Distribution and patterns of emergence of suprabenthic and pelagic crustaceans in a shallow temperate estuary (Ria de Aveiro,Portugal)

Ecology and dynamics of the zooplankton in the Ria de Aveiro, Portugal, were researched to understand the correlation between the patterns of abundance of suprabenthic and zooplanktonic crustaceans and its tidal, day/night and lunar cycles. This study involved the collection of a series of samples during nine 25 h sampling cycles, separated weekly, at a fixed station located near the mouth of the lagoon. In each 25 h cycle, the samples were collected every 2 h with a 500 μm net, at three depths, at the surface, mid-water and over the bottom. The overall effect of the phases of tide and day and depth on the vertical position of the organisms was further investigated through a three-way analysis of variance (ANOVA), in which the phase of the tide (ebb, flood), the phase of the day (day, night) and depth (surface, mid-water, bottom) constituted the three factors in the analysis. The distribution of the species in the water column exhibited distinct tidal, day/night, and lunar patterns, reflecting the ecological particularities of the different species in relation to their seasonal occurrence and behavioural relationships. This distribution showed that for the dominant species, there was a significant interaction between behavioural strategies and cyclic changes of environmental conditions.     

具休眠体淡水水生生物分布假说的研究进展

淡水水生生物的分布模式一直是进化和生物地理学研究中的热点问题, 特别是对具休眠体水生生物的研究, 早在达尔文时代就开始探讨具休眠体淡水水生生物分布的原因。根据对文献的整理, 可以将具休眠体淡水水生生物分布模式归纳为三种假说: 随机分布(Everything is Everywhere, EIE)假说、独占(Monopolisation)假说和局域特有(Moderate Endemicity, ME)假说。通过对三种假说的定义, 研究进展和形成不同模式的原因等进行分析, 提出了在研究时需注意是否存在隐存种、季节变化引起基因型变化、研究区域是否覆盖了本物种的所有分布范围、地理环境及地质事件与分布模式之间的关系等问题, 以便更准确的研究具休眠体淡水水生生物分布模式。     

Temporal and spatial structure in the suprabenthic community of a shallow estuary (western Portugal: Mondego river estuary)

The suprabenthic fauna of the Mondego river estuary (western Portugal) was sampled monthly between June 1996 and June 1997. Quantitative samples were taken, with a suprabenthic 500-μm mesh size net, at regularly spaced stations covering the entire south arm of the estuary. The diversity of the samples and the distribution of the species were assessed. Suprabenthic communities were identified using a divisive multivariate statistical technique and species composition, density and biomass of the dominant species of each community were compared among communities. Diversity was highest in the mouth of the estuary where density and biomass were lowest. Diversity decreased upstream and was lowest in the mid and inner estuary where density and biomass reached maximal values. Suprabenthic animals, mainly the mysid Mesopodopsis slabberi, reached high densities in inner and upstream stations, whereas the more seaward stations had lower densities but a higher number of species. The spatial patterns dominated over the temporal patterns.     

Phylogenetic status and matrilineal structure of the biting midge, Culicoides imicola, in Portugal, Rhodes and Israel

The biting midge Culicoides imicola Kieffer (Diptera: Ceratopogonidae) is the most important Old World vector of African horse sickness (AHS) and bluetongue (BT). Recent increases of BT incidence in the Mediterranean basin are attributed to its increased abundance and distribution. The phylogenetic status and genetic structure of C. imicola in this region are unknown, despite the importance of these aspects for BT epidemiology in the North American BT vector. In this study, analyses of partial mitochondrial cytochrome oxidase subunit I gene (COI) sequences were used to infer phylogenetic relationships among 50 C. imicola from Portugal, Rhodes, Israel, and South Africa and four other species of the Imicola Complex from southern Africa, and to estimate levels of matrilineal subdivision in C. imicola between Portugal and Israel. Eleven haplotypes were detected in C. imicola, and these formed one well-supported clade in maximum likelihood and Bayesian trees implying that the C. imicola samples comprise one phylogenetic species. Molecular variance was distributed mainly between Portugal and Israel, with no haplotypes shared between these countries, suggesting that female-mediated gene flow at this scale has been either limited or non-existent. Our results provide phylogenetic evidence that C. imicola in the study areas are potentially competent AHS and BT vectors. The geographical structure of the C. imicola COI haplotypes was concordant with that of BT virus serotypes in recent BT outbreaks in the Mediterranean basin, suggesting that population subdivision in its vector can impose spatial constraints on BT virus transmission.     

