Nucleotide substitutions in Staphylococcus aureus strains, Mu50, Mu3, and N315.

A specific phenotype of Staphylococcus aureus strains Mu50 and Mu3 is characterized by thickened cell wall and moderate resistance to vancomycin. The N315 strain is a prototype of methicillin-resistant S. aureus (MRSA), but it is methicillin susceptible, despite carrying the mecA resistance gene. Here, we revised differences in the sequences of Mu50 and N315, referencing that of Mu3 which were assumed to be of one lineage. The 362 ORFs diverse between Mu50 and N315 were picked up, and the corresponding ones in three strains were re-sequenced. This defined 213 ORFs diverse between Mu50 and N315, and 9 between Mu50 and Mu3. The fixed diversities of 174 ORFs (except for 39 silent ORFs from 213), including nucleotide substitution (NSs), frame shift, and truncation were grouped into three major functional categories, which were transport (14.9% in the 174 diverse ORFs), metabolism of carbohydrates (5.7%), and RNA synthesis (9.6%). The other gene categories had small diversities. These gene categories seemed to be functionally decisive for the Mu50-specific characters, the thickened cell wall and moderate vancomycin resistance. All of the diverse genes and the high quality sequence of Mu50 can be viewed at the web site (http://133.5.48.239/VRSA/).     

Evaluation of a forced-air-ventilated micro-isolation system for protection of mice against Pasteurella pneumotropica.

Studies to date have established that the physical environment inside cages can be controlled adequately by setting the intra-cage ventilation at 60 air changes per hour in a forced-air-ventilated micro-isolation system (FVMIS). In this study, the capability of FVMIS to prevent inter-cage transmission of microorganisms was evaluated using Pasteurella pneumotropica as a reference microorganism. One FVMIS rack and a conventional rack were used, and cages with mice positive for P. pneumotropica and those with P. pneumotropica-free mice were housed on both racks. The mice were examined for P. pneumotropica contamination every 4 weeks after initiating the experiment for 12 weeks using a polymerase chain reaction method. Some P. pneumotropica-free mice housed in open air cages in the conventional rack became positive for P. pneumotropica (four of 28 animals after 4 weeks; eight of 28 animals after 12 weeks), but all P. pneumotropica-free mice housed in the FVMIS cages remained negative for the bacterium throughout the experiment. The results demonstrate that FVMIS can prevent inter-cage transmission of P. pneumotropica when proper cage handling practice is under taken.     

Life history adaptations and stress tolerance of four domestic species of Drosophila

Life history traits and stress tolerance were studied in four domestic species of Drosophila–D. melanogaster, D. simulans, D. auraria and D. immigrans– to understand how they adapt to their environments. In all species, larval weight approximately doubled in 1 day. The relative egg weight (egg weight : pupal weight) was smaller and the larval period was longer in D. immigrans than in the other three species. The pupal period was the longest in D. auraria. However, the adaptive significance of these differences in larval and pupal periods was not clear. The pupal case was generally thicker in the larger species, probably to support the larger pupal body. The start of oviposition was earliest and reproductive effort was greatest in female D. simulans, followed by female D. melanogaster. In contrast, starvation tolerance and the increase in bodyweight after eclosion was greater in D. immigrans and D. auraria than in the other two species. Pupal desiccation tolerance was greatest in D. melanogaster and lowest in D. auraria, and the less tolerant species seemed to select more humid sites for pupation. Adult tolerance to desiccation was greatest in D. melanogaster and lowest in D. simulans. In contrast, adult cold tolerance was greater in D. auraria and adult heat tolerance was lower in D. immigrans than in the other species. These differences in life history traits and stress tolerance represent the Drosophila species differential adaptations, and are assumed to allow coexistence of the species.     

