Lanternflies (Hemiptera: Fulgoridae) of Taiwan.

The family Fulgoridae belongs to the order Hemiptera, superfamily Fulgoridea, with approximately 770 described species worldwide. Their extraordinary appearance attracts the attention of both entomologists and the public. In addition to the evolutionary issue of their special appearance, certain species are also considered notorious pests (e.g., Lycorma delicatula). Several issues widely exist in previous taxonomic studies of lanternflies: (1) application of uncertain morphological characters leads to synonymy or misidentification; (2) descriptions of male genitalia are incomplete; (3) information of nymphal morphology is insufficient. Therefore, this study aims to provide a comprehensive taxonomic study of Fulgoridae from Taiwan. In this study, eight species in six genera from Taiwan were reported, of which Limois westwoodii was recorded for the first time from Taiwan. Lycorma olivacea was proposed as a new junior synonym of L. meliae. The fifth-instar nymph of Saiva formosana was described for the first time. Detailed descriptions of these lanternflies and an identification key to adults of Fulgoridae from Taiwan were also included.

Chronobiological Effect on the Reproductive Behavior of Chrysomya megacephala (Diptera: Calliphoridae).

The most widely used entomological method of determining the time since death (minimum postmortem interval, mPMI) has been calculating the developmental time of blow flies (Diptera: Calliphoridae) on the deceased body. However, because blow flies are known to be diurnal, nocturnal oviposition has been excluded from standard mPMI calculations. This has been challenged by recent studies demonstrating nocturnal oviposition due to an unknown reason. Therefore, this study investigated the role of chronobiology. We recorded the locomotion amount and pattern of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) under different chronobiological conditions and examined whether Ch. megacephala can oviposit under nighttime conditions in field and laboratory settings. Subjects were found to have a daily activity pattern under normal darkness conditions (12:12 L:D) and under continuous darkness (DD), but they exhibited no pattern under continuous light (LL). Free-running period was approximately 1,341 min/d (22.35 h/d). In the field, no flies were observed during nighttime. Oviposition occurred in the laboratory setting during daytime with no lights and during nighttime with artificial lights. Free-running subjects oviposited in both active and resting periods, with more eggs laid during active than resting periods. The result of this study indicates it is possible to induce oviposition behavior during evening hours on Ch. megacephala. However, this was only observed in the laboratory setting and could only happen during the flies' subjective day.

Origin and Potential Expansion of the Invasive Longan Lanternfly, Pyrops candelaria (Hemiptera: Fulgoridae) in Taiwan.

The longan lanternfly Pyrops candelaria is a new invasive species on the main island of Taiwan. The introduction of an invasive species may negatively influence the native fauna, flora and environment. Thus, this study aimed to infer the invasion history, predict habitat suitability and potential expansion and assess the risk to crop cultivation areas in Taiwan. Genetic structures of P. candelaria from the main island of Taiwan and related regions were analyzed based on partial COI and ND2 sequences. Additionally, machine learning MaxEnt was utilized to study habitat suitability. The results suggested that the Taiwanese populations may originate from the Kinmen Islands and the plain areas of Taiwan are considered to have high habitat suitability. Furthermore, most of the cultivation areas of longan and pomelo crops showed high habitat suitability.

A new millipede-parasitizing horsehair worm, Gordius chiashanus sp. nov., at medium altitudes in Taiwan (Nematomorpha, Gordiida).

Gordius chiashanus sp. nov., a newly described horsehair worm that parasitizes the Spirobolus millipede, is one of the three described horsehair worm species in Taiwan. It is morphologically similar to G. helveticus Schmidt-Rhaesa, 2010 because of the progressively broadening distribution of bristles concentrated on the male tail lobes, but it is distinguishable from G. helveticus because of the stout bristles on the mid-body. In addition, a vertical white stripe on the anterior ventral side and areoles on the inside wall of the cloacal opening are rarely mentioned in other Gordius species. Free-living adults emerged and mated on wet soil under the forest canopy in the winter (late November to early February) at medium altitudes (1100-1700 m). Mucus-like structure covering on the body surface, which creates a rainbow-like reflection, might endow the worm with high tolerance to dehydration. Although Gordius chiashanus sp. nov. seems to be more adaptive to the terrestrial environment than other horsehair worm species, cysts putatively identified as belonging to this hairworm species found in the aquatic paratenic host, Ephemera orientalis McLachlan, 1875, suggest the life cycle of Gordius chiashanus sp. nov. could involve water and land. The free-living adults emerged from the definitive hosts might reproduce in the terrestrial environment or enter an aquatic habitat by moving or being washed away by heavy rain instead of manipulating the behavior of their terrestrial definitive hosts.

