Removal of wastewater pathogen indicators in a constructed wetland in Leon, Spain

This paper studies the seasonal presence and removal of the pathogenous micro-organisms Escherichia coli, total coliforms (TC), Clostridium perfringens (Cp), faecal streptococci (FS), Giardia cysts, Cryptosporidium oocysts and helminth eggs, in a constructed wetland treatment system. The removal efficiency of this system with respect to the indicator micro-organisms achieved maximum values in spring and autumn at 99.9% for E. coli and TC, respectively, in winter at 97.0% for FS, in summer at 100% for Clostridium and throughout the year, also at 100%, in the case of Giardia cysts, Cryptosporidium oocysts and helminth eggs. In general, very low protozoan and helminth egg counts were found, and the system demonstrated efficient reduction of the wastewater indicator pathogens.     

New species and new combinations in Peruvian Orchidaceae

seventeen new species and combinations are proposed in the generaChondrorhyncha, Cischweinfia, Cochlioda, Eloyella, Encyclia, Kefersteinia, Koellensteinia, Macroclinium, Rodriguezia, Solenidiopsis, andStenia. All new species are illustrated. A key is provided for 2-flowered species ofMacroclinium, PeruvianSigmatostalix, and PeruvianStenia. Solenidium (Solenidiopsis) peruvianum Schltr. is lectotypified.     

Phylogenetic relationships in theCampanulales based onrbcL sequences

Phylogenetic relationships within the angiosperm orderCampanulales were investigated by comparative sequencing of the chloroplast generbcL. CompleterbcL sequences were obtained for ten species in six families within the order. These data were analyzed along with previously publishedrbcL sequences from other taxa (for a total of 117 species) within the subclassAsteridae and outgroups, producing 32 equally parsimonious trees. A subset consisting of 44 of these taxa was then chosen and more rigorous analyses performed, resulting in four equally parsimonious trees. Results indicate that two major clades roughly corresponding to traditionally circumscribedAsterales andCampanulales exist as sister taxa. In particular, therbcL trees indicate thatSphenoclea is not a member ofCampanulales orAsterales, thatPentaphragma is more closely allied toAsterales thanCampanulales, that theCyphiaceae are not monophyletic, thatCampanulaceae andLobeliaceae are not sister taxa, and thatStylidiaceae are correctly placed withinCampanulales.     

Inbreeding in a french peasant population

Genealogical pedigrees were collected from 976 farming couples who lived in a canton of central France in the XIXth and XXth century. Several methods (genealogical, isonymic and probability of origin of genes) were used to appraise the evolution of the biological structure of this population, which has gone through two phases. The first phase from 1800 to the World War I, shows gene flows between the farming population of the canton and those of neighbouring rural districts which are linked to an increase in consanguineous marriages. The second phase, from 1920 onwards, shows important gene flow from distant areas. This trend implies a decrease in consanguineous marriages.     

Analysis of twin of eyeless regulation during early embryogenesis in Drosophila melanogaster

The Drosophila Pax6 homolog twin of eyeless (toy) is so far the first zygotically expressed gene involved in eye morphogenesis in Drosophila. The study of its expression during embryogenesis is therefore informative of the initial events of eye development in Drosophila. We have analyzed how the initial expression domain of toy at cellular blastoderm is regulated. We show that the three maternal patterning systems active in the cephalic region (the anterior, terminal and dorsal-ventral systems) cooperate with zygotically activated gap genes to shape the initial expression domain of toy. Whereas Bicoid, Dorsal and Torso signaling synergistically act as activators, Hunchback, Knirps and Decapentaplegic act as repressors.     

Vermicomposting in the management of pig-waste in Hong Kong

The treatment and disposal of pig-waste in Hong Kong has received much attention in recent years but, following any of the presently used treatment processes, solids remain to be further stabilized. Vermicomposting is a waste stabilization technique which converts waste into potentially recyclable materials such as worm protein and worm casts. The earthworm, Pheretima asiatica, can stabilize most of the solids arising from the treatment of pig-waste, including raw pig manure, suggesting that vermicomposting has a high potential as a unit process in the management of pig-waste in Hong Kong.S.H. Wong is with the Environmental Protection Department, Hong Kong; and D.A. Griffiths is with the Department of Botany, University of Hong Kong, Hong Kong.     

