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Social insects nesting in soil environments are in constant contact with entomopathogens but have evolved a range of defence mechanisms, resulting in both individual and social immunity that reduce the chance for epizootics in the colony, as in the case of subterranean termites. Coptotermes formosanus uses its faeces as building material for its nest structure that result into a 'carton material', and here, we report that the faecal nest supports the growth of Actinobacteria which provide another level of protection to the social group against entomopathogens. A Streptomyces species with in vivo antimicrobial activity against fungal entomopathogens was isolated from the nest material of multiple termite colonies. Termite groups were exposed to Metarhizium anisopliae, a fungal entomopathogen, during their foraging activity and the presence of Streptomyces within the nest structure provided a significant survival benefit to the termites. Therefore, this report describes a non-nutritional exosymbiosis in a termite, in the form of a defensive mutualism which has emerged from the use of faecal material in the nesting structure of Coptotermes. The association with an Actinobacteria community in the termite faecal material provides an extended disease resistance to the termite group as another level of defence, in addition to their individual and social immunity.
1. Introduction
The Formosan subterranean termite, Coptotermes formosanus, builds underground nests with extensive foraging galleries for up to 150 m; a colony can contain more than 1 million individuals and it is considered as an important urban pest because of the damage it can do to wood structures [1]. While the inner lining of foraging galleries are coated with a termite faecal envelop, the voids created at foraging sites and the core of the nest are filled with a sponge-like structure composed of chewed wood particles mixed with faecal material, usually referred to as 'carton material' (figure 1). It was suggested to have first emerged in a common ancestor of Coptotermes and Macrotermes as a result of subterranean nest-building behaviour [3]. This building behaviour reinforces the nest structure and helps to maintain homoeostatic conditions inside the nest by reducing the temperature and moisture fluctuations from the surrounding soils [4] and the faecal envelop can be the host of a diverse microbial community [5]. By maintaining relatively high temperatures and humidity inside the nest, it was suggested that termites, as social insects living in highly dense colonies, would face an increased risk of epizootic events within the nest [6]. However, subterranean termites have evolved a range of disease resistance mechanisms, preventing harmful pathogens from spreading within the group [7,8]. Such mechanisms involve the individual cellular and humoral immune defence of each termite [9,10] but also includes hygienic behaviours which results in a social immunity [11–13]. In addition to the collective effort of the colony to limit the risk of epizootics of fungal entomopathogens, for example Metarhizium anisopliae [14], endogenous termite secretions may help to create a termite nesting environment poorly conducive for such fungi [15–19]. However, the description of the termite–Metarhizium relationship as a simple bipartite association appeared reductive [20]. We therefore propose that an exogenous protection may complete the termite's defence repertoire, here provided by a mutualistic association within the termite nest.
Defensive mutualism involving insects and Actinobacteria have been extensively described [21–23]. Some ants [24–29], wasps [30–32] and beetles [33,34] are associated with Actinobacteria that are using the insect environment or the insect itself as a microniche, and can provide direct or indirect benefits for the insect against parasites or pathogens. Because the niche of such insects can be so unique, some of the Actinobacteria associated with these insects can be uncommon and may represent an untapped source for undiscovered antibiotics [32,35]. In subterranean termites, the carton material and the faecal lining in the foraging galleries provides a niche for the colonization of microorganisms [5,36] and specific Actinobacteria can provide secondary metabolites that benefit the termites against common soil entomopathogens. However, it can be difficult to provide evidence on the nature of an association between termites and microorganisms [20,37]. Therefore, to properly demonstrate a case of mutualism (see the electronic supplementary material, discussion), we established in this study the interaction of each organism in relation to each other (termites, fungal entomopathogens, Actinobacteria; figure 2), tested with in vivo conditions to confirm the reciprocal benefits that both the termites and Actinobacteria can obtain from this association.
2. Material and methods
(a) Actinobacteria isolation from termite nest material
Carton material of five C. formosanus field colonies were collected from 'bucket traps' as described by Su & Scheffrahn [39]. Bucket traps were installed at active C. formosanus foraging sites (Broward County, FL, USA) and an autoclaved bundle of spruce wood was placed in the bucket. After two weeks in the ground, the partially consumed bundles of wood with voids filled with carton material were retrieved. Carton material samples were then collected under sterile conditions in the laboratory. To isolate Actinobacteria from termite carton material that would preferentially be associated with such a microniche, we followed the suggestions of Kaeberlein et al. [40] that poorly cultivable microorganisms may easily grow in culture if provided with the nutritional components of their natural environment, and of Shlatter et al. [41] that soil amended with cellulose and lignin can host higher densities of Streptomyces than soil poor in such resources. The termite faecal material integrated in the carton material is essentially composed of partially degraded plant material and may provide a natural selective media for termite associated Actinobacteria. We therefore used termite carton material as a nutritional base for a novel selective medium, termite faecal agar (TFA). It was prepared by adding 60 g of ground carton material from a C. formosanus laboratory nest to 1 l of water agar and then autoclaved. The use of a laboratory termite colony allowed us to obtain large quantities of carton material with homogeneous quality, which were necessary to prepare enough media for this large isolation protocol. To prevent the growth of fast growing fungi, 100 mg of cycloheximide per litre of agar was added to the TFA after sterilization. For each termite colony of origin, three subsamples of fresh carton material were subjected to serial dilutions (10–4 to 10−6) using deionized water, which were plated on the TFA medium (four replicates per dilution), and incubated in the dark at 28°C for 9 days.
Bacterial colony forming units (CFU) with an Actinobacteria morphotype were individually isolated and subcultured on 1/5 strength potato dextrose agar (1/5 PDA). In order to prevent a systematic bias in the isolation process, half of the replicates of the dilution plates were processed by T.C., whereas the other half was processed by C.A.E. Pure Actinobacteria colonies (500 + isolates) were archived in storage and a subset of representatives (based on morphology and antibiotic activity profile described below) from each nest of origin was selected for sequencing. The 16SrDNA sequence (1200 bp) was obtained by using the microbial sequencing service provided by Laragen (Culver City, CA, USA). 16SrDNA sequences were edited using DNA Baser v. 2.9 (Heracle software). Sequence alignment and phylogeny trees were computed with Mega v. 5.0, using neighbour-joining method with default parameters. Sequences were deposited in GenBank database under the accession numbers KC111822-1873.
(b) Antimicrobial assay
Actinobacteria isolates obtained from C. formosanus carton materials were tested for their antimicrobial activity against two fungal entomopathogens, M. anisopliae (ATCC 98094) and Beauveria bassiana (undescribed isolate). In addition, isolates were tested against a range of microorganisms to determine a basic profile of their overall antimicrobial activity. These microorganisms represent species of Gram-negative and Gram-positive bacteria, as well as yeast and other fungi. Test species included Bacillus subtilis (ATCC 6633), Serratia marcescens (ATCC 43862) Staphylococcus aureus (NCIMB 9518), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 25619), Xantophomonas maltophilia (ATCC 13636), Klebsiella pneumonia (BAA-2146), Aspergillus nomius (AsFL-07), Penicillum sp. (undescribed isolate) and Saccharomyces cerevisiae (ATCC 2601). Some of these microbial species are infectious to immune-compromised individuals in humans and are known to be problematic in hospitals as nosocomial diseases, owing to their acquired drug resistance to commonly used antibiotics [42]. Therefore, this preliminary screening not only aimed to provide a basic antimicrobial profile, but also to prospect for novel antibiotic agent.
The antimicrobial screening was accomplished by growing Actinobacteria isolates individually on 1/5 PDA, centred in the plate, for 4 days to allow diffusion of secondary metabolites through the plate (two replicates per isolate). The culture was then overlaid with 10 ml of water agar containing a concentrated cell or conidia suspension of the test organism (100 #CFU per ml). The incubation for a given test organism was done at their optimal growth temperatures [43] and measurements of the growth inhibition were done 24 h after overlay for bacteria and yeasts, and 72 h after overlay for fungi. The zone of growth inhibition was established by measuring the average distance (four orthogonal measurements) between the edge of the Actinobacteria colony and the edge of the zone of inhibition. Zones more than 40 mm indicated that the inhibition was complete over the entire plate. We are aware that testing for antimicrobial activity only in a 1/5 PDA medium (pH = 5.25) is a reductive approach as some Actinobacteria may require a different nutritional or pH environment to produce particular antimicrobial secondary metabolites, however, in this study, we restricted the screening for practical purpose and to focus our effort on a thorough termite bioassay.
(c) Metarhizium anisopliae preparation
Because a vast majority of virulence studies and defence mechanisms studies in termites were performed with M. anisopliae as a model entomopathogen [7,8,37,38], this fungus species was selected as a representative for further in vitro and in vivo bioassays. Conidia of M. anisopliae were spread on 1/5 PDA and incubated at 28°C in the dark. After 48 h, single conidia colonies were transferred to 1/5 PDA containing one worker termite, previously killed and surface-sterilized for 1 h in a vial saturated with chloroform vapours [44]. The use of the sterilized termite is to provide proper nutrition for the fungus, which helps maintain its virulence through subculture. Inoculated plates were then incubated at 28°C for 14 days in the dark. Fresh conidia were harvested from these plates with a 0.05% Tween80 solution (for conidia suspension) and a stock suspension of 5 × 107 viable conidia per millilitre was prepared (determined by conidia germination on 1/5 PDA). Conidia stock suspensions were stored at 4°C and used for dilutions (with sterile deionized water) within 15 days of the experiments described herein (97% viable conidia after 15 days).
(d) Streptomyces #2338 as a representative
The isolate Streptomyces #2338 was used as a representative of a ribotype sampled from all five termite nests (see electronic supplementary material, figure S1). This ribotype also showed strong in vitro antifungal activity against the entomopathogenic fungus M. anisopliae and it was considered as a potential candidate for mutualism in C. formosanus. However, the observation of such antibiotic activity in a highly artificial environment may have little relevance biologically [45] and may not represent the antimicrobial activity that the isolate could display in a termite nest environment. Therefore, we first tested the interaction of Streptomyces #2338 and M. anisopliae in a termite nest-like environment, that is in the presence of termite carton material, before proceeding to the termite bioassay. A Streptomyces #2338 stock solution of 109 viable cells per millilitre was obtained from 1/5 PDA subcultures and suspended with a 0.05% Tween80 solution.
