Digestive amylase and pectinase activity in the larvae of alfalfa weevil Hypera postica (Coleoptera: Curculionidae)

The alfalfa weevil Hypera postica is a serious economic pest in most alfalfa grown in many countries worldwide. Digestive α-amylase and pectinase activities of larvae were investigated using general substrates. Midgut extracts from larvae showed an optimum activity for α-amylase against starch at acidic pH (pH 5.0). α-Amylase from larval midgut was more stable at mildly acidic pH (pH 5–6) than highly acidic and alkaline pH. The enzyme showed its maximum activity at 35°C. α-Amylase activity was significantly decreased in the presence of Ca²⁺, Mg²⁺ and sodium dodecylsulfate. On the contrary, K⁺ and Na⁺ did not significantly affect the enzyme activity. Zymogram analysis revealed the presence of one band of α-amylase activity in in-gel assays. Pectinase activity was assayed using agarose plate and colorimetric assays. Optimal pH for pectinase activity in the larval midgut was determined to be pH 5.0. Pectinase enzyme is more stable at pH 4.0–7.0 than highly acidic and alkaline pH. However, the enzyme was more stable at slightly acidic pH (pH 6.0) when incubation time increased. Maximum activity for the enzyme incubated at different temperatures was observed to be 40°C. Optimum pH activity for α-amylase and pectinase is not completely consistent with the pH prevailing in the larval midgut. This is the first report of the presence of pectinase activity in H. postica.     

An Immunoassay Method for Detecting Microctonus aethiopoides (Hymenoptera: Braconidae: Euphorinae) Parasitism in the Alfalfa Weevil, Hypera postica (Coleoptera: Curculionidae)

A simple immunological assay was developed as an alternative to the dissection/visualization method for detecting the presence of the parasitoid, Microctonus aethiopoides Loan, in the alfalfa weevil, Hypera postica (Gyllenhal). The dot-blot assay was validated using laboratory-reared and field-collected adult weevils of known parasitization status. The dot-blot assay was also used to estimate the developmental stage of the parasitoid within parasitized adult hosts. The assay results can be used to forecast parasitoid emergence dates and estimate the parasitism rate of M. aethiopoides in alfalfa weevil populations.     

Coexistence of two mitochondrial DNA haplotypes in Japanese populations of Hypera postica (Col., Curculionidae)

Abstract:  In Japan, the alfalfa weevil, Hypera postica , was first recorded in 1982 from Fukuoka and Okinawa Prefectures and has been spreading to many other prefectures. The weevil seriously infests the Chinese milk vetch, Astragalus sinicus , one of the most important honey resources for honeybees in Japan. Direct sequencing of partial mitochondrial DNA and PCR-RFLP data for alfalfa weevil individuals indicated the coexistence of two haplotypes at various localities in Japan. Molecular phylogenetic analysis for H. postica haplotypes and strains indicated that the two Japanese haplotypes had not derived from a single genetic origin. Based on the results, special comments are made on biological control measures using introduced parasitic waSPS.     

The attachment and stylostome of Trombidium newelli (Acari: Trombidiidae), an ectoparasitic mite on adults of alfalfa weevil, Hypera postica (Coleoptera: Curculionidae)

Larvae of the mite Trombidium newelli Welbourn and Flessel are ectoparasitic on adult alfalfa weevil, Hypera postica (Gyllenhall), an insect pest of alfalfa. The mite larvae are found under the elytra, attached to the dorsal surface of weevils abdomen. T. newelli larvae use their chelicerae to penetrate the pliable and weakly sclerotized areas of the hosts integument, and to hold on firmly to the host. The attachment sites associated with larval mites of different degrees of engorgement were examined using both light and scanning electron microscopy. The ventral inside surface of parasitized host tergites revealed a characteristic injury as spots varying in color from white-yellow to golden-brown depending on the engorged size of the mite. In addition, spots associated with fully engorged mite larvae showed an aggressive dendritically radiating mass expanding outward from the site of cheliceral penetration. This mass, known as feeding tube or stylostome was branched, with each branch ending in a cluster of closed bulbs. Stylostomes appear to exist independently in the hosts tissue. Stylostome mass corresponded with the engorgement state of the mite, increasing as the mite larva increased in size. The possible nature and mechanism of stylostome formation is discussed.     

