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Ultrasonography is widely used to examine reproductive systems in chelonians. Previous studies have described how follicular size is associated with the reproductive stages of yellow-margined box turtles (YMBTs), but our understanding of follicular morphology remains limited. This study describes the different ultrasonographic appearances of preovulatory follicles of these turtles in Taiwan. Randomly selected 28 adult captive female turtles were examined under ultrasonography, weekly from November 2012 to November 2013. The females were divided into ovulatory and non-ovulatory groups with equal number coincidentally based on the whole-year ovulation records. The ovulatory group had significantly larger follicles than the non-ovulatory group (average size of largest follicle, [Formula: see text][Formula: see text]mm versus [Formula: see text][Formula: see text]mm; [Formula: see text]). Across both groups, preovulatory follicles exhibited five different morphologies: previtellogenic, homogeneous, heterogeneous, atretic, and multilayered. The multilayered follicles were only observed in the ovulatory group during the late preovulatory (February) and early ovulatory periods (March), resulting in a significant correlation with ovulatory stage ([Formula: see text]). During the preovulatory period (from December to February), homogeneous follicles were the predominant follicles in both groups. During the ovulation period, heterogeneous follicles were the main follicles, but only in the ovulatory group. Finally, atretic follicles were predominant in the non-ovulatory group during the early latent period (August). This study provides detailed ultrasonographic data clarifying the correlation between turtle follicular morphology and reproductive cycle. A better understanding of how such morphology is associated with reproductive stages should improve breeding programs in zoos and thus facilitate conservation efforts.

The incidence of invasive fungal infections (IFIs) has recently increased, and early and accurate diagnosis of IFIs is important for the rational selection of antifungal drugs with high efficacy. We developed a method for rapid and accurate clinical diagnosis of IFIs and provide a reference for personalized drug treatment. We designed and screened fungal internal transcribed spacer regions with universal primers and designed 8 TaqMan detection probes to establish a multi-channel real-time fluorescent polymerase chain reaction (PCR) melting curve analysis (MCA) method. The sensitivity, specificity, and reproducibility of this method were investigated using standard fungal strains and clinical isolates. Candidemia was detected using the MCA method. The limit of detection and assay cut-off (melting temperature [Tm]) for Candida albicans were 0.05 pg/μL and 66.50 °C; Candida glabrata were 0.1 pg/μL and 66.25 °C; Candida tropicalis were 0.1 pg/μL and 60.15 °C; Candida krusei were 0.1 pg/μL and 72.15 °C; Candida parapsilosis were 0.2 pg/μL and 63.10 °C; Candida guilliermondii were 0.1 pg/μL and 61.84 °C; Cryptococcus neoformans were 0.1 pg/μL and 65.50 °C; Aspergillus flavus were 0.05 pg/μL and 71.50 °C; Aspergillus terreus, Aspergillus fumigatus, and Aspergillus niger were 0.05 pg/μL and 76.80 °C. Analytical specificity was evaluated using 13 clinical pathogens including Streptococcus pneumoniae, Staphylococcus aureus, and Haemophilus influenzae, etc. No false-positive results were obtained for any of these samples. The MCA method can detect and identify different candidemia simulations. The limit detection concentration of C albicans was 44 cfu/mL, C glabrata was 73 cfu/mL, C tropicalis was 29 cfu/mL, C parapsilosis was 21 cfu/mL, C krusei was 71 cfu/mL, and C guilliermondii was 53 cfu/mL. The multi-channel real-time fluorescence PCR melting curve analysis displayed high sensitivity and specificity in detecting various clinically invasive fungi. Furthermore, it simultaneously detected the genera Candida, Cryptococcus, and Aspergillus and identified Candida at the species level. Our method can facilitate early and accurate clinical diagnosis and personalized medication regimens.

Carbapenem-resistant Acinetobacter baumannii (CRAB) is a common nosocomial bacterial pathogen with limited treatment options. CRAB outbreaks are disastrous for critically ill patients. This study investigated carbapenemase-produced A. baumannii outbreaks in a tertiary hospital. Although multiple outbreaks were suggested by pulse-field gel electrophoresis, the genetic lineages and evolution between these isolates were not clear. To investigate the genomic epidemiology of these outbreaks and to reveal possible transmission routes, whole genome sequences (WGS) were compared and analyzed. From the WGS data, thirty isolates had the same sequence type (ST208); acquired resistance genes and chromosome resistant genes were detected and were responsible for multidrug resistance. A phylogenetic tree of single-nucleotide polymorphisms (SNPs) compared to the earliest index isolate found that three outbreaks had emerged and disseminated simultaneously. Of these

