Microbial colonization in the seagrass Posidonia spp. roots

The pattern of colonization by microorganisms on root surfaces from three species of seagrass belonging to the genus Posidonia was assessed. Microbial abundance on roots was measured by two electronic microscope techniques. Trends in microbial colonization between species and root order were defined. In addition, eutrophication status of the sampling sites and physiological status of Posidonia oceanica (L.) Delile roots have been taken into account. Our results show high microbial abundance in the Mediterranean species P. oceanica, in comparison with the low rates of colonization found in the Australian species P. australis Hook f. and P. sinuosa Cambridge et Kuo. Microbial density tended to decrease as root order increased, and living roots always showed higher microbial abundance than dead ones. Colonization of P. oceanica roots at the three sites with different environmental status follows different trends according to root order. It is suggested that root age influences the rate of microbial colonization of seagrass roots and that colonization of root surface by microorganisms is associated with organic exudates from the roots rather than with decaying root tissues.     

Occurrence of okadaic acid in the feeding grounds of dugongs (Dugong dugon) and green turtles (Chelonia mydas) in Moreton Bay, Australia

Okadaic acid (OA) is a diarrhetic shellfish poison (DSP) produced by a number of marine organisms including the benthic dinoflagellate Prorocentrum lima, which are often found on seagrass. As seagrass forms the basis of the diet of dugong (Dugong dugon) and green turtle (Chelonia mydas), these herbivores may potentially be exposed to OA through ingestion of P. lima found on the seagrass. In this study, the abundance of epiphytic P. lima, on seagrass, and the concentration of OA produced by these epiphytic dinoflagellates was measured in Moreton Bay, Queensland, Australia. P. lima and OA were found on all four species of seagrass collected. OA was detected in epiphytic material collected from seagrass, with a maximum of 460 ng OA/kg_(wwtSG) found on Halophila spinulosa. From this information, the estimated maximum daily intake (DI) of OA by an adult dugong consuming 40 kg_(wwtSG)/day was 18,400 ng/day, and an adult turtle consuming 2 kg_(wwtSG)/day was 920 ng/day. Analysis by HPLC/MS/MS of 54 stranded dugongs and 19 stranded turtles did not yield OA above the detection limit of 10,000 ng/kg_{(animal tissue)}. OA was found on seagrass, however it was not detected in the tissue samples of dugongs and turtles.     

Protists and detrital particles as prey for the first larval stage of the brachyuran crab, Hemigrapsus oregonensis

Newly-hatched larvae of the brachyuran crab, Hemigrapsus oregonensis, were raised in the laboratory on an autotrophic dinoflagellate (Prorocentrum micans), a heterotrophic dinoflagellate (Noctiluca milaris), a green alga (Dunaliella tertiolecta), an unfed control, and a fed control of Artemia sp. nauplii. Larvae also were fed preparations of seagrass detritus that had been cultured both to promote microbial colonization and to discourage it. Detrital diets were used both alone and in combination with sub-optimal applications of Artemia sp. nauplii. Larvae raised on P. micans showed survival to zoeal stage II equal to those raised on the Artemia sp. nauplii control, although development was delayed. Larvae raised on N. milaris showed substantial (34.7%) survival to zoeal stage II; however survival was lower and development slower than for Artemia sp. nauplii-fed larvae. Survival on D. tertiolecta was less than 3%. Larvae fed microbially-enriched detritus showed a delay in mortality as compared to unfed controls. No larvae fed solely on detritus survived to zoeal stage II. When larvae were fed a sub-optimal diet of Artemia nauplii, supplemented by detrital particles, survival to zoeal stage II increased, although not to the level shown by Artemia-fed larvae in optimal application. Development was not accelerated over the sub-optimal diet in either treatment. The potential for larval crabs to utilize a wide variety of potential prey immediately upon hatching is significant given their susceptibility to early starvation. Such omnivory also suggests a trophic link between carbon sources of the microbial loop and crab larvae.     

