Shih-Hsiung Liang

Application of real-time PCR, DGGE fingerprinting, and culture-based method to evaluate the effectiveness of intrinsic bioremediation on the control of petroleum-hydrocarbon plume.

Real-time polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and the culture-based method were applied in the intrinsic bioremediation study at a petroleum-hydrocarbon contaminated site. The genes of phenol hydroxylase (PHE), ring-hydroxylating toluene monooxygenase (RMO), naphthalene dioxygenase (NAH), toluene monooxygenase (TOL), toluene dioxygenase (TOD), and biphenyl dioxygenase (BPH4) were quantified by real-time PCR. Results show that PHE gene was detected in groundwater contaminated with benzene, toluene, ethylbenzene, xylene isomers (BTEX) and methyl tert-butyl ether (MTBE), and this indicates that intrinsic bioremediation occurred at this contaminated site. Results from DGGE analyses reveal that the petroleum-hydrocarbon plume caused the variation in microbial communities. In this study, MTBE degraders including Pseudomonas sp. NKNU01, Bacillus sp. NKNU01, Klebsiella sp. NKNU01, Enterobacter sp. NKNU01, and Enterobacter sp. NKNU02 were isolated from the contaminated groundwater using the cultured-based method. Results from MTBE biodegradation experiment show that the isolated bacteria were affected by propane. This indicates that propane may influence the metabolic pathway of MTBE by these bacteria. Knowledge and comprehension obtained from this study will be helpful in evaluating the occurrence and effectiveness of intrinsic bioremediation on the remediation of petroleum-hydrocarbon contaminated groundwater.

Treatment of TCE-Contaminated Groundwater Using Fenton-Like Oxidation Activated with Basic Oxygen Furnace Slag

The industrial solvent trichloroethylene (TCE) is among the ubiquitous chlorinated organic compounds found in groundwater contamination. The objective of this study was to evaluate the potential of applying basic oxygen furnace (BOF) slag as the catalyst to enhance the Fenton-like oxidation to remediate TCE-contaminated groundwater. Results indicate that TCE oxidation via the Fenton-like process can be enhanced with the addition of BOF slag. Results from the X-ray powder diffraction analysis reveal that the major iron type of BOF slag/quartz sand media was iron oxyhydroxide ( α -Fe2 O3 ) . Approximately 81% of TCE removal was observed (with initial TCE concentration of approximately 5 mg  L−1 ), with the addition of 1,000 mg  L−1 of H2 O2 and 10 g  L−1 of BOF slag. Results also show that TCE concentrations dropped from 5 to 1.1 mg  L−1 , and chloride concentrations increased from 0 to 2.7 mg  L−1 after 60 min of reaction with the presence of H2 O2 and BOF slag. This indicates that the depletion of TCE cor...

Treatment of Fuel-Oil Contaminated Soils by Biodegradable Surfactant Washing Followed by Fenton-Like Oxidation

Among petroleum-hydrocarbon pollutants, fuel-oil is more difficult to treat compared to gasoline and diesel fuel. The objectives of this bench-scale study were to: (1) develop a two-stage remedial system consisting of surfactant washing followed by Fenton-like oxidation process to remediate fuel-oil contaminated soils; (2) evaluate the effects of residual surfactant and soil organic matter (SOM) on the efficiency of Fenton-like oxidation; (3) evaluate the effect of potassium dihydrogen phosphate ( KH2 PO4 ) addition on the stability of H2 O2 and oxidation efficiency; and (4) evaluate the possible oxidation products after the oxidation process. In the surfactant washing stage, biodegradable surfactant, Simple Green (SG) ( 50 g  L−1 ) , was applied to flush fuel-oil contaminated soils with initial total petroleum-hydrocarbons (TPHs) concentration of 50,000 mg  kg−1 . Results show that approximately 90% of TPH could be removed after washing with 45 pore volumes (PVs) of SG followed by 25 PVs of deionized wat...

