Toxicity studies for indigenous Bacillus thuringiensis isolates from Malang City, East Java on Aedes aegypti larvae.

OBJECTIVE: To investigate the toxicity of indigenous Bacillus thuringiensis (B. thuringiensis)isolates from Malang City for controlling Aedes aegypti (Ae. aegypti) larvae. METHODS: Soil samples were taken from Purwantoro and Sawojajar sub-districts. Bacterial isolation was performed using B. thuringiensis selective media. Phenotypic characteristics of the isolates were obtained with the simple matching method. The growth and prevalence of spores were determined by the Total Plate Count method, and toxicity tests were also performed on the third instar larval stage of Ae. aegypti. The percentage of larval mortality was analysed using probit regression. The LC50 was analysed by ANOVA, and the Tukey HSD interval was 95%. RESULTS: Among the 33 selected bacterial isolates, six were obtained (PWR4-31, PWR4-32, SWJ4-2b, SWJ4-4b, SWJ-4k and SWJ5-1) that had a similar phenotype to reference B. thuringiensis. Based on the dendrogram, all of the bacterial isolates were 71% similar. Three isolates that had a higher prevalence of reference B. thuringiensis were PWR4-32, SWJ4-4b and SW5-1, of which the spore prevalence was 52.44%, 23.59%, 34.46%, respectively. These three indigenous isolates from Malang City successfully killed Ae. aegypti larvae. The PWR4-32 isolates were the most effective at killing the larvae. CONCLUSIONS: Six indigenous B. thuringiensis isolates among the 33 bacterial isolates found in the Sawojajar and Purwantoro sub-districts were toxic to the third instar larvae of Ae. aegypti. The PWR4-32 isolates were identical to the reference B. thuringiensis and had 88% phenotype similarity. The PWR4-32 isolates had the highest spore prevalence (52.44%), and the early stationary phase occurred at 36 h. The PWR4-32 isolates were the most effective at killing Ae. aegypti larvae (LC50-72 h=2.3×10(8) cells/mL).

Toxicity studies for indigenous Bacillus thuringiensis isolates from Malang city, East Java on Aedes aegypti larvae

Objective: To investigate the toxicity of indigenous Bacillus thuringiensis (B. thuringiensis) isolates from Malang City for controlling Aedes aegypti (Ae. aegypti) larvae. Methods: Soil samples were taken from Purwantoro and Sawojajar sub-districts. Bacterial isolation was performed using B. thuringiensis selective media. Phenotypic characteristics of the isolates were obtained with the simple matching method. The growth and prevalence of spores were determined by the Total Plate Count method, and toxicity tests were also performed on the third instar larval stage of Ae. aegypti. The percentage of larval mortality was analysed using probit regression. The LC50 was analysed by ANOVA, and the Tukey HSD interval was 95%. Results:Among the 33 selected bacterial isolates, six were obtained (PWR4-31, PWR4-32, SWJ4-2b, SWJ4-4b, SWJ-4k and SWJ5-1) that had a similar phenotype to reference B. thuringiensis. Based on the dendrogram, all of the bacterial isolates were 71%similar. Three isolates that had a higher prevalence of reference B. thuringiensis were PWR4-32, SWJ4-4b and SW5-1, of which the spore prevalence was 52.44%, 23.59%, 34.46%, respectively. These three indigenous isolates from Malang City successfully killed Ae. aegypti larvae. The PWR4-32 isolates were the most effective at killing the larvae. Conclusions:Six indigenous B. thuringiensis isolates among the 33 bacterial isolates found in the Sawojajar and Purwantoro sub-districts were toxic to the third instar larvae of Ae. aegypti. The PWR4-32 isolates were identical to the reference B. thuringiensis and had 88%phenotype similarity. The PWR4-32 isolates had the highest spore prevalence (52.44%), and the early stationary phase occurred at 36 h. The PWR4-32 isolates were the most effective at killing Ae. aegypti larvae (LC50-72 h=2.3í108 cells/mL).

Water quality monitoring system for determination of ionic nutrients by ion-exclusion chromatography with spectrophotometric detection on cation- and anion-exchange resin columns using water eluent.

