Habitat requirements and niche selection of Maculinea nausithous and M. teleius (Lepidoptera: Lycaenidae) within a large sympatric metapopulation

Identifying the factors determining the distribution and abundance of species is a fundamental question in population ecology. Based on habitat characteristics, we sought to identify factors affecting the abundance of two vulnerable large blue butterflies, Maculinea teleius and Maculinea nausithous. Both species often occur in sympatry and thus seem to have very similar requirements with regard to their wetland habitats. Even though, M. nausithous is usually far more widespread and abundant than M. teleius in Central Europe. For understanding this difference despite a similar ecology, more data is needed on the niche selection of both species. We have recorded Maculinea numbers, vegetation structure, food-plant abundance and ant community for 120 patches within a large sympatric metapopulation of both species in the Westerwald area (Germany). Unexpectedly (based on previous data), both species were equally common in the study area, reaching high densities of up to 61 (M. teleius) and 58 (M. nausithous) individuals per 50 m2. In 116 out of 120 patches both species occurred sympatrically. The preferred habitat was extensively managed grassland, being mown or grazed twice a year. The abundance of both Maculinea species was positively related to vegetation height, food-plant and Myrmica rubra abundance. There was no association between M. teleius and its putative main host-ant Myrmica scabrinodis, indicating that M. rubra may serve as an important host in the Westerwald area. The positive effect of food-plant abundance was more pronounced for M. teleius, coinciding with a higher food-plant abundance at sites dominated by M. teleius compared to those dominated by M. nausithous. Thus, M. teleius seems to be slightly more demanding than M. nausithous in terms of food-plant availability.     

Incipient sympatric speciation via host race formation in Phengaris arion (Lepidoptera: Lycaenidae)

Organisms Diversity & Evolution - The plausibility of sympatric speciation is still debated despite increasing evidence, such as host races in insects. This speciation process may be occurring...     

Cochylis Treitschke in China: one new species and five new records (Lepidoptera,Tortricidae, Cochylini)

Six species of Cochylis Treitschke, 1829 are recorded for China. Among them, Cochylis triangulasp. n. is described as new; Cochylis atricapitana (Stephens, 1852), Cochylis discerta Razowski, 1970, Cochylis dubitana (Hübner, [1799]), Cochylis faustana (Kennel, 1919) and Cochylis posterana hyrcana (Toll, 1948) are recorded for the first time for China. The female of Cochylis discerta Razowski, 1970 is described for the first time. Adults and genitaliaare illustrated, a key is given for the identification of Chinese species based on male genitalia.     

Analysis of the Territorial, Courtship and Coupling Behavior of the Hemipterophagous Butterfly, Spalgis epius (Westwood) (Lepidoptera: Lycaenidae)

In this study we examined the territorial, courtship and coupling behavior, mating interruptions and influence of sunlight on mating behavior in Spalgis epius (Westwood). We constructed an ethogram of territorial and courtship behavior, and catalogued the sequence of behavioral acts associated with mating behavior. Mating behavioral acts were divided into four repertoires i.e. pre-courtship, pre-coupling, coupling and post-coupling behaviors. Totally 22 behavioral acts were recorded from four repertoires. Tree canopy, canopy height and sunlight conditions are the important factors that influence copulation in S. epius. The courtship activity led to successful copulation in 71.9 % pairs. Incidence of different types of courtship and copula interruptions in S. epius was also recorded.     

Microsatellite markers for the large blue butterflies Maculinea nausithous and Maculinea alcon (Lepidoptera: Lycaenidae) and their amplification in other Maculinea species

We developed microsatellite markers for Maculinea nausithous and Maculinea alcon, two of five species of endangered large blue butterflies found in Europe. Two separate microsatellite libraries were constructed. Eleven markers were developed for M. nausithous and one for M. alcon. The primers were tested on both species as well as on the three other European Maculinea species. The number of alleles per locus ranged from two to 14. These markers will be useful tools for population genetic studies of Maculinea species.     

