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PubMed2006

Simultaneous radioassays of bacterial production and mercury methylation in the periphyton of a tropical and a temperate wetland.

Laboratory radioassays were made to study mercury (Hg) methylation together with bacterial production in the periphyton of two aquatic macrophytes, the submerged Myriophyllum spicatum, from a constructed wetland in Sweden and the floating Eichhornia crassipes, from a eutrophied tropical lake in Brazil. Time course incubations were made by addition of (203)HgCl(2) and the methylmercury formed was extracted at pre-defined time intervals. Bacterial production ((14)C-leucine incorporation) was measured at the same time intervals, with plants removed from parallel incubations made with and without addition of cold HgCl(2). For E. crassipes, higher methylmercury production was observed at elevated bacterial production, whereas for M. spicatum, the bacterial production was significantly lower, and Hg methylation was below the detection limit. The combined results confirm the importance of microbial processes for Hg methylation, although other factors are known to influence this process in complex ways. The addition of Hg did not significantly influence bacterial production, while the incubation temperatures used (25 and 35 degrees C) resulted in different methylation rates. Radiotracer techniques for measurements of bacterial production such as (14)C-leucine uptake can provide useful insights into the Hg cycle in aquatic environments, and our data suggest that they may be used as a proxy of mercury methylation potentials.

Bacteria↗
PubMed2006

Tidal transport and dispersal of marine toxic microalgae in a shallow, temperate coastal lagoon.

A numerical model was used to predict toxic microalgal transport and dispersion in Ria de Aveiro in Portugal. A previously developed Lagrangean particle tracking model coupled to a calibrated two-dimensional hydrodynamic model of Ria de Aveiro was used. Microalgae were regarded as passive particles and the methodology used allowed the determination of their trajectories, as induced by the tidal currents predicted by the hydrodynamic model. The model assumes Ria de Aveiro as vertically homogeneous and does not take into account the vertical distribution patterns of microalgae. Simulations were carried out during extreme spring and neap tides, with microalgal released at the mouth of the lagoon at the local flood. The maximum and minimum areas affected during the occurrence of toxic microalgal blooms were estimated to evaluate the suitability of the distribution of the sampling stations included in the local monitoring program. It was found that the tidal currents greatly determine the microalgal horizontal distribution and dispersal in the lagoon. The results confirmed that the locations of water and bivalve shellfish sampling stations, postulated by INIAP/IPIMAR, in the context of the local harmful algal bloom (HAB) program, were appropriate, although some possible refinements were identified.

Animals↗
PubMed2005

Influence of small scale conditions on the diversity of wood decay fungi in a temperate, mixed deciduous forest canopy.

Studies on fungal richness and ecology have been largely disregarded since the first intensive efforts to investigate organismal diversity in forest canopies. We used the Leipzig Canopy Crane research facility to sample wood-decaying fungi in a mixed deciduous forest canopy 10-30 m in height. The structural complexity of the canopy was analysed using different methods, including meteorological measurements. With respect to temperature and relative humidity, marked differences existed between forest floor and upper canopy layers that persisted on smaller scales. Of the 118 taxa found in 128 sample units, pyrenomycetes and corticioid fungi outnumbered other macrofungal groups. Fungal communities showed distinct variations both in species richness and composition with respect to substrate (tree species), height in the canopy, stage of decay, and branch diameter. Pyrenomycetes and their anamorphs dominated the mycobiota on thin, exposed twigs at great heights, indicating their ability to overcome extended periods of drought and high levels of solar irradiance. Other taxa of Tremellales (Exidia spp.), Orbiliales (Hyalorbilia inflatula, Orbilia spp.) or Agaricales (Episphaeria fraxinicola, Cyphellopsis anomala, Lachnella spp.) also exhibited features that enabled them to develop in lesser protected habitats within tree crowns.

Cluster Analysis↗
PubMed2006

Simulation of the treatment performance of outdoor subsurface flow constructed wetlands in temperate climates.