A unified ion-exclusion chromatography (IEC) system for monitoring anionic and cationic nutrients like NH4+, NO2-, NO3-, phosphate ion, silicate ion and HCO3- was developed and applied to several environmental waters. The IEC system consisted of four IEC methodologies, including the IEC with ultraviolet (UV) form connected with detection at 210 nm for determining NH4+ on anion-exchange separation column in OH anion-exchange UV-conversion column in I- form in tandem, the IEC with UV-detection at 210 nm for determining simultaneously NO3- and NO3- on cation-exchange separation column in H+ form, the IEC with UV-detection at 210 nm for determining HCO3- on cation-exchange separation column in H+ form connected with anion-exchange UV-conversion column in I- form in tandem, and the IEC with visible-detection based on molybdenum-blue reaction for determining simultaneously silicate and phosphate ions on cation-exchange separation column in H+ form. These IEC systems were combined through three manually-driven 6-port column selection valves to select each separation column to determine selectively the ionic nutrients. Using this sequential water quality monitoring system, the analytical performances such as calibration linearity, reproducibility, detection limit and recovery were also tested under the optimized chromatographic conditions. This novel water quality monitoring system has been applied successfully for the determination of the ionic eutrophication components in sub-urban river waters.

Water quality monitoring of Jialing-River in Chongqing using advanced ion chromatographic system.

The water quality monitoring operation to evaluate the water quality of polluted river is an extremely important task for the river-watershed management/control based on the environmental policy. In this study, the novel, simple and convenient water quality monitoring of Jialing-River in Chongqing, China was carried out using an advanced ion chromatography (IC) consisting of ion-exclusion/cation-exchange chromatography (IEC/CEC) with conductivity detection for determining simultaneously the common anions such as SO4(2-), Cl(-), and NO3(-) and the cations such as Na+, NH4+, K+, Mg2+, and Ca2+, the ion-exclusion chromatography (IEC) with visible detection for determining simultaneously the nutrient components such as phosphate and silicate ions, and the IEC with the enhanced conductivity detection using a post column of K+-form cation-exchange resin for determining HCO3(-)-alkalinity as an inorganic-carbon source for biomass synthesis in biological reaction process under the aerobic conditions. According to the ionic balance theory between the total equivalent concentrations of anions and cations, the water quality evaluation of the Jialing-River waters taking at different sampling sites in Chongqing metropolitan area was carried out using the advanced IC system. As a result, the effectiveness of this novel water quality monitoring methodology using the IC system was demonstrated on the several practical applications to a typical biological sewage treatment plant on Jialing-River of Chongqing.

Ion-exclusion/cation-exchange chromatography with dual detection of the conductivity and spectrophotometry for the simultaneous determination of common inorganic anionic species and cations in river and wastewater.

Simultaneous determinations of common inorganic anionic species (SO(4)(2-), Cl(-), NO(3)(-), phosphate and silicate) and cations (Na(+), NH(4)(+), K(+), Mg(2+) and Ca(2+)) were conducted using an ion-chromatography system with dual detection of conductivity and spectrophotometry in tandem. The separation of ionic species on a weakly acidic cation-exchange resin was accomplished using a mixture of 100 mM ascorbic acid and 4 mM 18-crown-6 as an acidic eluent (pH 2.6), after which the ions were detected using a conductivity detector. Subsequently, phosphate and silicate were analyzed based on derivatization with molybdate and spectrophotometry at 700 nm. The detection limits at S/N = 3 ranged from 0.11 to 2.9 µM for analyte ionic species. This method was applied to practical river water and wastewater with acceptable criteria for the anion-cation balance and comparisons of the measured and calculated electrical conductivity, demonstrating the usefulness of the present method for water quality monitoring.

Simultaneous determinations of Cr(VI) and Cr(III) by ion-exclusion/cation-exchange chromatography with an unmodified silica-gel column.

In order to characterize the ion-exclusion and cation-exchange properties of an unmodified silica-gel column, the retention behaviors of Cr(VI) and Cr(III) ions were investigated using a Develosil 30-5 (150 x 4.6 mm i.d.) in the acidic region. Cr(VI) was separated from other anions by an ion-exclusion and ion-adsorption mechanism, and Cr(III) was separated from other cations with a cation-exchange mechanism. When using 2.0 mM oxalic acid (pH 2.6) as an eluent, a good separation of Cr(VI) and Cr(III) was obtained using conductimetric detection in 12 min. The method was successfully applied to the simultaneous determinations of Cr(VI) and Cr(III) added into tap-water and river-water samples.