Snails as Biomonitors of Oil-Spill and Bioremediation Strategies

Aquatic and pulmonate snails were evaluated for their suitability as biomonitors of habitat recovery following an experimental oil spill in a freshwater marshland. The mystery snail, Viviparus georgianus, and the mimic pondsnail, Pseudosuccinea columella, were used as sediment quality biomonitors for a controlled oil-spill experiment at a wetland site along the St. Lawrence River (Ste. Croix, Quebec) to assess the impacts of crude oil, rates of natural recovery, and the efficacy of bioremediation treatments to enhance the bacterial degradation of residual oil in the sediments. Sediments from control sites and oiled sites with or without the application of fertilizers as bioremediation strategies, were evaluated both in situ and under controlled laboratory conditions at various time intervals. Snail survival, growth, and histopathological changes were monitored. While V. georgianus proved to be good biomonitors, P. columella appeared unaffected by the treatments. The differing sensitivity may depend on the gastropods' feeding habits. V. georgianus being a detritivore assimilated contaminants from the sediments, while P. columella, being an herbivore, did not directly assimilate contaminants. Nevertheless, snails show potential as important and ideal “tools” for testing environmental conditions because of their abundance, ease of collection, wide distribution, and relatively sedentary nature.     

Varying foraging strategies of Labridae in seagrass habitats: Herbivory in temperate seagrass meadows?

The diets of five species of Labridae in south-western Australia were examined to determine whether: (1) grazing of seagrass and epiphytic algae is a prominent feature of the food web within the deeper seagrass meadows of this temperate region; (2) levels of grazing differ among different seagrass systems; and diets differ among these closely-related species. Fish were collected seasonally from three seagrass habitats mainly comprising either Posidonia sinuosa, Posidonia coriacea or Amphibolis griffithii between the summer of 1996/97 and spring of 1997. Consumption of considerable amounts of algae and seagrass by Odax acroptilus and seagrass by Haletta semifasciata indicates that macrophyte grazing by fish is a component of the trophic dynamics of south-western Australian seagrass meadows. O. acroptilus and H. semifasciata were both omnivorous, feeding on a range of epifauna, infauna and flora, whereas Siphonognathus radiatus, Neoodax balteatus and Notolabrus parilus were carnivorous, feeding predominantly on motile epifauna, such as molluscs and crustaceans. The level of macrophyte grazing is likely to be underestimated in temperate offshore meadows of P. sinuosa and A. griffithii where omnivorous labrids, monacanthids and terapontids are abundant. Stable isotope data for O. acroptilus from the study region suggest that animal prey is more important to tissue maintenance than macrophyte material. Macrophytes may be grazed to acquire attached animal prey or for fulfilling energy requirements. Based on the distribution of prey, it appears that species in A. griffithii meadows forage within and below the seagrass canopy, whilst species in P. sinuosa meadows are likely to forage towards the basal area of this seagrass.     

Natural recolonisation of seagrasses at a disused sewage sludge outfall

Digested sludge from a sewage outfall that operated for 15 years adjacent to Adelaide in South Australia caused total seagrass loss in a 365 ha area around the outfall. Eight years after its closure, an underwater survey was conducted to determine the extent of seagrass recovery in a 2 ha area immediately adjacent to the disused outfall. Total seagrass cover was estimated to be 28% comprising 23% Halophila australis, 3% Posidonia angustifolia, 1% Posidonia sinuosa, and less than 1% each for Zostera tasmanica and Amphibolis antarctica. The recovery of seagrasses at the outfall site is probably due to recolonisation by propagules from a distant source. While results from this study suggest that seagrasses can return to a severely polluted site if the pollution source is removed and that Posidonia can be a primary coloniser of disturbed sites, they also suggest that it will take many decades for the seagrass community to recover to its former state.     

