Rearing Costelytra zealandica (Coleoptera: Scarabaeidae)

Interaction between larvae can be a cause of mortality when scarabaeid larvae are concentrated in a confined volume of soil. Larvae of Costelytra zealandica (White), the New Zealand grass grub, were held for 30 days at population densities between 1 and 50 larvae per 200 ml of medium, and the effect of crowding on their survival and weight change was assessed. Larval density had no effect on survival in soil plus chopped sheep dung (3 :1), with up to 10 2nd‐instar larvae or 5 3rd‐instar larvae per 200 ml of medium. Larval survival and weight gain were increased by adding germinating ryegrass seed (Grasslands Ruanui) to the medium base, but the effect of density did not diminish. Weight gain of 3rd‐instar larvae decreased as their density increased. When the amount of germinating ryegrass seed in the medium was varied, there was no significant difference between the survival of larvae held in treatments with 5, 25, and 50 g of seed per litre of medium. Survival was significantly lower with 0 and 100 g of seed per litre of medium. It is concluded that, in rearing conditions, the final density of fully grown 3rd‐instar larvae should be a maximum of about 20 per litre of medium to prevent mortality caused by larval interaction—provided that adequate food is available.     

Chemical attractants for larvae of Costelytra zealandica (Coleoptera: Scarabaeidae)

In controlled laboratory tests, using shallow glass chambers containing a coarse river sand substrate between ‘blank’ and ‘test’ sample cages, 3rd‐instar larvae of Costelytra zealandica (White) were attracted to parsnip and carrot root, freshly ground nutmeg, and ß‐pinene (a constituent common to these materials). The terpenol a‐fenchol was strongly attractive to the larvae. Ryegrass root, α‐pinene, and limonene were not significantly attractive, and limonene may indeed have some repellent effect.     

Influence of low temperature on pupation induction in Costelytra zealandica (Coleoptera: Scarabaeidae)

Costelytra zealandica (White) lives within the soil throughout its life cycle, except for brief, crepuscular excursions to the surface by the adults to mate and feed. Larvae collected more than 18 weeks before 50% field pupation did not pupate unless they were artificially chilled. Such chilling had no significant effect on the percentage pupation of larvae collected after this time; it is assumed that these larvae had received sufficient chilling from their environment to induce pupation. Larger larvae were more likely to pupate than smaller larvae during the period 13–7 weeks before 50% field pupation. It is postulated that larvae remain quiescent until the soil temperature increases in the spring, and pupate only then. Such a process would explain how the adults of a population emerge at about the same time.     

Electroantennogram responses of male Costelytra zealandica (Coleoptera: Scarabaeidae) to phenol and related compounds

Electroantennograms recorded from male New Zealand grass grub beetles, Costelytra zealandica (White), were used to compare the activity of phenol with that of closely related phenolics. These compounds had no masking effect on the antennal response. A comparison of activity between phenol and a crude female extract indicated that phenol is the sole pheromone.     

The male reproductive system of Costelytra zealandica (White)(Coleoptera: Scarabaeidae: Melolonthinae)

Abstract

The morphology and histology of the male internal reproductive organs of Costelytra zealandica show many similarities to other Scarabaeoidae, and are particularly close to other Melolonthinae and to Rutelinae. Testes follicles of C. zealandica have the usual structure for Melolonthinae with basal lobes surrounding the ends of the vasa efferentia. Epithelial cells of the vasa efferentia, vasa deferentia, and vesiculae seminales have similar densely basophilic cytoplasm but muscle layers are best developed around the latter. Accessory glands lack muscle and are not differentiated histologically into regions but do differ from their reservoirs. The ejaculatory duct has a cuticular intima and is differentiated into anterior and posterior regions. Both are surrounded by a muscular sheath which expands in the posterior region to enclose fluid. This forms a hydraulic mechanism for everting the internal sac during intromission. The parameres hook into the female’s genital chamber during copulation and have no pincer action. Probable homologies are listed between muscles of the external genitalia and anus of C. zealandica and other Scarabaeoidea.     

An iridescent virus and a rickettsia from the grass grub Costelytra zealandica (Coleoptera: Scarabaeidae)

Natural iridescent virus and rickettsia infections of Costelytra zealandica (White) and Odontria sp. indet. larvae were studied at a site in the upper Pareora Gorge scenic reserve, S. Canterbury. By sequentially sampling the site, it was found that neither the iridescent virus nor the rickettsiae appear to give rise to host mortalities that significantly alter the population density. Many larvae were found that appeared healthy, but carried inapparent iridescent virus infections. These diseases are not suitable for biological control of the grass grub.     