Changes of dung beetle communities from rainforests towards agroforestry systems and annual cultures in Sulawesi (Indonesia)

Little is known about how tropical land-use systems contribute to the conservation of functionally important insect groups, including dung beetles. In a study at the margin of Lore Lindu National Park (a biodiversity hotspot in Central Sulawesi, Indonesia) dung-beetle communities were sampled in natural forest, young secondary forest, agroforestry systems (cacao plantations with shade trees) and annual cultures (maize fields), each with four replicates (n = 16 sites). At each site we used 10 pitfall traps, baited with cattle dung, along a 100 m transect for six 3-day periods. The number of trapped specimens and species richness at the natural forest sites was higher than in all land-use systems, which did not significantly differ. Each land-use system contained, on average, 75% of the species richness of the natural forest, thereby indicating their importance for conservation. However, a two-dimensional scaling plot based on NESS indices (m = 6) indicated distinct dung beetle communities for both forest types, while agroforestry systems and annual cultures exhibited a pronounced overlap. Mean body size of dung beetles was not significantly influenced by land-use intensity. Five of the six most abundant dung beetle species were recorded in all habitats, whereas the abundance of five other species was significantly related to habitat type. Mean local abundance and number of occupied sites were closely correlated, further indicating little habitat specialisation. The low dung beetle diversity (total of 18 recorded species) may be due to the absence of larger mammals in Sulawesi during historical times, even though Sulawesi is the largest island of Wallacea. In conclusion, the dung beetle fauna of the lower montane forest zone in Central Sulawesi appears to be relatively robust to man-made habitat changes and the majority of species did not exhibit strong habitat preferences.     

Genetics of esterase isoenzymes in Malus

Summary Three main zones of esterase activity (EST-I, EST-III, EST-IV) identified in leaf extracts of cultivated apple and Malus species were determined by the genes EST-1, EST-3 and EST-4, respectively. In addition to earlier reported alleles of EST-1 (a, b) three further bands c, d and f were identified in the EST-I zone of which c was found to be determined by an allele, c. Two alleles, a, b, and a null allele were found for both the genes EST-3 and EST-4. Differences in allelic frequency were observed between cultivars, rootstocks and Malus species. Allele EST-1a was rare amongst the rootstocks. The examination of Malus species and derivatives showed a geographical relationship. Allele EST-1c was confined to species of Asian origin, and EST-1d was confined to American species.     

Comparison between the allergenic airborne pollen in Trieste and at Lozzo di Cadore (Italy) in 1989

Summary A comparative study was carried out between aeroallergenic pollen spectra in Trieste and at Lozzo di Cadore. The two localities were found to be different on a five-day running mean basis as to the 8 pollen taxa monitored by the National Aerobiological Monitoring Network managed by the Italian Association of Aerobiology (AIA). In the zone of Lozzo the pollination curve of the spring flowering taxa showed a lag of about one month due to the colder climate. Lozzo di Cadore showed a poor airborne pollen content,Corylus and Graminaceae being prevailing. Trieste has a higher airborne pollen diversity and longer pollination times because of its higher floristic and vegetational complexity. In Trieste the allergenic pollen spectrum showed great quantities ofOstrya, Quercus, Oleaceae, Graminaceae and of ruderal taxa, widespread over the area, due to man's influence.     

Callus,shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants

We analyzed the impact of ethylene and auxin disturbances on callus, shoots and Agrobacterium rhizogenes-induced hairy root formation in tomato (Solanum lycopersicum L.). The auxin low-sensitivity dgt mutation showed little hairy root initiation, whereas the ethylene low-sensitivity Nr mutation did not differ from the control Micro-Tom cultivar. Micro-Tom and dgt hairy roots containing auxin sensitivity/biosynthesis rol and aux genes formed prominent callus onto media supplemented with cytokinin. Under the same conditions, Nr hairy roots did not form callus. Double mutants combining Rg1, a mutation conferring elevated shoot formation capacity, with either dgt or Nr produced explants that formed shoots with little callus proliferation. The presence of rol + aux genes in Rg1 hairy roots prevented shoot formation. Taken together, the results suggest that although ethylene does not affect hairy root induction, as auxin does, it may be necessary for auxin-induced callus formation in tomato. Moreover, excess auxin prevents shoot formation in Rg1.     

Incubation temperature critical to successful stimulation of in vitro zygotic embryo growth in four Australian native Cyperaceae species

Many species of Western Australian Cyperaceae (sedges) are vital components of the indigenous flora but commonly display low seed set, poor seed quality and intractable seed dormancy. We report the effects of incubation temperature and in vitro growth media on whole seed germination compared with extracted zygotic embryo growth in Tetraria capillaris, T. octandra, Lepidosperma drummondii and L. tenue. No germination was observed from intact whole seeds of all test species regardless of the treatment evaluated. In contrast, excised zygotic embryos of all study species exhibited significant increases in growth when cultured at 15°C compared to embryos incubated at 25°C; however, optimal media for embryo growth were genera specific. Extracted embryos of T. capillaris and T. octandra exhibited maximum percentage growth (30 and 40%, respectively) at 15°C on ½ MS medium with no plant growth regulators required. In the case of L. drummondii and L. tenue 1 μM thidiazuron was a necessary addition to the ½ MS medium resulting in 40 and 77% growth of embryos (at 15°C), respectively. Incubation of extracted embryos at 25°C (regardless of medium treatment) resulted in <10% embryo growth for T. octandra and L. tenue, while the remaining two species (L. drummondii, T. capillaris) showed no embryo growth at 25°C on any medium treatment.     

