Establishment of the exotic parasiteTetrastichus haitiensis [Hym.: Eulophidae] and recovery of a newTrichogramma [Hym.: Trichogrammatidae] from root weevil egg masses in Florida

The exotic root weevil egg parasite,Tetrastichus haitiensis Gahan, imported from Puerto Rico and released in Florida between 1969 and 1971, was found established on citrus root weevil egg masses in 1978 near Oakhill, Florida. It was also recovered there from field-collected weevil egg masses from June to August 1978 and from citrus bouquets containing egg masses placed in the grove during July and August 1978. In addition, a new species ofTrichogramma was discovered. In the laboratory, this species parasitized egg masses of 4 species of weevils that attack citrus in Florida. These areDiaprepes abbreviatus (L.),Pachnaeus litus (Germar),P. opalus (Olivier) andArtipus floridanus Horn.     

Food Web Responses to Augmenting the Entomopathogenic Nematodes in Bare and Animal Manure-Mulched Soil

Factorial treatments of entomopathogenic nematodes (EPN) and composted, manure mulches were evaluated for two years in a central Florida citrus orchard to study the post-application biology of EPN used to manage the root weevil, Diaprepes abbreviatus. Mulch treatments were applied once each year to study the effects of altering the community of EPN competitors (free-living bactivorous nematodes) and antagonists (nematophagous fungi (NF), predaceous nematodes and some microarthro-pods). EPN were augmented once with Steinernema riobrave in 2004 and twice in 2005. Adding EPN to soil affected the prevalence of organisms at several trophic levels, but the effects were often ephemeral and sometimes inconsistent. EPN augmentation always increased the mortality of sentinel weevil larvae, the prevalence of free-living nematodes in sentinel cadavers and the prevalence of trapping NF. Subsequent to the insecticidal effects of EPN augmentation in 2004, but not 2005, EPN became temporarily less prevalent, and fewer sentinel weevil larvae died in EPN-augmented compared to non-augmented plots. Manure mulch had variable effects on endoparasitic NF, but consistently decreased the prevalence of trapping NF and increased the prevalence of EPN and the sentinel mortality. Both temporal and spatial abundance of NF were inversely related to the prevalence of Steinernema diaprepesi, whereas Heterorhabditis zealandica prevalence was positively correlated with NF over time. The number of weevil larvae killed by EPN was likely greatest in 2005, due in part to non-target effects of augmentation on the endemic EPN community in 2004 that occurred during a period of peak weevil recruitment into the soil.     

Feeding preferences of the weevils Sitona gressorius and Sitona griseus on different lupin genotypes and the role of alkaloids

The weevils Sitona gressorius and Sitona griseus are specialist herbivores on lupins in Europe. The adult weevils feed on the leaves, and the larvae on the root nodules of the plants. This causes severe damage to lupin crops. In the present study, the feeding preferences of lupin weevil adults on different lupin genotypes were examined with respect to a possible effect of lupin alkaloids on host selection. A total of 12 genotypes from the species Lupinus albus, L. angustifolius, L. luteus, and L. nanus were grown in a field experiment and the feeding damage on the leaves caused by naturally occurring lupin weevil adults was estimated. Additionally, a feeding choice test with S. gressorius adults was performed to examine feeding preferences under laboratory conditions. A gas chromatographic analysis provided information on the alkaloid content and profiles in the leaves of the tested lupin genotypes. In the field experiment, significant differences in the extent of the feeding damage within the 12 lupin genotypes were observed. The dual-choice feeding bioassay did not show discrimination of lupin species, but two L. angustifolius genotypes were significantly less affected than the standard L. luteus “Bornal”. The alkaloid analysis revealed large contrasts in alkaloid concentrations and profiles in the leaves of the tested genotypes. Correlation analysis with the results from the field and laboratory did not indicate a significant influence of the total foliar alkaloid content on the extent of weevil feeding.     