Identification and biological role of the endosymbionts Wolbachia in rice water weevil (Coleoptera: Curculionidae)

Wolbachia spp. are obligate intracellular bacteria present in reproductive tissues of many arthropod species. Wolbachia infection status and roles in host reproduction were studied in the rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera, Curculionidae), an introduced species in China. We examined Wolbachia infection status in five populations in China where it reproduces parthenogenetically, and one native population in Southeast Texas, where it reproduces bisexually. All populations were infected by Wolbachia, and all specimens in each population were infected by Wolbachia of a single strain. Phylogenetic analyses based on multilocus sequence typing system indicated that Wolbachia in non-native L. oryzophilus weevils diverges evidently from those in native weevils. After treatments with tetracycline, parthenogenetic weevils reduced the fecundity significantly and eggs were not viable. Our results suggest that Wolbachia are necessary for oocyte production in L oryzophilus.     

Wolbachia infection in the Korean endemic firefly,Luciola unmunsana (Coleoptera: Lampyridae)

Wolbachia is one of the most prevalent endosymbiontic bacteria of arthropods. The bacterium induces sex ratio distortions in various host insects through processes such as cytoplasmic incompatibility, feminization, male killing, and parthenogenesis. We investigated if the Korean endemic firefly, Luciola unmunsana was infected with the bacterium because the species had an abnormal sex ratio in the field. The results show that some individuals are infected with the bacterium. Phylogenetic analyses showed that the bacterial strain infecting the firefly is closely related to strains that infect phylogenetically distant hosts.     

New Weevils (Coleoptera: Curculionidae) from Rovno Amber

Paleontological Journal - A new genus Rovnoslonik Legalov, Nazarenko and Perkovsky gen. nov. (type species Rovnoslonik damzeni Legalov, Nazarenko and Perkovsky sp. nov.; tribe Stromboscerini,...     

Mating behavior ofBathyplectes curculionis [Hym.: Ichneumonidae] a parasitoid of the alfalfa weevil,Hypera postica Col.: Curculionidae]

We describe the courtship behavior of male and femaleBathyplectes curculionis (Thomson). FemaleB. curculionis are polygamous and sexually receptive for up to 2 days after eclosion. The high activity levels of virgin females combined with their polygamy and long period of sexual receptivity allow for post-dispersal mating which may facilitate maximum gene flow between localB. curculionis populations. This high genetic diversity may be partially responsible for the widespread occurrence ofB. curculionis in North America.

---

Résumé Nous décrivons le comportement sexuel deBathyplectes curculionis (Thomson) male et femelle. Les femelles deB. curculionis sont polygames et réceptives 2 jours après l'éclosion. L'activité des femelles vierges et leur polygamie associées à la longue période de réceptivité sexuelle permettent un maximum d'échanges génétiques entre les populations locales. Cette grande diversité génétique peut être en partie responsable de l'étendue de la distribution deB. curculionis en Amérique du Nord.

---

---

Partially supported by funds from both the Department of Entomology and the Graduate School of The Ohio State University.

---

Submitted to the Graduate School of The Ohio State University in partial fulfillment of the requirements of the degree of Doctor of Philosophy. Present address: Agricultural Research Center, University of Florida, 3205 S. W. 70th Avenue, Ft. Lauderdale, Florida, U.S.A., 33314.     

Grandlure dosage and attraction of boll weevils (Coleoptera: Curculionidae)

The effects of grandlure dosage on of boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), attraction were assessed. Traps collected more boll weevils under field and laboratory conditions as the amount of grandlure in laminated plastic strips was increased from 0 to 10, 30, and 60 mg. Spreading the point source of the lure by cutting the strip into quarters and positioning each quarter on separate corners of the large capacity trap to create an expanded source for the pheromone plume, however, resulted in fewer trap captures than traps with quartered lures all positioned on a single corner. The large capacity trap with the quartered lure on one corner also caught more weevils than the traps with an intact lure fastened to one corner. Although aging lure strips for three weeks reduced emissions of the four pheromone components and their attractiveness to boll weevils, cutting the aged lure into quarters resulted in greater emissions and attraction than lures that were aged intact or as quarters. Some pheromone components volatilized faster than others, resulting in time-related changes in blend ratios, but the underlying factor in boll weevil attraction to grandlure strips was dosage, the amount of volatilized pheromone available for interacting with an adult boll weevil.     

