Bao-Sen Shieh

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.

Biodegradation of methyl tert-butyl ether (MTBE) by Enterobacter sp. NKNU02

We previously isolated and identified Enterobacter sp. NKNU02 as a methyl tert-butyl ether (MTBE)-degrading bacterial strain from gasoline-contaminated water. In this study, tert-butyl alcohol, acetic acid, 2-propanol, and propenoic acid were detected using gas chromatography/mass spectrometry when MTBE was degraded by rest cells of Enterobacter sp. NKNU02 cells. We also found that biodegradation of MTBE was decreased, but not totally inhibited in mixtures of benzene, toluene, ethylbenzene and xylene. The effects of MTBE on the biology of Enterobacter sp. NKNU02 were elucidated using 2D proteomic analysis. The cytoplasmic proteins isolated from these MTBE-treated and -untreated cells were carried out for proteomic analysis. Results showed that there were 6 differential protein spots and 8 differential protein spots, respectively, as compared to their corresponding control (without MTBE addition), at the indicated incubation times when 40% and 60% of 100 mg/L of MTBE had been removed, Among these proteins, nine were successfully identified with matrix-assisted laser desorption ionization-time of flight-mass spectrometry. Proteins identified included extracellular solute-binding protein, periplasmic-binding protein ytfQ, cationic amino acid ABC transporter, isocitrate dehydrogenase, cysteine synthase A, alkyl hydroperoxide reductase (AhpC), transaldolase, and alcohol dehydrogenase. Based on these differential proteins, we discuss the bacterial responses to MTBE at the molecular level.

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.

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.

Burkholderia pseudomallei infection induces the expression of apoptosis-related genes and proteins in mouse macrophages.

BACKGROUND/PURPOSE In this study, we addressed whether the production of apoptosis-related genes and proteins is induced in mouse macrophages infected with Burkholderia pseudomallei cells. METHODS Mouse macrophages were infected with B. pseudomallei cells at 0.5 hours, 1 hour, 2 hours, 4 hours, and 6 hours, respectively, followed by real-time polymerase chain reaction (PCR) array analysis. The amount of apoptosis-related proteins (caspase-3, caspase -8, caspase -9, Bax, and Bcl-2) was confirmed by Western blot. RESULTS After infection, an increase of these proteins was observed. The expression levels of other apoptosis-related genes were also determined by PCR array. Experimental results revealed that the messenger RNA levels of tumor necrosis factor ligand (e.g., tnfsf10 and tnfrs10b) and fas were increased, whereas the expression levels of some antiapoptosis genes such as Birc5, Hells, and Bnip3 were decreased. CONCLUSION Our study results demonstrate that the apoptosis-related genes and proteins in mouse macrophages were modulated by B. pseudomallei.

Development and assays estradiol equivalent concentration from prawn (p-EEQ) in river prawn, Macrobrachium nipponense, in Taiwan.

The present study established a fast and convenient bioassay method for aqueous ecosystems using the prawn estradiol equivalent concentration (p-EEQ) of male Macrobrachium nipponense, which produce vitellogenin (VTG) after exposure to xeno-estrogens. This method was then used to determine the concentrations of xeno-estrogen pollutants in the rivers of Taiwan. To establish the calibration curve for the concentrations based on the p-EEQ, the induced VTG content was determined using the alkali-labile phosphate method after male M. nipponense were exposed to 0, 10, 100, 1,000 and 10,000ng/L of 17β-estradiol for 1, 3, 5, 7, 10 and 14 days, respectively. The results of the experiments showed that the induced VTG content in all of the experimental groups stabilized after 10 days, except for the 10,000ng/L experimental group, in which the induced VTG content decreased after 10 days. A 17β-estradiol-VTG10day response curve was then established based on the induced VTG content in the 0, 10, 100 and 1000ng/L experimental groups at day 10. After establishing the curve, male M. nipponense were captured from the upper, middle and lower reaches of the Chuo-shui River, the Beigang River, the Jishui River, the Agongdian River and the Sichong River in Taiwan, and the VTG content in these prawns was determined. In addition, the p-EEQ in the waters was determined based on the VTG content, and the estradiol equivalent concentration (EEQ) in the waters was also measured immediately after sampling using the solid-phase extraction-enzyme-linked immunosorbent assay (SPE-ELISA) method. The results showed that the p-EEQ in the middle and lower reaches of the rivers in certain parts of Taiwan ranged from 38 to 400ng/L, and the detection rate was 100%. Moreover, the EEQ ranged from 7.9 to 92.9ng/L, and the detection rate was 42.9%, indicating that most of the middle and lower reaches of the rivers in Taiwan were polluted by xeno-estrogens. The 17β-estradiol concentrations determined based on the p-EEQ were all higher than those based on the EEQ (SPE-ELISA method). The results of the present study showed that the use of M. nipponense to determine the p-EEQ in environmental waters provided advantages that included a high detection rate, high sensitivity and convenience. However, the p-EEQ cannot be used in waters that do not contain M. nipponense.