Association of the Red Ring Nematode and Other Nematode Species with the Palm Weevil,Rhynchophorus palmarum

The palm weevil, Rhynchophorus palmarum (L.), was collected in cocoons from red ring-diseased coconut palms (Cocos nucifera L.) in Trinidad and Tobago. Juveniles of five species of nematodes were extracted from the genitalia and macerated bodies of newly emerged adults of the palm weevil: Rhadinaphelenchus cocophilus (Cobb) Goodey (the red ring nematode), Teratorhabditis sp., Diplogasteritus sp., Mononchoides sp., and Bursaphelenchus sp. Over 90% of newly emerged weevil females and males were infested internally with red ring nematode juveniles, and over 47% of the weevils contained more than 1,000 red ring nematodes each. There was no significant correlation between weevil body length and the number of red ring nematodes carried internally by each weevil. Teratorhabditis sp. and Diplogasteritus sp. were extracted from over 50% of the palm weevils, and Monochoides sp. and Bursaphelenchus sp. were found in a small proportion of the weevils. Field-collected adult weevils were also internally and externally infested with a Rhabditis sp., which was not observed in or on weevils allowed to emerge from field-collected cocoons.     

Modifying orchard planting sites conserves entomopathogenic nematodes,reduces weevil herbivory and increases citrus tree growth,survival and fruit yield

In Florida, a root weevil pest of citrus, Diaprepes abbreviatus, is more damaging and attains higher population density in some orchards on fine textured, poorly drained “flatwoods” soils than in those on the deep, coarse sandy soils of the central ridge. Previous research revealed that sentinel weevil larvae were killed by indigenous entomopathogenic nematodes (EPNs) at significantly higher rates in an orchard on the central ridge, compared to one in the flatwoods. We hypothesized that filling tree planting holes in a flatwoods orchard with sandy soil from the central ridge would provide a more suitable habitat for EPNs, thereby reducing weevil numbers and root herbivory. Fifty trees were planted in oversized planting holes filled with coarse sand and 50 trees were planted in native soil in a split plot design where whole plots were species of introduced EPNs and split plots were soil type. Each of Steinernema diaprepesi, Steinernema riobrave, Heterorhabditis indica, Heterorhabditis zealandica, or no EPNs were introduced into the rhizospheres in 10 plots of each soil type. During four years, EPN numbers in soil samples and the relative abundance of seven species of nematophagous fungi associated with nematodes were measured three times using real-time PCR. The efficacy of EPNs against sentinel weevil larvae was also measured three times by burying caged weevils in situ. EPN species richness (P = 0.001) and diversity (P = 0.01) were always higher in sand than native soil. Soil type had no effect on numbers of EPNs in samples, but EPNs were detected more frequently (P = 0.01) in plots of sandy soil than native soil in 2011. Two nematophagous fungi species, Paecilomyces lilacinus and Catenaria sp. were significantly more abundant in nematode samples from sandy soil on all three sampling dates. Efficacy of EPNs against weevil larvae was greater in sandy soil inoculated with S. diaprepesi (P = 0.03) in June 2010 and in all treatments in sandy soil in May 2011 (P = 0.03). Sixty-eight percent more adult weevils (P = 0.01) were trapped emerging from native soil during two years than from sandy soil. By May 2011, the cumulative number of weevils emerging from each plot was inversely related (P = 0.01) to the numbers of EPNs detected in plots and to EPN efficacy against sentinels. Three trees in sandy soil died as a result of root herbivory compared to 21 trees in native soil. Surviving trees in sandy soil had trunk diameters that were 60% larger (P = 0.001) and produced 85% more fruit (P = 0.001) than those in native soil. Although it is not possible to characterize all of the mechanisms by which the two soil treatments affected weevils and trees, substitution of sand for native soil was an effective means of conserving EPNs and shows promise as a cultural practice to manage D. abbreviatus in flatwoods citrus orchards with a history of weevil damage to trees.     

Biological Role of Nardonella Endosymbiont in Its Weevil Host

Weevils constitute the most species-rich animal group with over 60,000 described species, many of which possess specialized symbiotic organs and harbor bacterial endosymbionts. Among the diverse microbial associates of weevils, Nardonella spp. represent the most ancient and widespread endosymbiont lineage, having co-speciated with the host weevils for over 125 million years. Thus far, however, no empirical work on the role of Nardonella for weevil biology has been reported. Here we investigated the biological role of the Nardonella endosymbiont for the West Indian sweet potato weevil, Euscepes postfasciatus. This insect is an experimentally tractable pest insect that can easily be reared on a natural diet of sweet potato root as well as on an agar-based artificial diet. By larval feeding on an antibiotic-containing artificial diet, Nardonella infection was effectively eliminated from the treated insects. The antibiotic-treated insects exhibited significantly lighter body weight and lower growth rate than the control insects. Then, the antibiotic-treated insects and the control insects were respectively allowed to mate and oviposit on fresh sweet potatoes without the antibiotic. The offspring of the antibiotic-treated insects, which were all Nardonella-negative, exhibited significantly lighter body weight, smaller body size, lower growth rate and paler body color in comparison with the offspring of the control insects, which were all Nardonella-positive. In conclusion, the Nardonella endosymbiont is involved in normal growth and development of the host weevil. The biological role of the endosymbiont probably underlies the long-lasting host-symbiont co-speciation in the evolutionary course of weevils.     