Fungi associated with Scolytogenes birosimensis (Coleoptera: Curculionidae) infesting Pittosporum tobira

The bark beetle Scolytogenes birosimensis Niijima is suspected to be involved in the decline of Pittosporum tobira (Thunb. ex Murray) Aiton in the coastal areas of Japan. We isolated fungi from adult S. birosimensis in nine different localities in Japan to assess their potential association and predict their contribution to the success of the beetle. Results from morphological identification of associated fungi showed that the beetle was associated with Fusarium solani and Candida spp. Furthermore, molecular analysis showed that F. solani was most closely related to the plant pathogenic fungus F. solani f. sp. mori. Fungal isolation from surface-sterilized, dissected beetles and scanning electron miscroscope (SEM) observation of the body surface suggested that the associated fungi were carried in the pits on the beetles' elytra. These findings contribute to the understanding of the relationships between S. birosimensis and its associated fungi.     

Captures of boll weevils (Coleoptera: Curculionidae) in traps associated with different habitats

Programs to eradicate the boll weevil, Anthonomus grandis grandis Boheman, from cotton, Gossypium hirsutum L., in the United States rely heavily on pheromone traps for monitoring weevil populations in both active and posteradication maintenance programs. Modifications to trapping protocols that increase trap effectiveness should contribute to this eradication effort. Between October 1996 and May 1997 and between September 1997 and April 1998, we compared trap effectiveness, indicated by the numbers of captured weevils, in relation to selected habitat types. Each study period was divided into fall, winter, and spring seasons. Traps were closely associated with seven habitat types, including four types with prominent erect vegetation (brush-lined irrigation canal, brush, sugarcane, and resaca or ox-bow lake) and three types with only low-growing or sparse erect vegetation (irrigation drainage canal, unimproved pasture, and fallow fields). Captures of male and female weevils were statistically similar regardless of season or trapping habitat. Although captures differed significantly among habitats, these differences varied among seasons. Trapping habitats with prominent vegetational features generally produced higher weekly captures of weevils than habitats lacking these features. Also, captures in traps associated with prominent vegetation indicated seasonal differences in weevil activity, with highest captures occurring during the fall. Traps associated with habitats lacking prominent vegetation did not statistically demonstrate seasonal differences. Our results indicate that immediate trap surroundings strongly influence the effectiveness of the boll weevil pheromone trap. These results also suggest that effectiveness of current trapping programs may be improved through purposeful association of traps with selected vegetational features.     

Size-dependent feeding and reproduction by boll weevil (Coleoptera: Curculionidae)

The considerable variation in adult size of the boll weevil, Anthonomus grandis grandis Boheman, has been well documented, but the influences of adult size on reproductive rate are not known. We examined the relationship between the size of boll weevils and their feeding and oviposition. Weevils weighed to the nearest milligram were grouped into five categories based on pupal weight: < or =5, 6-10, 11-15, 16-20, and >20 mg. Numbers of lifetime punctures produced in flower buds (squares) of cotton, Gossypium hirsutum L., by both sexes of adults tended to increase with pupal weight. Boll weevil females with pupal weights >10 mg produced progeny with significantly higher survival to adulthood and also produced a higher percentage of female progeny than those with pupal weights < or =10 mg. The population growth indices for females having pupal weights >10 mg averaged 1.8-fold higher than those of females weighing < or =10 mg. Survivorship of adults of both sexes also tended to increase with pupal weight. The percentage of females laying eggs on any given day averaged 2.1 times higher when their pupal weights were >10 mg than when their pupal weights were < or =10 mg. Although small size negatively affected female reproductive potential, even extremely small females produced some viable offspring. However, the penalties of small adult size, in terms of longevity and reproductive potential, suggest that cultural practices that result in the production of small adults may be used to impact weevil populations.     

Internal Spatiotemporal Population Dynamics of Infection with Three Wolbachia Strains in the Adzuki Bean Beetle, Callosobruchus chinensis (Coleoptera: Bruchidae)

The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 × 10⁸, 2 × 10⁸, and 5 × 10⁷ wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.     