Genetic variation and Wolbachia infection status of the lycaenid butterfly Zizina emelina (Lepidoptera,Lycaenidae) from Korean populations

The lycaenid butterfly Zizina emelina emelina (de l’Orza) (previously Zizina otis emelina; Lepidoptera, Lycaenidae) has populations distributed across Japan and Korea. Zizina emelina is listed on Japan’s Red List as an endangered species, owing to the loss of its principal plant food and habitat. Researchers previously investigated the genetic diversity and structure in Japanese populations by using mitochondrial and nuclear DNA analyses alongside their Wolbachia infection status to consider the conservation strategy. In this study, we investigated the genetic variation and Wolbachia infection status of Korean populations and compared our findings with these traits in the Japanese populations. Four novel mitochondrial haplotypes were found in the Korean populations, whereas nuclear DNA analysis revealed one new haplotype and another that matched the one reported previously in Japan. The two known strains of Wolbachia in the Japanese populations were also found in the Korean ones. Mitochondrial DNA analysis demonstrated that no gene flow had occurred between the Japanese and Korean populations. The fact that they share the same nuclear haplotype and Wolbachia strains, however, indicates a low level of population differentiation between the two.     

P. falciparum and P. vivax Epitope-Focused VLPs Elicit Sterile Immunity to Blood Stage Infections

In order to design P. falciparum preerythrocytic vaccine candidates, a library of circumsporozoite (CS) T and B cell epitopes displayed on the woodchuck hepatitis virus core antigen (WHcAg) VLP platform was produced. To test the protective efficacy of the WHcAg-CS VLPs, hybrid CS P. berghei/P. falciparum (Pb/Pf) sporozoites were used to challenge immunized mice. VLPs carrying 1 or 2 different CS repeat B cell epitopes and 3 VLPs carrying different CS non-repeat B cell epitopes elicited high levels of anti-insert antibodies (Abs). Whereas, VLPs carrying CS repeat B cell epitopes conferred 98% protection of the liver against a 10,000 Pb/Pf sporozoite challenge, VLPs carrying the CS non-repeat B cell eptiopes were minimally-to-non-protective. One-to-three CS-specific CD4/CD8 T cell sites were also fused to VLPs, which primed CS-specific as well as WHcAg-specific T cells. However, a VLP carrying only the 3 T cell domains failed to protect against a sporozoite challenge, indicating a requirement for anti-CS repeat Abs. A VLP carrying 2 CS repeat B cell epitopes and 3 CS T cell sites in alum adjuvant elicited high titer anti-CS Abs (endpoint dilution titer >1x10⁶) and provided 80–100% protection against blood stage malaria. Using a similar strategy, VLPs were constructed carrying P. vivax CS repeat B cell epitopes (WHc-Pv-78), which elicited high levels of anti-CS Abs and conferred 99% protection of the liver against a 10,000 Pb/Pv sporozoite challenge and elicited sterile immunity to blood stage infection. These results indicate that immunization with epitope-focused VLPs carrying selected B and T cell epitopes from the P. falciparum and P. vivax CS proteins can elicit sterile immunity against blood stage malaria. Hybrid WHcAg-CS VLPs could provide the basis for a bivalent P. falciparum/P. vivax malaria vaccine.     

Wolbachia Enhances West Nile Virus (WNV) Infection in the Mosquito Culex tarsalis

Novel strategies are required to control mosquitoes and the pathogens they transmit. One attractive approach involves maternally inherited endosymbiotic Wolbachia bacteria. After artificial infection with Wolbachia, many mosquitoes become refractory to infection and transmission of diverse pathogens. We evaluated the effects of Wolbachia (wAlbB strain) on infection, dissemination and transmission of West Nile virus (WNV) in the naturally uninfected mosquito Culex tarsalis, which is an important WNV vector in North America. After inoculation into adult female mosquitoes, Wolbachia reached high titers and disseminated widely to numerous tissues including the head, thoracic flight muscles, fat body and ovarian follicles. Contrary to other systems, Wolbachia did not inhibit WNV in this mosquito. Rather, WNV infection rate was significantly higher in Wolbachia-infected mosquitoes compared to controls. Quantitative PCR of selected innate immune genes indicated that REL1 (the activator of the antiviral Toll immune pathway) was down regulated in Wolbachia-infected relative to control mosquitoes. This is the first observation of Wolbachia-induced enhancement of a human pathogen in mosquitoes, suggesting that caution should be applied before releasing Wolbachia-infected insects as part of a vector-borne disease control program.     