Description of the third-instar larva of Aphodius bimaculatus (Laxmann) (Coleoptera, Scarabaeidae)

The third-instar larva of Aphodius bimaculatus (Laxmann) is described from the material collected in Belarus. The larva differs from other species of the subgenus Acrossus in the convex clypeus with a complex structure and in the coloration of the head.     

Transformation and mineralization of soil organic nitrogen by the humivorous larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae)

In common bean (Phaseolus vulgaris L.), Fusarium root rot (caused by Fusarium solani f. sp. phaseoli) disease severity is increased by environmental factors that stress the plant. The current study used reciprocal grafting techniques with the resistant cultivar FR266 and the susceptible cultivar Montcalm to determine if the genetic control of resistance is conferred by the rootstock (root genotype) or the scion (shoot genotype) and if root vigor played a role in resistance. The influence of a compacted layer on root and shoot genotype response and root rot resistance was studied. Root rot resistance was found to be controlled by the root genotype, such that on a scale of 1 to 7 (severe disease) the FR266 root had an average score of 2.3 and the Montcalm root had an average score of 4.4. However, when grafted plants were grown in the presence of a compacted layer, the FR266 root and/or shoot genotype in any graft combination with the susceptible Montcalm had reduced root rot (score = 2.4 average) than the Montcalm self graft (score = 4.5). Root mass was shown to be controlled by the root genotype in the absence of compaction such that the FR266 root was 26% larger that the Montcalm root when grafted onto a FR266 shoot or a Montcalm shoot. When a compacted layer was present the root and shoot genotype both contributed to root mass. Average root diameter was controlled by the shoot genotype, as the FR266 shoot grafted to Montcalm or FR266 roots had thicker roots (average diameter 0.455 mm) than the Montcalm shoot (average diameter 0.418 mm). This study shows evidence that root vigor in the presence of Fusarium disease pressure should be evaluated to effectively develop common bean lines resistant to Fusarium root rot across a range of environments.     

Structure and Topology of Microbial Communities in the Major Gut Compartments of Melolontha melolontha Larvae (Coleoptera: Scarabaeidae)

Physicochemical gut conditions and the composition and topology of the intestinal microbiota in the major gut compartments of the root-feeding larva of the European cockchafer (Melolontha melolontha) were studied. Axial and radial profiles of pH, O₂, H₂, and redox potential were measured with microsensors. Terminal restriction fragment length polymorphism (T-RFLP) analysis of bacterial 16S rRNA genes in midgut samples of individual larvae revealed a simple but variable and probably nonspecific community structure. In contrast, the T-RFLP profiles of the hindgut samples were more diverse but highly similar, especially in the wall fraction, indicating the presence of a gut-specific community involved in digestion. While high acetate concentrations in the midgut and hindgut (34 and 15 mM) corroborated the presence of microbial fermentation in both compartments, methanogenesis was confined to the hindgut. Methanobrevibacter spp. were the only methanogens detected and were restricted to this compartment. Bacterial 16S rRNA gene clone libraries of the hindgut were dominated by clones related to the Clostridiales. Clones related to the Actinobacteria, Bacillales, Lactobacillales, and γ-Proteobacteria were restricted to the lumen, whereas clones related to the β- and δ-Proteobacteria were found only on the hindgut wall. Results of PCR-based analyses and fluorescence in situ hybridization of whole cells with group-specific oligonucleotide probes documented that Desulfovibrio-related bacteria comprise 10 to 15% of the bacterial community at the hindgut wall. The restriction of the sulfate-reducer-specific adenosine-5′-phosphosulfate reductase gene apsA to DNA extracts of the hindgut wall in larvae from four other populations in Europe suggested that sulfate reducers generally colonize this habitat.     

Histopathological effects of the Yen-Tc toxin complex from Yersinia entomophaga MH96 (Enterobacteriaceae) on the Costelytra zealandica (Coleoptera: Scarabaeidae) larval midgut

Yersinia entomophaga MH96, which was originally isolated from the New Zealand grass grub, Costelytra zealandica, produces an orally active proteinaceous toxin complex (Yen-Tc), and this toxin is responsible for mortality in a range of insect species, mainly within the Coleoptera and Lepidoptera. The genes encoding Yen-Tc are members of the toxin complex (Tc) family, with orthologs identified in several other bacterial species. As the mechanism of Yen-Tc activity remains unknown, a histopathological examination of C. zealandica larvae was undertaken in conjunction with cultured cells to identify the effects of Yen-Tc and to distinguish the contributions that its individual subunit components make upon intoxication. A progressive series of events that led to the deterioration of the midgut epithelium was observed. Additionally, experiments using a cell culture assay system were carried out to determine the cellular effects of intoxication on cells after topical application and the transient expression of Yen-Tc and its individual components. While observations were broadly consistent with those previously reported for other Tc family members, some differences were noted. In particular, the distinct stepwise disintegration of the midgut shared features associated with both apoptosis and necrotic programmed cell death pathways. Second, we observed, for the first time, a contribution of toxicity from two chitinases associated with the Yen-Tc complex. Our findings were suggestive of the activities encoded within the subunit components of Yen-Tc targeting different sites along putative programmed cell death pathways. Given the observed broad host range for Yen-Tc, these targeted loci are likely to be widely shared among insects.     

