Relationships among filamentous microorganisms in rotating biological contactors

The existing relationships were studied among the different types of filamentous microorganisms that appear in the biofilm of a biological contactor system. Using the hierarchical cluster analysis it was observed that, in all the stages, Beggiatoa sp. and the Eikelboom’s types 0803 and 1863 always appeared associated, while Sphaerotilus natans was always associated with the morphological type 021N. The remaining microorganisms were associated in variable forms in the plants. In addition, different association models were obtained according to the season of the year and the stage-season interaction. It has also been observed that a significant correlation exists among the filamentous microorganisms we have studied and the different physical-chemical parameters.     

Isolation and characterization of filamentous bacteria present in bulking activated sludge

Summary Several types of filamentous microorganisms were observed and identified in samples of poorly settling (bulking) activated sludge. The major types encountered and the frequency (percentage) of appearance in the total of all treatment plants sampled were: Eikelboom type 0041 (60), type 1701 (45), Haliscomenobacter hydrossis (35), type 021N (30), Thiothrix spp. (20), and Sphaerotilus natans (20). Isolation techniques and culture media were developed and used to recover 42 axenic strains of filamentous bacteria from sludge samples collected. The isolates were identified as strains of Thiothrix, Beggiatoa, S. natans, and Eikelboom types 021N, 1701, 0041, and 0803. Nutritional and differential characterization of the bacteria was important to the differentiation of groups which could not be easily distinguished on the basis of morphology. Although certain treatment plant operating parameters (organic loading) seemed associated with the presence of specific filamentous organism types, possible interaction among factors precluded definite establishment of a cause and effect relationship for most of the treatment plant characteristics and organisms observed.     

Genetic Diversity in Musa acuminata Colla and Musa balbisiana Colla and some of their natural hybrids using AFLP Markers

Genetic diversity and relationships were assessed in 28 accessions of Musa acuminata (AA) Colla and Musa balbisiana (BB) Colla, and some of their natural hybrids, using the amplified fragment length polymorphisms (AFLP) technique. Fifteen AFLP +3 primer pairs produced 527 polymorphic bands among the accessions. Neighbor-joining and principal co-ordinate (PCO) analyses using Jaccard's similarity coefficient produced four major clusters that closely corresponded with the genome composition of the accessions (AA, BB, AAB and ABB). The AFLP data distinguished between the wild diploid accessions and suggested new subspecies relationships in the M. acuminata complex that are different from those based on morphological data. The data suggested that there are three subspecies within the M. acuminata complex (ssp. burmannica Simmonds, malaccensis Simmonds, and microcarpa Simmonds). 'Tjau Lagada' (ssp. microcarpa), 'Truncata' [ssp truncata (Ridl.) Shepherd] and 'SF247' [ssp. banksii (F.Muell) Simmonds] clustered very closely with 'Gros Michel' and 'Km 5', indicating that more than one M. acuminata subspecies may be involved in the origin of triploid AAA bananas. 'Calcutta 4' (ssp. burmannicoides De Langhe &; Devreux) and 'Long Tavoy' (ssp. burmannica) were closely related and could be together in the same subspecies. This study also showed that there is much more genetic diversity within M. balbisiana that was split into two groups: (1) 'I-63' and 'HND' and (2) 'Los Banos', 'MPL' (Montpellier), '10852', 'Singapuri', 'Etikehel', and 'Butohan 1' as the other.     

Narrow hybrid zone between two subspecies of big sagebrush (Artemisia tridentata: Asteraceae)