Dam removal has the potential to efficiently solve the problems caused by fragmented stream habitats but may simultaneously cause negative impacts on biotic communities. To conserve the critically endangered Formosan landlocked salmon (Oncorhynchus masou formosanus), a 15-m-tall check dam was partially removed from the Chichiawan Stream at the end of May 2011, before the flood season. Using this dam removal experience, we aimed to cast dam removal as an action comparable to a natural flood event. We applied a before-after-control-impact (BACI) design and quantified the environmental factors and major biotic communities at four sampling sites in the stream bimonthly before (2010) and after (2012 and 2013) the dam removal and monthly in the year of the dam removal (2011). After the dam removal, a faster current velocity and more turbid water were observed at the downstream sites, and the area’s deposition consisted of small-grained sediments. Despite this, our results show that the dam removal was performed during a suitable period. There was no obvious influence on tadpoles as they metamorphosed into adult frogs and left the stream before the dam removal. Fish exhibited a greater resistance to the alteration in flow resulting from the dam removal. An increase in fish abundance at the upstream sites after the dam removal suggests that the corridors created by the dam removal allowed access to more habitats for the fish. In particular, the periphyton biomass and aquatic insect densities decreased markedly at the downstream sites after the dam removal, but they recovered within a year, demonstrating the resilience of these taxa. Coleoptera, Plecoptera and Trichoptera were more resistant than the periphyton, Diptera and Ephemeroptera after the dam removal and an extreme flood event. In conclusion, the responses of stream communities to dam removal were similar to the responses to an extreme flood event. To mitigate the impacts caused by dam removal, our results suggest that stream communities may respond to dam removal as a natural flow alteration if the timing of the dam removal occurs just before the flood season.

Abstract ContextUnderstanding the relationships between land use and anuran habitat selection would help make conservation-management and habitat-restoration decisions. However, only a few studies have investigated all the available amphibian habitats in terrestrial and aquatic environments. AimsThe objective of the present study is to assess the associations between different land-use types and individual anuran species in a traditional agricultural area. We also hypothesise that connectivity and land-use heterogeneity are important in determining species richness. MethodsWe conducted monthly roadside calling surveys at 71 sites in the Sun Moon Lake (SML) region in central Taiwan during 2008–2010. Non-metric multidimensional scaling (NMDS) with envfit was used to investigate the relationships between land-use characteristics and anuran assemblage. We use a site-occupancy model to assess the associations between land use and occupancy rate of each species. Generalised linear regression with Poisson distribution was used to assess the relationships among connectivity, land-use heterogeneity and species richness. Key resultsIn total, 19 species in five families were detected by roadside calling surveys. The NMDS results showed anuran assemblages in the SML region being distributed through an environmental gradient from those most dominated by agricultural land to forest-dominant sites. According to site-occupancy models, the occupancy rates of anurans were mainly correlated with proportion of agricultural land, grassland and distance to river. The species richness was positively correlated with land-use heterogeneity. ConclusionsThe results showed that the distributions of anurans largely depended on surrounding land-use types in the terrestrial environment. The major trend of anuran assemblages in the traditional agricultural area was dependent on the transition from forest to agricultural land. ImplicationsThe data suggested that land-use heterogeneity should be a primary consideration for increasing amphibian species richness.

Bloodstream infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) strains have been a severe problem with high clinical costs and high mortality rates. The bla KPC-2 -producing CRKP strain XPY20 was collected from the blood of a patient. The genome characteristics and antimicrobial resistance mechanisms were determined using next-generation sequencing.

Although co-amplification of polymorphic microsatellite markers is the current gold standard for preimplantation genetic diagnosis (PGD) of single-gene disorders (SGD), this approach can be hampered by the lack of availability of informative markers. We recently (2011) devised a novel in-house assay for PGD of aromatic l -amino acid decarboxylase deficiency, based on an amplification refractory mutation system and quantitative PCR (ARMS-qPCR). The objective of the present study was to verify ARMS-qPCR in a cohort of 20 PGD cycles with a diverse group of SGDs (15 couples at risk for 10 SGDs). Day-3 cleavage-stage embryos were subjected to biopsy and genotyping, followed by fresh embryo transfer (FET). The diagnostic rate was 82.9%; unaffected live births were achieved in 9 of 20 FET cycles (45%), with only one false negative (among 54 transferred embryos). Overall, the ARMS-qPCR had frequent allele-dropout (ADO), rendering it inappropriate as the sole diagnostic method (despite a favorable live-birth rate). Regardless, it has the potential to complement the current gold-standard methodology, especially when trophectoderm biopsy becomes a preferred option and genotyping needs to be timely enough to enable FET.