Taxonomic revision of the genus Ponera Latreille, 1804 (Hymenoptera: Formicidae) of Taiwan and Japan, with a key to East Asian species.

Ponera is a widespread genus of litter and soil ants. The highest diversity of the genus is found in Asia, with Taiwan and Japan being two of the most species-rich regions. Here, we systematically review the taxonomy of the 16 Taiwanese and Japanese Ponera species, two of which are new species from Taiwan: Ponera terayamai sp. n. and P. wui sp. n. A new key for Ponera species of East Asia is presented.

Characterisation of the complete mitochondrial genome of Lucanus chengyuani (Coleoptera: Lucanidae).

We sequenced and assembled the complete mitochondrial genome of Lucanus chengyuani, from the Alishan, Chiayi County, Taiwan. The length of the complete mitogenome of L. chengyuani is 16,926 bp and the mitogenome contains 13 protein-coding, 22 tRNA, and 2 rDNA genes. Nucleotide compositions of the whole mitogenome of L. chengyuani are 38.37% for A, 27.96% for T, 23.03% for C, and 10.637% for G. The AT and GC skewness of mitogenome sequence are 0.157 and -0.368, showing the A-skew and C-skew. The reconstructed phylogenetic relationships of 9 Lucanidae species based on 13 mitochondrial protein-coding genes are highly supported. The clade including Neolucanus maximus and Odontolabis cuvera is sister to the rest of the stag beetle clades, which contains L. chengyuani and L. mazama. Mitogenomic data from this study will provide useful information for further studies for the population genetics, speciation, biogeography, and conservation of L. chengyuani in the future.

A new orthopteran-parasitizing horsehair worm, Acutogordius taiwanensis sp. n., with a redescription of Chordodes formosanus and novel host records from Taiwan (Nematomorpha, Gordiida).

A description of a new species of horsehair worm, Acutogordius taiwanensissp. n., a redescription of Chordodes formosanus, and novel host records for the latter are provided. Acutogordius taiwanensissp. n. is morphologically similar to A. protectus with moderately flat areoles on its tail tips, but is distinguishable by small mid-body ornamentations. Despite the distinct differences in the post-cloacal crescents between 14 male samples, their conspecific status, along with that of nine female samples, was upheld by a phylogenetic comparison of partial cytochrome oxidase subunit I (COI) sequences. Chordodes formosanus is another common horsehair worm species in Taiwan, which was previously believed to specifically parasitize Hierodula mantids. However, in this study, five C. formosanus were observed emerging from an Acromantis mantid, and two long-horned grasshopper hosts (Leptoteratura sp. and Holochlora japonica). These five worms showed high degrees of similarity in COI sequences and morphology, but one of these individuals bore abnormal crowned areoles, which has never been observed in C. formosanus, and may be attributed to the incomplete development of this particular individual.

Annual Survey of Horsehair Worm Cysts in Northern Taiwan, with Notes on a Single Seasonal Infection Peak in Chironomid Larvae (Diptera: Chironomidae).