Identification of a binding region on Escherichia coli heat-stable enterotoxin to intestinal guanylyl cyclase C

Summary The heat-stable enterotoxin (ST), produced byEscherichia coli, causes acute diarrhea in infants and domestic animals by activation of the intestinal membrane-bound receptor, guanylyl cyclase C. We have investigated a region on the ST molecule, which is recognized by the receptor, by introducing a photochromophore,p-azidophenylalanine (Pap), into three different regions of STp(4–17), which has the full toxic activity. Each ST analog bound to the receptor, but only STp(4–17) containing a Pap residue at position 11 in the central portion of the ST molecule, showed a high efficiency in the reaction which cross-linked with the receptor by UV radiation. These data clearly demonstrate that the region of the ST molecule encompassing the Asn¹¹ residue directly interacts with the receptor.     

The electrogenic proton pumping from parenchyma symplast into xylem—direct demonstration by xylem perfusion

Abstract We have devised an experimental system for simultaneous measurement of the activity of the xylem electrogenic ion pump, which is located on the inner cell membrane between the parenchyma symplast (p) and the xylem (x). and pH of the xylem exudate of a hypocotyl segment of Vigna unguiculata under pressurized xylem perfusion. Anoxia caused immediate depolarization of the inner cell membrane followed by alkalization of the xylem exudate several minutes later. Activity of the xylem pump was recovered by reaeration and acidification of the xylem exudate took place. These results indicate that the xylem pump is the respiration-dependent electrogenic proton-pump extruding proton from the parenchyma symplast into the xylem.     

Further evidence for the transformation ofVinca rosea protoplasts byAgrobacterium tumefaciens spheroplasts

Protoplasts ofVinca rosea were transformed by spheroplasts ofAgrobacterium tumefaciens harboring nopalinetype Ti plasmids according to the procedure of Hasezawa et al. (1981). These transformants frequently differentiated tracheids, but further differentiation to teratomata has not so far been observed. Transformation was confirmed by the improved detection of nopaline synthase, where the sensitivity and specificity of the enzyme reaction was increased by employing¹⁴C--ketoglutaric acid and³H-arginine as substrates. The nopaline synthase activity was identified by the comigration of these two radioisotopes in the cnromatogram. Furthermore, the T-DNA structure of one of these transformants was examined by Southern hybridization according to Thomashow et al. (1980) and compared with that ofVinca rosea crown gall.Abbreviations PEG Polyethylene glycol - PVA polyvinyl alcohol - TLC thin-layer chromatography     

Evaluation of global warming impacts on the carbon budget of terrestrial ecosystems in monsoon Asia: a multi-model analysis

This study assesses the potential impacts of future global warming on the carbon budget of terrestrial ecosystems across monsoon Asia using the Inter-Sectoral Impact Model Intercomparison Project (ISI-MIP) dataset. We used simulation results of two emission pathways (RCP2.6 and RCP8.5), climate projections of five climate models, and seven terrestrial biome models to analyze the changes in net primary production and carbon stocks in the South, Southeast, and East Asian subregions during the period 1981–2099. The simulations indicated that by the end of the 21st century, net primary production would increase by 9–45 % and ecosystem carbon storage would increase by 42–86 Pg C. The clearest climatic impacts were found when using the adaptation-oriented emission scenario (RCP8.5), which assumes a greater CO₂ increase and a larger change in climatic conditions. Substantial disparities in temporal trajectories and spatial patterns were found in the estimated changes, owing to the uncertainties in the emission scenarios, climate projections, and ecosystem models. We attempted to derive consistent patterns throughout the simulations to specify potential hotspots of climatic impacts (e.g., soil carbon change in the southern Tibetan Plateau). Finally, we discuss changes to the climatic characteristics in the study region (e.g., a change in the rainy season), the implications for ecosystem services, and the need for collaborative field monitoring studies.     