The harvest mouse (Micromys minutus) is a small rodent species with a wide range of vertical distribution in Taiwan, extending from the sea level to 3100 m altitude. This species has recently suffered from habitat loss in high-altitude areas due to orchard cultivation, which may have resulted in mouse migration from high to low altitude. To investigate whether there is any physiological mechanism involved in altitude acclimation, rat cDNA microarray was used to compare transcriptomic patterns of the skeletal muscle tissues taken from individuals native to the high-altitude environment and those transferred to the low-altitude captive site. Of the 23,188 genes being analyzed, 47 (33 up-regulated and 14 down-regulated) were found to have differential expression (fold change &gt, 4 or &lt, &minus, 4, ANOVA p &lt, 0.05). However, after multiple testing correction with a false discovery rate (FDR), only the result for Tnfrsf12a was found to be statistically significant (fold change = 13, FDR p &lt, 0.05). The result was confirmed by quantitative polymerase chain reaction (q-PCR). The expression of Tnfrsf12a possibly relates to the skeletal muscle biology and thus can be correlated with altitude acclimation. However, finding only one gene transcript with significant alteration suggests that transcriptomic response may not play a major role in high- to low-altitude acclimation in harvest mouse.

The harvest mouse, Micromys minutus (MMIN), has a very wide range of distribution (from the British Isles across the Euroasian continent to Japan and Taiwan). We studied an isolated population of MMIN in Taiwan, which is at the southeastern margin of the species' geographic distribution, and compared its genetic complement with those of the same species previously reported from other geographic locations and with two model rodent species, the house mouse (Mus musculus) and the brown Norway rat (Rattus norvegicus). The diploid number (2N) of MMIN was 68, consistent with that reported for other populations. However, variations were noted in the fundamental number (FN) and the shape and banding patterns of the individual chromosomes among populations. The FN of MMIN was estimated to be 72, including 2 bi-armed autosomes, 31 one-armed autosomes, and one pair of one-armed sex chromosomes. Here, we propose the first ideogram for MMIN. C-banding, Ag-NOR, and the locations of 18S rRNA gene sequences (MMIN chromosomes no. 10, 14, 19, 29, 31, 33, and X) mapped by fluorescence in situ hybridization (FISH) are also reported. Additionally, we compared the 18S rDNA sequences and performed cross-species X chromosome painting (FISH) for M. minutus, M. musculus, and R. norvegicus. The results indicate that both genetic elements are rather conserved across species. Thus, implications for the phylogenetic position of Micromys were limited.

Using the Ecopath approach and a network analysis, a mass-balance trophic model was constructed to analyze the structure and energy flows of a high-altitude subtropical headwa- ter stream, Chichiawan Stream, the only habitat of the Critically Endangered Formosan land- locked salmon Oncorhynchus formosanus. Natural abundances of carbon and nitrogen isotopes (δ 13 C and δ 15 N) were used as a complementary method to stomach content analysis to trace food sources of consumers in the stream. Riparian C3 plants were the major food source of dominant gatherer and filterer aquatic insects, while diatoms were the major food source of scraper aquatic insects. Lindeman trophic analysis indicated that the flows of detritivory and herbivory were quantitatively equally important as food sources for consumers in the stream, suggesting a close terrestrialaquatic linkage. The trophic model of Chichiawan Stream comprised 13 compartments and 4 integer trophic levels with the highest trophic level of 3.12 for birds. The pedigree index for the trophic model was 0.71, indicating a high-quality model. The mixed trophic impacts indicated that the biomass of Formosan salmon was unlikely constrained by food sources or predation pres- sure by birds. In a comparative analysis with other trophic models of temperate mountain streams, the total consumer biomass and mean trophic transfer efficiency were relatively low in this sub- tropical mountain stream, which is characterized by high precipitation, steep channels, rapid cur- rent velocity, and changing flow regime. To conserve the Formosan salmon, the first priorities are agricultural runoff control and maintaining a well-vegetated riparian zone.