Numerical models are a means to increase the understanding of the processes occurring in the "black box" constructed wetland. Once reliable models for constructed wetlands are available they can be also used for evaluating and improving existing design criteria. The paper shows simulation results for outdoor experimental subsurface vertical flow constructed wetlands using CW2D, a multi-component reactive transport module developed to simulate transport and reactions of the organic matter, nitrogen and phosphorus in subsurface flow constructed wetlands. The surface area of the experimental vertical flow bed was 20 m(2). The organic load applied was 27 g COD m(-2) d(-1) (corresponding to a specific surface area of 3 m(2) per person). The aim of the work is to calibrate the model for temperature dependency that has been implemented in CW2D. Water temperature during the investigation period varied between 4 degrees C and 18 degrees C. The measured effluent concentrations during summer could be simulated using the standard CW2D parameter set when the flow model was calibrated well. However, the increasing effluent concentrations at low temperatures could not be simulated with the standard CW2D parameter set where temperature dependencies are considered only for maximum growth, decay, and hydrolysis rates. By introducing temperature dependencies for half-saturation constants for the hydrolysis and nitrification processes it was possible to simulate the observed behaviour. The work presented is a step on the way to validate the CW2D module. Model validation is a necessary step before numerical simulation can be finally used in practice, e.g. for checking existing design guidelines.

Computer Simulation↗
PubMed2005

Diverse rhizobia associated with woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the temperate zone of China.

Fifty-nine bacterial isolates from root nodules of the woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the central and eastern regions of China were characterized with phenotypic analysis, PCR-based 16S and 23S rRNA gene RFLP, Box PCR and 16S rRNA gene sequencing. Seven main phena were defined in numerical taxonomy, which corresponded to distinct groups within the genera Agrobacterium, Bradyrhizobium, Mesorhizobium and Rhizobium in 16S and 23S rRNA gene PCR-RFLP. The phylogenetic relationships of the 16S rRNA genes supported the grouping results of PCR-RFLP. Most of the isolates from Amorpha fruticosa were classified into two groups closely related to Mesorhizobium amorphae. Seventeen of the 21 isolates from Wisteria sinensis were identified as two groups related to Rhizobium and Agrobacterium. Six out of 10 isolates from Cercis racemosa were identified as a group related to Bradyrhizobium. Our results indicated that each of the investigated legumes nodulated mainly with one or two rhizobial groups, although isolates from different plants intermingled in some small bacterial groups. In addition, correlation between geographic origin and grouping results was found in the isolates from Amorpha fruticosa. These results revealed that the symbiotic bacteria might have been selected by both the legume hosts and the geographic factors.

Alphaproteobacteria↗
PubMed2004

Seasonal development of Cyathostominae larvae on pasture in a northern temperate region of the United Kingdom.

A study following the development of Cyathostominae from egg to the infective larval third stage was conducted from April to December 2001 in west central Scotland. Duplicate samples (1 kg) of naturally infected faeces were placed on a 78 cm2 plot each week on a cyathostomin-free pasture. Subsamples of the grass surrounding the faecal plot were collected weekly on four occasions and the number of larvae obtained determined. Few larvae were recovered in the first week of development of individual plots, followed by a rise in the numbers of larvae in second, third and fourth weeks of development of each sample. The climatic conditions were seen to have an effect on the rate of development. Specifically, from multilevel, multivariable linear regression models it was evident that the factors associated with numbers of infective larvae recovered from pasture were the time since the faeces samples were laid down, the average temperature and rainfall during the previous week, as well as the interaction between temperature and rainfall. Conversely, from the model, the number of larvae recovered from pasture was associated with neither the number of eggs within the faeces samples placed on the plots nor with the viability of these eggs.

Animals↗
PubMed2005

The possibilities and limitations of evasive grazing as a control measure for parasitic gastroenteritis in small ruminants in temperate climates.