Nectary structure and nectar characteristics in someBignoniaceae

The nectary structure and chemical nectar composition of 15 species belonging to 12 genera ofBignoniaceae are analyzed. All taxa bear a conspicuous nuptial nectary surrounding the ovary base. The secretory tissue is mostly supplied by phloem branches. The stomata are located in the middle and upper part of the nectary epidermis with an homogeneous distribution. The nuptial nectary is proportionally large in relation to the ovary (15–30%), disregarding the nectary volume. Most species have extranuptial nectaries in both inner and outer surfaces of the calyx. Both kinds of nectaries lack a vascular tissue that straightly supplies them. Nuptial nectar concentration (wt/wt) ranges from 19 to 68%. Sugars and amino acids are found in all species. Half of the species have hexose predominant nectars, the remaining sucrose predominant. Phenols are detected in only three species, whereas reducing acids exclusively inTecoma stans. Alkaloids and lipids were never detected. Extranuptial nectar chemical composition is analyzed in two species:Dolichandra cynanchoides andPodranea ricasoliana. Bees constitute the main flower visitors of the species studied whereas hummingbirds were seen visiting three species. A correlation analysis is performed with the data obtained. There are a few significant correlations which indicate a parallel increase of three parameters: the longer the flower length, the more voluminous the nectary and the higher stomata number, independently of the floral biotype. Phenograms are obtained using 24 floral characters including nectary and nectar data. The clusters obtained do not reflect taxonomic relationships but are useful in the understanding of animal-plant interactions when the flower biotype is considered.This paper is based on a chapter of a doctoral thesis presented at the University of Córdoba (Argentina).     

Thioglycosides as glycosylating agents in oligosaccharide synthesis

A review is given of the use of thioglycosides as glycosyl donors in oligosaccharide synthesis. Both indirect use, by conversion of the thioglycoside into a glycosyl halide and direct use, by electrophilic activation of the thioglycoside, are discussed.Abbreviations DMTST dimethyl(methylthio)sulfonium trifluoromethanesulfonate - Bz benzoyl - Bn benzyl - pNBz p-nitrobenzoyl - Phth phthallyl - Ph phenyl     

Adaptive ecology of traditionally derived agroforestry in China

This research was designed to test the following hypothesis: Among traditional (nonliterate and minimally literate) people residing in their home environment, traditionally derived knowledge of specific cases of ecological processes can approximate scientifically derived knowledge of those same cases. After locating three minimally literate peasant family lineages in Lijiayang and Linfengkeng villages, Shouning County, Fujian Province, China, not applying science-based systems of agroforest management, a decision-tree model of an indigenous agroforest management system was constructed from extensive interviews. The system was centered around shamu (Cunninghamia lanceolata),an important timber species with a long history of management in China. A major part of the management system is agricultural intercropping with a wide variety of cereal, cash, medicinal, and oil-producing crops. Intercropping is practiced in the initial phases of afforestation; is an integral aspect of site preparation and the tending of the young stand; and contributes to tree establishment, growth, and survival. The cases presented demonstrate that local peasant knowledge of the effects of burning, site preparation, crop selection, crop tending, and intercropping duration on soil quality and the survival and development of shamu is strongly analogous to the knowledge derived through systematic scientific research. This process of deriving scientifically valid ecological knowledge through traditional means is called protoscience.     

Intralineage variation in the pattern ofrbcL nucleotide substitution

Variation in chloroplastrbcL sequences was studied in representative species of four different lineages: the tribeRubieae (Rubiaceae), and the generaDrosera (Droseraceae),Nothofagus (Nothofagaceae) andIlex (Aquifoliaceae). Each lineage has its particular non-overlapping set ofrbcL polymorphic sites, indicating that common unconstrainedrbcL sites are not shared. Large differences in the rate and pattern of nucleotide substitution are observed among the four lineages. The genusIlex has the lowest rate of substitution, the lowest transition/transversion ratio, the lowest synonymous/replacement ratio and the lowest number of substitutions at the third codon position. An apparent relationship of these measures to the age of the lineages is observed. The A + T content and codon use among the four lineages are very similar and, apparently, cannot account for the observed differences in patterns of nucleotide substitution. However, the A + T content of the two bases immediately flanking the polymorphic sites is higher inIlex than in the other lineages. This could be correlated with the transversion/transition bias observed inIlex. The particularly low synonymous/replacement ratio found inIlex could also be explained by the small population sizes of species in this genus.     

