous asconoid sponges associated with coral rubble.