Simultaneous spectrophotometric determination of orthophosphate and silicate ions in river water using ion-exclusion chromatography with an ascorbate solution as both eluent and reducing agent, followed by postcolumn derivatization with molybdate.

Ion-exclusion chromatography was examined for the simultaneous spectrophotometric determinations of orthophosphate and silicate ions in river water using an ascorbate solution as both an eluent and a reducing agent, followed by postcolumn derivatization using molybdate. The detector responses for both ions increased with increased ascorbic acid concentration in the eluent, but peak tailing was observed for the orthophosphate ion. This suggests that the amounts of undissociated orthophosphate ions increased with decreased eluent pH, resulting in the penetration of the phosphate to the Donnan's membrane formed on the resin surface. Using a neutral sodium ascorbate solution as an eluent, the peak shape was improved. With optimized separation and derivatization conditions (eluent, 20 mM sodium ascorbate; color-forming reagent, 10 mM sodium molybdate-60 mM sulfuric acid; flow rates of eluent and color-forming reagent, 0.4 and 0.2 mL min(-1); coil length, 6 m), the detection limits of orthophosphate and silicate ions were 0.9 and 1.0 microg L(-1), respectively. This method was successfully applied to the determination of orthophosphate and silicate ions in Kurose River water and the quantitative evaluations of the effects of water intake to a reservoir and discharge from a biological sewage treatment plant on the fluxes of these ions in the river.

Shaping of genetic structure along Pleistocene and modern river systems in the hydrochorous riparian azalea, Rhododendron ripense (Ericaceae).

To determine the effects of hydrochory on the formation of the present range of a species and the spatial distribution of genetic variation, we assessed the rangewide genetic structure of a hydrochorous riparian Japanese species (Rhododendron ripense) using four nuclear microsatellite loci. The patterns of isolation by distance and Bayesian clustering analyses of 33 populations suggested that the present range, characterized by both localized and disjunct distributions across the sea, arose from two contrasting colonization events: (1) primary colonization along two Pleistocene rivers that have been submerged and become partly isolated by marine transgression by 6000 years ago, and (2) additional range expansions from these rivers into unconnected neighboring rivers as a result of river captures. Along the Pleistocene rivers, frequent gene flow by hydrochory resulted in the retention of considerable genetic diversity within each population and genetic homogenization among populations. Within unconnected neighboring rivers, genetic diversity was also retained by the simultaneous redistribution of many individuals as a result of river captures, whereas restricted gene flow within a river resulted in genetic divergence among the river populations. Thus, the evolutionary history of hydrochorous R. ripense appears to have been strongly shaped by both ancient and modern rivers.

Simultaneous spectrophotometric determination of phosphate and silicate ions in river water by using ion-exclusion chromatographic separation and post-column derivatization.

The simultaneous spectrophotometric determination of phosphate and silicate ions in river water was examined by using ion-exclusion chromatography and post-column derivatization. Phosphate and silicate ions were separated by the ion-exclusion column packed with a polymethacrylate-based weakly acidic cation-exchange resin in the H(+)-form (TSKgel Super IC-A/C) by using ultra pure water as an eluent. After the post-column derivatization with molybdate and ascorbic acid, so-called molybdenum-blue, both ions were determined simultaneously by spectrophotometry. The effects of sulfuric acid, sodium molybdate and ascorbic acid concentrations and reaction coil length, which have relation to form the reduced complexes of molybdate and ions, on the detector response for phosphate and silicate ions were investigated. Under the optimized conditions (color-forming reactant, 50 mM sulfuric acid-10 mM sodium molybdate; reducing agent, 50 mM ascorbic acid; reaction coil length, 6 m), the calibration curves of phosphate and silicate ions were linear in the range of 50-2000 microg L(-1) as P and 250-10,000 microg L(-1) as Si. This method was successfully applied to water quality monitoring of Kurose-river watershed and it suggested that the effluent from a biological sewage treatment plant was significant source of phosphate ion in Kurose-river water.

Effect of total cholesterol, glucose and blood urea nitrogen on embryo quality in post-partum superovulated suckling Japanese Black cattle.