Establishment of Peristenus digoneutis and P. relictus (Hymenoptera: Braconidae) in California for the control of Lygus spp. (Heteroptera: Miridae)

Lygus hesperus Knight is native to the western United States and is a perennial pest of numerous crops in California. It is responsible for triggering the early season application of insecticides on cotton, Gossypium hirsutum L., and strawberries, Fragaria L. Despite several surveys conducted in alfalfa (Medicago sativa L.) grown in central California, nymphal parasitoids associated with L. hesperus and L. elisus have not been found. Two exotic parasitoids were released into California beginning in 1998. Peristenus relictus (Ruhte), formerly P. stygicus Loan, and P. digoneutis Loan were collected from several locations in southern Europe and released at up to six locations over a 6-year period. At the original release site in Sacramento, a 0.25-ha plot of alfalfa, parasitism by P. digoneutis and P. relictus combined increased from zero to 90%, 3 years after the last releases were made. Parasitoids have been recovered from vacant fields of weedy annuals within 2 km of this site. Recoveries at more southerly release sites in central California have been poor.     

Detrital stocks and dynamics of the seagrass Posidonia oceanica (L.) Delile in the Spanish Mediterranean

Previous studies have shown that most leaf production (>90%) of the seagrass Posidonia oceanica is shed after senescence and that a substantial percentage (up to 80%) may thereafter be exported off the seagrass meadows by waves and currents. It has also been reported that P. oceanica meadows can accumulate large stocks of belowground detritus due to slow decomposition rates. However, the generality of these results across broad spatial scales is poorly known. In this report, we examine the fate of leaf production and the magnitude and dynamics of belowground detritus in 16 P. oceanica meadows distributed along the Spanish Mediterranean. Herbivores removed a small percentage of leaf production in all the meadows (≤13%), with most leaf production (>85%) being shed after senescence. Most shed leaves (>90%) were exported off the meadows by physical agents, such as waves and currents. The amount of belowground detritus stored within 10–15 cm from the sediment surface varied from ca. 70 to 7500 g DW m⁻² among the meadows examined, and they accumulated at rates ranging from ca. 65 to 650 g DW m⁻² per year. These values are large when compared to other communities of aquatic and terrestrial macrophytes. Our results show that P. oceanica meadows in the Spanish Mediterranean support high values of secondary production in other systems by exporting large amounts of leaf detritus as well as acting as substantial carbon sinks by accumulating large reservoirs of belowground detritus. Therefore, the increasing anthropogenic threats on P. oceanica could entail an important loss of secondary production and carbon storage in Mediterranean coastal ecosystems.     

基于ITS和matK基因对牡丹组序列分析及其亲缘关系的研究

以芍药属牡丹组全部9个野生种、5个紫斑牡丹栽培品种及3个中原牡丹品种为试材,进行核糖体内转录间隔区（ITS）和叶绿体成熟酶K （matK）基因片段测序分析,探讨ITS和matK序列为牡丹组所有野生种种间关系提供分子证据。从GeneBank中选取了1个牡丹及3个外类群芍药、川赤芍和草芍药的ITS及matK序列。对试验样品进行DNA提取、PCR扩增并双向测序得到44条序列,人工校正后将所得44条序列进行比对;计算碱基组成频率、变异位点、简约信息位点数、转换/颠换比率、种内及种间遗传距离,以邻接法进行系统发育分析。结果表明,牡丹组所有个体ITS序列长度在750~800 bp,含有86个多态位点,74个简约信息位点,转换/颠换比率（R）为1.2;而matK序列含有20个简约信息位点,转换/颠换比率（R）为1.7。ITS序列分析将牡丹组野生种分为两大枝,稷山牡丹、紫斑牡丹、卵叶牡丹和杨山牡丹聚为一枝,狭叶牡丹、滇牡丹、黄牡丹和大花黄牡丹聚为另一枝,这两枝分别与革质花盘亚组和肉质花盘亚组相对应,而四川牡丹位于革质花盘亚组最底端,支持前人研究将四川牡丹归为革质花盘亚组。matK序列分析的牡丹组野生种间遗传距离结果不理想,未能清晰的表明野生种之间的亲缘关系。由此说明,ITS序列更适合牡丹组野生种间亲缘关系的研究分析。     