一种濒危大疣蛛的潜在分布区域以及气候变暖的影响

本研究的目的是i)确定伊比利亚半岛一种大疣蛛(Macrothele calpeiana)分布的气候相关性以预测其潜在分布,ii)详细阐述该物种在伊比利亚半岛的分布假说,iii)通过推断模型预测确认该物种在北非和整个地中海地区的适合区域,iv)预测气候变暖对蜘蛛潜在分布区的影响。基于物种的存在、远离目前环境条件下的可能缺失以及其它的气候参数,使用广义线性模型发展了潜在分布的可预测模型。蜘蛛在伊比利亚半岛上的潜在分布远大于目前已知的分布区,延伸到尚未发现蜘蛛分布的葡萄牙广大地区。本文提出了该种大疣蛛在适合分布区域内没有分布的历史因素。北非具有适合该物种的条件但却没有该属物种的分布支持了大疣蛛(Macrothele)祖先的东方起源假说。对地中海地区蜘蛛分布的推断突出显示了阿根廷地区有合适的分布区,该地区也发现有另一种欧洲大疣蛛。气候变暖将对现存于伊比利亚的M.calpeiana种群产生负面影响,因为气候变暖将减少并破碎化蜘蛛在北非的潜在栖息地。目前,急需确认在葡萄牙广大地区是否存在蜘蛛物种,发展对该属的系统发育研究以确定大疣蛛属起源和扩散史的理论。     

Detection of diverse new Francisella-like bacteria in environmental samples

Following detection of putative Francisella species in aerosol samples from Houston, Texas, we surveyed soil and water samples from the area for the agent of tularemia, Francisella tularensis, and related species. The initial survey used 16S rRNA gene primers to detect Francisella species and related organisms by PCR amplification of DNA extracts from environmental samples. This analysis indicated that sequences related to Francisella were present in one water and seven soil samples. This is the first report of the detection of Francisella-related species in soil samples by DNA-based methods. Cloning and sequencing of PCR products indicated the presence of a wide variety of Francisella-related species. Sequences from two soil samples were 99.9% similar to previously reported sequences from F. tularensis isolates and may represent new subspecies. Additional analyses with primer sets developed for detection and differentiation of F. tularensis subspecies support the finding of very close relatives to known F. tularensis strains in some samples. While the pathogenicity of these organisms is unknown, they have the potential to be detected in F. tularensis-specific assays. Similarly, a potential new subspecies of Francisella philomiragia was identified. The majority of sequences obtained, while more similar to those of Francisella than to any other genus, were phylogenetically distinct from known species and formed several new clades potentially representing new species or genera. The results of this study revise our understanding of the diversity and distribution of Francisella and have implications for tularemia epidemiology and our ability to detect bioterrorist activities.     

Occurrence of Salmonella spp in estuarine and coastal waters of Portugal

The presence of Salmonella and its relationship with indicator organisms of fecal pollution, such as total coliforms, fecal coliforms and fecal streptococci, was studied at two marine zones in Portugal. Seventeen different Salmonella serotypes were isolated and identified, S. virchow was the most frequently isolated (21.6%). In addition, a high percentage (35.1%) was recorded for some Salmonella serotypes of clinical significance, namely S. enteritidis, S. infantis, S. typhimurium and S. virchow. In any of the samples from the two zones Salmonella was not detected in the absence of any of the indicator organisms. However, the incidence of Salmonella as a function of indicator concentration intervals established by the EEC standards was 0, 10 and 19.3% at guide values of total coliforms, fecal coliforms and fecal streptococci, respectively in the Faro samples (south of Portugal). In contrast, Salmonella incidence rates of 37.5, 36.4 and 33.3% were recorded at the corresponding guide values the Caminha samples (north of Portugal). No significant correlations (p>0.005) were obtained between Salmonella and the indicators at the sampling stations; however, total coliforms and fecal streptococci were the indicators most closely related to Salmonella in Caminha and Faro samples, respectively. Survival experiments in Escherichia coli, Enterococcus faecalis and S. typhimurium, using diffusion chambers, were performed to verify whether the lack of correlation between indicators and Salmonella was due to different inactivation rates in seawater. The results indicate that survival percentages of the three microorganisms tested were similar after 48 h of exposure to seawater.     