Several species of gall-forming insects specialize on big sagebrush (Artemisia tridentata), a species that shows much clinal and subspecific variation throughout its geographic range. Two of those subspecies, basin big sagebrush (A. t. ssp. tridentata) and mountain big sagebrush (A. t. ssp. vaseyana), form a narrow hybrid zone at Salt Creek, Utah. Reciprocal transplant experiments have shown that the hybrid big sagebrush at Salt Creek are more fit than either parental subspecies, but only in the hybrid zone. Do genotype and environment influence the density and distribution of galls on big sagebrush? We counted galls on parental and hybrid big sagebrush in three reciprocal transplant gardens at Salt Creek. Gardens were in each of the two parental zones and in the hybrid zone. Transplanted seedlings came from five source populations: two parental and three hybrid populations. We identified seven kinds of gall-forming flies (Rhopalomyia midges and Eutreta fruitflies) that produced identifiable galls. Densities of galls varied among the three gardens and five source populations, and there was also a significant garden by source interaction in gall density. In general, variation in gall density among gardens (i.e., environments) was much greater than the variation among source populations (i.e., genotypes). Nevertheless, significant genotype-environment interactions were observed for five of the seven kinds of galls. Overall density of galls, mostly due to Rhopalomyia ampullaria, was greatest in the high-elevation (mountain) garden and least in the low-elevation (basin) garden. To the best of our knowledge, this is the first reciprocal transplant experiment addressing herbivore richness in a hybrid zone.     

A novel method of enumerating Nocardia amarae in foaming activated sludge

Various experimental investigations were carried out in order to develop a novel method of enumerating Nocardia amarae in foaming activated sludge using n-octadecane as a carbon source (OD medium). Colonies of N. amarae isolate appeared about 3 d earlier on OD medium than on MS medium. N. amarae counts on OD medium with 20 mg/l of nalidixic acid, using both synthetic and practical activated sludges, were always higher than those on MS medium. It is concluded that OD medium with nalidixic acid was more effective for the count of N. amarae in activated sludge than MS medium.     

Insects as vectors of plant pathogens: mutualistic and antagonistic interactions

Interactions between plants and their herbivores and pathogens are mostly analysed separately, thereby neglecting mutualistic or antagonistic interactions between these antagonists and possible joint effects on the host. We studied interactions between the weed Cirsium arvense, the rust fungus Puccinia punctiformis and three herbivorous insects, the aphids Aphis fabae ssp. cirsiiacanthoidis and Uroleucon cirsii, and the beetle Cassida rubiginosa. All three insect species mechanically transported spore material and significantly increased rates of P. punctiformis infection in healthy thistles. The interaction between C. rubiginosa and the fungus was antagonistic. Although C. rubiginosa transferred spores, biomass of adults was significantly reduced, development of adults tended to be prolonged and mortality increased when feeding on plants infected with P. punctiformis. In contrast, the relationship between the aphid U. cirsii and P. punctiformis was mutualistic: U. cirsii profited by fungal infection and formed significantly larger colonies on fungus-infected plants. Although the differences in insect performance suggest that aphids may be better vectors than the beetle, infection rates were similar. This is the first study to demonstrate that the relationship between herbivores, which increase the dispersal of a pathogen, and the pathogen itself can be mutualistic or antagonistic, depending on the species.     

Influence of forest type and tree species on canopy ants (Hymenoptera: Formicidae) in Budongo Forest, Uganda

The arboreal ant fauna was investigated in Budongo Forest, a seasonal rain forest in Uganda, using the insecticidal fogging technique. Ants were collected from 61 trees, between 7 and 33 m in height, belonging to four tree species. Trees were growing in adjacent plots of forests characterized by different use and structure: an old primary forest, a primary swamp forest along a small river, and a secondary forest where selective logging was carried out for 30 years. A total number of 37,065 ants, belonging to 161 species in 30 genera were collected. Considering the high number of species found only once, the completeness of the canopy ant fauna was relatively high and of relatively similar magnitude as samples from the Neotropics or the Oriental region. Up to 37 ant species on a single tree, with an average of 18.2 species per tree, were found. Forty-four ant species (28.1%) were found only once, less than ten individuals were found for each of 88 species (54.7%), but 64.0% of all individuals belonged to one of five species. Considering the high numerical dominance of a few ant species like a Pheidole sp., Tetramorium aculeatum (Mayr) and a Crematogaster sp., there is some evidence for an ant mosaic in the lower canopy of the Budongo Forest. Individual numbers of ants were strongly correlated with nests in the fogged tree, though the ants were not homogeneously distributed in the tree crowns. Diversity measures that strongly depend on individual numbers such as the Morisita-Horn index or rarefaction methods were calculated, but results were not concordant with those of incidence-based estimates such as jack-knife calculations. Differences in ant species richness and faunal composition between tree species were low, but more significant between forest types. The ant fauna in the secondary forest was less diverse with 12.6% fewer species compared to the primary forest sites. The average number of ant species per tree was significantly lower in the secondary forest (<20% of the species; F=8.03, df=59, P<0.01) than in the undisturbed forest types. Cataulacus, Leptothorax, Tetraponera, and Polyrhachis, which are typical canopy-dwelling ant genera, had a significantly higher diversity and frequency in the two primary forest types (F=4.17, df=53, P<0.05). Secondary forest trees are often younger, lacking dead branches and epiphytes which are important requisites for ant colonization on trees.     