Two new species of rhacophorid tree frog were identified in Taiwan. In both new taxa, derived reproductive characteristics of laying eggs in tree holes and oophagous tadpoles are shared with Kurixalus eiffingeri, but they are divergent from each other in molecular genetics, mating calls, and tadpole and adult morphology. The morphological characteristics and the molecular phylogenetic evidence support the hypothesis that the two new species, Kurixalus berylliniris sp. n. and Kurixalus wangi sp. n., are both monophyletic lineages.

Summary 1. River food webs rely on two major food sources: autochthonous primary production within the river and allochthonous organic matter transferred to the river. We characterised the consumer communities and assessed the food sources of dominant consumers along a subtropical mountainous river (the Lanyang River of north-eastern Taiwan) at the catchment scale from the headwater to the estuary using natural abundances of stable carbon and nitrogen isotopes. 2. The downstream transport of fine particulate organic matter (FPOM) was two orders of magnitude greater than that of coarse particulate organic matter (CPOM). Transport of both materials increased from the headwater and reached a maximum in the midstream reach. CPOM composition exhibited a gradual shift from leaves and branches in the headwater, an area characterised by high canopy cover, to algae in the midstream reaches and marsh plants in the downstream reaches. 3. Consumer communities can be classified into two regional categories: the upland category in the headwater and upstream and midstream reaches and the lowland category comprised of samples from the downstream reach and estuary. The upland category revealed a clear and gradual seasonal shift in community composition, but a seasonal shift was not apparent for the lowland category. Nutrient concentrations and water temperature were the main factors explaining longitudinal and seasonal variations. 4. The use of sources of organic matter by dominant consumers along the Lanyang River was primarily determined by their availability. Riparian C3 plants were the major food sources in the headwater, upstream reach and estuary, but the contribution of periphyton increased in the upper midstream reach where the river flows through an agricultural area. In the lower midstream and downstream reaches, the contribution of riparian C4 plants became dominant. 5. The trophic transfer of organic materials in the Lanyang River may be influenced by the fast current velocity and by sewage nutrient loading in the river, both of which have important implications for predicting how the functioning of subtropical river food webs will respond to human-related changes in land use.

The objective was to apply a novel modification of a genome-wide, comparative cytogenetic technique (comparative genomic hybridization, comparative genomic hybridization (CGH)), to study species belonging to the myrmecophagous (ant/termite eating) mammalian orders/superorders (Pholidota, Tubulidentata, Carnivora, and Xenarthra), as a model for other applications in mammalian systematics and conservation biology. In this study, CGH was applied to high-quality metaphase spreads of pangolin (Pholidota), using probes of sloth and canine (Xenarthra and Carnivora, respectively) genomic DNA labeled with different fluorophores, thereby facilitating analysis of the visible color spectrum on pangolin karyotypes. Our results posited that pholidotes are closer to carnivores than to xenarthrans, which confirmed the current consensus that myrmecophagy in these mammalian lineages was more likely because of homoplasy (convergent evolution) than being an ancestral character. Since the modified CGH technique used is genome-wide, has chromosome-level resolution, and does not need full genome sequencing, it has considerable potential in systematics and other fields.

Many studies indicated that small mammals are important reservoirs for Bartonella species. Using molecular methods, several studies have documented that bats could harbor Bartonella. This study was conducted to investigate the relationship of Bartonella spp. identified in bats and small mammals living in the same ecological environment. During May 2009 and March 2010, a total of 102 blood specimens were collected. By whole blood culture and molecular identification, a total of 6 bats, 1 rodent and 9 shrews were shown to be infected by Bartonella species. After sequencing and phylogenetic analyses of the sequences of gltA, ftsZ, rpoB and ribC genes, these specific isolates from bats were not similar to the known Bartonella species (the similarity values were less than 91.2%, 90.5%, 88.8%, and 82.2%, respectively); these isolates formed an independent clade away from other known Bartonella type strains. The Bartonella spp. isolated from small mammals, which were closely related to Bartonella tribocorum, Bartonella elizabethae, Bartonella grahamii, Bartonella rattimassiliensis and Bartonella queenslandensis, were similar to the findings in previous studies worldwide. Therefore, the results implied that the species of Bartonella strains isolated from small mammals were different from those identified in bats. Our results strongly suggested that the bat isolate could be a new Bartonella species. This study is also the first one to isolate Bartonella organisms from Asian gray shrews, Crocidura attenuata tanakae.