Application of a simulation model of hydrobiological survey in complex analysis of in situ data

The model realizes the algorithm simulating the motions of a group of organisms with mutually independent, stochastic paths in a homogeneous medium, which results in the random distribution of the organisms in local areas in space (sampling units). Substitution of the mean abundance values of individual species found in the actual hydrobiological samples into the sample as one of its parameters produces a given set of artificial samples similar to the in situ sample with its elements possessing complete stochastic independence. They are further used to test the reliability of results obtained by statistical methods of data analysis and computations of threshold values for some indices and coefficients employed in ecology.     

Chromosome numbers and geographic distribution of Ulex gallii and U. minor (Leguminosae)

FERNANDEZ PRIETO, J. A., NAVA, H. S., VERA, M. L., ALVAREZ MARTINEZ, M. J., DIAZ, T. E., FERNANDEZ CASADO, M. A., FERNANDEZ-CARVAJAL, M. C. & GUTIERREZ VILLARIAS, M. I., 1993. Chromosome numbers and geographic distribution of Ulex gallii and U. minor (Leguminosae). Chromosome numbers from 188 samples of the Ulex gr. gallii-minor from Portugal, Spain, France and Great Britain are reported. The gametic and somatic chromosome numbers are n = 16 and In = 32, respectively, in all samples of U. minor. Three levels of ploidy (diploid, tetraploid and hexaploid) have been detected in samples of U. galli, sensu lata from N Spain. In samples from Brittany and Great Britain, only the tetraploid level has been found. New data are compared with all previous reports for the species of this group of gorses.     

Gamma-proteobacteria Aquicella lusitana gen. nov., sp. nov., and Aquicella siphonis sp. nov. infect protozoa and require activated charcoal for growth in laboratory media

Several isolates, belonging to two new species of the same novel genus of gamma-proteobacteria, were recovered from drilled well (borehole) and spa water at S?o Gemil in central Portugal. These organisms are phylogenetically most closely related to the strictly intracellular uncultured species of the genus Rickettsiella, which cause disease in arthropods, and to the facultatively intracellular species of the genus Legionella, some of which cause Legionnaires' disease and Pontiac fever. The S?o Gemil strains grew only on media containing charcoal, as is also true of the species of the genus LEGIONELLA: Unlike the vast majority of Legionella isolates, the new isolates did not require L-cysteine or ferric pyrophosphate for growth but like the legionellae had an absolute requirement for alpha-ketoglutarate. Strains SGT-39(T) and SGT-56 grew consistently between 30 and 43 degrees C, while strains SGT-108(T) and SGT-109 grew between 30 and 40 degrees C. The pH ranges for growth of these organisms were surprisingly narrow: strains SGT-39(T) and SGT-56 grew between pH 6.3 and 7.3, while strains SGT-108(T) and SGT-109 grew between pH 6.3 and 7.0. Both organisms proliferated in the amoeba Hartmannella vermiformis but did not grow in U937 human cells. Based on 16S rRNA gene sequence analysis and physiological, biochemical, and chemical analysis we describe two new species of one novel genus; one species is represented by strain SGT-39(T), for which we propose the name Aquicella lusitana, while strain SGT-108(T) represents a second species of the same genus, for which we propose the name Aquicella siphonis.     