(e) Interaction between Metarhizium anisopliae and Streptomyces #2338 in a termite nest-like nutritional environment
Acetone-washed and triple-rinsed white sand (150–500 µm particle size) was autoclaved and used as a neutral soil matrix for all experiments. Two types of soil environment were tested on microbial growth: pure sand (oligotrophic, as a negative control), and sand amended with sterile termite carton material (75% sand, 25% carton). For each treatment, 10 g of sand or sand–carton mix were placed in a 10 cm sterile glass Petri dish and moistened with 2 ml of sterile deionized water. There were four treatments per type of soil (three replicates per treatment): control treatment (deionized water), Streptomyces #2338 only (106 cell per gram of dry soil), M. anisopliae (105 conidia per gram of dry soil) and Streptomyces #2338 + M. anisopliae (concentrations as above). Microbial growth was monitored every 7 days for 42 days (except at 35 days) using the following protocol: for each Petri dish, three subsamples of 0.6 g (0.5 g dry weight equivalent) were subjected to serial dilutions (three replicate for each subsamples) and plated on 1/5 PDA for all treatments from both soil environments to count for CFUs. However, for the Streptomyces #2338 + M. anisopliae treatments, the serial dilutions of the subsamples were replicated and plated separately on 1/5 PDA amended with cycloheximide (100 mg l−1) to monitor only Streptomyces #2338 growth, and on 1/5 PDA amended with chloramphenicol (100 mg l−1) to monitor only M. anisopliae growth. CFUs were counted 3 days after plating the serial dilutions (n = 27 per treatment, per dilution).
(f) Inhibition of Metarhizium anisopliae by Streptomyces #2338: exclusion and/or chemical interference?
After the inhibition of M. anisopliae by Streptomyces #2338 was determined in a termite nest-like environment, the type of inhibition between both microorganisms was investigated. In a subterranean termite faecal niche, the observed inhibition can be owing to three possible mechanisms: (i) exclusion, where Streptomyces #2338 outcompetes M. anisopliae by dominating the niche with a high density, to reduce the access to nutrients by M. anisopliae, (ii) chemical interference, where Streptomyces #2338 produces antifungal secondary metabolites that reduce the growth of M. anisopliae and (iii) a combination of both mechanisms. To test the origin of the inhibition, an identical protocol as above (interaction in a termite nest-like environment) was repeated, but instead of using Streptomyces #2338, the isolate Streptomyces #2345 was used. This isolate was used for comparison with #2338 because they were both isolated from the same termite carton material, both belong to the same 16S subclade, but contrary to #2338, the isolate #2345 did not display any antibiotic activity in vitro (see electronic supplementary material, figure S1). If the growth of isolate #2345 was similar to #2338 in a termite faecal environment, without affecting the growth of M. anisopliae, then according to the in vitro assay, this would suggest that the inhibition of M. anisopliae by #2338 in a carton material environment was owing to chemical interference.
(g) Termite preparation for the arena bioassay
Termites were collected from five field colonies of the Formosan subterranean termite C. formosanus in Broward County, FL, USA and two colonies were collected in New Orleans, LA, USA using the 'bucket trap' method [39] processed in the laboratory according to Tamashiro et al. [46] and kept in groups of at least 2000 for 10–15 days in containers at 28°C before use in the bioassay. For each colony, groups of 50 termites (45 workers + 5 soldiers) were prepared. The groups of termites were then introduced into a planar arena (described below). In our study, the planar arena has the advantage over other standard protocols (Petri dish, jar) because it provides a soil environment enabling termites to forage and establish their own tunnel structure, while allowing continuous monitoring. The advantages of the use of this protocol have been fully discussed [47]. For each of the four microbial treatments discussed below, 14 replicates (two per termite colony per microbial treatment) were prepared, for a total of 56 arenas (2800 termites).
(h) Soil treatments for the arena bioassay
Four soil treatments were prepared for the arena bioassay (table 1). For each arena, 15 g of sterile sand (dry weight) was placed in a sterile glass Petri dish, and mixed with 2.6 ml of a 15% sterile termite carton material in water suspension. This suspension aims to provide enough nutrients for microorganisms to establish a stable density and to express their metabolism as in a field termite nest. This carton material treatment of the sand likely approximates a termite gallery environment in which faecal deposition is present. For the Streptomyces #2338 only treatment and mixture of Streptomyces #2338 plus M. anisopliae treatment, 0.2 ml of 0.05% Tween80 solution containing Streptomyces #2338 was added to the wet sand–carton suspension in the Petri plates. This treatment simulates a termite gallery with the Actinobacteria community already established. An equivalent amount (0.2 ml) of 0.05% Tween80 solution was added to the control treatment Petri plates (no Streptomyces and no M. anisopliae) and to the future M. anisopliae treatment Petri plates. The Petri plates with the mixtures were kept at 28°C in the dark for 15 days. Finally, after 15 days of incubation, depending on the treatment, Petri dishes received 0.2 ml of a 0.05% Tween80 solution or a M. anisopliae conidia suspension in 0.05% Tween80 solution. Table 1 summarizes the soil microbial treatments. The control treatment contains only the sand, termite carton nest material water suspension and Tween80 solution.
Suspension densities of M. anisopliae and Streptomyces #2338 were previously adjusted by dilution in 0.05% Tween80 to obtain the final concentrations. Expected densities of viable conidia and cells per gram of soil were confirmed by plating serial dilutions of 0.6 g of soil samples from each preparation on 1/5 PDA and counting CFUs after 3 days, with a ±5% margin of error. Within 2 h after the 0.2 ml of Tween80 control solution or M. anisopliae suspension was added, the final sand mixture treatments (15 g dry sand + 3 ml of Tween80, with or without microbial suspensions) were introduced into individual planar arenas as described below. In this study, the logic behind establishing the Streptomyces population in the sand–carton mixture for 15 days prior to introducing termites was to simulate a termite tunnel condition where Streptomyces had the opportunity to be established and produce its secondary metabolites within its environment. Metarhizium anisopliae was inoculated just prior to the termite introduction to simulate the encountering of the fungus by the termites during foraging in the soil.
(i) Arena bioassay set-up
Planar arenas (see electronic supplementary material, figure S2) as described in Chouvenc et al. [47] were composed of two sheets of transparent Plexiglas (12 × 12 × 0.2 cm in thickness) separated from each other by Plexiglas laminates (2 cm in width and 0.2 cm in thickness) on the four sides, creating a 10 × 10 × 0.2 cm space inside the arena. A 0.8 × 0.8 × 0.2 cm spacer was placed in the centre of the arena. The two sheets of Plexiglas and the central spacer were held together by a 3 mm-diameter screw, in order to maintain the 0.2 cm space layer evenly throughout the entire arena. A 0.4 mm diameter hole was drilled on the top Plexiglas sheet, 2 cm away from the spacer–screw hole, for adding liquids with the help of a syringe and to allow a small amount of air flow. A 5-mm-diameter hole was provided in one corner for the introduction of the termites into the arena chamber. Before assembly, all elements were washed with soap, immersed in bleach (3% sodium hypochlorite solution) for 2 h and rinsed three times with sterile deionized water. A sterile cellulose absorbent pad (45 mm in diameter, 2 mm thick) was placed below the central spacer, centred with the 0.4 mm hole. The arena was filled with 18 g of wet sand–carton (15 g of sterile dry sand, 150–500 µm size and 3 ml of treatment solution, see 'soil treatment' above), leaving a band of 10 by 2.5 cm empty on one border, exposing the cellulose absorbent pad to the empty space, which allow termites to find a food source immediately. The arena pieces were held together with eight 1-cm binder clips and the mounted arena was set horizontally. The four sides of the arena were sealed by hot glue, in order to prevent sand desiccation. One ml of sterile deionized water was injected via the 0.4 mm centre hole onto the absorbent pad. A group of 50 termites (45 workers + 5 soldiers) was introduced into the arena with the help of a small funnel, and the introduction hole was sealed with a thin transparent plastic cover after all termites entered the arena. All 56 arenas were placed at 28°C in the dark and termite mortality was monitored daily for 60 days by taking digital pictures with brief illumination from beneath with LED lights. Dead termites (determined by their lack of movement and absence of reaction when in contact with nest-mates) were not removed as they were buried or cannibalized. During the daily census, cannibalized and buried cadavers were counted as 'dead' in this study, as this aspect was previously discussed in [14].
(j) Statistical analysis
The effect of the presence of a microorganism on the growth of another microorganism in sand–carton material mixture was established by comparing the number of CFUs at 42 days in the two treatments (presence or absence of a competitor) with a Student t-test. Because of the growth rates of the Streptomyces isolates, the data were log-transformed. The absence of growth of microorganisms over time in plain sand was confirmed by the absence of positive correlation between time and number of CFUs (Pearson correlation).
A Cox proportional-hazard regression analysis (using the program R-Project for statistical computing, v. 2.4; http//cran.r-project.org/) was performed on all individual termites and a Wald statistic was generated. The factors tested were: colony of origin, fungal treatment and Actinobacteria treatment. The resulting hazard function defines the instantaneous rate of death at a particular time. Pairwise comparisons of the hazard ratio of death among treatments were adjusted by the Holm–Bonferroni method (α = 0.05).
3. Results and discussion
(a) Isolation of Actinobacteria from termite nest materials
More than 500 Actinobacteria isolates were obtained from five C. formosanus colonies, 37 ribotypes (16S rDNA) were identified and 70% of isolates showed in vitro antimicrobial activity against a range of Gram-positive and Gram-negative bacteria, yeast and fungi (see electronic supplementary material, figure S1). Streptomyces isolates were the most common Actinobacteria genus found in all termite nest materials (62%), but species of Kitasatospora, Pseudonocardia, Nocardioides, Nocardia, Gordonia, Mycobacterium and Kribbella were also isolated. A particular Streptomyces 16S ribotype was found in the nest material of all five termite colonies at 106 cells and higher per gram of carton material (all isolates were obtained from the 10−6 serial dilutions), and was the most bioactive isolate against entomopathogenic fungi such as M. anisopliae and B. bassiana in vitro. After this preliminary antimicrobial screening, we used Streptomyces #2338 isolate as a representative to determine whether an in vivo mutualistic relationship could be established between termites and this Streptomyces isolate.