Mile-a-minute weed (Persicaria perfoliata) and weevil (Rhinoncomimus latipes) response to varying moisture and temperature conditions

The combined effects of herbivory and water stress on growth and reproduction of mile-a-minute weed (Persicaria perfoliata (L.) H. Gross) were investigated in greenhouse trials over two years, with well-watered or water-limited plants either exposed or not exposed to herbivory by the mile-a-minute weevil (Rhinoncomimus latipes Korotyaev). Moisture limitation and weevil herbivory significantly reduced the number of seeds produced by P. perfoliata, with the fewest seeds produced when both factors were present. Seed weight was reduced by moisture limitation and weevil herbivory the second year, and seed viability was reduced by herbivory both years. Plant biomass was lower both years under conditions of water limitation, with an additional effect of herbivory the second year. Well-watered plants the second year also produced substantially more weevils than water-limited plants by the end of the season. Results are consistent with field observations suggesting that years of high rainfall allow resurgence of P. perfoliata plant populations that were previously suppressed by R. latipes. An additional environmental chamber trial assessed the interaction between the weed and weevil at two different temperatures. Here, plant mortality occurred only at the higher temperature with weevil herbivory, suggesting that herbivory has a greater negative effect on P. perfoliata under warm conditions. Additional studies on temperature effects are needed for a more complete understanding of interactions between P. perfoliata and R. latipes under different abiotic conditions.     

Hypera postica [Col.: Curculionidae] and its natural enemies in Maryland and West Germany — 1971

A comparative study of field populations ofHypera postica (Gyllenhal), the alfalfa weevil, and its natural enemies was conducted in Maryland and West Germany in 1971. Populations of alfalfa weevil larvae peaked at 231/.09 m² in Maryland versus 45/.09 m² in West Germany. The ichneumonid parasiteBathyplectes curculionis (Thomson), and the eulophid,Tetrastichus incertus (Ratzeburg), were recovered in both Maryland and West Germany.Bathyplectes anurus (Thomson) was recovered only in West Germany, and the braconid,Microctonus colesi Drea, was recovered only in Maryland. A review of the agricultural practices and climatological conditions indicated that these factors may contribute to the regulation of the alfalfa weevil in West Germany and the United States. This paper is the 2nd in a series of 3 studies on the biology, ecology and biological control of the alfalfa weevil from 1966–1979.     

Resource limitation in natural populations of phytophagous insects. A long-term study case with the chestnut weevil

The chestnut weevil, Curculio elephas (Gyll.), is a non-outbreaking species whose populations and food resources, the European chestnut, Castanea sativa, can be precisely defined. Thirteen and 17 generations of this insect were studied in two isolated sites. Field observations and experiments allowed us to estimate the absolute abundance, availability and use of chestnuts for weevil oviposition, and the number of weevil females emerging per site. Unavailable chestnuts were defined as the fruits either infested first by the chestnut moth (Cydia splendana) larvae (because of competition between the two species) or those avoided by chestnut weevil females when selecting their egg-laying sites, independently of chestnut moth presence. From a third to a half of the chestnuts were not available on the average for weevil infestation. Only one-fourth, on the average, of those available for oviposition were actually used by chestnut weevil females. Regardless of year and site, the number of available chestnuts per weevil female was higher than that of weevil-infested fruits per female, considering global food resources independently of their temporal variation in quality. However, realized fecundity of weevil females was positively correlated with the mean number of available chestnuts per female. We concluded that food resources can be limiting without being fully exploited by females because of temporal variation in chestnut quality.     