The life cycle of the freshwater horsehair worm typically includes a free-living phase (adult, egg, larva) and a multiple-host parasitic phase (aquatic paratenic host, terrestrial definitive host). Such a life cycle involving water and land can improve energy flow in riparian ecosystems; however, its temporal dynamics in nature have rarely been investigated. This study examined seasonal infection with cysts in larval Chironominae (Diptera: Chironomidae) in northern Taiwan. In the larval chironomids, cysts of 3 horsehair worm species were identified. The cysts of the dominant species were morphologically similar to those of Chordodes formosanus. Infection with these cysts increased suddenly and peaked 2 mo after the reproductive season of the adult horsehair worms. Although adult C. formosanus emerged several times in a year, only 1 distinct infection peak was detected in September in the chironomid larvae. Compared with the subfamily Chironominae, samples from the subfamilies Tanypodinae and Orthocladiinae were less parasitized. This indicates that the feeding behavior of the chironomid host likely affects horsehair worm cyst infections; however, bioconcentration in predatory chironomids was not detected.

Morphological allometry and intersexuality in horsehair-worm-infected mantids, Hierodula formosana (Mantodea: Mantidae).

Parasitic castration is a strategy used by parasites to minimize damage to the host by consuming its reproductive system, which results in the morphological alteration of the host. We determined that the forewing shape and density of the antennal sensilla of field-collected adult male mantids (Hierodula formosana), infected by horsehair worms (Chordodes formosanus) was partially feminized (intersexuality), and both male and female mantids infected by horsehair worms exhibited allometric changes in their wings and walking legs. In addition, the testes of most infected male adults disappeared or reduced in size, whereas the number of ovarioles in infected female adults was unaffected. The infection mainly influenced the structures related to host reproduction and locomotion, suggesting unbalanced energy exploitation and the reduction of parasitic virulence. In addition, the intersexuality of infected male adults indicated that sexual differentiation in insects, which researchers have considered to be an autonomous process, was influenced by the infection. The similarity of the antennae of infected male adults with those of last-instar female nymphs suggested that parasitic juvenilization may cause such feminization, but the mechanism of parasitic influence on insect sex characteristics should be studied further.

Keys to the blow flies of Taiwan, with a checklist of recorded species and the description of a new species of Paradichosia Senior-White (Diptera, Calliphoridae).

BLOW FLIES (DIPTERA: Calliphoridae) show a great diversity in behavior and ecology, play important roles in ecosystems, and have medical and forensic importance to humans. Despite this, the taxonomy and classification of Taiwan's Calliphoridae have rarely been studied. In this study, specimens of Taiwanese calliphorids were collected and carefully studied, and all 76 species recorded in Taiwan are listed following the identification keys. Dichotomous keys to all subfamilies, tribes, genera, and species of blow flies recorded in Taiwan are provided, including 16 species that are newly recorded from Taiwan. In addition, one new species of the genus Paradichosia Senior-White is described and illustrated. We also discuss the morphological differences between the specimens of Silbomyia hoeneana Enderlein collected from China and Taiwan, a species that has only been found previously in Southern China.

Life history data on the fly parasitoids Aleochara nigra Kraatz and A. asiatica Kraatz (Coleoptera: Staphylinidae), and their potential application in forensic entomology.

Knowledge of the developmental time of the immature stages of necrophagous flies has been the main tool for estimating minimum post-mortem intervals (min PMIs) in forensic entomology. Many parasitic insects can alter the development of immature stages of flies and thus affect min PMI estimates. The larvae of most species of Aleochara rove beetles are ectoparasitoids of the pupae of cyclorrhapha flies. Among them, some species that parasitise necrophagous flies may have forensic importance. Two Taiwanese Aleochara species, A. nigra and A. asiatica, which visit carrion sites were studied herein. All five necrophagous (Hemipyrellia ligurriens, Lucilia cuprina, Chrysomya megacephala, C. rufifacies and sarcophagid sp.) and one non-necrophagous fly species (Bactrocera dorsalis) we examined have the potential to be parasitised by these two Aleochara species, but differences among the acceptability and suitability of these hosts to rove beetle species suggested that rove beetles may prefer specific hosts. Each stage of the beetle life history was recorded to estimate developmental durations at six different temperatures. The larval stage together with the pupal stage of both beetle species was longer than the pupal stages of their hosts, implying the possibility of elongating the min PMI estimation. In addition, the host weight and larval duration of these two Aleochara beetles were positively correlated; thus, potential applications can be expected when using parasitised fly pupae in min PMI estimations.