Between 1999 and 2003 studies were done at Utrecht University on the population dynamics of gastrointestinal nematodes and the potential to use evasive grazing for the control of nematode infections in small ruminants. On most pastures grazed in May and June it took at least 3 weeks before high pasture infectivity levels for Haemonchus contortus developed while it took at least 2 weeks on pastures grazed in July, August and September. Occasionally, larvae emerged earlier on pasture when high temperatures coincided with rain. Pasture infectivity levels for H. contortus subsequently increased and the highest levels were found between 5 and 9 weeks on pastures grazed in May-June and between 3 and 9 weeks on pastures grazed in July, August and September. Pasture infectivity usually had decreased again to low levels after approximately 3 months. Larvae of the other trichostrongylids emerged later on pasture. However, patterns of Teladorsagia circumcincta, Trichostrongylus spp. and Cooperia curticei were basically similar to H. contortus. Strongyloides papillosus larvae emerge within 2 weeks on pasture and survival is short. On the other hand Nematodirus larvae took a lot longer to emerge on pasture than the other trichostrongylids. The results imply that only on a small proportion of farms (mixed dairy cattle/sheep farms; some organic dairy goat farms) evasive grazing can be recommended as only control measure for parasitic gastroenteritis. However, the present data offer possibilities to combine evasive grazing with other methods in an integrated control scheme.

Animal Husbandry↗
PubMed2004

Molecular and transcriptional analysis of the temperate lactococcal bacteriophage Tuc2009.

The genome of bacteriophage Tuc2009 consists of 38347 base pairs on which 57 open reading frames (ORFs) were identified, divided in two oppositely transcribed regions. The leftward-transcribed region harbors three ORFs, two of which are involved in the establishment of lysogeny. The rightward-transcribed region contains 54 ORFs, which are assumed to be required for the lytic life cycle. An exception to the above organization is ORF 10, of unknown function, located within the rightward-transcribed region that has an orientation opposite to the ORFs surrounding it. Transcriptional analysis of the Tuc2009 genome following infection of a sensitive host revealed that most ORFs are transcribed in a sequential manner. ORFs that are presumed to form (part of) the genetic switch along with the superinfection exclusion-encoding gene are transcribed immediately after infection, followed by transcription of the presumed replication region. Subsequent to this, several small transcripts could be identified followed by a single 24-kb transcript. This latter transcript was shown to specify most of the identified structural proteins as well as two proteins required for host lysis. Interestingly, the 24-kb mRNA was shown to undergo splicing through the activity of a type I intron whose removal from the mRNA resulted in the formation of an ORF specifying a major structural protein. Primer extension analysis was employed to identify the 5' ends of mRNA transcripts and the genome and transcriptional data are discussed in relation to other lactococcal bacteriophages.

Bacteriophages↗
PubMed2004

An orthologue of the cor gene is involved in the exclusion of temperate lambdoid phages. Evidence that Cor inactivates FhuA receptor functions.

A new set of lambdoid phages (mEp) classified into different immunity groups was previously described. Phages mEp213, mEp237, and mEp410 were unable to grow in mEp167 lysogenic cells, presumably due to an exclusion mechanism expressed constitutively by the mEp167 repressed prophage. In this work, to analyze the exclusion phenomenon, we constructed a genomic library from mEp167 phage in a pPROEX derivative plasmid. A DNA fragment containing an open reading frame for a 77 amino acid polypeptide was selected by its ability to confer resistance to heteroimmune phage infection. This ORF shows high amino acid sequence identity with putative Cor proteins of phages HK022, phi80 and N15. Cells expressing the mEp167 cor gene from a plasmid (Cor(+) phenotype) excluded 13 of 20 phages from different infection immunity groups. This exclusion was observed in both tonB(-) and tonB(+) cells. Lambdoid mEp phages that were excluded in these cells were unable to infect cells defective in the outer membrane FhuA receptor (fhuA(-)). Thus, Cor-mediated exclusion was only observed in fhuA(+) cells. Phage production after DNA transfection or the spontaneous induction of mEp prophage in Cor(+) cells was not blocked. In addition, ferrichrome uptake, which is mediated by FhuA, was inhibited in Cor(+) cells. Our results show that not only phage infection via FhuA but also a FhuA transport activity (ferrichrome uptake) are inhibited by Cor, presumably by inactivation of FhuA.

Amino Acid Sequence↗
PubMed2005

Complete genomic nucleotide sequence and analysis of the temperate bacteriophage VWB.

The entire double-stranded DNA genome of the Streptomyces venezuelae bacteriophage VWB was sequenced and analyzed. Its size is 49,220 bp with an overall molar G + C content of 71.2 mol%. Sixty-one potential open reading frames were identified and annotated using several complementary bioinformatics tools. Clusters of functionally related putative genes were defined, supporting a refined version of the modular theory of phage evolution.