Early Root Herbivory Impairs Arbuscular Mycorrhizal Fungal Colonization and Shifts Defence Allocation in Establishing Plantago lanceolata

Research into plant-mediated indirect interactions between arbuscular mycorrhizal (AM) fungi and insect herbivores has focussed on those between plant shoots and above-ground herbivores, despite the fact that only below-ground herbivores share the same part of the host plant as AM fungi. Using Plantago lanceolata L., we aimed to characterise how early root herbivory by the vine weevil (Otiorhynchus sulcatus F.) affected subsequent colonization by AM fungi (Glomus spp.) and determine how the two affected plant growth and defensive chemistry. We exposed four week old P. lanceolata to root herbivory and AM fungi using a 2×2 factorial design (and quantified subsequent effects on plant biomass and iridoid glycosides (IGs) concentrations. Otiorhynchus sulcatus reduced root growth by c. 64%, whereas plant growth was unaffected by AM fungi. Root herbivory reduced extent of AM fungal colonization (by c. 61%). O. sulcatus did not influence overall IG concentrations, but caused qualitative shifts in root and shoot IGs, specifically increasing the proportion of the more toxic catalpol. These changes may reflect defensive allocation in the plant against further attack. This study demonstrates that very early root herbivory during plant development can shape future patterns of AM fungal colonization and influence defensive allocation in the plant.     

Dual inoculation of Fusarium oxysporum endophytes in banana: effect on plant colonization,growth and control of the root burrowing nematode and the banana weevil

The burrowing nematode (Radopholus similis (Cobb) Thorne) and the banana weevil (Cosmopolites sordidus Germar, Coleoptera: Curculionidae) are major pests of banana (Musa spp.) in the Lake Victoria basin region of Uganda. Among biological options to control the two pests is the use of non-pathogenic Fusarium oxysporum Schltdl.: Fries endophytes of banana. We investigated the ability of endophytic F. oxysporum isolates Emb2.4o and V5w2 to control the banana weevil and the burrowing nematode, alone and in combination. Plant colonization by the endophytes was determined by inoculating their chemical-resistant mutants separately and in combination, onto banana roots. Plant growth promotion was determined by measuring plant height, girth, number of live roots and fresh root weight at harvest, and control of the nematode and weevil was determined by challenging endophyte-inoculated plants with the pests 8 weeks after endophyte inoculation. Endophytic root colonization was highest in plants inoculated with both endophytes, compared with those inoculated with only one of the endophytes. Root colonization was better for isolate V5w2 than Emb2.4o. Dually inoculated plants showed a significant increase in height, girth, fresh root weight and number of functional roots following nematode challenge. Nematode numbers in roots were reduced 12 weeks after challenge of 8-week-old endophyte-inoculated plants. Significant reductions in weevil damage were observed in the rhizome periphery, inner and outer rhizomes, compared with endophyte non-inoculated controls. We conclude that dual inoculation of bananas with endophytic isolates Emb2.4o and V5w2 increases root colonization by the endophytes, reduces nematode numbers and weevil damage, and enhances plant growth in the presence of nematode infestation.     

A fungal metabolite masks the host plant odor for the pine weevil (Hylobius abietis)

The pine weevil is one of the most important pest insects of conifer reforestation areas in Europe. Female pine weevils cover their eggs with chewed bark and feces (frass) resulting in avoidance behavior of feeding conspecifics towards egg laying sites. It has been suggested that microorganisms present in the frass may be responsible for producing deterrent compounds for the pine weevil. The fungi Ophiostoma canum, O. pluriannulatum, and yeast Debaryomyces hansenii were isolated from aseptically collected pine-weevil frass. The isolated fungi were cultured on weevil frass broth and their volatiles were collected by SPME and identified by GC–MS. D. hansenii produced methyl salicylate (MeS) as a major compound, whereas, in addition, O. canum and O. pluriannulatum produced 6-protoilludene. In a multi-choice lab bioassay, MeS strongly reduced pine weevil's attraction to the Pinus sylvestris volatiles. Thus, a fungal metabolite was found that strongly affects the pine weevil host-odor search.     

Feeding on roots in the humus layer by adult pine weevil, Hylobius abietis

1 The consumption by adult pine weevil, Hylobius abietis, of the bark of roots present in the humus layer was assessed in a field study conducted in southern Sweden during two years (1998 and 2002). The study sites were divided into two areas: (i) a shelterwood where 80–100 mature Scots pine trees per hectare remained after cutting and (ii) a clearcut where no trees were left. 2 On average, 3741 m² per hectare of root bark was present in the humus layer, of which 135 m² was not coniferous but comprised species such as bilberry and broadleaved trees. 3 The mean area debarked by pine weevils was 2.9 m² per hectare; 2.6 m² of conifer roots and 0.3 m² of bilberry roots. Roots of broadleaved trees were almost never consumed. No clear preferences for roots of a specific level of vitality were observed. 4 No consistent difference between the shelterwood and clearcut was found, either in the amount of root bark area available or in the extent of root feeding by pine weevil. 5 A weak negative correlation between debarked areas on roots and seedlings was found, indicating that root feeding may have reduced damage to seedlings. 6 It is concluded that conifer roots in the humus layer constitute a major food source for the pine weevil and can be utilized for a considerable period in both clearcuts and shelterwoods.     