(b) In vivo interaction between Streptomyces and Metarhizium anisopliae
First, we determined that termite carton material (after sterilization) provides proper nutritional requirements to maintain a stable population of Streptomyces #2338 within the termite nest (see electronic supplementary material, figure S3a). When M. anisopliae was introduced into the carton material with Streptomyces #2338, the Streptomyces growth was not affected (t-test, n = 54, p = 0.76). Second, M. anisopliae was able to thrive when alone in the carton material, but not when associated with Streptomyces #2338 (see electronic supplementary material, figure S3b) as the fungal growth was reduced by 69% after six weeks (t-test, n = 54, p < 0.001). Therefore, in a termite carton material nutritional environment, Streptomyces #2338 had a direct negative impact on the growth of M. anisopliae. Third, to determine whether the growth reduction of M. anisopliae in a carton environment was the result of exclusion competition or chemical interference [48] by Streptomyces #2338, a different isolate of Streptomyces (#2345) with no in vitro antifungal activity was used in an identical protocol. We found that Streptomyces #2345 grew in a carton material environment as previously observed with Streptomyces #2338, but that M. anisopliae growth was not affected by the presence of Streptomyces #2345 (see electronic supplementary material, figure S4). This result supports the hypothesis that the inhibition of M. anisopliae by Streptomyces #2338 in a nutritional environment similar to the termite nest is by chemical interference, with the production of antimicrobial compounds.
(c) Effect of Streptomyces on Coptotermes formosanus survival
The direct impact of Streptomyces #2338 on termite survival when exposed to entomopathogens remained to be determined. We used termite 'mini-nests' to maintain groups of termites in their own gallery system for extended time [47]. Groups of 50 termites foraged in the mini-nest for 60 days, where the sterile sand–carton material in the mini-nest was preliminarily treated with a combination of microorganisms and their survivorship was recorded daily (figure 3). The termite colony of origin was not a significant predictor of survival but both Streptomyces #2338 and M. anisopliae were independent predictors of termite survival (0.675 and 3.03 hazard ratios of death, respectively, Wald = 221.6, d.f. = 2, p < 0.001). Termite groups that were able to forage and establish their tunnel system in sand–carton material already containing Streptomyces #2338 had similar survival to the control groups (0.81 times the hazard ratio of death of the controls, Wald = 2.39, d.f. = 1, p = 0.12). Such results implied that Streptomyces #2338 had no virulence toward the termites when incorporated within the tunnel walls. Groups of termites that were exposed to M. anisopliae showed a significantly lower survival than the control groups (3.37 times the hazard ratio of death, Wald = 3.4, d.f. = 1, p < 0.001), confirming the virulence of the fungus on termites at the given density (104 conidia g−1, ≈LD50 at 60 days [14]) in their foraging environment. Finally, termite groups that were kept in mini-nests treated with both Streptomyces #2338 and M. anisopliae, had higher survival than the groups treated with M. anisopliae only (0.63 times the hazard ratio of death, Wald = 29.8, d.f. = 1, p < 0.001). Therefore, the presence of Streptomyces #2338 in the termite galleries, once established in the termite faecal material, provided direct protection to the termites against a soil entomopathogenic fungus.
(d) Extended disease resistance
Our study demonstrated that the presence of Streptomyces #2338 in the termite carton material reduced the pathogenic pressure from M. anisopliae against C. formosanus and represents a case of defensive mutualism. It was previously established that subterranean termites and other termites have evolved a range of defence mechanisms which resulted in an efficient interaction of individual and social immunity, reducing the chances of fungal epizootics in the colony [7,8,14,19,44]. We report here that the association of termites with beneficial microorganisms within their nest material provides additional protection to termites against the risks of epizootic, which may help to explain the repeated failures of biological control attempts against subterranean termites [37]. A analogous beneficial association was recently described in ants [49,50]; although in such cases, the mutualistic Actinobacteria forms a protective biofilm on the surface of specialized structures of the ants' cuticle. In the absence of such structures in termites, an analogue association emerged from the termites nest structure, considered the extended phenotype of the colony [47,51]. The overall termite nest environment is coated with a layer of faecal-based material [4], which favours the growth of potentially beneficial Actinobacteria, for instance Streptomyces #2338, and reduces the pressure on termites from soil pathogens as an extended disease resistance (figure 4), which could be part of an extended immunity.
(e) Origin and nature of the mutualism
Termites (Isoptera) are successful lignocellulose decomposers owing to the acquisition of diverse nutritional mutualists through their evolutionary radiation [52]. The transition between lower and higher termites (Coptotermitinae–Macrotermitinae) represents a key change in the biology of termites as the digestion process moved from an internal rumen with mutualistic protozoans, to an external rumen with the mutualistic fungus, Termitomyces [53]. This transition involved the loss of protozoans in all higher termites, and in the most derived Termitidae, the rumen was later re-internalized with the use of symbiotic bacteria as a base for cellulose digestion [53]. It was suggested that the origin of the fungal comb in a Macrotermes ancestor was the result of a primitive carton nest structure being invaded by Termitomyces [3]. Despite the presence of a carton material in Coptotermes, this genus, like all lower termites, never lost its protozoan endosymbionts, and apparently does not rely on an external rumen for nutritional purposes [54]. However, in the absence of the cultivation of a mutualistic fungus in Coptotermes nests, this carton material still represents a niche opportunity for various microorganisms [5,36] and we here showed that some of these microorganisms can provide a non-nutritional benefit to the termites in the form of an exogenous source of protection against diseases.
While our study was limited to a bioassay with a particular Streptomyces isolate (#2338), the presence of various Streptomyces strains and other Actinobacteria with a range of antimicrobial activity in all sampled termite nests, implies that multiple strains may be involved in such beneficial associations. For example, our in vitro result with Streptomyces #2345 does not fully exclude the potential survival benefit for the termites by a mechanism of exclusion competition in vivo. In addition, many other isolates should be tested individually or in association with other isolates (to explore eventual synergistic activity among isolates) to quantify their role in an active or potentially passive protection of termites against entomopathogens. This raises questions about the evolutionary traits of such an association in Coptotermes, considering the potential facultative nature of such symbiosis [55].
In attine ants, the mutualism with Pseudonocardia is a well-supported case of coevolution [29,56,57] with an adaptation of the ant cuticular structures to host the Actinobacteria biofilm [58,59]. However, recent studies [60–64] have suggested a potential recruitment of mutualists as a dynamic association, as an alternative to strict coevolution through obligatory vertical transmission. In the case of C. formosanus, a soil dwelling social insect also in recurrent contact with soil microorganisms during foraging, it can easily spread newly acquired microbes throughout the gallery system. This would result into a potentially dynamic symbiont turnover, as termites do not possess a specialized structure like attine ants [59]. In our study, Streptomyces isolates with identical 16S ribotype and similar antimicrobial activity to Streptomyces #2338 were present in all five C. formosanus nests surveyed. In addition, this 16S ribotype was also identical to many isolates from soil samples when compared to sequences in GenBank. However, the five carton materials evaluated were all sampled within Broward County, FL, USA and C. formosanus is an invasive species in southeastern United States that went through a strong bottleneck effect [65]. In addition, it is important to mention that the resolution of 16S gene may not provide sufficient evidence to confirm the soil recruitment hypothesis [22,56]. Therefore, it has yet to be established if the mutualistic association is the result of the recruitment of specific Streptomyces from local surrounding soils, if it is owing to vertical transmission by inheritance from the native distribution of the species, or a combination of both acquisition models [60,64].
While our current results may suggest that an assemblage of Actinobacteria, essentially originating from a community of free-living Streptomyces, invaded the termite tunnel system owing to the nutritional nature of the termite carton material, it remains to be confirmed. On-going surveys of the Actinobacteria community associated with the termite carton nest will provide valuable information: (i) a survey of the Actinobacteria community of termite nests from the current worldwide C. formosanus distribution will provide an overview of the Actinobacteria commonly associated with C. formosanus and determine whether this association can be maintained as a stable mutualism, (ii) the profiling of the Actinobacteria community from the surrounding soil of these termite nests will provide additional information that will help us refining the understanding of the mode of acquisition of this association and (iii) the sampling of the nest material, the gut content and the culticular microbial load of individuals from two generations of C. formosanus will cast light on the importance of both vertical and horizontal transmission of known isolates. Finally, the screening of the antimicrobial activity of all obtained isolates will provide unique information on their role in C. formosanus extended disease resistance, and could provide an alternative model for the study of evolution of defence mutualism in insects.
Funding statement
This study was supported in part by a grant from USDA-ARS under the grant agreement no. 5-6435-8-276 to N.-Y.S. and a research opportunity SEED fund from the University of Florida under the grant agreement no. 00094648 to N.-Y.S., M.L.E. and T.C.
We thank R. Pepin, A. Mullins and S. Osorio for technical assistance. We are grateful to R. Rosengaus and an anonymous reviewer for providing helpful comments in a previous version of this manuscript.
- Received July 19, 2013.
- Accepted August 29, 2013.
© 2013 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0/, which permits unrestricted use, provided the original author and source are credited.


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- Explains available features and their benefits
According to General Motors, most full-size truck owners use their vehicles for towing. To meet their needs, the 2014 GMC Sierra is completely redesigned and available with a range of features for towing an RV, boat or other trailer. James Bell, GM's head of consumer affairs, demonstrates these features in a new video featuring the 2014 Sierra and Airstream trailers. Bell shows how the Sierra tows smoothly and predictably, even when executing sudden lane changes or other abrupt maneuvers. Those with towing experience know sudden moves can upset a trailer's motion and cause hazardous results. Available towing features for the Sierra are designed to prevent these issues.
In the first video, Bell pilots a 2014 GMC Sierra with the Airstream in tow and executes a double-lane change followed by a panic stop. The maneuver mimics what drivers may encounter in the real world, swerving around an obstacle in the roadway. Available trailer sway control and integrated trailer brake control in the Sierra keep the truck and trailer composed. Without these features, the trailer could begin swinging from side to side in this exercise. Once that sway starts, getting everything back under control can be difficult -- or, for drivers without much towing experience, downright scary.
Other Sierra features that improve the full-size pickup's towing capability include a backup camera to ease getting your trailer hitched and two helpful brake functions: Hill Start Assist and Trailer Grade Braking. Hill Start Assist keeps the truck from rolling forward or backward as the driver moves his foot from brake pedal to accelerator pedal. That's ideal for low-speed trailering, such as on a boat ramp.
Trailer Grade Braking is great for drivers who take to the hills for work or play. This feature allows the GMC Sierra to select a lower gear automatically to slow the vehicle while descending steep hills. This keeps the truck and trailer under control, and the brakes cool and ready to apply full braking power.
If you're curious about towing with the 2014 GMC Sierra or want to see how the truck performs with an Airstream hitched up, check out the series of four videos. To learn more about the new Sierra, check out our first drive review.
What it means to you: GM says six out of 10 full-size truck owners use their vehicles for towing. A new video series demonstrates the technology in the GMC Sierra to make towing easier, safer and more fun.