Aim: This study was conducted to examine the effect of blood metabolites on embryo quality in post-partum suckling Japanese Black cattle. Methods: Blood samples were taken from 23 cows 30 days before, at and 30 days after parturition. Cows were synchronized 40 or 41 days after calving (day 0) and divided into three groups: control (n = 6), gonadotropin-releasing hormone ([GnRH]n = 10) and estradiol benzoate ([EB]n = 7). All groups received a controlled internal drug release (CIDR) device intravaginally together with 2 mg EB i.m. on day 0 and superovulation was induced in all groups from days 5-7 with a gradually decreasing dose of follicle-stimulating hormone (FSH). Two milligrams of EB was given on day 8 and GnRH (0.1 mg) was given on day 9 of insertion of the CIDR in the EB and GnRH groups. Cows were inseminated twice after the onset of estrus and embryos were recovered 7-8 days after artificial insemination. Results: The number of corpus luteum detected by ultrasonography in the EB group was significantly higher (P < 0.05) than that in the GnRH group. The number and rate of transferable and freezable embryos did not differ significantly among the groups. Regardless of the treatments, the total cholesterol level from parturition until 30 days after parturition was significantly higher (P < 0.01) in the good category than in the poor category of cows. Conclusions: The number of transferable embryos produced by post-partum superovulated suckling Japanese Black cattle was affected by the level of total cholesterol from parturition until 30 days after parturition. Moreover, administration of EB in CIDR-treated cows increased the numbers of corpus luteum and yielded better rates of transferable and freezable embryos. (Reprod Med Biol 2008; 7: 55-62).

Predicting herbicides concentrations in paddy water and runoff to the river basin.

This study was conducted to investigate the dissipation pattern and runoff of herbicides to the river basin from the paddy fields. Pesticide paddy field model (PADDY) was applied to predict herbicide concentration in paddy fields. A field study was conducted in a paddy farm of Higashi Hiroshima City, Hiroshima Prefecture, Japan in the year of 2003 paddy season. The herbicides were mefenacet, thiobencarb, and bensulfuron methyl. The sample water was analyzed by using gas chromatography and HPLC after solid phase extraction. Predicted dissipation rate of thiobencarb in paddy water was higher (DT50 = 4.36) than that measured, with a lower k value (-0.069). Two weeks after application no thiobencarb was detected in the drainage channel and down stream. In the down stream, thiobencarb was detected until 3 d after application, with a range of 0.02% to 0.08% of applied herbicide. The predicted dissipation rate (k) and half-life (DT50) of mefenacet was not significantly different from that of measured. In the drainage channel, upstream and downstream mefenacet was found during the whole study period. In downstream, the maximum concentration of mefenacet was present 0.61% of applied in the paddy field on 3DAH. The dissipation rate (k) of BSM varied from -0.0860 to -0.1059 to with half-life (DT50) 3.5 and 2.84 d. In upstream water, no BSM was detected except trace amounts (0.01 microg/L) at 3 d after application. However, in the drainage channel 8%, 6% and 1.58% of applied BSM was present at 0, 1 and 3 d after application respectively. In the down stream, the highest concentration was 1.06%, shortly after application.

River system in Japan from a landscape ecological aspect.

The objective of this study was to elucidate characteristics associated with rivers by classification of major rivers in Japan into several types based on riparian conditions. As the results of principal component analysis (PCA) with use of parameters reflecting form and artificial alterations of respective rivers, four major components such as "comprehensive riparian size", "intactness of water front", "continuity of streams" and "simplicity of landform in river basin" were extracted. Subsequently, cluster analysis was performed based on principal component scores, leading to successful classification of major rivers into 6 types. These findings disclosed that (1) the extracted principal components provide effective viewpoint for classification of rivers; (2) distribution of respective classes indicates area properties; and (3) the employed quantitative procedures were found effective for classification of major rivers.

The impact of urban planning on land use and land cover in Pudong of Shanghai, China.