Flavonoid data and populational observations in support of hybrid status for Populus acuminata

Flavonoid distribution and populational composition at the collection sites of several samples of Populus acuminata strongly suggest that this taxon is an inter-sectional hybrid between P. angustifolia, sect. Tacamahaca and any one of two or three different species of the section Aigeiros, when the entire range is considered. In the vicinity of the type locality, the Aigeiros parent is P. sargentii. Within any population, foliage of trees of P. acuminata is characterized by morphological intermediacy and an essentially additive flavonoid profile, as compared to the two parental taxa present. In habit the trees resemble those of P. sargentii, and habitats at the collection sites were found to be somewhat intermediate. Fifteen flavonoid compounds were consistently present in samples of the putative hybrid. Twelve of these had diagnostic value because they were absent in one of the parental species. One compound, apigenin 7-O-diglucoside, was found only in P. acuminata. In part, our identifications correspond to compounds reported by Crawford in a morphological and chemical study of P. acuminiata.     

Seagrass Transplanting in Cockburn Sound, Western Australia: A Comparison of Manual Transplantation Methodology Using Posidonia sinuosa Cambridge et Kuo

Seagrass transplants (plant units [PUs]) were established to examine the feasibility of seagrass rehabilitation in Cockburn Sound, Western Australia. Five hundred and twenty plant units (plugs and sprigs) of Posidonia sinuosa Cambridge et Kuo were placed at seven locations at depths of 3, 5, and 9 m and monitored to determine the influence of transplant method, location, and depth upon survival and growth over 2 years. Depending on the site, more plugs had survived at the completion of the trial (mean survival 41%) than sprigs (mean survival 15%). Plug and sprig survival differed significantly with transplant depth, decreasing overall with increasing depth. Forty‐five percent of surviving plugs and 50% of sprigs exhibited horizontal rhizome extension. Mean rhizome extension after 2 years was 9.5 cm/plug (1–23 cm) and 18.3 cm/sprig (0.5–31 cm). Declines in PU survival and variable growth correlate with site‐specific variability in light climate. Plug transplantation was deemed the most suitable method for further manual seagrass rehabilitation, exhibiting higher survival across all sites and conditions; however, they are costly to deploy. Sprig PUs have greatest potential in shallow water with fine sands, moderate water movement, and maximum light availability. The low cost of deploying sprigs may outweigh their lower survival compared to plugs; further efforts should be directed to enhancing survival of sprig PUs under a wider range of conditions. Suitable locations for future rehabilitation efforts in Cockburn Sound were the Eastern and Western Banks and shallow areas off Woodman Point and Mangles Bay.     

水杉原生种群核心种质资源的繁殖特性

水杉(Metasequoia glyptostroboides)原生种群天然更新困难, 为探究其原因是否与繁殖特性有关, 本文以其核心种质资源(40株个体)为对象, 通过对生境、母树个体和种子性状的调查, 并结合田间播种试验, 分析其繁殖特性。以方差分析检测不同母树个体间种子性状、发芽率和幼苗生长特征的差异; 采用相关性分析和多元线性回归模型分析个体性状对其种子、发芽率及幼苗生长的影响; 通过Logistic模型拟合, 分析幼苗生长的节律参数。结果显示: (1) 40株母树个体仅有7株产种, 产种率仅为17.50%, 且不同母树个体间产种量差异显著(P < 0.05)。(2)不同母树个体间的种子千粒重、种子发芽率、幼苗存活率和基径生长量均存在显著差异(P < 0.05)。(3) 7株母树个体种子的平均发芽率为19.73%, 幼苗平均存活率为73.42%, 不同母树之间差异显著(P < 0.05)。(4)母树个体的胸径与种子千粒重、种子发芽率呈显著负相关(P < 0.05), 种子发芽率与幼苗存活率呈极显著正相关(P < 0.001), 树高与幼苗存活率呈显著正相关(P < 0.05)、与幼苗基径呈极显著正相关(P < 0.001)。(5)母树树高对幼苗存活率有显著正效应(P < 0.05), 胸径对种子发芽率有显著负效应(P < 0.05)。(6)苗高、基径的生长符合“S”型生长曲线(R ² ≥ 0.928)。水杉原生种群产种量低, 种子发芽率低、幼苗存活率较低, 且不同母树之间差异显著可能是造成其天然更新困难的原因。     