Review of the genus Cidariplura Butler, 1879 (Lepidoptera, Erebidae, Herminiinae) in Taiwan with descriptions of four new species.

The genus Cidariplura Butler, 1879 from Taiwan is reviewed. A total of seven species are confirmed to occur in this old landbridge island, and four new species are described: C. shanmeii Wu & Owada sp. nov., C. maraho Wu & Owada sp. nov., C. atayal Wu & Owada sp. nov. and C. ilana Wu & Owada sp. nov. The Taiwanese C. bilineata (Wileman & South, 1919) is superficially similar to the Indian and Nepalese C. brevivittalis (Moore, 1867) but their genitalia show distinct differences. Elyra albifascia Hampson 1929 is regarded as a junior synonym of C. brevivittalis (syn. nov.). All diagnostic characteristics of Cidariplura from Taiwan and its adjacent areas are illustrated.

Oviposition preferences of two forensically important blow fly species, Chrysomya megacephala and C. rufifacies (Diptera: Calliphoridae), and implications for postmortem interval estimation.

Necrophagous blow fly species (Diptera: Calliphoridae) are the most important agents for estimating the postmortem interval (PMI) in forensic entomology. Nevertheless, the oviposition preferences of blow flies may cause a bias of PMI estimations because of a delay or acceleration of egg laying. Chrysomya megacephala (F.) and C. rufifacies (Macquart) are two predominant necrophagous blow fly species in Taiwan. Their larvae undergo rather intense competition, and the latter one can prey on the former under natural conditions. To understand the oviposition preferences of these two species, a dual-choice device was used to test the choice of oviposition sites by females. Results showed when pork liver with and without larvae of C. rufifacies was provided, C. megacephala preferred to lay eggs on the liver without larvae. However, C. megacephala showed no preference when pork liver with and without conspecific larvae or larvae of Hemipyrellia ligurriens (Wiedemann) was provided. These results indicate that females of C. megacephala try to avoid laying eggs around larvae of facultatively predaceous species of C. rufifacies. However, C. rufifacies showed significant oviposition preference for pork liver with larvae of C. megacephala or conspecific ones when compared with pork liver with no larvae. These results probably imply that conspecific larvae or larvae of C. megacephala may potentially be alternative food resources for C. rufifacies, so that its females prefer to lay eggs in their presence. When considering the size of the oviposition media, pork livers of a relatively small size were obviously unfavorable to both species. This may be because females need to find sufficient resources to meet the food demands of their larvae. In another experiment, neither blow fly species showed an oviposition preference for pork livers of different stages of decay. In addition, the oviposition preferences of both species to those media with larvae were greatly disturbed in a dark environment. If we removed the larvae that had previously fed on the pork liver and let the females choose, no oviposition preference was observed; but both species still showed a preference for the larger media in the dark. This suggests that female blow flies can use visual cues to recognize larvae on the media and decide on their oviposition site. Our studies point out the effects of some biotic and abiotic factors which were previously overlooked, and remind us to reevaluate these effects on oviposition, especially when using insect developmental data to estimate PMIs.

A new horsehair worm, Chordodes formosanus sp. n. (Nematomorpha, Gordiida) from Hierodula mantids of Taiwan and Japan with redescription of a closely related species, Chordodes japonensis.

A new species of horsehair worm, Chordodes formosanussp. n., is described and compared to a closely related species, Chordodes japonensis. Although both species possess the same six cuticular structures of areoles on the surface, the significantly longer filaments on the female crowned areoles can be used as diagnostic characters for the new species. The different taxonomic status of these two species was also confirmed after analyzing the partial cytochrome oxidase subunit I sequence, and the mantid hosts, which are respectively limited to the genus Tenodera for Chordodes japonensis and Hierodula for Chordodes formosanussp. n. In addition, the immature stages of eggs and larvae of the new species are also described and discussed in detail.