Acoustic adaptations to anthropogenic noise in the cicada Cryptotympana takasagona Kato (Hemiptera: Cicadidae)

Anthropogenic noise produced by human activities affects acoustic communication in animals living in urban habitats. We recorded the calling songs of the cicada Cryptotympana takasagona in the Kaohsiung metropolitan areas of southern Taiwan to investigate possible acoustic adaptations to anthropogenic noise. C. takasagona did not call more in noise gaps. Acoustic features (peak frequency, quartile 25%, quartile 50%, and quartile 75%) of calling songs significantly increased with ambient noise levels. C. takasagona shifted the energy distribution of calling songs to higher frequencies in the presence of higher noise levels. We suggest that the acoustic adaptation by which song frequencies increase with levels of anthropogenic noise in C. takasagona may result from a size-dependent calling strategy in which small-sized males call more in noise conditions or large-sized males adjust their song frequency by changing their abdominal cavities.

Cloning, expression, and characterization of cadmium-induced metallothionein-2 from the earthworms Metaphire posthuma and Polypheretima elongata.

In this study we report the sequences of MT-2 cDNA from two species of Megascoleidae earthworms, Metaphire posthuma and Polypheretima elongata, by mRNA differential display after exposure of the organisms to cadmium. Complementary (c)DNA was verified as the MT-2 gene by the characteristics of its predicted translation product, namely a high cysteine content, conserved CXC motifs, and a molecular weight of around 8 kDa. Amino acid sequence alignment revealed a conserved TKCCG in the cloned MT-2 of both megascolecid earthworms instead of the corresponding conserved TQCCG found in lumbricid earthworms. The cDNAs corresponding to the two megascolecid MT-2 genes were expressed, and the MT-2 proteins were purified for biochemical characterization. The binding of Cu2+ exhibited monophasic kinetics and those of Zn2+ and Cd2+ biphasic kinetics. The proteins bound more tightly to Cd2+ than to Zn2+ and more tightly still to Cu2+. Zn-MT and apo-MT were the most effective at scavenging free radicals, followed by Cd-MT. In conclusion, MT-2s from M. posthuma and P. elongata showed unique sequence features compared to those of lumbricid earthworms. These earthworms could be used to evaluate heavy-metal pollution in soil due to the inducible MT-2 by cadmium exposure.

Enhancement of microbial 2,4,6-trinitrotoluene transformation with increased toxicity by exogenous nutrient amendment

In this study, the bacterial strain Citrobacter youngae strain E4 was isolated from 2,4,6-trinitrotoluene (TNT)-contaminated soil and used to assess the capacity of TNT transformation with/without exogenous nutrient amendments. C. youngae E4 poorly degraded TNT without an exogenous amino nitrogen source, whereas the addition of an amino nitrogen source considerably increased the efficacy of TNT transformation in a dose-dependent manner. The enhanced TNT transformation of C. youngae E4 was mediated by increased cell growth and up-regulation of TNT nitroreductases, including NemA, NfsA and NfsB. This result indicates that the increase in TNT transformation by C. youngae E4 via nitrogen nutrient stimulation is a cometabolism process. Consistently, TNT transformation was effectively enhanced when C. youngae E4 was subjected to a TNT-contaminated soil slurry in the presence of an exogenous amino nitrogen amendment. Thus, effective enhancement of TNT transformation via the coordinated inoculation of the nutrient-responsive C. youngae E4 and an exogenous nitrogen amendment might be applicable for the remediation of TNT-contaminated soil. Although the TNT transformation was significantly enhanced by C. youngae E4 in concert with biostimulation, the 96-h LC50 value of the TNT transformation product mixture on the aquatic invertebrate Tigriopus japonicas was higher than the LC50 value of TNT alone. Our results suggest that exogenous nutrient amendment can enhance microbial TNT transformation; however, additional detoxification processes may be needed due to the increased toxicity after reduced TNT transformation.

Acoustic and temporal partitioning of cicada assemblages in city and mountain environments.