Mating tactics and courtship behavior inCleogonus rubetra (Fabricius) (Coleoptera: Curculionidae)

Females of the tropical weevilCleogonus rubetra oviposit into fruits of the leguminous treeAndira inermis, and larvae develop in the pulp and seed of these fruits. We hypothesized three alternative tactics by which males might secure matings. By using focal observations of males and by evaluating predictions specific to each hypothesis, we demonstrate that males search within aggregations of conspecifics for receptive females, and upon finding a suitable partner, males mount and perform courtship behavior consisting of stroking the eyes and sides of the female's abdomen. Males also stridulate and emit a sequence of short buzzing sounds. While mounted, males actively prevent rival males from mating with their partner. Males defend their mates for a mean duration of 3.7 h (including copulation). As predicted, paired males were larger than solitary individuals, although the difference was marginally nonsignificant. The overabundance of fruits relative to males, the prolonged period during which females are active, and the probability of last male sperm precedence are factors that may have contributed to the evolution of this female-defense tactic by males. Paired females were significantly larger than solitary females. We observed no competition among females for mates, and the correlation between elytron length of paired males and females was not significant.     

萧氏松茎象的早期寄主研究

本文以调查的事实材料为基础,对萧氏松茎象的早期寄主进行了研究,说明该虫最先在马尾松树种上发生,而后逐渐蔓延到从国外引进的湿地松、火炬松和国外松上猖獗危害。 萧氏松茎象（ＨｙｌｏｂｉｔｅｌｕｓｘｉａｏｉＺｈａｎｇ）是江西省吉安地区 １９８８年发现的国外松新害虫。十多年来,该虫蔓延十分迅速,危害国外松特别严重。据１９９９年调查,江西省４０余县（市）有分布．发生面积１万公顷以上,成为国外松的主要害虫。究竟此虫最初寄主是何树种,它为什么发展那么快呢？弄清这一问题十分重要,不仅对制订正确的防范措施,而且对研…     

Microstructure and mechanical properties of rostrum in Cyrtotrachelus longimanus (Coleoptera: Curculionidae)

The microstructure, composition and mechanical properties of the rostrum in Cyrtotrachelus longimanus (JHC Fabre) were studied utilizing light, fluorescent, scanning electron microscopy (SEM) and energy-dispersive spectroscopy. SEM images show the morphological characteristics of rostrum’s cross section; it is a typical lightweight multilayer structure – one rigid exocuticle layer and dense endocuticle layers, which construct unevenly overlapping fiber structures. The composition analysis of the rostrum shows that it is mainly composed of C, H, N, O, as well as some metal elements and microelements, such as Mg, Si, Zn, Ca and Na, which contribute to its mechanical performance. The mechanical properties of the rostrum were tested by the electronic universal testing machine, which shows it has high-specific strength and is almost the same as that of the stainless steel. The results may provide a biological template to inspire biomimetic lightweight structure design.     

Dynamics ofSitotroga cerealella (olivier) (lepidoptera: gelechiidae) andSitophilus zeamais motschulsky (Coleoptera: Curculionidae) populations in a small bulk of stored corn

The dynamics of Angoumois grain moth, Sitotroga cerealella (Olivier ), and maize weevil, Sitophilus zeamaisMotschulsky , populations breeding in a small bulk (initially 5.36 t) of shelled corn were studied over an 8-year period by monthly sampling. The weevil population showed wide fluctuations in density superimposed on a general decline with time. The moth population showed no upward or downward trend for the first 60 months, although it fluctuated widely. Following a decline that occurred between 56 and 60 months, the moth population fluctuated within a much narrower range, and there was a general decrease in density with time. The decline of the weevil population paralleled deterioration of the corn as did that of the moth population after ca 60 months, and the decline of both species probably resulted from increasing scarcity of suitable breeding sites. Both populations exhibited seasonal variation in density with minima in late summer and early fall, following periods of adversely high temperatures in the storage shed. The populations increased during the fall, leveled off or declined slightly during the winter months, and then increased to maximum levels in late spring or early summer. It thus appears that high temperatures had a greater adverse effect on the populations than low temperatures. The grain moth and the maize weevil both tended to be randomly dispersed at low population levels and moderately aggregated at intermediate and high levels, although the degree of aggregation was not correlated with population density when low population levels were considered separately, and the maize weevil showed a greater tendency for aggregation than did the grain moth. Analysis of individual samples at fixed points in time showed a conspicuous bias for negative correlation between numbers of the two species within sampling quadrats, suggesting a tendency for the two species to segregate within the grain mass. This process could have resulted from behavioral differences or from the destruction of one species by the other. Competitive displacement of the grain moth by the maize weevil has been demonstrated in laboratory experiments but has rarely been observed under natural conditions, and in our study the two species coexisted for 8 years in a relatively small grain bulk.