subset: commoncrawl


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Barbecued Salmon with Smoked Ratatouille
|Seafood, Vegetables
|Day Part
|Cuisine Type
Source: Recipe from Chef Greg Markey
This salmon is grilled with a luscious maple glaze and served on a bed of creamy mashed potatoes. On the side, ratatouille prepared with smoked tomatoes adds great flavor.
2 1⁄2 tbsp. canola oil
2 yellow onions
3⁄4 cup maple syrup
3⁄4 cup cider vinegar
2 1⁄2 tbsp. Worcestershire sauce
1 1⁄4 cup ketchup
1 1⁄2 tsp. red chile flakes
2 1⁄2 cups chicken stock
2 tbsp. Dijon mustard
1 lb. eggplant, 1⁄2-in. slices
1⁄2 cup olive oil
1 lb. tomatoes, peeled and cold smoked, 1⁄2-in. dice
1 lb. zucchini
1 lb. yellow zucchini
1 lb. red pepper
1 lb. yellow pepper
1 lb. yellow onion, peeled and cold-smoked, 1⁄2-in. dice
1⁄8 cup garlic, minced
1⁄8 cup parsley, minced
1 tbsp. thyme, minced
15 6-oz. salmon fillets
1. For glaze, heat oil in a large pot and sauté onions 2-3 min. Add maple syrup and reduce to a glaze. Add cider vinegar, simmer for 2 min., and add remaining ingredients except mustard. Simmer 40-50 min. until thickened. Add mustard and strain.
2. To prepare ratatouille, season eggplant, brush with olive oil, grill until tender, cut into 1⁄2-in. dice. Cut smoked tomatoes in half and reserve juice.
3. In a large hot pan, sauté the vegetables (except smoked tomato) separately, combine all the sauté vegetables with the eggplant, tomato, and reserved tomato juice. Bake in 325° F. oven for 20 min. Add herbs and season with salt and pepper to taste.
4. Season fillet and grill to desired temperature on wood grill.
5. Brush with maple glaze. Mount salmon on mashed potatoes and arrange ratatouille around plate.


text:
United's DeLeon to miss six weeks
Washington, D.C. – D.C. United winger Nick DeLeon is expected to miss the next six weeks because of a hamstring problem, the club has announced.
DeLeon suffered the injury in the first half of United's 2-1 defeat to Columbus last weekend and had to be replaced by Kyle Porter midway through the opening half.
The 22-year-old tallied six goals and four assists in 28 games during his rookie campaign, and his absence is another blow to an already sputtering United attack which has managed to score only two goals in its first four games this season.


text:
COPPEROPOLIS - Manteca police reported late Monday night that they had recovered three stolen Tillamook cheese microbuses in a Copperopolis storage unit.
COPPEROPOLIS - Manteca police reported late Monday night that they had recovered three stolen Tillamook cheese microbuses in a Copperopolis storage unit.
The vans, a truck and a cargo trailer were stolen from a Manteca hotel parking lot early Saturday morning. The vintage Volkswagen vans, each worth about $100,000, were undamaged. The California Highway Patrol came across the Tillamook cheese truck and trailer stolen along with the vans on fire in a rural area of Tuolumne County on Sunday. Prior to that discovery, the truck and trailer had been spotted near Copperopolis Town Square earlier in the day.
Manteca police have identified several suspects in the theft and will be providing that information shortly, according to Manteca Police Chief Nick Obligacion.
Police notified Tillamook shortly after the discovery. "They're ecstatic that all three were found so quickly," he said.
The storage locker, at 49 Cosmic Court in Copperopolis, had been "very recently rented," Obligacion said. A tip led to the location, and he said the tipster would likely be eligible for a $10,000 reward the company had offered previously. Several regional agencies were involved with the search warrant, including the auto theft task force and CHP, Lodi police, the San Joaquin County Sheriff's and District Attorney's offices, and the county probation department.
"If not for the efforts of the Delta Regional Auto Theft Task Force, this quick of a recovery would not have been possible," said Obligacion, whose department has been fielding calls from across the country in connection to the theft.
Visit recordnet.com throughout the day Tuesday for more news on this breaking story.


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...To live is to die. To die is to live. My name is Kikyō. I am a priestess who is unable to leave this world and reach the sacred place. No, that is not entirely the truth. Once I was a priestess. Now I am but a corpse with a cursed, false existence. No matter how much I desire to do good and help others, the truth is I live off the souls of the dead. Pure is Impure. Impure is pure. Good is evil. Evil is Good. Or is it?
|Name meaning
Chinese Bellflower
St. Hijiri
Biographical information
Killed by Naraku
Physical information
169 cm (5'6")
57 kg (126 lb.)
|Eye color
|Hair color
|Skin color
Skills information
Spiritual powers
|Manga Debut
|InuYasha Anime
|Movie Debut
|Game Debut
Voice Actors
|Japanese Seiyū
|English VA
The InuYasha Wiki has 138 related images
Fifty years after the incident, she is resurrected by the demon ogress Urasue, using grave soil and the soul of her reincarnation, Kagome Higurashi. Shortly after, Kagome calls her soul back and Kikyō is forced to rely on the souls of the dead, collected by her Shinidamachū, to sustain her fake body. Though she was initially cold-hearted and harbored deep hatred towards Inuyasha, this hostility gradually disappears and her love for Inuyasha returns. As she is already dead, she resolves to not rekindle her relationship with Inuyasha and pours all of her resources into helping others, destroying Naraku and purging the Shikon no Tama from the world. In the end, however, Naraku's darkness proves too powerful and overcomes Kikyō's power, completely corrupting the jewel. He strikes her down, wounding her fatally. After sharing one last kiss together with Inuyasha, her soul finally finds peace and her body disappears into a bright cluster of souls and light. The Shinidamachū then carries Kikyō's soul into the star-filled sky.
During their youth, Kikyō and her younger sister Kaede (whose parents died when they were younger) traveled through villages as a part of their training to become priestesses, and killed many demons as a part of it. While on their journey, they met Tsubaki, who would later become a dark priestess. Kikyō's great spiritual power and purity became well-known, and thus the Shikon no Tama was entrusted to her, much to Tsubaki's dismay. It became Kikyō's duty to spend her life guarding the jewel, protecting it from the wicked humans and demons who sought it for their own selfish means. However, her duty as the jewel's protector also meant that she could not show any signs of weakness, lest demons or evil men should take advantage of her.
As a priestess, Kikyō spent her life fending off demons who attempted to steal the Shikon Jewel, as well as protecting the village from disease and famine. However, the one demon whom she refused to kill was Inuyasha because she sensed that he was only a hanyō (half-demon), and has lived an abnormal life of solitude, much like herself. She repeatedly warned Inuyasha to stay away from her if he valued his life but he repeatedly refused. During their first non-violent encounter, Kikyō developed feelings for him both from having common lives and from him saving Kaede from Mistress Centipede. She also tended to Onigumo, a paralyzed bandit with burn scars all over his body. Later on, Kikyō confronted Inuyasha and told him about how she had to hide her real self in order to protect the jewel, and Inuyasha sympathized with her. Eventually, Inuyasha and Kikyō fell in love. Sometime during this period, Tsubaki attempted to attack Kikyō, assuming that she was weaker after falling in love. However, Kikyō reflected the curse back at Tsubaki, and then ordered her to leave.
During her time with Inuyasha, Kikyō and Inuyasha had once sailed to an island that only appeared every fifty years. Kikyō suggested using the Shikon Jewel to turn Inuyasha into a human, an offer to which Inuyasha agreed. If this plan had in fact worked, it might have been possible for Kikyō to carry on the life of an ordinary woman alongside Inuyasha, and for him to finally feel like he belonged somewhere as a human. She promised to deliver the Shikon Jewel to Inuyasha on a certain day. However, the thief Onigumo, who desired both Kikyō and a corrupted jewel, decided to use that day to gain control of both. He offered his body to a horde of demons, and thus, the demon shape-shifter Naraku was born. Naraku disguised himself as Inuyasha and struck down Kikyō, and then attacked Inuyasha while disguised as Kikyō. He set the both of them against each other.
Thinking that Inuyasha had betrayed her, Kikyō summoned the last ounce of her strength and spiritual power to stop Inuyasha, who attacked the village for the Shikon Jewel. However, unable to kill the man she loved with her sacred arrow, Kikyō instead shot a sealing arrow at Inuyasha to pin him to the Goshinboku, where Inuyasha remained in slumber for fifty years. She then ordered her younger sister Kaede to burn her body along with the Shikon Jewel so that its immeasurable power would never be used for evil again. Because Kikyō was full of bitter hatred for Inuyasha at the time of her death, the Shikon Jewel became corrupted. However, because of her love, Kikyō wished to see Inuyasha again and the Shikon Jewel used this desire to return to the world five centuries later through Kagome, Kikyō's reincarnation.
During the storyEdit
Fifty years later, an Oni witch named Urasue stole Kikyō's remains and some graveyard soil to create a new body for her out of clay and bones. However, Urasue was disturbed to find out that the body she created would not entirely animate without its spirit and soul. She quickly realized that it was because her spirit and soul had already been reincarnated into another body, and that Kagome's soul, being the reincarnation of Kikyō, was needed in order to completely animate Kikyō's clay body. When the lifeless body was infused with Kagome's soul, Kikyō was revived, immediately turning on Urasue and burning her to death. Having yet to discover what truly happened fifty years before, her immediate priority was to kill Inuyasha. She nearly succeeded in this task until Kagome's body began recalling the soul back to her own body, which effectively stopped Kikyō from completing her goal. Kikyō managed to retain a part of her soul though, and thus was able to escape into the mist, where she was pursued by Inuyasha. Quickly thereafter, Kikyō fell off a nearby cliff, presumed dead by the group.
Kikyō survived the fall and moved to a small village, tending to the wounded and curing the sick. She was loved by the children and the villagers respected her. However, she was faced with an internal struggle with what she had become, not a living woman, nor a dead person, but the walking dead. She wanted to live peacefully in the village, but was troubled by the thought that even such a simple existence may be forbidden. A curious monk sensed something abnormal about the resurrected priestess and intended to find out her true nature. He spotted her absorbing the souls of deceased women as a means to sustain her clay body, using her soul collectors, the shinidamachū. The monk tried to lay her soul to rest by attacking her with one of his spiritual demons called the "Demon binding spell", but Kikyō used her spiritual power to blast the demon to pieces. The monk was killed by a piece of the demon that struck him in the neck. Before he died, the monk asked Kikyō why she continued to wander this earth as the walking dead. He told her that time continues for the living, but it does not continue for the dead, thus the dead do not belong in such a place, and such an existence is tragic. While Kikyō pondered on what the monk had said, she discovered that Sayo, a child who was very fond of Kikyō, had watched the entire scene and was now terrified of her. Saddened, Kikyō gave up on living in the village and left, apologizing to Sayo for scaring her.