Molecular Detection,Isolation, and Physiological Characterization of Functionally Dominant Phenol-Degrading Bacteria in Activated Sludge

DNA was isolated from phenol-digesting activated sludge, and partial fragments of the 16S ribosomal DNA (rDNA) and the gene encoding the largest subunit of multicomponent phenol hydroxylase (LmPH) were amplified by PCR. An analysis of the amplified fragments by temperature gradient gel electrophoresis (TGGE) demonstrated that two major 16S rDNA bands (bands R2 and R3) and two major LmPH gene bands (bands P2 and P3) appeared after the activated sludge became acclimated to phenol. The nucleotide sequences of these major bands were determined. In parallel, bacteria were isolated from the activated sludge by direct plating or by plating after enrichment either in batch cultures or in a chemostat culture. The bacteria isolated were classified into 27 distinct groups by a repetitive extragenic palindromic sequence PCR analysis. The partial nucleotide sequences of 16S rDNAs and LmPH genes of members of these 27 groups were then determined. A comparison of these nucleotide sequences with the sequences of the major TGGE bands indicated that the major bacterial populations, R2 and R3, possessed major LmPH genes P2 and P3, respectively. The dominant populations could be isolated either by direct plating or by chemostat culture enrichment but not by batch culture enrichment. One of the dominant strains (R3) which contained a novel type of LmPH (P3), was closely related to Valivorax paradoxus, and the result of a kinetic analysis of its phenol-oxygenating activity suggested that this strain was the principal phenol digester in the activated sludge.Many scientists have used the rRNA approach (29, 30) to detect microbial populations and to describe the structures of microbial communities in various environments without isolating the component microorganisms. These studies have shown that most 16S ribosomal DNA (rDNA) sequences directly amplified from environmental samples are different from the sequences of comparable laboratory strains. Workers have concluded from such observations that many bacteria that are predominant in the natural environment have not been isolated in the laboratory yet and that the microbial diversity in the natural environment is much greater than the diversity of the bacteria that have been isolated (2, 7, 13, 25, 35, 36, 39, 40).Currently, one important aspect of microbial ecology studies is functional dissection of microbial communities based on structural information obtained by the approach mentioned above. An analysis of a population shift accompanied by a change in the function of a community yields information useful for identifying functionally dominant populations (2, 3, 42), although information concerning the function (activity) of each population can never be obtained by this kind of approach. Hence, workers have emphasized that pure-culture experiments are indispensable for detailed analysis of the functions of each population and that isolation of the functionally dominant populations in a microbial community is quite important.Phenol and its derivatives are some of the major hazardous compounds in industrial wastewater (1, 31, 43), and for this reason biodegradation of phenol has attracted keen attention (34, 46). However, since most studies of phenol biodegradation have been carried out under laboratory conditions with arbitrarily selected phenol-degrading bacteria, phenol biodegradation in the environment is not well understood yet. In the present study, to better understand phenol degradation in activated sludge, we isolated and characterized the phenol-degrading bacteria that were identified by the rRNA approach to be the dominant population in phenol-digesting activated sludge. Physiological and genetic differences between the dominant phenol-degrading bacteria isolated in this study and representative phenol-degrading bacteria characterized previously in several laboratories are discussed below.     

Observations on five species of philometrid nematodes from marine fishes in Japan

Five species of philometrid nematodes (subfamily Philometrinae) are redescribed on the basis of newly collected materials from marine fishes from Japan. The species Philometra sciaenae Yamaguti, 1941, considered by Rasheed (1963) a synonym of P. lateolabracis (Yamaguti, 1935), is re-established as a valid species and the male of this nematode, collected from the ovary of the type-host, Argyrosomus argentatus, is described for the first time; the latter is characterised mainly by the length of equally long spicules (0.108–0.126 mm) and the length ratio of the gubernaculum and spicules (1:2.25). Other species redescribed on females include Philometra inimici Yamaguti, 1941 and P. sebastisci Yamaguti, 1941 from the ovary of their type-hosts, Inimicus japonicus and Sebastiscus marmoratus, respectively, and also Philometroides seriolae (Ishii, 1931) from the musculature of Seriola quinqueradiata (type-host), and Clavinema mariae (Layman, 1930) from the subcutaneous tissue of Pleuronectes schrenki and Acanthogobius flavimanus (a new host record). For the first time, cephalic ends of some of these species were studied under the scanning electron microscope (SEM) and the character of their cephalic papillae was described. The importance of a knowledge of male morphology for the taxonomy of philometrids is stressed.