Constructed wetlands are often built for wastewater treatment to mitigate the adverse effects of organic pollution in streams and rivers caused by inputs of municipal wastewater. However, there has been little analysis of biodiversity and related factors influencing the ecosystem functioning of constructed wetlands. The purpose of this study was to evaluate the biodiversity of two free-water-surface integrated constructed wetlands in subtropical Taiwan by analyzing the water quality, habitat characteristics, and biotic communities of algae, macrophytes, birds, fish, and aquatic macroinvertebrates in the treatment cells. Our results indicated that the two integrated constructed wetlands (Hsin-Hai II and Daniaopi Constructed Wetlands) achieved good performance in reducing the concentrations of total nitrogen (TN) and total phosphorus (TP), and loadings of biochemical oxygen demand (BOD) and chemical oxygen demand (COD) from municipal sewage. In total, 58 bird species, 7 fish species, and 34 aquatic macroinvertebrate taxa were recorded in the two wetlands. The results of stepwise multiple regressions showed that the richness, abundance, and diversity of birds increased with wetland area. Fish richness and abundance respectively increased with wetland area and dissolved oxygen, while the diversity decreased with increases in TP concentrations. The richness and density of aquatic macroinvertebrates increased with the cover of aquatic macrophytes, while the diversity increased with wetland area. Ordination analyses indicated that variations in the community structures of birds, fishes, and aquatic macroinvertebrates were respectively best explained by water temperature, wetland area, and species richness of fish. Our results suggest that wetland area, cover of aquatic macrophytes, and water quality were the most important factors governing the diversity in the constructed wetlands, and that the factors influencing community structures varied among different taxonomic groups. In addition to improving water quality, this study implied that the biodiversity of constructed wetlands for wastewater treatment can be enhanced through proper design and management.

Animals may build multiple structures to provide benefits to counter the costs of building. Many orb web spiders add multiple structures, e.g., barricading barrier webs and silk decorations, to their webs and these structures have been hypothesized to function to deter predators or attract prey. The heavily armored spiny spiders construct barrier webs around their orb webs and decorate them with conspicuous silk tufts. Why these organisms, already well protected by a thick cuticle and spines, make the extra investment of building barrier webs and adding conspicuous silk decorations is not known. We predicted that these structures function to both attract prey and deter predators. Field experiments were conducted in two consecutive years using orb webs built by the East Asian spiny spider Thelacantha brevispina. We either (1) concealed the decoration, (2) removed the barrier webs, or (3) left the decorations and barrier webs intact. We found year and treatment to interactively influence prey interception rates. In 2010, but not in 2009, we found prey interception with T. brevispina webs to be greater when the decorations were conspicuous than when they were concealed suggesting that the decorations may lure prey. Prey interception was lower when the barrier webs were present without decorations compared to when they were absent without decorations. The prey-attracting function of the decorations thus may counter the reduction in prey interception incurred by adding a barrier web. Predatory wasp interactions were not influenced by any of our treatments, probably because the spiders’ thick cuticle is the primary means of protection from wasps. Bird predation events, while rare, occurred only when decorations were concealed or the barrier webs were removed. It is therefore plausible that the tuft decorations both lure prey and deter birds.

The systematic status of Pholidota has been a matter of debate, particularly regarding the apparent inconsistency between morphological and molecular studies. The Sry gene, a master regulator of male sex determination in eutherian mammals, has not yet been used for phylogenetic analyses of extant mammals. The objective of the present study was to clone and characterize the complete gene (1300 base pairs; bp) and amino acid sequences (229 residues) of Sry from the Formosan pangolin (Manis pentadactyla pentadactyla), a member of Pholidota. The Sry amino acid identity between pangolin and other reported species ranged from 42.5% (mouse, Mus musculus) to 84.1% (European hare, Lepus europaeus). Sequence conservation was primarily in the high motility group (HMG) box (234 bp), whereas homology outside the HMG box was low. The cloned Sry was mapped to the pangolin Y chromosome by fluorescence in situ hybridization (FISH); this was confirmed to be the first Y-borne molecular marker identified in Pholidota. Based on Bayesian phylogenetic analysis for Sry HMG sequences from 36 representative taxa, including the Formosan pangolin, Pholidota was more closely related to Carnivora than to Xenarthra, consistent with the emerging molecular tree inferred from markers not located on the Y chromosome. In conclusion, this study characterized the gene structure of Sry of the Formosan pangolin and provided insights into the phylogenetic position of Pholidota.