After leaving the village, Kikyō tried to spend the night alone in a forest, placing a barrier around her to prevent being noticed. However, Kagome was able to cross Kikyō's barrier. Filled with jealousy towards her living reincarnation, Kikyō paralyzed Kagome with her touch and bound her to a tree so that Inuyasha could neither see nor hear Kagome when he arrived. When Kagome tried to explain to Kikyō the true circumstances of her death, Kikyō dismissed her, saying whoever caused her death is unimportant, as it cannot bring her back to life. A dead person like herself wished only to live again, which cannot be granted, so she instead wished not to be forgotten. This can be achieved through deepening bitterness, enabling the dead to live in the heart of the living. Before Kagome could convince Kikyō otherwise, Inuyasha arrived. Kikyō told Inuyasha he must despise her for collecting souls to drive her vengeance, but Inuyasha told her he could not stop loving her, even if she hates him. Moved, Kikyō kissed him and professed her love in his arms. Sensing that Kikyō's body lacks warmth, Inuyasha realized that he could not help her and wished time stopped, to which Kikyō responded by attempting to take him with her to the Netherworld. However, Kagome was able to reach Inuyasha with her voice in time, and Inuyasha left Kikyō to free Kagome from the tree. Kikyō, heart-broken, asked if Kagome was more important to him than herself, and left with her soul collectors, telling Inuyasha not to forget that her feelings when they kissed were true. Afterwards, she visited Kaede and learned that Naraku was responsible for her death and separation from Inuyasha.
Kikyō then moved to a temple, healing the wounds of soldiers. When men from the palace asked her to heal their master's disease, Kikyō unwillingly accompanied them to the palace, only to discover a dark aura around the castle. Kikyō was disturbed to find that the master of the castle was already dead from the neck and down. As it turned out, the master of the castle was the man who once called himself Onigumo before he was born as Naraku. Kikyō tried to leave the palace, but couldn't and thus, Naraku erected a barrier that prevented her Shikigami from accessing it, which rendered her powerless.
Under the assumption that he could control Kikyō, Naraku attached a corrupted fragment of the Shikon Jewel to a soul and placed it within the priestess before sending her after Kagome. The corrupted shard is not enough to control Kikyō, however, and she destroys Naraku's golem. When Kagome arrives, she is subdued by Kikyō's Shikigami. Kikyō tells Kagome that Naraku fears her and wished to control Kikyō and force her into fighting Kagome, but that she had not fallen so low that she, the keeper of the Jewel of Four Souls, would be controlled by a mere fragment. Kikyō then forcefully takes Kagome's Jewel shards, and pushes her down a hole onto the roots of the trees that reveals a person's darkest fears. Once Inuyasha arrived to save Kagome, she left while laughing at Inuyasha, as he could not fight a woman he still loved. Kikyō returns to Naraku's castle, breaking the barrier he had erected to keep Inuyasha away. She then mocks him, explaining to him that his petty feats of minor sorcery does not work on her, and willingly hands over the shards that she had stolen from Kagome, further mocking him by saying that he needs the power of the jewel shards more than anyone due to being a half demon. Kikyō, now finally feeling the freedom that she did not have in her past life, stated that once Naraku gained all of the shards of the sacred Shikon Jewel, she would purify him along with the Jewel and send his evil and corrupt spirit to the Netherworld.
When Naraku confronted Kikyō again and threatened to kill her, she announced that he could not kill her, as Naraku still had the heart of Onigumo and Onigumo loved her with all his wicked heart. Naraku sent a giant soul collector to steal the souls that Kikyō used to sustain herself in order to kill her indirectly, but Kikyō managed to escape, and Inuyasha found and saved her. When Kikyō told Inuyasha that Naraku still harbored feelings for her, a disgusted Inuyasha takes her into his arms and tells her he is the only one who could love and protect her. Though Kikyō pretended to comply, she later wielded a knife to his neck and left after telling Inuyasha that she would use Naraku's weak spot in order to purify him and the Shikon Jewel. Watching Inuyasha call after her, Kikyō thought to herself that the string of fate (with Inuyasha) could not be rejoined since it had already been cut, implying that she could no longer return to him.[3]
Later, Naraku made a second attempt to kill Kikyō by prematurely getting rid of his human heart, but he stopped himself, realizing that he was much weaker without his human heart. Some time afterwards, Naraku left to Mount Hakurei in order to strengthen himself, and succeeded in both completely removing his human heart and becoming much stronger. He confronted Kikyō, and when she asked him what his true objective was, he responded by wounding her chest, belittling her inability to bleed, and pushed her into a river of his poisonous miasma, presumably killing her.
Much later, villagers told Inuyasha's group of a mysterious person known as Saint Hijiri who was rumored to be extremely powerful. Inuyasha encounters her with two Shikigami that had human form: Kochō and Asuka, who told him that Kikyō survived but her body was ravaged by miasma and her voice was lost. Kochō and Asuka assisted Kikyō by bringing Kagome to a waterfall, where Kikyō's severely injured and weakened body lay submerged under the water. They told Kagome that she was the only one able to save Kikyō, and stated that by rubbing some of the soil from Kikyō's grave site into her wounds, she would successfully save Kikyō's life. Kagome agreed to perform this task, and in doing so, witnessed in a dream the events that occurred between Inuyasha and Kikyō fifty years previously. She passed out during the process of purifying the wounds Naraku had inflicted upon Kikyō, but seemingly saved Kikyō by temporarily purifying the wounds in Kikyō's chest. Recovering, Kikyō had Kochō and Asuka bring Inuyasha to her, giving him an arrow covered in Onigumo's cave dirt to be used against Naraku. She would later be forced to create a Mayose spell to kill demon rats released by Hakudōshi to drive her out for Naraku once he realized she survived the attack at Mount Hakurei.
Kikyō gave her shikigami a lock of her hair to find Naraku's heart; they nearly succeeded, only for Kohaku to destroy them before they could reach the Infant. She saw his heart was his own, yet followed Naraku's orders. Questioning him, Kikyō came face-to-face with Naraku, who she couldn't sense approaching thanks to the Fuyōheki. She was nearly decapitated by Kohaku on Naraku's order, but Inuyasha's arrival forced him to retreat. Both she and Inuyasha exchanged intel about Naraku, and she left to find the Infant. She came across it and Kanna, realizing the Fuyōheki was the blue stone the Infant held. However, Mōryōmaru attacked her, and left with her targets. Kikyō would later come across Mōryōmaru again, when he was trying to take Kōga's shards (in the anime), firing an arrow that severed his lower body. When the demon took off, Kikyō noticed two things: Mōryōmaru seemed to have a soul now, and that she was exhausted after firing a single arrow. This meant something was wrong. At night, she was amazed to see two novice monks get past the barrier she had conjured up while recovering. She collapsed, realizing she didn't have much time.
In truth, Kagome's purification powers proved insufficient and Kikyō's wounds reopened. This drove Kikyō to fuse the soul of Midoriko, the all-powerful priestess who created the sacred Shikon jewel, into her own body to close her wounds and hold out the miasma until she could defeat Naraku before she died. Inuyasha arrived to witness the wounds in Kikyō's chest healing very quickly, and she explained to him that since Midoriko was also a priestess who died fighting yōkai, she would understand Kikyō's desire to defeat Naraku. She also told Inuyasha that Naraku could not be killed with the Tessaiga; the only way to completely destroy Naraku was by purifying both his soul and the Shikon Jewel the instant Naraku completes the entire jewel. Kohaku, arriving at the scene by Midoriko's will, overheard what Kikyō told Inuyasha and decided to follow Kikyō in order to aid Kikyō's plan to defeat Naraku. She left promptly saying she had no time hesitate, leaving Inuyasha to think to himself that Kikyō's plan involved removing Kohaku's life-sustaining shard, and that Kikyō could not be capable of doing such a thing.
Kohaku later offered Kikyō his shard to use against Naraku, and Kikyō allowed him to travel by her side, but wished to herself that they had met on different terms so that she could have nurtured his scarred soul and encouraged him to live. While traveling with Kohaku, she kept the Shikon Shard in Kohaku's neck extremely pure, rendering it untouchable to Naraku. During her journey, she saved Kōga and asked that he hand over his shards to her, telling him her plan to purify the Shikon Jewel once it is complete, but Kōga refused. She later encountered Sango and Kagome, who begged Kikyō not to take Kohaku's life and trust in Inuyasha, who was continually working to strengthen his sword, to defeat Naraku.
Naraku tried capturing her in spider webs, resorting to capturing an innocent child in them to force Kikyō to touch them. She sent Kohaku away with her shikigami to keep Naraku's contamination from defiling his shard. She attempted purifying the webs, but found herself unable to while being shown the day she died repeatedly. Meanwhile, Miroku and Inuyasha battled Naraku, during which Miroku opened his Wind tunnel and sucked in an enormous amount of miasma. Miroku was near death, but Kikyō saved him by absorbing and purifying the miasma in her body, although this greatly worsened her own body's condition.
Kikyō and Kohaku encountered Naraku's spiderwebs during their travels together. He used these spiderwebs as a way to contaminate and weaken Kikyō. However, Kikyō had already realized Naraku's plan and remained under her own incredibly strong and powerful spiritual barrier. However, she contaminated herself when she attempted to save a child from the webs. She summoned her Shikigami Kochō and Asuka to place a barrier over Kohaku in order to protect him from Naraku's reach, however, due to the fact that she was entangled in Naraku's spiderwebs, she became defiled and the barrier over Kohaku was broken by one of Naraku's incarnations, Byakuya and Kochō and Asuka is destroyed by Byakuya. She told Kagome, who had also been defiled by Naraku's webs, that the only way to save her life was by acquiring the longbow from the sacred Mount Azusa, but only if it was truly what she wished. Kagome traveled to Mount Azusa, where she was tested by an illusion cast upon her by the Spirit Guardian of the sacred mountain. After passing the test, Kagome and Inuyasha returned to find that Naraku had already captured Kikyō.