基底颜色对两种沙蜥体色变异的影响

生存在不同基底颜色环境下的爬行动物种群通常表现出丰富的体色地理变异, 其体色变化的潜在机制具有多样性。变色沙蜥(Phrynocephalus versicolor)和草原沙蜥(P. frontalis)具有较近的遗传关系, 曾被认为与荒漠沙蜥(P. przewalskii)组成同一系统发育种组。本文应用光纤光谱仪(AvaSpec-2048), 通过记录沙蜥背部体表12个部位的皮肤光反射率, 定量比较在黑化环境下的深色变色沙蜥与非黑化环境下的浅色草原沙蜥自然体色变异, 研究其种群体色变异是否具有时间可逆性, 并探讨基底颜色对沙蜥体色的影响机制。研究结果表明, 黑化生境下的变色沙蜥体色显著深于非黑化枯黄色生境下的草原沙蜥。此外, 对黑化与非黑化样本开展的生境互换移植围栏实验, 即把枯黄色生境中非黑化的草原沙蜥移植于黑色的基底环境中饲养, 把黑化生境中黑化的变色沙蜥移植于枯黄色生境中饲养。结果表明, 饲养1周后黑化群体背部6个检测部位的光反射率显著变大, 其他部位均无显著变化; 而非黑化群体只有左后肢和背部右上方2个部位的皮肤光反射率发生显著变化, 其他部位反射率无显著变化。结果表明, 变色沙蜥体色变异能力比草原沙蜥强, 体色表型可能已经在两个近缘沙蜥物种中稳定遗传, 基底生境颜色的短期变化在统计学上能引起肉眼难以识别的轻微的体色变异, 个体发育相关的一些遗传因素可能对体色变异起控制 作用。     

洮河上游紫果云杉种群结构特征

该研究以洮河上游尕海-则岔自然保护区、卡车林区和冶力关林区的紫果云杉(Picea purpurea)天然种群为研究对象, 通过样地调查和数据统计, 绘制种群结构图, 编制静态生命表, 拟合并分析存活曲线, 运用数量化方法研究种群动态, 揭示种群生存现状, 预测种群发展趋势, 以期为该物种的保护、管理及结构恢复提供理论依据。结果显示: 3个林区紫果云杉种群自然更新能力强, 幼苗、幼树储量丰富, 幼小龄期死亡率普遍偏高; 尕海-则岔种群存活曲线符合Deevey-III型, 种群稳定结构完整, 卡车林区和冶力关林区种群存活曲线均符合Deevey-II型, 且均出现了局部衰退; 3个林区种群动态指数均大于0, 说明种群均属于增长型, 增长潜力为尕海-则岔>卡车>冶力关; 受随机干扰时卡车林区紫果云杉最敏感, 冶力关次之, 尕海-则岔种群最稳定。该研究表明: 竞争和自疏作用是造成紫果云杉幼小龄级个体存活率偏低的普遍因素, 3个林区不同的生存状况反映了紫果云杉种群在不同生境及生活史下生存能力的差异, 保护幼苗生存环境并提高幼苗质量和存活率是种群更新和发展的关键。尕海-则岔紫果云杉生存良好, 种群生存状况主要受自身生物学特性和环境因子的影响; 卡车林区主要受人为影响, 种群结构遭到破坏; 冶力关林区受分布限制, 造成种群结构不稳定, 须采取一定的人工措施来促进种群更新与增长。     