Amino Acid Sequence↗
PubMed2005

Identification of DNA-binding sites for the activator involved in late transcription of the temperate lactococcal phage TP901-1.

Alt, encoded by the lactococcal phage TP901-1, is needed for late transcription. We identify Alt as a DNA-binding protein, and footprint analysis shows that Alt binds to a region containing four imperfect direct repeats (ALT boxes) located -76 to -32 relative to the P(late) transcriptional start site. The importance of the ALT boxes was confirmed by deletion of one or two ALT boxes and by introducing mutations in ALT boxes 1 and 4. Alt is proposed to act as a tetramer or higher multimer activating transcription of TP901-1 late genes by binding to the four ALT boxes, and bending of the DNA may be important for transcriptional activation of P(late). Furthermore, our results suggest that DNA replication may be required for late transcription in TP901-1. Additionally, we identify gp28 of the related lactococcal phage Tuc2009 as an activator and show that the activators required for late transcription in TP901-1 and Tuc2009 are interchangeable.

Amino Acid Sequence↗
PubMed1992

"Well-tempered" diuresis renography: its historical development, physiological and technical pitfalls, and standardized technique protocol.

Differentiating the various causes of hydronephrosis from that of obstruction can be very difficult. Diuretic renography has been adopted as a noninvasive clinical management tool to assist in this differentiation. However, the correlation of the results of diuresis renography, the Whitaker test, and the surgical results has only been between 40% and 85%. This is believed to be due to the many physiological factors and technical pitfalls of the technique. These include variable renal function, variable compliance of the collecting system, the effect of back pressure from a full bladder, the state of hydration, the choice of radiopharmaceutical, and the timing of the diuretic injection. Even the technique of measuring the clearance half-time (T 1/2) is controversial, with at least eight different methods defined. In order to diminish the effect of these variables, a standardized protocol for diuretic renography has been proposed by a consortium of members of the Society for Fetal Urology and the Pediatric Nuclear Medicine Club of the Society of Nuclear Medicine. These include a standardized hydration with a dilute glucose solution, bladder catheterization and measurement of urine output response, uniform radiopharmaceutical choice, and diuretic dose with specific timing of the diuretic injection. The various methods for T 1/2 calculation are illustrated, and various stereotypical renogram and diuresis response curves are offered as aids in the interpretation of the study. Diuresis renography is one of the most complex functional studies in nuclear medicine today. A thorough understanding of the physiological basis for diuresis renography and the pitfalls of the technique is required for its appropriate use in the management of patients suspected of urinary tract obstruction.

Diuretics↗
PubMed1992

Delimitation of cohesive ends site (cos) of Streptomyces temperate bacteriophage R4.

The cohesive ends site (cos) of actinophage R4 was delimitated by assaying the in vivo packaging activity of cosmid derivatives in Streptomyces lividans. A region of 66 bp from -30 to +36 from the center of cohesive ends was required for the basal level of packaging activity. Two additional regions outside the basal sequences from -39 to -31 and from +37 to +97 were necessary for the high level of activity, defined as the accessory sequences. Direct- or inverted-repeat sequences were found within the delimitated region, which might be involved in the recognition by the terminase of actinophage R4.

Bacteriophages↗
PubMed2002

The F286Y mutation of PrlA4 tempers the signal sequence suppressor phenotype by reducing the SecA binding affinity.

SecYEG forms the protein-conducting channel of the Escherichia coli translocase. It binds the peripheral ATPase SecA that drives the preprotein translocation reaction. PrlA4 is a double mutant of SecY that enables the translocation of preproteins with a defective or even missing signal sequence. The effect of the individual mutations, F286Y and I408N, was studied with SecYEG proteoliposomes. SecY(I408N) is responsible for the increased translocation of preproteins with a defective and normal signal sequence, and exhibits a stronger prl phenotype than PrlA4. This activity correlates with an elevated SecA-translocation ATPase and SecA binding affinity. SecY(F286Y) supports only a low SecA binding affinity, preprotein translocation and SecA translocation ATPase activity. These results suggest that the second site F286Y mutation reduces the strength of the I408N mutation of PrlA4 by lowering the SecA binding affinity.

Adenosine Triphosphatases↗