The strong connection between Kagome and Kikyō was shown when Kagome was successfully able to tell Kikyō's thoughts by shooting her with the longbow and arrow she had obtained from Mount Azusa at the right time. Kikyō, shot by the sacred longbow from Mount Azusa, fired the sacred arrow from her own body at Naraku. The Jewel was purified before it entered Naraku's body, but Naraku won against Kikyō's powers and successfully contaminated the Shikon Jewel, rendering Kikyō completely powerless and on the verge of death.
Though Naraku declared his victory and Kikyō's defeat, Kikyō thinks to herself that Naraku will find out about that after he dies, as she had let a sliver of purifying light in the Shikon Jewel. While Kagome grieves that her powers were insufficient to save Kikyō, Kikyō told Kagome in her spirit that she was able to save her soul.
The wounds Kikyō sustained from saving Miroku from Naraku's miasma spread, and she lay in Inuyasha's arms until night fall. They talked about their past, and how Naraku poisoned their love for each other. She then saw something she had never seen before: Inuyasha was crying. Inuyasha told Kikyō she was the first person he ever cared for or loved, making him feel terrible for being unable to save her. However, Kikyō tells him that the fact he came is all that matters. They share a final kiss before she dies, turning into a luminous sphere of light. Rather than return to Kagome's body, Kikyō's spirit was gathered by her Soul Collectors and ascended to the twinkling stars, finally able to rest in peace.
The speck of purity Kikyō had left in the defiled Shikon Jewel remained after her death, and this prevented Naraku from taking the last shard out of Kohaku's neck, until Magatsuhi defiled it and sealed Kagome's spiritual powers, preventing her from purifying it again. He was possessed by this demon and left in order to give Naraku the last shard, meanwhile being forced to relive his painful past through an illusion in his sleep. In this illusion, one of Kikyō's soul-collectors lead Kohaku out of his past, to his sister, who asks for his help and encourages him. He was successfully brought out of this illusion and released from Magatsuhi's possession, and Kohaku gained the strength to stop running from his past and continue to live. However, after a struggle with Naraku, Kohaku has his shard removed, killing him. Naraku mocks Inuyasha's inability to prevent the deaths of both Kikyō and Kohaku, and Sango holds him in tears. Had Naraku absorbed the completed Shikon Jewel, Kikyo's light would have purified it and killed Naraku. However, Kikyō decides to leave the task of defeating Naraku to Inuyasha's group and transfers her speck of purity from the complete Shikon Jewel into Kohaku, so that he may live.
Personality Edit
Kikyō was known as a compassionate, kind, caring and considerate woman before her death who was very sympathetic toward everyone, even her enemies. This was seen when she tended to Onigumo's well-being despite him being a bandit. She also had compassion toward Kansuke, and sympathy toward Inuyasha. She offered help to anyone in need. Kikyō was also very good with children during her lifetime; no matter what village she went to, the children were always fond of her. However, Kikyō, after her resurrection, was not always as kind as she once was. She had developed a new-found hatred that she never had when she was alive. This freedom to hate satisfied her, as she never was granted this when she was alive. At first, she wanted revenge on Inuyasha, believing him to be her murderer. However, she eventually found out the truth and turned her hatred toward Naraku. Slowly, she started to become more like her former self. Despite her compassion, before meeting Inuyasha, she rarely, openly showed any emotions, always maintaining a calm, seemingly apathetic composure, akin to Sesshōmaru. Unlike him however, Kikyō did not hide the fact that she cared for others, especially her younger sister, Kaede.
When she was alive, Kikyō was completely devoted to her duties as a priestess and was completely uncorruptible, not even using the Shikon no Tama to save her life when she could have, instead instructing Kaede to burn it along with her body to prevent it from falling in the hands of those that would abuse it.
Kikyō was also a very wise woman for her age, and quite deceptive. Kikyō devised cunning plans to destroy Naraku, although she went about it in strange ways. For a time, Kikyō seemed to be on Naraku's side since she handed him Kagome's Shikon jewel shards, to which this action caused Inuyasha to accuse Kikyō of betrayal. However, she explained that she gave him the shards to cast Naraku toward death and that she would never allow him to take Inuyasha's life. She wanted revenge just as much as he did and desired to end Naraku's life. She figured out that Naraku was a half-demon and that he still possessed Onigumo's heart, whose owner still had feelings for her. Kikyō found his feelings for her ironic and ludicrous, and felt much contempt toward him. As a Specter, Kikyō has shown more of a sadistic side of her than she was alive as she laughs at Naraku and taunting him, in regards to Naraku's possession of Onigumo's heart, whenever she gets the chance as Naraku spies on her. She was never intimidated by Naraku and was bold whenever she faced him.
After Kikyō's resurrection, she developed a strong attachment to life that she didn't have when she was alive. Refusing to die alone, she decided she would take Inuyasha with her to hell. However, this goal eventually faded as she learned more about the circumstances behind her death. Kikyō's feelings for Inuyasha were complex and strained. At first, Kikyō's pure love for him turned to deep hatred (although deep down, part of her still had some feelings for him). As the series progressed, Kikyō's hateful feelings toward Inuyasha disappeared and it soon became clear to everyone that Kikyō's love for Inuyasha had returned. Although Kikyō and Inuyasha's feelings were mutual, their relationship was awkward and strained because of Kagome, her 20th century reincarnation. Kikyō was well aware of Kagome's love for Inuyasha (as well as his for Kagome), and the situation did not sit well with her. Because of this, Kikyō was jealous of Kagome, believing that she was replacing her, and as a result, was not very fond of her. Despite her hidden discomfort over this, Kikyō came to accept the fact that Kagome was taking her place in this world and in the anime, would go as far as to protect Kagome since Inuyasha would be heartbroken if she died. She has been surprised by Kagome's unflinching kindness towards her, despite Kikyō's cold demeanor towards her.
Physical description Edit
Kikyō was often described as a very beautiful woman. She had flawless alabaster-white skin and long, rich black hair that fell past her slim waist, which was often tied into a loose ponytail with a ribbon and had big, brown eyes. She stood around five foot six.
Outfit Edit
Kikyō wore the traditional dress of a miko, which has remained essentially unchanged to this day except for the lack of the slit-shoulder seam. Kikyō wore a white jacket with sode-kukuri (cords) through the sleeves and open shoulders (similar to Inuyasha's and Jaken's outfits). Strings called muna-himo were attached to each lapel and tied in front to keep the garment closed. Kikyō's kosode would be very much like a modern Kendo or Naginata Keikogi, with the sleeve fitting somewhat closely like a modern loose shirt and extending slightly past the wrist. The slits on the side of her hakama, and the openings at the sleeves and shoulders of her hitoe (jacket) showed the white kosode she sported. This was a deliberate fashion effect. Kikyō wore a bright red nagabakama (a very long hakama), which included the small board on the lower back. Like a majority of other characters, she also was perpetually barefoot throughout the series, however this was changed in the anime where Kikyō wore common tabi (socks) and rice straw sandals. Kikyō sported a red Obi, or belt, over all her other garments.
Powers & Abilities Edit
- Master Archer: Coming from an era of endless wars, Kikyō has fully mastered the art of archery. She can easily and swiftly shoot enemies from long or short distances and has the uncanny ability to strike demons from much longer and greater distances. Kikyo can even accurately hit Inuyasha near the heart with one arrow despite being fatally injured which would normally impede such an activity even when Inuyasha was moving at high-speed through the air. Kikyo was able to teach her younger sister, Kaede, the skill of archery that led to her being a powerful priestess and archer herself.
- Master Healer: Kikyō is an extremely skilled and experienced healer and is able to treat and save even those on the brink of death. She is able to see the beings from the Netherworld and can dispose of them with a single touch by purifying them.
- Immense Spiritual Power: Kikyō is a highly respected priestess with tremendous skills and powers. Her sister Kaede noted that Kikyō is exceptionally powerful even for a priestess. She was one of the only people Naraku was scared of, going to extreme lengths to kill Kikyō. She is able to erect unusually strong and powerful spiritual shields and perform several mystic spells or enchantments that can be used for both offensive and defensive purposes. Likewise, Kikyō has the ability to break seals, shields, barriers, spells, and detect fragments of the Shikon Jewel wherever they may be. Kikyō can also combine Shikon jewel shards into one shard. Her spiritual powers manifest as a light lavender glow of light in color though it is light blue on a few occasions.
- Immense Purification: These powers and abilities come in a variety of different forms when used by Kikyō. When she was resurrected, she used her purification powers to strike Urasue as well as Inuyasha with light purple-colored glowing bolts of purification. These powers killed Urasue and injured Inuyasha during this occasion along with being able to temporarily purify Tessaiga itself. When confronted by the priest Seikai she was able to create a shockwave to destroy the dragon from Seikai's Soul Binding Spell. During her attempt to drag Inuyasha to Hell she was able to fire a bolt that could destroy a tree in order to intimidate Kagome who was bound to it. On another occasion, when Kikyō and Kagome were trapped alone in a cave together, Kikyō used these abilities to create a large glowing orb of light purple-colored light from her hands in order to destroy insects that were attacking Kagome. She can also purify the Shikon Jewel Shards by just touching them. Her purification are as such that the light imbued into Kohaku's Shikon Jewel shard is still effective enough to threaten Naraku despite having already died and only the likes of Magatsuhi (the evil of the Shikon Jewel) could corrupt it.
- Spiritual Immobilization: Kikyō is able to stun someone and make them invisible to everyone else but her, she does this by touching someone with two of her index fingers. She used this skill on Kagome.
- Mayose: The Mayose is a high-level mystical spell cast on a twig that can only be performed by a highly skilled and experienced priest or priestess with exceedingly strong and powerful spiritual power and knowledge. The twig grows into a huge tree that reaches the clouds once it is enchanted with a special spell. It is used to lure demons of a specified type and purify them when the demons climb to the top.
- Shikigami Creation: She is able to create human-like Shikigami that she uses to perform various tasks for her, which she used most when she hid herself; these specific Shikigami are called Kochō and Asuka. Another Shikigami Kikyō had created was Saint Hijiri, who much like herself, contained immense spiritual power. She used Hijiri as a distraction so that she could take time to fully recover from the miasma-filled wounds Naraku had inflicted upon her at Mount Hakurei. Kochō and Asuka originally stayed by Hijiri's side, but after St. Hijiri was destroyed by Kagura, they went on to help Kikyō directly.
- Resurrection: Kikyo's residual spiritual powers that were imbued into Kohaku's Shikon Jewel shard was capable of resurrecting him when Naraku took it back despite Sesshōmaru's mother stating that even Tenseiga couldn't revive him when his life is sustained by the shard.