大兴安岭兴安落叶松和樟子松径向生长对气候变化的响应差异

大兴安岭是我国气候变化最为显著的地区之一,兴安落叶松和樟子松是该地区最为重要的树种,研究它们径向生长对气候变化的响应差异,可以为预测气候变化下我国北方森林动态提供科学依据。在大兴安岭地区选择6个样点共采集兴安落叶松树轮和樟子松树轮样芯451个,建立了12个标准年表。比较了1900年以来树木径向生长趋势,利用Pearson相关分析法分析各样点兴安落叶松和樟子松生长对气候因子的响应,运用线性混合模型探讨温度和降水对兴安落叶松和樟子松年径向生长的影响,通过滑动相关对比两个树种生长-气候关系的时间稳定性。结果表明: 兴安落叶松径向生长与3月平均温度呈负相关,与上一年冬季和当年7月降水呈正相关。樟子松径向生长与当年8月温度呈正相关,与当年生长季(5—9月)降水呈正相关。冬季降雪对兴安落叶松径向生长起到重要的促进作用,夏季过多降水对樟子松径向生长起到显著的限制作用。兴安落叶松和樟子松生长对气候变化的响应存在明显差异,因此,气候变化可能会影响北方森林生态系统的树木生长、物种组成以及空间分布等。     

Edge effects in patchy seagrass landscapes: The role of predation in determining fish distribution

Predation is often described as an underlying mechanism to explain edge effects. We assessed the importance of predation in determining edge effects in seagrass using two approaches: a video survey to sample predators at small scales across seagrass edges, and a tethering experiment to determine if predation was an underlying mechanism causing edge effects. Underwater videos were placed at four positions: middle of seagrass patches; edge of seagrass; sand immediately adjacent to seagrass and sand distant from seagrass. Fish abundances and the time fish spent in view were measured. The main predatory fish (Australian salmon, Arripis spp.) spent more time over adjacent sand than other positions, while potential prey species (King George whiting, Sillaginodes punctata (Cuvier), recruits) were more common in the middle of seagrass patches. Other species, including the smooth toadfish, Tetractenos glaber (Freminville), and King George whiting adults, spent more time over sand adjacent to seagrass than distant sand, which may be related to feeding opportunities. King George whiting recruits and pipefish (Stigmatopora spp.) were tethered at each of the four positions. More whiting recruits were preyed upon at outer than inner seagrass patches, and survival time was greater in the middle of shallow seagrass patches than other positions. Relatively few pipefish were preyed upon, but of those that were, survival time was lower over sand adjacent to seagrass than at the seagrass edge or middle. Video footage revealed that salmon were the dominant predators of both tethered King George whiting recruits and pipefish. The distribution of predators and associated rate of predation can explain edge effects for some species (King George whiting) but other mechanisms, or combinations of mechanisms, are determining edge effects for other species (pipefish).     