Functional zones in cities constitute the most conspicuous components of newly developed urban area, and have been a hot spot for domestic and foreign investors in China, which not only show the expanse of urban space accompanied by the shifts both in landscape (from rural to urban) and land use (from less extensive to extensive), but also display the transformation of regional ecological functions. By using the theories and methods of landscape ecology, the structure of landscape and landscape ecological planning can be analyzed and evaluated for studying the urban functional zones' layout. In 1990, the Central Government of China declared to develop and open up Pudong New Area so as to promote economic development in Shanghai. Benefited from the advantages of Shanghai's location and economy, the government of Pudong New Area has successively built up 7 different functional zones over the past decade according to their functions and strategic goals. Based on the multi-spectral satellite imageries taken in 1990, 1997 and 2000, a landscape ecology analysis was carried out for Pudong New Area of Shanghai, supported by GIS technology. Green space (including croplands) and built-up area are the major factors considered in developing urban landscape. This paper was mainly concerned with the different spatial patterns and dynamic of green space, built-up areas and new buildings in different functional zones, influenced by different functional layouts and development strategies. The rapid urbanization in Pudong New Area resulted in a more homogeneous landscape. Agricultural landscape and suburban landscape were gradually replaced by urban landscape as the degree of urbanization increased. As consequence of urbanization in Pudong, not only built-up patches, but also newly-built patches and green patches merged into one large patch, which should be attributed to the construction policy of extensive green space as the urban development process in Pudong New Area. The shape of green area of 7 functional zones became more and more regular because of the horticultural needs in Shanghai urban planning. Some suggestions were finally made for the study of future urban planning and layout.

The effects of drainage basin geomorphometry on minimum low flow discharge: the study of small watershed in Kelang River Valley in Peninsular Malaysia.

This study investigate the relationships between geomorphometric properties and the minimum low flow discharge of undisturbed drainage basins in the Taman Bukit Cahaya Seri Alam Forest Reserve, Peninsular Malaysia. The drainage basins selected were third-order basins so as to facilitate a common base for sampling and performing an unbiased statistical analyses. Three levels of relationships were observed in the study. Significant relationships existed between the geomorphometric properties as shown by the correlation network analysis; secondly, individual geomorphometric properties were observed to influence minimum flow discharge; and finally, the multiple regression model set up showed that minimum flow discharge (Q min) was dependent of basin area (AU), stream length (LS), maximum relief (Hmax), average relief (HAV) and stream frequency (SF). These findings further enforced other studies of this nature that drainage basins were dynamic and functional entities whose operations were governed by complex interrelationships occurring within the basins. Changes to any of the geomorphometric properties would influence their role as basin regulators thus influencing a change in basin response. In the case of the basin's minimum low flow, a change in any of the properties considered in the regression model influenced the "time to peak" of flow. A shorter time period would mean higher discharge, which is generally considered the prerequisite to flooding. This research also conclude that the role of geomorphometric properties to control the water supply within the stream through out the year even though during the drought and less precipitations months. Drainage basins are sensitive entities and any deteriorations involve will generate reciprocals and response to the water supply as well as the habitat within the areas.

Relatedness structure in Rhododendron metternichii var. hondoense revealed by microsatellite analysis.

The relatedness structure of Rhododendron metternichii Sieb. et Zucc. var. hondoense Nakai was analysed in a 150 x 70-m quadrat in Hiroshima Prefecture, western Japan. The population of R. metternichii occurred as three subpopulations at the study site (A1-A3). Pairwise relatedness based on microsatellite genotypes at eight loci and Mantel tests revealed a hierarchical structure of relatedness within and among subpopulations: (i) relatedness between individuals within 10 m of one another was significantly positive; (ii) relatedness between individuals in the same subpopulation was significantly positive, but negative between individuals in distant subpopulations; and (iii) relatedness was not significantly different from zero among neighbouring subpopulations. In detail, however, relatedness within each subpopulation was significantly positive in subpopulation A1, relatively weak but significantly positive in subpopulation A2, and not significantly different from zero in subpopulation A3. Relatedness within each subpopulation was inversely related to correlations between interindividual distance and relatedness. The aggregation of related individuals at short distances from one another may lead to decreasing relatedness within subpopulations as a whole. Moreover, negative correlations between interindividual distance and relatedness corresponded to high flowering densities at less than 10-m distance, implying that high flowering densities reduce pollinator foraging distance and lead to stronger genetic structure within subpopulations. Small individuals (< 2.0 m in height) showed stronger genetic structure compared with that of large individuals (> or = 2.0 m in height). The different relatedness structure within and among subpopulations may be caused by various degrees of gene flow affected by distribution patterns of individuals and population density.