- Shinidamachū Command: These serpent-like creatures collect the souls of the recently departed to sustain Kikyō's body. They can also steal the souls of the living, making them her mindless slave. Additionally, soul collectors enhance Kikyō's spiritual powers by delivering souls to her.
- Enhanced Eyesight: Kikyō is able to see things that are or may seem invisible to the human eye.[citation needed]
- Immense Endurance: Despite being fatally injured by Naraku disguised as Inuyasha, Kikyo was able to muster the strength in order to seal Inuyasha before ultimately dying in spite of having to walk a considerable distance with the wounds being open and bleeding profusely. When resurrected by Urasue, Kikyo returned in her injured state but was still able to put up a considerable fight against Inuyasha despite bleeding profusely from her open wounds.
- Immortality: Upon being resurrected by Urasue, Kikyo gained a form of immortality since being a specter of a deceased human she was unable to die by normal means. As such Kikyo is unaffected by diseases, poisons, and other ordinary fatalities for humans as she survived falling from a great height right after being resurrected though strong poisons such as Naraku's improved miasma can impede her. This is shown by the fact that despite having been blown into a river of Naraku's powerful miasma and having it infect her body, she managed to survive (albeit she was incapacitated until temporarily purified by Kagome). However the drawback to this immortality of Kikyo is that she must periodically absorb the souls of deceased maidens through her Shinidamachu or else her existence can't be maintained in the mortal world as Kikyo finally died when she willingly gave up her remaining life force to the Shikon Jewel in an attempt to kill Naraku.
- Invisibility: Kikyo can turn herself and others invisible but to a miko like Kagome.
Weapons Edit
- Bow and Arrows: Kikyō often carries a traditional red Saigu-Yumi and a quiver of Hamaya. She has the power to imbue each arrow with great amount of spiritual power and has destroyed, disabled, purified or sealed many demons.
- Sacred arrows: The sacred arrow is an arrow used by high-level priestesses like Kikyō and Kagome. It can easily break through barriers, destroy spells, obliterate demons, and even deal fatal blows to powerful demons. Kikyō's Sacred Arrows are too powerful and contains enough spiritual powers to penetrate Naraku's Barrier and damage him with ease.
- Arrow of Sealing: The Arrow of Sealing is a sacred arrow that places a demon under a spell of eternal sleep. It is said that these arrows are meant to seal demons that are utterly invincible. If a demon is sealed onto the Goshinboku (the sacred tree or Tree of Ages) the demon's body with remain perfectly intact. They will only awaken when the one who shot the arrow desperately wishes for the demon to be revived.
- Reflection: This ability was once used on Tsubaki the dark priestess when Kikyō was still alive. She infuses her spiritual power into her longbow, which will reflect any oncoming demonic energy back at the demon or person.
Relationships Edit
Friends Edit
Kikyō and Inuyasha met when she first started guarding the jewel. At first they battled each other, but as time went on they both fell in love. Wanting to be a normal human, Kikyō asked Inuyasha if he would use the Shikon no Tama to become a human and to be with her. Naraku then takes action by causing Inuyasha and Kikyō to turn against other ending in Inuyasha being sealed to the Tree Of Ages and Kikyō's death. However, both Inuyasha and Kikyō would meet again after the demon Urasue brings Kikyō back to life and the seal on Inuyasha is broken by her modern reincarnation, Kagome. All Kikyō wanted in the end was to live together with Inuyasha as a normal woman, but in the end, she was killed yet again by Naraku. However, after a long period of suffering, she said her soul was saved by Kagome, so she shared a final kiss with Inuyasha and died in his arms. Although she died, her light saved Kohaku instead of destroying Naraku after her death. Inuyasha suffered a heartbroken depression of losing Kikyō, as he wanted to protect her forever; he suffered from self-inflicted insomnia for some time because he did not want to constantly see Kikyo in his dreams, which he found too painful.
Kagome is Kikyō's reincarnation, even though their personalities are very different. Kikyo is vengeful, distrustful, and stoic, while Kagome is forgiving, open-hearted, cheerful, and kind. At first, Kagome's body is used by a witch to revive Kikyō. However, not much later after Kikyō is reanimated, Kagome begins to call back her soul. Kikyō eventually escapes the scene with a small fragment of her soul intact. Early into the series, Kikyō has a confrontation with Kagome. Kikyō tried to kill Kagome (which she tells to Inuyasha afterwards). Both of them care and are in love with Inuyasha and several times they've work together and helped each other out and both have saved one another on numerous occasions throughout the series. Kagome and Kikyō have jealousy of one another as both share a special bond with Inuyasha, although Kikyo has shown her jealousy to be greater. Right before Kikyō's death, Kikyō gives Kagome her own bow and wishes Kagome well, asking her not to cry as Inuyasha takes her off. Kikyō also entrusts Kagome with the duty of destroying Naraku and the jewel, as well as keeping the "light" in Kohaku's shard. Unable to save Kikyō from Naraku's spiderweb's wounds, Kagome sheds tears and wishes she could have somehow done more to save her; Kikyo consoles her that she did manage to purify her soul, ensuring her a peaceful afterlife. Kikyō also has a high opinion of Kagome, telling Tsubaki her curse would not affect Kagome and would not interfere with Tsubaki's attempt to kill Kagome, apparently wanting her to get stronger from the experience.
Kikyō was fond of her younger sister Kaede. The two were very close at the time that Kikyō was alive. During the time she was young, Kikyō accidentally shot a demon within close proximity of Kaede and a part of the demon flew at Kaede, blinding her in one eye. At the time of Kikyō's resurrection, Kikyō could not recognize Kaede (mainly due to her now being an older lady) but as the story progresses, Kikyō visits Kaede occasionally. At Kikyō's final death, Kikyō's soul visits Kaede and bids her last apologies and farewell to her younger sister, showing her great love for her younger sister.
Kohaku was found sometime later that year by Kikyō who felt pity for the boy. During her journey, Kikyō would try purifying Kohaku. During the last season, Kohaku would be Kikyō's travel companion after being freed from Naraku's corruption. Kohaku, believing that his shard would help in destroying Naraku, decides to sacrifice his shard and dies. As her last act of compassion for Kohaku, Kikyō withdrew her spirit and power from the now-contaminated Shikon Jewel and instead use it in resurrecting Kohaku.
Enemies Edit
Naraku is the man who tricked Kikyō into betraying Inuyasha. While Kikyō harbors an obvious hatred and disgust for him, Naraku is mixed between hatred and love for Kikyō due to his human heart that once belonged to a bandit named Onigumo. Onigumo had once been in love with Kikyō when she tended to him out of pity. Throughout the series, Kikyō and Naraku interact on several occasions. At one point, Kikyō gives Naraku sacred jewel shards. It is misunderstood at the time that Kikyō had formed an alliance with Naraku, however in truth, she had been planning to purify the Shikon jewel along with Naraku once it was completed. Naraku also attempts to kill Kikyō on many occasions because she is one of his largest weaknesses, eventually succeeding. At the end of the series, it is revealed that Naraku's true wish on the Sacred Jewel had been to have Kikyō's love but it was not granted.
Quotes Edit
- "I must show my weaknesses to no one. I must never have doubts or demons would overtake me. I am human yet I cannot be human. We are quite alike, you and I. You half-demon... That is why I could not kill you."
- ―Kikyō
- "It's possible. It's true that you are half-demon, but you are also half-human. If the Sacred Jewel of the Four Souls fell into the hands of a demon, their powers would undoubtedly increase. However, if it were used to turn you into a human, it would be purified. The Jewel of the Four Souls would probably cease to exist."
- ―Kikyō
- "Liar! I was a fool beyond compare in believing your farfetched tales and wishing to live together with you. I despise you with my last breath. My spirit cannot forget that all-consuming hatred. So as long as you live, my spirit cannot be freed."
- ―Kikyō
- "You are saying you wish for me to die, is that it? For if my soul were to return to the girl, then I would cease to exist forever. You must know that. Is that what you desire, Inuyasha? I refuse to die. My spirit cannot rest in peace until I see you dead!"
- ―Kikyō
- "This is the second time you've asked me to die, Inuyasha. So then why do you look so sad? Is it because you feel the heat from my hand? The heat of anger? The flames of vengeance?"
- ―Kikyō
- "Inuyasha, after meeting you I renounced my position as a shrine priestess. I became an ordinary woman. I longed to embrace you like this when I was alive. We are unable to turn back the hands of time. So please, allow me to embrace you a little longer."
- ―Kikyō
- "Inuyasha, never forget. Remember the feel of my lips against yours, for it was real. Never forget."
- ―Kikyō
- "If I had lived, it would have been I who tended to the wounds in his soul."
- ―Kikyō
- "Go, Naraku... Gather the shards of the Jewel. And once you've found them all, then I will send you to Hell. I am free to hate. My soul is so much freer now than it was then. Free to hate. Free to love."
- ―Kikyō
- "Inuyasha, Naraku will only grow stronger and stronger. You mustn't allow him to kill you. Until I am able to cast both Naraku and the Shikon Jewel from this world, your life belongs to me. No one else will have you."
- ―Kikyō
- "He is reluctant to admit it, but I am certain that he has a lingering desire for me. And in an effort to ease that feeling, he's trying to get rid of me."
- ―Kikyō
- "Tsubaki, do as you please with Kagome, I have no intention of interfering. But if you should ever do any harm to Inuyasha, I will personally see to your demise!"
- ―Kikyō
- "You can never be with Inuyasha again. Nor I, for I am one of the dead and as such... I too don't belong here."
- ―Kikyō
- "I choose to stay here. I want to stay close to those children for a while. Besides, even if I wish to, I cannot go. Inuyasha, you saw them. You saw my soul collectors being repelled by the aura of this land. It took everything I had, just to stand at the base of the mountain."
- ―Kikyō
- "The battle continues. The fact that I walk this path once more only serves to confirm my wretched fate. And what awaits me beyond this darkness. Once more, just once more, I must ascertain the whereabouts of those mortals resurrected from the grave."
- ―Kikyō
- "No! Why are these memories coming back to me now?"
- ―Kikyō
- " (After each were unknowingly deceived by Naraku) I believed in you! I trusted you"
- ―Kikyō
- "Nothing's stopping me. I will put an end to Naraku's life. That's what this means."
- ―Kikyō
- "I'm surprised you allowed yourself to be distracted by that abomination. How pathetic."
- ―Kikyō
- "I loathe all things. I despise every living creature who's bound to time."
- ―Kikyō
- "Naraku, I will not die now. Do you expect me to die in your filthy arms!?"