Physicochemical conditions and microbial activities in the highly alkaline gut of the humus-feeding larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae)

The soil macrofauna plays an important role in the carbon and nitrogen cycle of terrestrial ecosystems. In order to gain more insight into the role of the intestinal microbiota in transformation and mineralization of organic matter during gut passage, we characterized the physicochemical conditions, microbial activities, and community structure in the gut of our model organism, the humus-feeding larva of the cetoniid beetle Pachnoda ephippiata. Microsensor measurements revealed an extreme alkalinity in the midgut, with highest values (pH > 10) between the second and third crown of midgut ceca. Both midgut and hindgut were largely anoxic, but despite the high pH, the redox potential of the midgut content was surprisingly high even in the largest instar. However, reducing conditions prevailed in the hindgut paunch of all instars (E(h) approximately -100 mV). Both gut compartments possessed a pronounced gut microbiota, with highest numbers in the hindgut, and microbial fermentation products were present in high concentrations. The stimulation of hindgut methanogenesis by exogenous electron donors, such as H(2), formate, and methanol, together with considerable concentrations of formate in midgut and hemolymph, suggests that midgut fermentations are coupled to methanogenesis in the hindgut by an intercompartmental transfer of reducing equivalents via the hemolymph. The results of a cultivation-based enumeration of the major metabolic groups in midgut and hindgut, which yielded high titers of lactogenic, propionigenic, and acetogenic bacteria, are in good agreement not only with the accumulation of microbial fermentation products in the respective compartments but also with the results of a cultivation-independent characterization of the bacterial communities reported in the companion paper (M. Egert, B. Wagner, T. Lemke, A. Brune, and M. W. Friedrich, Appl. Environ. Microbiol. 69:6659-6668, 2003).     

Physicochemical Conditions and Microbial Activities in the Highly Alkaline Gut of the Humus-Feeding Larva of Pachnoda ephippiata (Coleoptera: Scarabaeidae)

The soil macrofauna plays an important role in the carbon and nitrogen cycle of terrestrial ecosystems. In order to gain more insight into the role of the intestinal microbiota in transformation and mineralization of organic matter during gut passage, we characterized the physicochemical conditions, microbial activities, and community structure in the gut of our model organism, the humus-feeding larva of the cetoniid beetle Pachnoda ephippiata. Microsensor measurements revealed an extreme alkalinity in the midgut, with highest values (pH > 10) between the second and third crown of midgut ceca. Both midgut and hindgut were largely anoxic, but despite the high pH, the redox potential of the midgut content was surprisingly high even in the largest instar. However, reducing conditions prevailed in the hindgut paunch of all instars (E_h ~ −100 mV). Both gut compartments possessed a pronounced gut microbiota, with highest numbers in the hindgut, and microbial fermentation products were present in high concentrations. The stimulation of hindgut methanogenesis by exogenous electron donors, such as H₂, formate, and methanol, together with considerable concentrations of formate in midgut and hemolymph, suggests that midgut fermentations are coupled to methanogenesis in the hindgut by an intercompartmental transfer of reducing equivalents via the hemolymph. The results of a cultivation-based enumeration of the major metabolic groups in midgut and hindgut, which yielded high titers of lactogenic, propionigenic, and acetogenic bacteria, are in good agreement not only with the accumulation of microbial fermentation products in the respective compartments but also with the results of a cultivation-independent characterization of the bacterial communities reported in the companion paper (M. Egert, B. Wagner, T. Lemke, A. Brune, and M. W. Friedrich, Appl. Environ. Microbiol. 69:6659-6668, 2003).     