Molecular techniques for the early warning of toxic cyanobacteria blooms in freshwater lakes and rivers

The aim of this work was to test the efficacy of molecular techniques for detecting toxigenic cyanobacteria in environmental water samples collected from freshwater lakes, rivers and reservoirs in Portugal. Of 26 environmental samples tested, 21 were found to contain Microcystis using a genus-specific polymerase chain reaction (PCR). Another primer pair was applied to the same DNA template to test for the presence of microcystin synthetase genes. This primer pair resulted in the formation of a PCR product in 15 of the samples containing Microcystis and one sample that did not give a positive result in the Microcystis genus-specific PCR. A restriction assay using the enzyme EcoRV was then applied to show that in most cases, the gene fragment was from toxigenic strains of Microcystis and, in one above-mentioned case, from a microcystin-producing strain of Planktothrix. All environmental samples were examined microscopically to confirm the presence of cyanobacteria species. Samples were also tested for the presence of microcystins using the ELISA plate assay. There was good agreement between the results obtained with molecular techniques and those obtained from microscopy and chemical methods. The PCR techniques applied in this paper were found to be useful, particularly when the concentration of the target organism was very low compared with other organisms. This technique can be used to detect inocula for cyanobacterial populations and therefore provide a useful tool for assessing under which conditions particular species can grow into bloom populations.     

Survey of Chickpea Rhizobia Diversity in Portugal Reveals the Predominance of Species Distinct from Mesorhizobium ciceri and Mesorhizobium mediterraneum

Several Mesorhizobium species are able to induce effective nodules in chickpea, one of the most important legumes worldwide. Our aims were to examine the biogeography of chickpea rhizobia, to search for a predominant species, and to identify the most efficient microsymbiont, considering Portugal as a case study. One hundred and ten isolates were obtained from continental Portugal and Madeira Island. The 16S ribosomal RNA gene phylogeny revealed that isolates are highly diverse, grouping with most Mesorhizobium type strains, in four main clusters (A–D). Interestingly, only 33% of the isolates grouped with Mesorhizobium ciceri (cluster B) or Mesorhizobium mediterraneum (cluster D), the formerly described specific chickpea microsymbionts. Most isolates belong to cluster A, showing higher sequence similarity with Mesorhizobium huakuii and Mesorhizobium amorphae. The association found between the province of origin and species cluster of the isolates suggests biogeography patterns: most isolates from the north, center, and south belong to clusters B, A, and D, respectively. Most of the highly efficient isolates (symbiotic effectiveness >75%) belong to cluster B. A correlation was found between species cluster and origin soil pH of the isolates, suggesting that pH is a key environmental factor, which influences the species geographic distribution. To our knowledge, this is one of the few surveys on chickpea rhizobia and the first systematic assessment of indigenous rhizobia in Portugal.     

Biogeography of Wood-Boring Crustaceans (Isopoda: Limnoriidae) Established in European Coastal Waters

Marine wood-borers of the Limnoriidae cause great destruction to wooden structures exposed in the marine environment. In this study we collated occurrence data obtained from field surveys, spanning over a period of 10 years, and from an extensive literature review. We aimed to determine which wood-boring limnoriid species are established in European coastal waters; to map their past and recent distribution in Europe in order to infer species range extension or contraction; to determine species environmental requirements using climatic envelopes. Of the six species of wood-boring Limnoria previously reported occurring in Europe, only Limnoria lignorum, L. quadripunctata and L. tripunctata are established in European coastal waters. L. carinata and L. tuberculata have uncertain established status, whereas L. borealis is not established in European waters. The species with the widest distribution in Europe is Limnoria lignorum, which is also the most tolerant species to a range of salinities. L. quadripunctata and L. tripunctata appear to be stenohaline. However, the present study shows that both L. quadripunctata and L. tripunctata are more widespread in Europe than previous reports suggested. Both species have been found occurring in Europe since they were described, and their increased distribution is probably the results of a range expansion. On the other hand L. lignorum appears to be retreating poleward with ocean warming. In certain areas (e.g. southern England, and southern Portugal), limnoriids appear to be very abundant and their activity is rivalling that of teredinids. Therefore, it is important to monitor the distribution and destructive activity of these organisms in Europe.