- ―Kikyō
Trivia Edit
- Of all his enemies, Naraku considers Kikyō his only serious threat, the only one who scares him.
- Kikyō's arrows quiver is always changing the amount of arrows from one scene to the next. Sometimes when she uses one, not long after, her quiver will still be full.
- Both Kikyō's and Kagome's spiritual aura changed from a blueish color, to a pinker one.
- Noriko Hidaka who voiced Kikyō also voiced Akane Tendo in Ranma ½, another series by Rumiko Takahashi.
- Willow Johnson also voiced Kasumi Tendo in Ranma ½.
- In the Korean dub, her name was changed to Geumgang.
- Rumiko Takahashi stated in an interview that Kikyō had died around the age of 17, but this wasn't a direct statement. Plus, it seemed to contradict her official character profiles book, which states she was 18.[4]
- Rumiko Takahashi stated that she chose Kikyō's name basing on the language of flowers, as the Chinese Bellflower's meaning is "unchanging love."
- In Affections Touching Across Time, a number of instances occur where Kikyō seems to have irrationally omniscient knowledge for the purposes of exposition. Firstly, Kikyō refers to Menōmaru by name, despite never meeting him or learning his name from anyone who had. Although it is possible she knew his name from before his resurrection, this is unlikely, as Menōmaru's presence in Japan was not known to humans and he had been sealed away for two whole centuries prior to his return. Confusingly, when referring to him she alternates between both "Menōmaru" and "Hyōga", suggesting she learned of his latter name shortly after he assumed it or even less likely, that she knew of the naming traditions of the Hyōga clan, which supposedly only Myōga and Sesshōmaru (both of whom were actually alive at the time of the Hyōga clan's invasion) were aware of. She also had extensive knowledge of how Menōmaru was using the Tree of Ages to increase his power, despite the tree never having been used in such a manner before, though it is somewhat possible she simply gained such insights through observation.
- In the same movie, Kikyō sends Kagome back to the future with a shard of the Sacred Jewel, intending for her reincarnation to remain there forever, away from Inuyasha. However, Kikyō's primary strategy throughout the entire series is to allow Naraku to gain all shards of the Sacred Jewel so that she can purify both Naraku and the jewel simultaneously. Yet, if Kagome had really been trapped in the future and unable to return to the Sengoku era, Kikyō's other plan would have been ruined. This was either a plot inconsistency on the part of the filmmakers, or else an instance of Kikyō being uncharacteristically illogical. Luckily for Kikyō's plans, though, her attempt to trap Kagome was rebuffed and the jewel was allowed to return to the past.
Media appearancesEdit
- Episode 1 (First appearance)
- Episode 14 (Resurrected)
- Episode 15
- Episode 22
- Episode 37 (Mentioned)
- Episode 43
- Episode 47
- Episode 98
- Episode 108
- Episode 109
- Episode 110 (Mentioned)
- Episode 8 (FA) (Deceased)
References Edit
- ↑ BS Manga Yawa (BS Manga Night Talk) [1]
- ↑ Zusetsutaizan Ōgikaiden, page 136
- ↑ InuYasha anime; Episode 47
- ↑ Rumiko Takahashi Backstage 1


text:
All degrees are voted by the Board of Trustees on the recommendation of the faculty. The faculty explicitly reserves the right to determine whom it will recommend for any degree, taking into account not only the candidate's fulfillment of minimum course requirements but also the quality, coherence, and integrity of the student's program as a whole as indicated by the reports of the assessments. A minimum grade point average of 2.00 (C) on a three-point scale is required for graduation in the Certificate, M.T.S., and M.A.M. programs and M.Div. programs. A student with a grade point average below 2.00 (C) is not making satisfactory progress in the Certificate, M.T.S. and M.A.M. programs and M.Div. programs and may be placed on academic probation. Candidates for the degree of Master of Divinity are reminded of the statement on "Readiness for Ministry" found in the academic catalog. Candidates for any McCormick masters degree must be registered for the full final year preceding graduation, unless special arrangements have been made with the concurrence of the Associate Dean for Student Academics.
Students registering for their final semester of work before qualifying for a degree will at the same time fill out an application for graduation and pay the established graduation fee. In order to be eligible to graduate, degree candidates must submit all course work to instructors no later than Friday two weeks prior to the last day of the spring term. All students must meet all financial obligations to the Seminary by April 15, including payment of rent in housing owned by other ACTS seminaries, settlement of all debts and obligations to JKM Library, other ACTS libraries, and the Regenstein Library. Diplomas will be withheld until all the graduate's accounts are reconciled, financial obligations have been satisfied, and various forms and requests for information by the seminary, including an Association of Theological Schools questionnaire, completed.


text:
Smart kit 360
Smart kit 360 - is an all in one application. This program is two times smaller of regular "offline tools", such as a compass, a translator or an application for making a call signal. In this way, you can save lots of space, time and won't be disappointed in daily...read more


text:
Rokita Distributes Voting Equipment Funds to Porter County
Contact: AJ Freeney-Ruiz
Indianapolis, IN - Indiana Secretary of State Todd Rokita today distributed election reform money to Porter County. This was part of the latest round of funding Rokita's office is giving to counties from the federal Help America Vote Act (HAVA) of 2002.
Porter County received $395,808 in federal and state money which was distributed by the Secretary of State's office. This latest round of reimbursements brings to $29.9 million the total distributed under HAVA to Indiana counties since Rokita's office began distributing funds to help pay for voting equipment upgrades resulting from the required phase-out of lever and punch card voting machines.
Rokita personally met with Porter County Clerk Dale Brewer to reimburse the county for the new ES&S optical scan voting machines that will be available to all of Porter County's approximately 93,000 voters in all 133 election precincts beginning with the 2006 May Primary Election. Porter County will switch from punch card ballots.
"The counties were very fortunate to have a proactive Secretary of State like Secretary Rokita helping us get through this process." Said Brewer. "I'm glad we can get rid of punch cards here in Porter County and move forward in technology with a system our taxpayers can still count on."
"It is great to see federal and state money being put to good use to help ensure fair and efficient elections for Hoosiers," Rokita said. "Porter County can now focus on other election matters."
Congress passed HAVA in response to issues in various parts of the country during the 2000 Presidential Election. HAVA requires states and counties to replace all punch card and lever machine voting systems by January 1, 2006. A number of Indiana counties will be eligible for additional funding in the coming months.


text:
- Part I
- Part II
- Part III (The 8th Tune)
- Part IV
- Part V
- Part I Material in D
- Part II Material in a Handsome Stack
- Part III Material with no Tricks
- Part IV Material in a Long Cadence
- Part I Material in Eb
- Part II Material in Sevenths
- Part III Material in Two Keys
- Part IV Material with Shifting Drones
- Drones in Large Cycles
What the press says
The EP is a lilting, sometimes arch set of modern classical composition – by no means intimidating to neophytes, and encoded with delightful little motifs.
FACT Magazine (May 23rd 2012) Read all reviews
Breaking down as a cycle of five efficiently short pieces, the results are captivating and continue to strengthen the case for Nico Muhly as one of the world's consistently brilliant young composers.
Boomkat (May 25th 2012) Read all reviews
Brubaker alternates between lovely ivory lines and frantic, freeform eighty-eights pounding, while Sirota tends to supply sublimely droning string swells in the background.
Exclaim! (May 25th 2012) Read all reviews
Drones & Piano is an expressive, enchanting, moving skein.
Lucy Jones — The Telegraph (May 28th 2012) Read all reviews
...it's real appeal is in savouring how Muhly creatively uses consonance and dissonance with his chosen drone.
Musical Toronto (May 31st 2012) Read all reviews
Don't assume this five-song collection sounds like your aunt singing Duran Duran whilst lugging a Dyson across the living room… the bedrock of these pieces becomes highly charged; stuck within a confined space they so desperately want to break free of.
Drowned in Sound (June 21st 2012) Read all reviews
For a large part of this quarter of an hour, Brubaker pokes the keys of his piano in an obsessive and compulsive way, and gives a strength and intensity to the story that at times abstracts any particular genesis or conception of the project, as if everything came to life on its own and full of meaning much beyond the constitution and starting point of the scores.
Playground Mag (June 22nd 2012) Read all reviews
...it is both playful and studied, something incredibly hard to achieve. Well worth a closer look.
Boomkat (August 7th 2012) Read all reviews
These are drones with an intellect.
Robinson Meyer — The Atlantic (August 20th 2012) Read all reviews
The sharp bites of fingers crashing against the piano's keys put against high pitched whirrs of the violin take you on a incredible journey through suspense and terror.
Francesca Davison — Dummy (August 30th 2012) Read all reviews
Nico Muhly has an intriguing creative mind; [Drones & Violin is] an interesting work and yet another addition to his diverse back catalogue.
The 405 (September 5th 2012) Read all reviews
Muhly's versatility has been commented on plenty at this point, and this series finale is yet more ammunition.
Boomkat (September 6th 2012) Read all reviews
Muhly challenges his compositional skills rather brilliantly and creates three intensely captivating series of compositions.
Bruno Lasnier — The Milk Factory (September 6th 2012) Read all reviews
What is noteworthy here is the way songcraft repeatedly emerges from tension. Muhly's explorations never fail to find something worthwhile.
Tobias Carroll — Dusted (October 5th 2012) Read all reviews
A compelling trio of EPs.
eMusic (November 12th 2012) Read all reviews
Drones is a necessary acquisition for anyone interested in Muhly's work outside pop.
AllMusic (November 19th 2012) Read all reviews
...Nico Muhly's pieces feel like a series of archly posed questions. In their formal inventiveness, love of blank space, and haiku-like neatness, they arouse the part of your brain that suspects it's being outsmarted…To feel your intellect being playfully, patiently tested, as if he is circling your mind and kicking its tires, can be a wonderfully maddening experience.
Jayson Greene — Pitchfork (November 29th 2012) Read all reviews
"...the best thing I've ever heard of Muhly's…"
Steve Hicken — Burning Ambulance (December 4th 2012) Read all reviews
One of the most impressive aspects of these pieces is the variety of relationships that unfold between the "solo" instruments and the drones…beautifully despondent.
The Boston Globe (December 22nd 2012) Read all reviews
"...the latest from this brilliant, boundary-pushing composer [...] is post-minimal virtuosity, sometimes rollicking in nods to the likes of Rzewski, sometimes static sculpture, sometimes rock-and-roll. It's, vitally, never timid."
Create Digital Music (December 26th 2012) Read all reviews
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