Autochthonous bacterial flora indicated by PCR-DGGE of 16S rRNA gene fragments from the alimentary tract of Costelytra zealandica (Coleoptera: Scarabaeidae)

Aims: To locate and identify putative autochthonous bacteria within the grass grub gut that may have a role in symbiosis. Methods and Results: Polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) fingerprinting was used to investigate bacterial diversity in the grass grub larval gut. The microbial community profiles from five geographically distinct populations were compared and the influence of feeding was analysed. Bacterial community in the midgut was highly variable between locations and was affected by feeding. The hindgut contained a more diverse but stable bacterial community that was less affected by external conditions. Forty-seven distinct DGGE bands, representing different bacterial genotypes, could be distinguished from all samples, with 34 different bands occurring in the hindgut. The 22 most common bands were isolated and DNA was sequenced. Sequence analysis revealed that most bacteria (16/22) were affiliated to the Clostridiales with the predominant bacteria affiliated to the genus Clostridium. The remaining bacteria were aligned to the Betaproteobacteria, Deltaproteobacteria and Bacteroidetes. Conclusions: The grass grub larva has an autochthonous microflora with predominance of Clostridium spp. in the hindgut. Significance and Impact of the Study: Occurrence of an autocthonous microflora in the grass grub hindgut suggests a symbiotic relationship which could help explain the ability of larval scarabs to feed on recalcitrant organic matter.     

Feeding behaviour of the grass grub Costelytra zealandica (White) (Coleoptera: Melolonthinae)

The feeding response of 3rd‐instar Costelytra zealandica larvae to 20 amino acids tested individually at 2 concentrations was assessed. Six of these compounds had a phagostimulatory effect; in particular, the ubiquitous amino acids L‐aspartic acid, L‐glutamic acid, and L‐serine induced considerable feeding by the grubs. Ascorbic acid was also a strong feeding stimulant and, in combination with sucrose, evoked an intense behavioural response. A synergistic effect of ‘Salt Mix W on the larval response to sucrose was demonstrated.     

Synopsis of Antitrogus Burmeister (Coleoptera: Scarabaeidae: Melolonthini)

Abstract The genus Antitrogus Burmeister is widely distributed in eastern Australia and, because some species are significant pests of sugarcane, pastures and pineapples, it has attracted considerable research interest. This paper reviews the systematics, biology, ecology and management of the genus. The morphology of the 22 known species is described and a further species, A. gubbi sp. n. from south-eastern Queensland which was previously confused with A. robertsi Britton, is described. These species clearly divide into two species-groups: (i) those from south-eastern Queensland, eastern New South Wales and south-eastern Australia, which sometimes have closely related species pairs; and (ii) a more diverse group of species from central and northern Queensland. The latter is linked to A. villosus Allsopp from western Victoria, and this may indicate that further species await discovery in western New South Wales. Known life cycles are of 1 or 2 years. Adults fly for short periods following rain in spring or summer and do not feed. Adult females tend to disperse poorly, and this, coupled with ephemeral and localised rainfall, has probably contributed to speciation. Larvae feed on the roots of grasses and other similar plants. Insecticides are widely used to control them in sugarcane, but are too expensive for use on pastures. Integrating new controls with an emphasis on cultural controls is seen as the best way of managing these pest species.     

Description of the larva of Siagona (Coleoptera: Carabidae)

The hitherto unknown larva of Siagona is described from a third instar specimen attributed to S.brunnipes Dejean. The unique features of this larva support the distinctiveness of the subfamily Siagoninae.     

Substituted phenols as repellents for male Costelytra zealandica (Goleoptera: Scarabaeidae) in mating flights

Methyl‐ and halogen‐substituted phenols were found to be strongly repellent to male Costelytra zealandica (White) beetles in mating flights. Phenols substituted in the 2‐position were stronger repellents than those substituted in the 3‐ or 4‐position. Some of these compounds reduced the effectiveness of a potent sex attractant source by > 90 %.     

Sexual and male horn dimorphism in Copris ochus (Coleoptera: Scarabaeidae)

Copris ochus (Coleoptera: Scarabaeidae), an endangered species, is the largest dung beetle in Japan. In C. ochus, males have a long head horn, while females lack this long horn (sexual dimorphism). Very large males of C. ochus have disproportionately longer head horns than small males, suggesting male horn dimorphism, although the dimorphism has not been investigated quantitatively. To clarify sexual and male horn dimorphism in C. ochus quantitatively, we examined the scaling relationship between body size (prothorax width) and head horn length in 94 females and 76 males. These beetles were captured during July 1978 from a natural population on Mt. Aso in southwestern Japan using a light trap. Although the horn length of the females and males scaled with prothorax width, the scaling relationship differed between the sexes, i.e., the relationship was linear in females and nonlinear in males. Statistical tests for dimorphism in male horn length showed a significant discontinuous relationship, thus indicating distinct sexual and male dimorphism in head horns. Long- and short-horned C. ochus males may have different reproductive behaviors, as described in other horned dung beetles.