Diurnal Timing of Bird Surveys Using an Acoustic Monitoring System in the Shan-Ping Forest Ecological Garden

In this study, we report how to select the best timing of diurnal bird surveys when using an acoustic monitoring system. Although the system has been operating since March 2009 in the Shan-Ping Forest Ecological Garden, we transcribed only 2 morning recordings in the breeding season of 2009 due to the difficulty in transcribing recorded data. We found that the species accumulation curve reached a plateau (80%) in 30 min of the first hour and was higher than those in the latter 4 h on both days, which implied that the dawn chorus is the best period for recording bird sounds. A further analysis was used to locate the timing of the dawn chorus. The highest accumulation curve in a 10-min period appeared when the starting point was set at 5 min and 20 or 15 min before sunrise on March 31 and May 4, respectively. The difference revealed that the peak of singing activities occurred at different times over the breeding season. We suggest recording bird sounds for 1 h from dawn and selecting a sampling period of 10~20 min, which would cover most of the dawn chorus, to reach the highest species richness at the study site.

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.

Interspecific comparison of traffic noise effects on dove coo transmission in urban environments

Most previous studies concerning avian adaptation to anthropogenic noise have focused on songbirds, but few have focused on non-songbirds commonly found in urban environments such as doves. We conducted field playback-recording experiments on the perch-coos of five dove species, including four native Taiwan species (the spotted dove, Spilopelia chinensis, the oriental turtle-dove, Streptopelia orientalis, the red collared-dove, Streptopelia tranquebarica, and the emerald dove, Chalcophaps indica) and one species not native to Taiwan (the zebra dove, Geopelia striata) to evaluate the detection and recognition of dove coos in habitats with differing levels of traffic noise. Our results suggest that traffic noise has selected dominant urban species such as the spotted dove to temporally and spatially adjust cooing to reduce the masking effects of traffic noise and rare urban species such as the emerald dove to avoid areas of high traffic noise. Additionally, although the zebra dove had the highest coo frequency among the study species, its coos showed the highest detection value but not the highest recognition value. We conclude that traffic noise is an important factor in shaping the distribution of rare and dominant dove species in urban environments through its significant effects on coo transmission.

Dominance rank and interference competition in foraging among six species of birds in a park in Kaohsiung City, Taiwan.

Through body size, social interaction, and foraging behavior, we investigated the dominance rank and interference competition among 6 bird species foraging at a park feeding site in Kaohsiung City, Taiwan. Social interactions and foraging behaviors of these birds were recorded in June to September 2009. Davids scores were calculated from an interspecific interaction matrix, and the score roughly increased with the body size of birds, but some exceptions were noted. Concerning foraging behavior, feral pigeons (Columba livia) and Spotted-necked Doves (Streptopelia chinensis) took over the food area once they appeared even though Tree Sparrows (Passer montanus) usually arrived first. A linear regression model indicated that the number of Tree Sparrows outside the food area was positively correlated with the number of feral pigeons and Spotted-necked Doves inside the food area. Feral pigeons and Spotted-necked Doves moved away as the food was gradually consumed, and smaller species accordingly increased their foraging in the food area. Nevertheless, the Tree Sparrow was also suppressed by other medium-sized birds, like the White-vented Myna (Acridotheres javanicus) and Chinese Bulbul (Pycnonotus sinensis), and they eventually occupied the food area in large numbers at a later stage. This study revealed that body size did matter and the Tree Sparrow was clearly the least dominant species among the 6. However, a discrepancy between the dominance status and interference competition in foraging was apparent. In addition to Davids score, we suggest incorporating body size, group size, and interference competition to reach a more-comprehensive dominance hierarchy in bird communities.