One of the predicted biological responses to climate warming is the upslope displacement of species distributions. In the tropics, because montane assemblages frequently include local endemics that are distributed close to summits, these species may be especially vulnerable to experiencing complete habitat loss from warming. However, there is currently a dearth of information available for tropical regions. Here, we present a preliminary appraisal of this extinction threat using the herpetological assemblage of the Tsaratanana Massif in northern Madagascar (the island’s highest massif), which is rich with montane endemism. We present meteorological evidence (individual and combined regional weather station data and reanalysis forecast data) for recent warming in Madagascar, and show that this trend is consistent with recent climate model simulations. Using standard moist adiabatic lapse rates, these observed meteorological warming trends in northern Madagascar predict upslope species displacement of 17–74 m per decade between 1993 and 2003. Over this same period, we also report preliminary data supporting a trend for upslope distribution movements, based on two surveys we completed at Tsaratanana. For 30 species, representing five families of reptiles and amphibians, we found overall mean shifts in elevational midpoint of 19–51 m upslope (mean lower elevation limit 29–114 m; mean upper elevation limit � 8t o 53 m). We also found upslope trends in mean and median elevational observations in seven and six of nine species analysed. Phenological differences between these surveys do not appear to be substantial, but these upslope shifts are consistent with the predictions based on meteorological warming. An elevational range displacement analysis projects complete habitat loss for three species below the 21C ‘dangerous’ warming threshold. One of these species is not contracting its distribution, but the other two were not resampled in 2003. A preliminary review of the other massifs in Madagascar indicates potential similar vulnerability to habitat loss and upslope extinction. Consequently, we urgently recommend additional elevational surveys for these and other tropical montane assemblages, which should also include, when possible, the monitoring of local meteorological conditions and habitat change.

In the recent past, the rate of alien species introduction has increased enormously, which is one of the main causes of the loss of biodiversity throughout the world. Pet trade is one of the important channels that lead alien species to invade local ecosystems. In Taiwan, alien reptiles have become popular pets in recent years that increased the risk of invasion. In order to evaluate the probability of alien reptile invasion, it is essential to know how many species of alien reptiles are present in the pet market. From March 2004 to February 2005, we investigated the alien pet reptiles in Taiwan. We checked the pet shops, aquaria and night markets to record the sales of reptiles in Northern, Central, and Southern Taiwan. We visited a total of 341 venders, some of which were visited more than once, and identified 239 species of alien reptiles. From this list, we suggested 10 popular, 14 dangerous, and 8 CITES I species for careful management and strict regulation. A small number of large pet shops carry most of the species, which suggests the sources of importation is limited to a few wholesalers, probably distributed in a few major ports within the island. It would be more efficient to focus on the upstream wholesalers in order to monitor the reptile trade markets. The sheer number of animals found in the pet trade poses the risk of invasion. Other factors, such as the similarity between the pet’s original and new environment, reproductive potential, habitat requirement, and diets of these reptiles, are suggested to be crucial in evaluating the risk of alien pets.

In this study, the situation with imported aquatic invertebrates via the live aquarium ornamental trade in Taiwan was investigated from March 2004 to February 2005. The scientific names, native geographic regions, amount, and trade pathways of imported invertebrates were recorded. Among the 108 imported invertebrate species recorded in this survey, 13 are freshwater species, while the others are marine ones, and most of them were from tropical zones. Through the assessments of invasive potential including the imported amount, the similarity of the native geographic regions to Taiwan, and its invasion record, three species from the class Anthozoa (Cribrinopsis crassa, Condylactis passiflora, and Alcyonium palmatum), one from the class Scyphozoa (Phyllorhiza punctata), three from the order Decapoda (Procambarus clarkia, Cherax destructor, and Neocaridina sp.), one from the class Bivalvia (Lima scabra), and two from the class Gastropoda (Pomacea bridge and Indoplanorbis exustus) were regarded as having high invasive potentials.

Pet trade is an important mechanism for introducing alien species. We surveyed a total of 434 pet shops in major cities of Taiwan and found 49 species of alien amphibians belonging to 14 families and 31 genera. Two of the alien species, Rana catesbeiana and Kaloula pulchra, have established in the fields and the other three, Bufo marinus, Xenopus laevis, and Dendrobates auratus, have invasion records in other countries. There were 16 CITES Appendix II species. The most frequently displayed species were the horned frogs, eratophrys spp. And the most abundant species was the American Bullfrog, Rana catesbeiana. We urge the authority of Taiwan establishing regulations on pet trade and enforcing the wildlife conservation law to reduce the risks of alien species invasions.