Effects of high nitrogen load on growth, photosynthesis and nutrient status of Cryptomeria japonica and Pinus densiflora seedlings

To evaluate the sensitivity of Japanese cedar (Cryptomeria japonica D. Don) and Japanese red pine (Pinus densiflora Sieb. et Zucc.) to high N deposition, 1-year-old seedlings were grown in brown forest soil treated with N as NH₄NO₃ at 0, 25, 50, 100 and 300 mg l^-1 fresh soil volume, equivalent to 0, 28, 57, 113 and 340 kg N ha^-1. Net photosynthetic rate and whole-plant dry mass of C. japonica seedlings were increased by the N treatment, whilst those of P. densiflora seedlings were significantly reduced by the highest N treatment. The reduction in the net photosynthesis of P. densiflora seedlings was mainly due to a depression of carboxylation efficiency accompanied by a decrease in concentration and activity of Rubisco in the needles. In P. densiflora seedlings, needle concentrations of P and Mg were decreased, and the concentrations of N and Mn were increased by the highest N treatment. The reductions in needle protein concentration and Rubisco activity were negatively correlated with the ratios of N/P and Mn/Mg in the needles. These results suggest that nutrient imbalances of these elements may be induced in P. densiflora seedlings grown under high N deposition. We conclude that P. densiflora is more sensitive to high N deposition than C. japonica, and that the relatively high atmospheric N deposition to Japanese forest ecosystems may adversely affect the health of N-sensitive tree species such as P. densiflora.     

Elevated CO2 influences nutrient availability in young beech-spruce communities on two soil types

The objectives of this study were to investigate how different soil types and elevated N deposition (0.7 vs 7 g N m^-2a^-1) influence the effects of elevated CO₂ (370 vs 570 µmol CO₂ mol^-1) on soil nutrients and net accumulation of N, P, K, S, Ca, Mg, Fe, Mn, and Zn in spruce (Picea abies) and beech (Fagus sylvatica). Model ecosystems were established in large open-top chambers on two different forest soils: a nutrient-poor acidic loam and a nutrient-rich calcareous sand. The response of net nutrient accumulation to elevated atmospheric CO₂ depended upon soil type (interaction soil 2 CO₂, P<0.05 for N, P, K, S, Ca, Mg, Zn) and differed between spruce and beech. On the acidic loam, CO₂ enrichment suppressed net accumulation of all nutrients in beech (P<0.05 for P, S, Zn), but stimulated it for spruce (P<0.05 for Fe, Zn) On the nutrient-rich calcareous sand, increased atmospheric CO₂ enhanced nutrient accumulation in both species significantly. Increasing the N deposition did not influence the CO₂ effects on net nutrient accumulation with either soil. Under elevated atmospheric CO₂, the accumulation of N declined relative to other nutrients, as indicated by decreasing ratios of N to other nutrients in tree biomass (all ratios: P<0.001, except the N to S ratio). In both the soil and soil solution, elevated CO₂ did not influence concentrations of base cations and available P. Under CO₂ enrichment, concentrations of exchangeable NH₄⁺ decreased by 22% in the acidic loam and increased by 50% in the calcareous sand (soil 2 CO₂, P<0.001). NO₃^- concentrations decreased by 10-70% at elevated CO₂ in both soils (P<0.01).     