基于线粒体基因组控制区序列的中国冰清绢蝶遗传分化和谱系生物地理研究

【目的】分析中国冰清绢蝶Parnassius glacialis遗传多样性和遗传分化情况及其系统发生关系,推测其起源及分歧时间,并探讨其历史生物地理分布格局的成因。【方法】以2008-2016年采集于中国13个地区的冰清绢蝶的325头及绢蝶属其他11个种的11头个体样品,对其线粒体基因组的AT富集区(控制区)序列进行PCR扩增和序列测定,采用MEGA 6.0, Dna SP 5.1和Arlequin 3.5等软件分析其遗传多样性和遗传分化情况;以其他近缘绢蝶种类作为外类群,采用PhyML3.0, MrBayes 3.2和BEAST V1.8.3等软件重建冰清绢蝶的系统发生树,并利用宽松分子钟和校准方法推测中国冰清绢蝶的起源和分歧时间;同时结合现今冰清绢蝶的地理分布特点和第四纪以来的地球环境事件,分析其扩散路径,探讨其历史生物地理分布格局及成因。【结果】冰清绢蝶AT富集区序列长度介于487～495 bp之间,平均为491 bp,其长度的差异主要体现在Poly(T)或Poly(A)的数目差异,A+T平均含量为95.76%。供试冰清绢蝶13个地理居群325头个体中,共检测出基于线粒体基因组AT富集区序列的239个单倍型,单倍型多样性Hd为0.9971;核苷酸多态性指数π值为0.02948, Theta (per site)Eta值为0.06594。系统发生分析和分子定年显示,冰清绢蝶和白绢蝶P.stubbendorfii为近缘姊妹种,二者在距今7.52百万年(Ma)的中新世晚期开始分化。冰清绢蝶的祖先可能起源于我国西南地区(现今横断山脉-喜马拉雅附近的青藏高原东北缘)距今1.53 Ma的更新世时期,先由起源地先期扩散到达小陇山、秦岭一带;后来,由于第四纪的冰期-间冰期轮回事件和扩散路径的不同而分化出2大支系,但两个支系之间存在少量的单倍型相互混杂现象;随后,2大支系继续向低海拔、低纬度地区扩散,即通过伏牛山脉向东北方向扩散到嵩山、泰山、昆嵛山一带,经大别山系向东南方向扩散到琅琊山、紫金山和天目山一带。【结论】中国13个居群的冰清绢蝶没有形成明显的地理分化格局,其遗传变异主要来自居群内部,居群间的遗传分化现象尚不明显;冰清绢蝶起源于更新世时期的较高海拔地区,由于第四纪的冰期-间冰期轮回事件以及扩散路径的不同而形成2大支系并向中国东部和南部低海拔和低纬度地区扩散。     

Food sources of two detritivore amphipods associated with the seagrass Posidonia oceanica leaf litter

This study focused on the ingestion and assimilation of Posidonia oceanica (L.) Delile litter by Gammarella fucicola Leach and Gammarus aequicauda Martynov, two dominant detritivore amphipods of the P. oceanica leaf litter. Scanning electron microscope observations indicated that leaf litter is highly colonized by diverse diatoms, bacteria and fungi, which may constitute a potential food source for the litter fauna. Gut content observations demonstrated that these species eat P. oceanica litter, and that this item is an important part of their ingested diet. Stable isotope analyses showed that the species do not experience the same gains from the ingested Posidonia. Gammarella fucicola displayed isotopic values, suggesting a major contribution of algal material (micro- and macro-epiphytes or drift macro-algae). On the other hand, the observed isotopic values of G. aequicauda indicated a more important contribution of P. oceanica carbon. The mixing model used agreed with this view, with a mean contribution of P. oceanica to approximately 50% (range 40-55%) of the assimilated biomass of G. aequicauda. This demonstrated that the two species, suspected to be detritus feeders, display in reality relatively different diets, showing that a certain degree of trophic diversity may exist among the detritivore community of the seagrass litter.     

Sequential activation of constitutive and inducible nitric oxide synthase (NOS) in rat cerebellar granule neurons by pseudomonas fluorescens and invasive behaviour of the bacteria

Previous studies have shown that Pseudomonas fluorescens and its lipopolysaccharide (LPS) exert dose-related cytotoxic effects on neurons and glial cells. In the present work, we investigated the time course effect of P. fluorescens MF37 and its LPS on cultured rat cerebellar granule neurons. The kinetics of binding of P. fluorescens to cerebellar granule neurons is rapid and reaches a mean of 3 bacteria/cell after 5 h. As demonstrated by measurement of the concentration of nitrite in the culture medium, P. fluorescens induces a rapid stimulation (3 h) of the nitric oxide synthase (NOS) activity of the cells. In contrast, LPS extracted from P. fluorescens requires a long lag phase (24 h) before observation of an activation of NOS. Measurement of the membrane resting potential of granule neurons showed that within 3 h of incubation there was no difference of effect between the action of P. fluorescens and that of its endotoxin. Two complementary approaches allowed to demonstrate that P. fluorescens MF37 presents a rapid invasive behaviour suggesting a mobilisation of calcium in its early steps of action. The present study reveals that P. fluorescens induces the sequential activation of a constitutive calcium-dependent NOS and that of an inducible NOS activated by LPS. Our results also suggest that in P. fluorescens cytotoxicity and invasion are not mutually exclusive events.