Objective: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. Methods: To measure the concentration of theophylline (n=122) and evaluate the assay. Results: The linear range of the CLIA method was 0.51:_40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (

The yellow-marginated box turtle (Cuora flavomarginata) is critically endangered and very few studies have been performed to aid the conservation effort. This study reports the first complete analysis of blood parameters for this species and will provide a reference for future conservation studies. Hematology and biochemistry reference range values were established from 52 (18 males and 35 females) clinically healthy yellow-marginated box turtles (C. flavomarginata). In order to investigate the gender and seasonal differences in these ranges, blood samples were collected from the jugular veins of the turtles in January, April, July, and October to represent the four seasons. To detect significant variation (P < 0.05), data were analyzed using repeated measurements of analysis of variance. Significant gender differences were observed in red blood cell mass parameters, absolute eosinophils, absolute monocytes, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, calcium, uric acid, cholesterol, creatine kinase, glucose, phosphorus (P), total protein, and triglycerides. Marked seasonal variations were noted in red blood cell mass parameters, absolute monocytes, albumin, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, uric acid, creatine kinase, creatinine, glucose, P, and triglycerides. Primary reasons for the differences between gender and season were associated with reproduction and temperature change. Cuora flavomarginata has lower alanine aminotransferase values compared with other chelonians.

Fetal chylothorax (FC) is a rare condition characterized by lymphocyte-rich pleural effusion. Although its pathogenesis remains elusive, it may involve inflammation, since there are increased concentrations of proinflammatory mediators in pleural fluids. Only a few hereditary lymphedema-associated gene loci, e.g. VEGFR3, ITGA9 and PTPN11, were detected in human fetuses with this condition; these cases had a poorer prognosis, due to defective lymphangiogenesis. In the present study, genome-wide gene expression analysis was conducted, comparing pleural and ascitic fluids in three hydropic fetuses, one with and two without the ITGA9 mutation. One fetus (the index case), from a dizygotic pregnancy (the cotwin was unaffected), received antenatal OK-432 pleurodesis and survived beyond the neonatal stage, despite having the ITGA9 mutation. Genes and pathways involved in the immune response were universally up-regulated in fetal pleural fluids compared to those in ascitic fluids. Furthermore, genes involved in the lymphangiogenesis pathway were down-regulated in fetal pleural fluids (compared to ascitic fluid), but following OK-432 pleurodesis, they were up-regulated. Expression of ITGA9 was concordant with overall trends of lymphangiogenesis. In conclusion, we inferred that both the immune response and lymphangiogenesis were implicated in the pathogenesis of fetal chylothorax. Furthermore, genome-wide gene expression microarray analysis may facilitate personalized medicine by selecting the most appropriate treatment, according to the specific circumstances of the patient, for this rare, but heterogeneous disease.

Objectives To develop a diagnostic platform for preimplantation genetic diagnosis (PGD) and prenatal genetic diagnosis (PND) to prevent births of aromatic L-amino acid decarboxylase deficiency (AADC) patients. Materials and Methods Five Taiwanese families carrying AADC were enrolled. A novel technique, amplification refractory mutation system-quantitative polymerase chain reaction (ARMS-qPCR), was developed for both of PGD and PND. For PGD, blastomere biopsies of day-3 cleavage-stage embryos were subjected to ARMS-qPCR. Villi, cultured amniocytes, or both were used to confirm the PGD result; this approach could also be used as the sole method for PND after in vivo conception). Results Unaffected live births were achieved in four of the five families, except one with ongoing PGD. The ARMS-qPCR correctly classified blastomeres (from day-3 cleavage-stage embryos) as affected (homozygous mutant), carrier (heterozygous for mutant and wild-type alleles), or normal (homozygous wild-type) within 1 working day. Conclusions To our knowledge, this is the first report of successful PGD of AADC. The molecular technique we devised (ARMS-qPCR) was applicable for PGD as well as PND of AADC. Furthermore, it has great potential for similar applications in other monogenic disorders.

In practice, when analyzing data from a capture-recapture experiment it is tempting to apply modern advanced statistical methods to the observed capture histories. However, unless the analysis takes into account that the data have only been collected from individuals who have been captured at least once, the results may be biased. Without the development of new software packages, methods such as generalized additive models, generalized linear mixed models, and simulation-extrapolation cannot be readily implemented. In contrast, the partial likelihood approach allows the analysis of a capture-recapture experiment to be conducted using commonly available software. Here we examine the efficiency of this approach and apply it to several data sets.