The diet of the South Georgia shag Phalacrocorax georgianus at South Orkney Islands in five consecutive years

The diet of the South Georgia shag Phalacrocorax georgianus at Laurie Island, South Orkney Islands, was studied by the analysis of 420 pellets (regurgitated casts) collected from the 1995 to 1999 breeding seasons. Demersal-benthic fish were by far the main prey, followed by molluscs (mainly octopods and bivalves) and polychaetes. Harpagifer antarcticus (in 1995, 1998, 1999) and Lepidonotothen nudifrons (1996, 1997) were the most frequent prey, followed by Gobionotothen gibberifrons or Trematomus newnesi. Harpagifer antarcticus, followed by L. nudifrons or T. newnesi, was the most numerous prey in all of the seasons and predominated in mass in 1995 and 1996, followed by Notothenia coriiceps and L. nudifrons. In 1997 and 1999, G. gibberifrons and N. coriiceps were the species that most contributed to the diet whereas N. coriiceps and H. antarcticus contributed most in 1998. Our results differ from those reported for the South Orkney Islands in previous studies. These differences could be due to the use of different diet-analysis methods and to shags-related and/or fisheries-related changes in the food availability around the colonies. These alternative explanations are analysed and discussed.     

Stable isotope enrichment (δ15N and δ13C) in a generalist predator (Pardosa lugubris, Araneae: Lycosidae): effects of prey quality

Analysis of the natural variations in stable isotope ratios in animal tissue may be a powerful tool to reveal the trophic position and feeding preferences of generalist predators such as lycosid spiders. In the present study, changes in ¹⁵N/¹⁴N and ¹³C/¹²C ratios in the lycosid spider Pardosa lugubris (Araneae: Lycosidae), fed with prey of different quality, were investigated. Experimental food chains included three trophic levels: prey media, prey organism and predator. In order to analyse the time course of stable isotope enrichment, different life stages of P. lugubris feeding on Drosophila melanogaster were studied. The ¹⁵N content of hatchlings of P. lugubris was significantly lower than that of their mothers, indicating the existence of nitrogen pools with different ¹⁵N signatures in female P. lugubris. With duration of feeding and progressing development, age and body weight, the ¹⁵N content in spiderlings increased. Starvation resulted in ¹⁵N and ¹³C enrichment in P. lugubris. Being fed with prey of different quality resulted in varying patterns of stable isotope enrichment. ¹⁵N but not ¹³C content consistently increased when fed on high quality prey (Heteromurus nitidus, D. melanogaster and a mixed diet consisting of H. nitidus and D. melanogaster). In contrast, low quality prey (Rhopalosiphum padi, Folsomia candida and a mixed diet consisting of H. nitidus, D. melanogaster and F. candida) resulted in deviations from postulated patterns of stable isotope enrichment with ¹⁵N content being similar to that of starving spiderlings. When fed on high quality prey, ¹⁵N enrichment of P. lugubris was close to ca. 3‰. It is concluded that the analysis of variations in the natural abundance of stable isotopes is particularly helpful in revealing the trophic structure of terrestrial food webs in which polyphagous feeders predominate, as is the case in litter and soil.     

Fungi isolated from Antarctic mosses

Microfungi were isolated from different moss species in Victoria Land. Twenty-eight taxa belonging to 18 genera were identified. New records for continental Antarctica were: Arthrobotrys superba, Conidiobolus sp., Penicillium minioluteum, Verticillium psalliotae and V. lamellicola. The most frequently isolated fungal species were: Cladosporium cladosporioides, Cryptococcus albidus, Cryptococcus laurentii, Geomyces pannorum var. pannorum, G. pannorum var. vinaceus, Mortierella antarctica, Cadophora malorum, Phoma herbarum and V. lecanii. Bryum pseudotriquetrum was the moss richest in fungal species. Within the Antarctic environment, moss is one of the microhabitats richest in microfungi, particularly in psychrophilic indigenous species.     