Molecular Evidence for Cryptic Speciation in the Cyclophorus fulguratus (Pfeiffer, 1854) Species Complex (Caenogastropoda: Cyclophoridae) with Description of New Species

A high degree of intraspecific variation, both genetic and in shell morphology, of the operculate land snail Cyclophorus fulguratus (Pfeiffer, 1854) suggests that its classification as a single species warrants reconsideration. We sequenced two nuclear (18S and 28S) and two mitochondrial (16S and COI) genes of 46 C. fulguratus specimens and used them to estimate the phylogeny and to determine the validity of species boundaries. Molecular phylogenetic analyses revealed the presence of three lineages corresponding to three geographically disjunctive populations of C. fulguratus in Thailand. Likelihood tests of topologies significantly supported the non-monophyly of the C. fulguratus–complex and Bayesian species delimitation analysis significantly supported the potential representation as distinct species of these three lineages. Discriminant function analysis based on geometric-morphometrics of shell shape allowed for significant distinction of these three candidate species, although they revealed a considerable degree of overlap of shell shape reflecting their crypsis morphologically. The diagnostic characters are provided by color pattern, pattern of protoconch and pattern of jaw. In conclusion, the results support that the C. fulguratus s.l., as currently recognized, consists of three distinct species in Thailand: C. fulguratus s.s., C. rangunensis and C. abditus sp.nov., which are described herein.     

Hexamermis cathetospiculae n. sp. (Mermithidae: Nematoda), a Parasite of the Rice Stemborer,Tryporyza incertulas (Wlk.) (Pyralidae: Lepidoptera) in Malaysia

Hexamermis cathetospiculae n. sp. (Mermithidae: Nematoda), a parasite of the rice stemborer, Tryporyza incertulas (Wlk.), is described. This mermithid was observed to control the stemborer in Malaysia. It is distinguished from other members of the genus by the shape of the vagina, the nearly straight spicules, and the form of the amphids.     

No seasonal trend in infection of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae), by Wolbachia

Wolbachia are Gram-negative bacteria that cause intracellular inherited infections in many invertebrates. They are extremely common, with 20–75 % of all insects being infected. Wolbachia belong to taxa of the Anaplasmataceae family, alpha proteobacteria. Because previous studies have shown that Wolbachia generally disappear from the host insect’s body under high-temperature conditions in laboratories, we investigated seasonal changes in infection of the pale grass blue butterfly, Zizeeria maha (Kollar, 1848) by Wolbachia, for 7 months of the year. Total annual infection was 86.7 % (n = 15) of females and 96.3 % (n = 81) of males. Statistical analysis showed that monthly infection was not significantly different among months. In addition, no significant difference was found between Wolbachia densities in cells of summer and winter butterfly samples by use of real-time PCR during the months examined. The results suggest that Wolbachia infection is not affected by seasonal factors, at least for Z. maha.     

Clinal Variation in Populations of the Common Blue Butterfly <Emphasis Type="Italic">Polyommatus icarus</Emphasis> Rott. (Lepidoptera,Lycaenidae)

Geographical trends in variation of wing pattern characters of the common blue butterfly Polyommatus icarus Rott. have been established. Peripheral populations were shown to be more rich and diverse phenotypically than those of the central parts of the species range. Phenotypic diversity of the populations increased from the center to the periphery of the range. The range boundaries were characterized by strong fluctuations of frequencies of wing pattern phenotypes. Hybrid areas were found on the part of the range examined.__________Translated from Genetika, Vol. 41, No. 8, 2005, pp. 1055–1067.Original Russian Text Copyright © 2005 by Artemyeva.     