Plant wound compounds from oviposition scars used in host discrimination by a stem-galling sawfly

The stem-galling sawfly Euura lasiolepisuses one or more plant wound compounds resulting from oviposition scars as cues in host discrimination (avoiding sites occupied by conspecifics). Four experiments were conducted to test hypotheses about how Euurapartitions resources. Experiment 1 demonstrated that Euuraavoids ovipositing on nodes with scars from previous ovipositions. Experiment 2 showed no evidence that the sawfly uses oviposition-deterring pheromones and indicated there is a time lag following oviposition before the oviposition scar becomes a deterrent. Experiment 3 showed that sawflies avoid artificially formed scars, demonstrating that a plant cue alone can lead to host discrimination. Experiment 4 showed that visual or tactile cues are not necessary for host discrimination and indicated that a plant wound compound functions as an oviposition deterrent. Both experimental results and field surveys showed that Euuraoviposition scars were more uniformly distributed than expected if sawflies were ignoring previous ovipositions.     

Biosynthesis of glycoconjugates in mitochondrial outer membranes

The results reported in this paper show two distinct ways for the incorporation ofN-acetylglucosamine into mitochondrial outer membranes. The first one is the glycosylation of dolichol acceptors, which is indicated by the inhibition of the synthesis of these products by the inhibitors of the dolichol intermediates (tunicamycin and GDP). The second one is the incorporation ofN-acetylglucosamine into protein acceptors directly from UDP-N-acetylglucosamine. This second way of glycosylation is only localized in mitochondria outer membranes.The existence of a direct route forN-glycoprotein biosynthesis has been based on the following evidence. First, the synthesis of theN-acetylglucosaminylated protein acceptors was not inhibited by tunicamycin or GDP. Second, the addition of exogenous dolichol-phosphate did not change the rate of biosynthesis of glycosylated protein material. Third, the sequential incorporation ofN-acetylglucosamine and mannose from their nucleotide derivatives in the presence of GDP and tunicamycin led to the synthesis of glycosylated protein material which entirely bound to Concanavalin A-Sepharose. The oligosaccharide moiety of the glycosylated protein material resulting from the direct transfer of sugars from their nucleotide derivatives to the protein acceptor is of theN-glycan type. On sodium dodecylsulphate polyacrylamide gel electrophoresis, this glycosylated material migrated as a marker protein with a molecular weight between 45 000 and 63 000. HPLC chromatofocusing analysis revealed that the fraction studied was anionic. The oligosaccharide moiety of the glycoprotein material can only be elongated by the incorporation ofN-acetylglucosamine and galactose from their nucleotide derivatives.     

Synthetic oligonucleotide probes for detection of mercury-resistance genes in environmental freshwater microbial communities in response to pollutants

Mercury-resistance genes were detected byin situ hybridization using new synthetic oligonucleotide probes specific formerA andmerB genes according to the published sequences of the corresponding enzymes. These DNA probes were used for the detection of specific mercury-resistant microorganisms isolated from the Rhine River which had been polluted 3 years previously in 1986. Mercuric reductase and organomercurial lyase genes persist in the bacterial genome even after the disappearance of the pollutant but are absent in axenic amoebae. A total of 49 bacterial isolates showed DNA homologies with the³²P-labelled DNA probes and 15 free-living amoebae were selected due to their harboured symbiotic mercury-resistant bacteria.     

Tracing of labile zinc in live fish hepatocytes using FluoZin-3

Intracellular zinc levels are homeostatically regulated and although most is bound, a pool of labile Zn(II) is present in cells. We show here that the zinc probe FluoZin-3 is useful to monitor zinc fluxes during fluorescent imaging of the trout hepatic cell line D11. Nuclei and bulk cytosol appeared to lack detectable labile zinc, while the punctuate staining pattern colocalized with a lysosome-specific probe. Applying extracellular zinc alone resulted in vesicular sequestration of the metal ion. Together with Na-pyrithione a delayed and toxic rise in cellular fluorescence was triggered. When using another ionophore, 4-Br A23187, a zinc buffering effect of the vesicular pools was evident. Secondly, N-ethylmaleimide induced a homogeneous fluorescence rise, which was strongly enhanced by addition of Zn-pyrithione and disappeared after TPEN washing. This suggests the involvement of thiol residues in controlling available cytosolic zinc. Our observations have implications for the interpretation of calculated intracellular Zn²⁺ concentrations.