Quantification of Gordona amarae Strains in Foaming Activated Sludge and Anaerobic Digester Systems with Oligonucleotide Hybridization Probes

Previous studies have shown the predominance of mycolic acid-containing filamentous actinomycetes (mycolata) in foam layers in activated sludge systems. Gordona (formerly Nocardia) amarae often is considered the major representative of this group in activated sludge foam. In this study, small-subunit rRNA genes of four G. amarae strains were sequenced, and the resulting sequences were compared to the sequence of G. amarae type strain SE-6. Comparative sequence analysis showed that the five strains used represent two lines of evolutionary descent; group 1 consists of strains NM23 and ASAC1, and group 2 contains strains SE-6, SE-102, and ASF3. The following three oligonucleotide probes were designed: a species-specific probe for G. amarae, a probe specific for group 1, and a probe targeting group 2. The probes were characterized by dissociation temperature and specificity studies, and the species-specific probe was evaluated for use in fluorescent in situ hybridizations. By using the group-specific probes, it was possible to place additional G. amarae isolates in their respective groups. The probes were used along with previously designed probes in membrane hybridizations to determine the abundance of G. amarae, group 1, group 2, bacterial, mycolata, and Gordona rRNAs in samples obtained from foaming activated sludge systems in California, Illinois, and Wisconsin. The target groups were present in significantly greater concentrations in activated sludge foam than in mixed liquor and persisted in anaerobic digesters. Hybridization results indicated that the presence of certain G. amarae strains may be regional or treatment plant specific and that previously uncharacterized G. amarae strains may be present in some systems.     

Feeding strategies and diet composition of four Antarctic cnidarian species

The diet of four species of Antarctic cnidarians, two hydroids and two anthozoans, was investigated. One hydroid, Tubularia ralphii, and one anthozoan, Anthomastus bathyproctus, seem to basically consume zooplankton whereas the other hydroid species, Oswaldella antarctica, has a diet mainly based on the fine fraction of seston. The last cnidarian investigated, the stoloniferan alcyonacean, Clavularia cf. frankliniana, feeds mainly on resuspended material. The wide range of diets of these Antarctic cnidarians indicates opportunistic behaviour by feeding on different sources and taking advantage of the available food sources. Data related to low C/N ratios in the sediment and high NO₂ concentrations in areas with dense communities of benthic suspension feeders, along with data on capture rates, lead us to hypothesize that these organisms play an important role in the recycling processes of organic matter in Antarctic benthic ecosystems.     

How plants shape the ant community in the Amazonian rainforest canopy: the key role of extrafloral nectaries and homopteran honeydew

Ant-plant interactions in the canopy of a lowland Amazonian rainforest of the upper Orinoco, Venezuela, were studied using a modified commercial crane on rails (Surumoni project). Our observations show a strong correlation between plant sap exudates and both abundance of ants and co-occurrence of ant species in tree canopies. Two types of plant sap sources were compared: extrafloral nectaries (EFNs) and honeydew secretions by homopterans. EFNs were a frequent food source for ants on epiphytes (Philodendron spp., Araceae) and lianas (Dioclea, Fabaceae), but rare on canopy trees in the study area, whereas the majority of trees were host to aggregations of homopterans tended by honeydew-seeking ants (on 62% of the trees examined). These aggregations rarely occurred on epiphytes. Baited ant traps were installed on plants with EFNs and in the crowns of trees from three common genera, including trees with and without ant-tended homopterans: Goupia glabra (Celastraceae), Vochysia spp. (Vochysiaceae), and Xylopia spp. (Annonaceae). The number of ant workers per trap was significantly higher on plants offering one of the two plant sap sources than on trees without such resources. Extrafloral nectaries were used by a much broader spectrum of ant species and genera than honeydew, and co-occurrence of ant species (in traps) was significantly higher on plants bearing EFNs than on trees. Homopteran honeydew (Coccidae and Membracidae), on the other hand, was mostly monopolised by a single ant colony per tree. Homopteran-tending ants were generally among the most dominant ants in the canopy. The most prominent genera were Azteca, Dolichoderus (both Dolichoderinae), Cephalotes, Pheidole, Crematogaster (all Myrmicinae), and Ectatomma (Ponerinae). Potential preferences were recorded between ant and homopteran species, and also between ant-homopteran associations and tree genera. We hypothesize that the high availability of homopteran honeydew provides a key resource for ant mosaics, where dominant ant colonies and species maintain mutually exclusive territories on trees. In turn, we propose that for nourishment of numerous ants of lower competitive capacity, Philodendron and other sources of EFNs might be particularly important.     