Chinese species of the genus Niviventer, predominantly distributed in the southeastern Tibetan Plateau and in Taiwan, are a diverse group and have not yet received a thorough molecular phylogenetic analysis. Here, we reconstructed the phylogenetic relationships of 32 specimens representing nine Chinese species of Niviventer, based on sequences of the complete mitochondrial cytochrome b gene. Maximum parsimony, maximum likelihood and Bayesian analysis resulted in three consistent trees, each supported by high bootstrap values. The results showed that the Niviventer species included here are monophyletic. The nine species were classified into three distinct clades: clade A with Niviventer brahma, N. confucianus, N. coxingi, N. culturatus, N. eha and N. fulvescens; clade B with N. andersoni and N. excelsior; clade C with N. cremoriventer. Our results also suggested that N. culturatus should be a valid species rather than a subspecies of N. confucianus. Divergence times among species were calibrated according to the middle-late Pleistocene (1.2-0.13 Mya) fossil records of N. confucianus. The results demonstrated that the first radiation event of the genus Niviventer occurred in early Pleistocene (about 1.66 Mya), followed by the divergence of clades A and B at about 1.46 Mya. Most of the extant Niviventer species appeared during early to middle Pleistocene (about 1.29-0.67 Mya). These divergence times are coincidental with the last uplift events of the Tibetan Plateau, Kun-Huang movement, Pleistocene glaciations and the vicariant formation of Taiwan Strait. Consequently geographical events and Pleistocene glaciations have played a great role in the diversification of Niviventer.

A new method for the calibration of wavelength in a measurement of high resolution molecular absorption spectra by referring the standard spectrum of iodine molecule is presented in this paper. Segment offset is employed in the calibration of wavelength, while the data smoothing, baseline determining and peak fitting are applied to preprocess the spectral data. A visual software is provided for automatically calibrating wavelength with a friendly style of mutual dialogue between human and computer. The software has been used to calibrate the absorption spectra of N2+ molecular ion in the visible region 16,422-17,896 cm-1, the absolute accuracy is about 0.006 cm-1.

Background: The tendency for organisms to be larger in cooler climates, the so-called Bergmann’s rule, has been widely observed in endotherms, but it is debatable in ectothermic anurans. Altitudinal variations in body size, age, and growth rate of the Sauter’s frog Rana sauteri were investigated with skeletochronology at six sites along the altitudinal gradients from 330 to 2,320 m in subtropical Taiwan. The aims of this study were to determine whether these life history traits vary with altitude and differ between males and females. Results: The body size of male R. sauteri followed a converse Bergmann’s cline; the size decreased with increase in altitude, while no significant altitudinal change was found for females. The size dimorphism was female-biased and became greater at higher altitudes because of the altitudinal decrease in male size. Ages ranged between 1 and 5 years for males and 1 and 6 years for females. There was no significant difference in the age structures between the two sexes, but both sexes had higher average ages at higher-altitude sites. According to von Bertalanffy’s model, the growth coefficients decreased with increase in altitude for males but not for females. The annual growth rates were greater for all age classes of females than males, suggesting that growth rate was a major factor underlying body size patterns of both sexes. Conclusions: Growth rate was a major factor underlying body size patterns in both sexes of R. sauteri. It could be affected by not only altitudinal temperature gradient but also sex-biased life history traits and environmental factors.

Specimens of Microhyla inornata were collected at the southern tip of Taiwan. Descriptions of adults and tadpoles are presented. We show that the separation of different color morphs of the species into subspecies is probably inappropriate. Tadpoles of M. inornata can be distinguished from M. ornata in mouth morphology, tail shape, and pigmentation. Tadpoles of this species may eat macrophytic plant material in the water, regardless of their beakless mouthparts, in addition to filter feeding on microor- ganisms. Microhyla inornata breeds during and immediately after the heavy rain. Duration of development (from eggs to froglets) is less than three weeks. Thus, this species is considered an explosive breeder. Mating calls of M. inornata consist of three frequency bands at 4.4-5.7 (fundamental), 8.8-10.0, andl4.0- 15.0 (two harmonics) kHz, respectively. The occurrence of Microhyla inornata Boulen- ger in Taiwan was first documented when Bou- lenger (1909) described the new species M. stei- negeri on the basis of five specimens collected