Various Chemical Strategies to Deceive Ants in Three Arhopala Species (Lepidoptera: Lycaenidae) Exploiting Macaranga Myrmecophytes

Macaranga myrmecophytes (ant-plants) are generally well protected from herbivore attacks by their symbiotic ants (plant-ants). However, larvae of Arhopala (Lepidoptera: Lycaenidae) species survive and develop on specific Macaranga ant-plant species without being attacked by the plant-ants of their host species. We hypothesized that Arhopala larvae chemically mimic or camouflage themselves with the ants on their host plant so that the larvae are accepted by the plant-ant species of their host. Chemical analyses of cuticular hydrocarbons showed that chemical congruency varied among Arhopala species; A. dajagaka matched well the host plant-ants, A. amphimuta did not match, and unexpectedly, A. zylda lacked hydrocarbons. Behaviorally, the larvae and dummies coated with cuticular chemicals of A. dajagaka were well attended by the plant-ants, especially by those of the host. A. amphimuta was often attacked by all plant-ants except for the host plant-ants toward the larvae, and those of A. zylda were ignored by all plant-ants. Our results suggested that conspicuous variations exist in the chemical strategies used by the myrmecophilous butterflies that allow them to avoid ant attack and be accepted by the plant-ant colonies.     

Male secondary sexual structures and the systematics of the Thereus oppia species group (Lepidoptera,Lycaenidae, Eumaeini)

The Thereus oppia species group includes species with and without a scent pad, which is a histologically and morphologically characterized male secondary sexual structure on the dorsal surface of the forewing. To assess the hypothesis that these structures are lost evolutionarily, but not regained (Dollo’s Law), the taxonomy of this species group is revised. Thereus lomalarga sp. n., and Thereus brocki sp. n., are described. Diagnostic traits, especially male secondary structures, within the Thereus oppia species group are illustrated. Distributional and biological information is summarized for each species. Three species have been reared, and the caterpillars eat Loranthaceae. An inferred phylogeny is consistent with the hypothesis that scent pads in the Thereus oppia species group have been lost evolutionarily twice (in allopatry), and not re-gained.     

Description of third instar larvae of Ceratitis fasciventris,C. anonae,C. rosa (FAR complex) and C. capitata (Diptera,Tephritidae)

Third instar larvae of members of the Ceratitis FAR complex, including Ceratitisfasciventris (Bezzi), Ceratitisanonae Graham, and Ceratitisrosa Karsch are described and compared with those of Ceratitiscapitata (Wiedemann). Diagnostic characters, such as presence vs. absence of a secondary tooth on the mandibles, previously used to separate Ceratitiscapitata from Ceratitisrosa, are shown to vary in each species. Significant variation in diagnostic morphological characters among populations of Ceratitisrosa from east and south Africa is documented; however, the differences are not simply congruent with the R1 and R2 designations based on other studies. Quantitative measures of numerous morphological characters are consistently smaller in the larvae of Ceratitisfasciventris and distinguish them from other species of the FAR complex. Larvae of Ceratitiscapitata can be distinguished from those of the FAR complex by characters such as absence of accessory plates of the oral ridges, the shape of the anterior spiracle, and the pattern of dorsal spinules. Previous studies indicated that absence of accessory lobes separate the genus Ceratitis from Bactrocera, but this is shown to be incorrect, as accessory lobes are in fact present in several species of Ceratitis.     

Feminizing Wolbachia in an insect,Ostrinia furnacalis (Lepidoptera: Crambidae)

Wolbachia, which forms a group of maternally inherited bacteria in arthropods, often cause reproduction alterations in their hosts, such as cytoplasmic incompatibility, parthenogenesis, male-killing, hybrid breakdown and feminization. To date, Wolbachia-induced feminization has been reported only in isopods. Here we report that a Wolbachia strain feminizes an insect host, Ostrinia furnacalis. Among 79 wild females of O. furnacalis examined, Wolbachia infection was detected in 13 females. Twelve of the 13 infected females produced all-female progenies, and this trait was maternally inherited. Tetracycline treatment of thelygenic matrilines resulted in the production of all-male progenies. The present findings indicate that the Wolbachia infection induces feminization of genetic males in O. furnacalis. Differences in the Wolbachia-induced feminization in O. furnacalis and that in isopods are discussed along with the differences in sex determination mechanisms between insects and isopods. Phylogenetic analysis of the wsp gene sequence of Wolbachia suggests independent evolutionary origins for the Wolbachia-induced feminizations in O. furnacalis and in isopods. Our findings over 5 years suggest that the infection has been maintained at a low prevalence in the O. furnacalis population.