In vitro germination and establishment of tissue cultures of Bulbine caulescens and of two Kniphofia species (Asphodelaceae)

A protocol for the in vitro germination and the establishment of tissue cultures of Bulbine caulescens Linn., Kniphofia pumila Kunth, and K. uvaria Hook has been developed. The members of the family Asphodelaceae contain interesting substances with antiplasmodial activity against several strains of Plasmodium falciparum. With respect to species of Bulbine, this is the first time that in vitro germination has been achieved. For seeds of K. pumila and K. uvaria, a facilitated germination was attained by an optimized pretreatment of the seeds. After in vitro germination, root explants of Kniphofia and bulb explants of B. caulescens were successfully transformed into callus cultures. From these, liquid cultures of the two species of Kniphofia were easily produced. Studies on shoot multiplication of Kniphofia in vitro were also performed.     

Effects of elevated CO<Subscript>2</Subscript> on foliar chemistry of saplings of nine species of tropical tree

This study examined the effects of elevated CO₂ on secondary metabolites for saplings of tropical trees. In the first experiment, nine species of trees were grown in the ground in open-top chambers in central Panama at ambient and elevated CO₂ (about twice ambient). On average, leaf phenolic contents were 48% higher under elevated CO₂. Biomass accumulation was not affected by CO₂, but starch, total non-structural carbohydrates and C/N ratios all increased. In a second experiment with Ficus, an early successional species, and Virola, a late successional species, treatments were enriched for both CO₂ and nutrients. For both species, nutrient fertilization increased plant growth and decreased leaf carbohydrates, C/N ratios and phenolic contents, as predicted by the carbon/nutrient balance hypothesis. Changes in leaf C/N levels were correlated with changes in phenolic contents for Virola (r=0.95, P<0.05), but not for Ficus. Thus, elevated CO₂, particularly under conditions of low soil fertility, significantly increased phenolic content as well as the C/N ratio of leaves. The magnitude of the changes is sufficient to negatively affect herbivore growth, survival and fecundity, which should have impacts on plant/herbivore interactions.     

Triacylglycerol lipase activities in tissues of Antarctic fishes

. The activity of the lipolytic enzyme, triacylglycerol lipase (TAG lipase; E.C. 3.1.1.3) was measured in heart ventricle, liver, oxidative skeletal muscle, and adipose tissue of four species of Antarctic fishes (Chaenocephalus aceratus, Notothenia coriiceps, Trematomus newnesi, Gobionotothen gibberifrons). Estimates of weight-specific activities at 0°C in tissues of the Antarctic fishes were broadly comparable to those reported for temperate-zone fish species assayed at their physiological temperature. Within each species, liver tissue displayed the highest weight-specific activity of TAG lipase in C. aceratus, T. newnesi, and G. gibberifrons, while adipose tissue of N. coriiceps showed the highest activity within that species. Combining these estimates with carefully measured somatic indices of the tissues, permitted assessment of the relative importance of different tissues to overall lipolytic capacity on an organismal level. On this basis, liver tissue displays the greatest capacity of TAG lipase per 100 g body weight in each of the species studied.     

Relations between efficiency of water transport and duration of leaf growth in some deciduous and evergreen trees

Shoot and leaf growth rate as well as shoot hydraulic conductance per unit leaf area (K_SL) were measured on three evergreen (Viburnum tinus L., Prunus laurocerasus L., Laurus nobilis L.) and three deciduous (Corylus avellana L., Juglans regia L., Castanea sativa L.) trees growing under the same environmental conditions. The times required to complete shoot growth (27 days for P. laurocerasus to 51 days for V. tinus) and leaf expansion (24 days for C. sativa to 42 days for C. avellana) were very different among the studied species. These species also differed in K_SL that ranged between 1.5 and 3.5 e-4 kg s^-1 m^-2 MPa^-1 in C. avellana and C. sativa, respectively, with intermediate values recorded in the other species. A strong, negative and statistically significant correlation was found to exist between K_SL and the time required for complete leaf expansion. This suggests that duration of leaf growth is shortened by the high hydraulic efficiency of the shoot. In contrast, no statistically significant relationship was found to exist between K_SL and shoot growth rate. Whether a high leaf growth rate can be interpreted as advantageous to plants or it is only an epiphenomenon of the high efficiency in the vertical water transport is discussed.