Larval competition of Chrysomya megacephala and Chrysomya rufifacies (Diptera: Calliphoridae): behavior and ecological studies of two blow fly species of forensic significance.

Chrysomya megacephala and Chrysomya rufifacies are two predominant necrophagous species in Taiwan. Larvae of the latter can prey on other maggots, including that of their own species as facultative food. This facultative characteristic of C. rufifacies may enhance its competitive advantage over other maggots and could also change the situation of other coexisting colonies. In this study, these two species were colonized in the laboratory, and the main objective was to try to understand the effect of competition on larval development. According to our results, intraspecific competition mostly occurred as competition for food; when the rearing density was increased, larvae pupated earlier, resulting in a lighter adult dry weight. The tendencies were similar in both species, but C. megacephala developed smaller viable adults and had higher survivorship at high densities. Although C. rufifacies could use the food resource of cannibalism, its survivorship was still low. Our results also showed there were significant interactions between intraspecific competition and the density factor. However, with interspecific competition, the first-instar larvae of C. rufifacies invaded maggot masses of C. megacephala to feed together. The third instars of C. rufifacies were able to expel C. megacephala larvae from food by using a fleshy protrusion on their body surface; C. megacephala was usually forced to pupate earlier by shortening its larval stages. The results indicated that a temporary competitive advantage could only be obtained by C. rufifacies under a proper larval density. In addition, the effects on different larval stages, the responses to different competition intensities, and the temperature-dependent effects on interspecific competition are also discussed. In general, under mixed-species rearing at different temperatures and densities, larval duration, adult dry weight, and survivorship of both species decreased. However, our results did not completely agree with previous studies, and we suspect that the difference was partially caused by different experimental designs and different biological characters of different blow fly colonies. Our results also suggest that both the predation ability and defense or escape activity should be taken into account when evaluating larval competitive advantages. The durations of larval stages of these two species could be decreased by approximately 54 h when a single species was reared alone and food was limited; the largest reduction in larval duration, approximately 25 h in C. megacephala and 34 h in C. rufifacies, caused by interspecific competition was under a high larval density. In conclusion, competition decreased the larval duration of these two species by up to 2 d; this also draws attention to justifying the postmortem interval estimation of using larval developmental data when larval competition exists.

Molecular identification of forensically important blow fly species (Diptera: Calliphoridae) in Taiwan.

Forensic entomology is a discipline that mainly uses insects collected in and around corpses to estimate the post-mortem interval in medicocriminal investigations. Among all scavenger and necrophagous insect groups that are related to corpses, blow flies (Diptera: Calliphoridae) are probably most important, not only because they occur in abundant numbers but also because they are one of the earliest groups to find corpses. However, most entomological evidence is strongly dependent on accurate species identification. Because identification allows the proper developmental data and distribution ranges to be applied in criminal investigations, species in Taiwan were surveyed from early 2000 and were identified using molecular data. Currently, eight species have been identified: Chrysomya megacephala (Fabricius), Chrysomya pinguis (Walker), Chrysomya rufifacies (Macquart), Hemipyrellia ligurriens (Wiedemann), Lucilia bazini Séguy, Lucilia cuprina (Wiedemann), Lucilia hainanensis Fan, and Lucilia prophyrina (Walker). We focused on classifying these blow fly species to establish a knowledge basis for further forensic entomological research in Taiwan. Because molecular data are helpful in identifying insect specimens, especially when no specimen of suitable condition for morphological identification is obtained, we extracted mitochondrial cytochrome oxidase subunit I (COI) DNA of the preceding blow fly species to study its application value for their differentiation. The cloning and sequencing of the COI gene (approximately 1,588 base pairs) of these eight species were completed, and the data were analyzed. Preliminary results revealed the high support of congeneric groupings of species by using COI data; these sequences were also shown to be highly conserved within the same species. To actually use the database of COI sequences under various specimen conditions, specific primers were also applied for different insect stages, different segments of adults, and specimens preserved for various times. A molecular primer key was ultimately constructed for the purpose of rapid and accurate species identification at the molecular level regardless of which stage or which part of a blow fly specimen is collected.