Comparing adaptations to noisy city environments with those to natural mountain environments on the community level can provide significant insights that allow an understanding of the impact of anthropogenic noise on invertebrates that employ loud calling songs for mate attraction, especially when each species has its distinct song, as in the case of cicadas. In this study, we investigated the partitioning strategy of cicada assemblages in city and mountain environments by comparing the acoustic features and calling activity patterns of each species, recorded using automated digital recording systems. Our comparison of activity patterns of seasonal and diel calling revealed that there was no significant temporal partitioning of cicada assemblages in either environment. In addition, there was no correlation between the acoustic distance based on spectral features and temporal segregation. Heterospecific spectral overlap was low in both city and mountain environments, although city and mountain cicada assemblages were subject to significantly different levels of anthropogenic or interspecific noise. Furthermore, for the common species found in both environments, the calling activity patterns at both seasonal and diel time scales were significantly consistent across sites and across environments. We suggest that the temporal calling activity is constrained by endogenous factors for each species and is less flexible in response to external factors, such as anthropogenic noise. As a result, cicada assemblages in city environments with low species diversity do not demonstrate a more significant temporal partitioning than those in mountain environments with high species diversity.

Detection of gentoxicity of benzidine and its derivatives with the Escherichia coli DJ 702 lacZ reversion mutagenicity assay

Aims:  The feasibility of Escherichia coli DJ 702 lacZ mutagenicity assay to detect genotoxicity of benzidine and its derivatives was evaluated.

Soil-eating behavior in Formosan macaques (Macaca cyclopis) at Shoushan Nature Park of southern Taiwan

In this first study of soil-eating behavior in Formosan macaques, we attempted to test the mineral supplementation hypothesis, and to document macaques using excavation sites at both the individual and social group levels. We observed macaques’ soil-eating behavior from October 1999 to January 2000 at Shoushan Nature Park in southern Taiwan, and compared the chemical compositions of soils from their excavation sites with those of randomly selected topsoils that were not eaten by macaques in the study area. Our results show that soils from excavation sites contained significantly higher levels of calcium than did topsoils taken from randomly selected sites. This provides evidence to support the mineral supplementation hypothesis. However, alternative hypotheses such as the treatment of diarrhea/endoparasites, detoxification, and behavioral tradition could not be excluded from explaining the occurrence of soil-eating behavior in Formosan macaques. At both the individual and social group levels, a behavioral tradition may play a particularly important role in affecting the choices of excavation sites where soil is eaten. Further research on the exact cues of excavation sites which stimulate soil-eating behavior is needed to clarify this suggestion.

Song frequency correlates with latitude and individual body size in the cicada Mogannia formosana Matsumura (Hemiptera: Cicadidae)

Analyses of acoustic variation between and within populations can help to clarify the evolution and diversification of cicada calling songs. In this study, we analyzed the acoustic variation in the calling song of Mogannia formosana within Taiwan and between Taiwan and Green Island to assess the effects of geographic locations. Furthermore, chorusing males in the Green Island population were recorded and collected from the same habitat site during the same time period to investigate the relationship between individual body size and the acoustic features of calling songs. Among populations of M. formosana, we found that most of the acoustic variation in M. formosana calling songs was associated with frequency parameters, in which six frequency parameters changed significantly with latitude on the island of Taiwan. In contrast, temporal parameters, which were associated with principal components corresponding to less acoustic variation than that of the first principal component, were also found to be significant among populations but did not show consistent trends of difference with latitude. However, the geographically isolated Green Island population exhibited the lowest number of short echemes in segment B, which is the diagnostic structure employed to separate M. formosana from other M. species. This finding suggests that the temporal pattern of segment B in the calling songs of M. formosana might be important for both population differentiation and interspecific recognition. In a chorus of the Green Island population, the sound frequency of the last short echeme was found to be significantly correlated with individual body size. The possible role played by sexual selection in shaping sound frequency as a result of its reliable association with body size was discussed. We suggest that, in comparison with temporal elements, the frequency elements of segment B in calling songs of M. formosana in chorus can serve as a more reliable indicator of body size for female mate choice.