Localization of acid phosphatase activity in maize root under phosphorus deficiency

The effect of phosphorus deficiency on acid phosphatase activity in the apical, middle and basal parts of the root of maize plants was followed. The supernatant obtained by centrifuging the homogenate of plant tissue at 1500 ×g was further centrifuged at 18 000 ×g, the sediment being marked as fraction II and the supernatant as fraction III. The results obtained document the fact that acid phosphatase activity of the two fractions of all analyzed root segments was higher in plants cultured in nutrient medium without phosphate than in those containing phosphorus in nutrient medium. In most cases this difference was significant to highly significant. The results of experiments proved unambiguously a higher enzymatic activity in all root segments in fraction III than in fraction II. In fraction III the highest acid phosphatase activity was found in the apical part, in fraction II in the basal part of the root.     

Killing power of the red imported fire ant [Hym.: Formicidae]: a key predator of the boll weevil [Col.: Curculionidae]

The red imported fire ant,Solenopsis invicta Buren, was identified as a key predator of the boll weevil,Anthonomus grandis Boheman. Although key factor analysis does not guarantee that the key factor is the causative agent, the evidence of boll weevil mortality was so clear that the fire ant could be identified as causing the mortality. Ant predation on immature boll weevils during the summer of 1981 averaged 84%. When this predation was combined with other natural mortality factors, the weevil population density throughout the season never exceeded levels that required control measures.     

Pleiotropic Impact of Endosymbiont Load and Co-Occurrence in the Maize Weevil Sitophilus zeamais

Individual traits vary among and within populations, and the co-occurrence of different endosymbiont species within a host may take place under varying endosymbiont loads in each individual host. This makes the recognition of the potential impact of such endosymbiont associations in insect species difficult, particularly in insect pest species. The maize weevil, Sitophilus zeamais Motsch. (Coleoptera: Curculionidae), a key pest species of stored cereal grains, exhibits associations with two endosymbiotic bacteria: the obligatory endosymbiont SZPE (“Sitophilus zeamais Primary Endosymbiont”) and the facultative endosymbiont Wolbachia. The impact of the lack of SZPE in maize weevil physiology is the impairment of nutrient acquisition and energy metabolism, while Wolbachia is an important factor in reproductive incompatibility. However, the role of endosymbiont load and co-occurrence in insect behavior, grain consumption, body mass and subsequent reproductive factors has not yet been explored. Here we report on the impacts of co-occurrence and varying endosymbiont loads achieved via thermal treatment and antibiotic provision via ingested water in the maize weevil. SZPE exhibited strong effects on respiration rate, grain consumption and weevil body mass, with observed effects on weevil behavior, particularly flight activity, and potential consequences for the management of this pest species. Wolbachia directly favored weevil fertility and exhibited only mild indirect effects, usually enhancing the SZPE effect. SZPE suppression delayed weevil emergence, which reduced the insect population growth rate, and the thermal inactivation of both symbionts prevented insect reproduction. Such findings are likely important for strain divergences reported in the maize weevil and their control, aspects still deserving future attention.     

Regional-scale impact of the weed biocontrol agent Mecinus janthinus on Dalmatian toadflax (Linaria dalmatica)

Dalmatian toadflax (Linaria dalmatica (L.) Mill.) is an important invasive plant on rangelands throughout western North America. In 1991, the stem-mining weevil, Mecinus janthinus Germar, was introduced into Canada from Europe as a classical biological control agent to reduce toadflax densities and improve rangelands, particularly in British Columbia. To determine if the program was a success at a regional level, this paper answers three key questions: (1) has M. janthinus spread throughout the study area, (2) is M. janthinus causing a decline in toadflax plant size or density at the regional scale, and (3) has the distribution of toadflax plants changed following M. janthinus introduction? These questions are answered by combining historical survey data and mensurative experimental data on plant and weevil densities across a 40,000 km² area in southern British Columbia. The results show that through a combination of intentional redistribution and natural dispersal weevils have spread throughout the study area. Stem densities at naturally colonized sites and historical release sites were equally low. Across weevil populations between 3 and 10 years old, weevil densities peaked in the eighth year, and there was a negative relationship between weevil density and stem length. Between 2000 and 2007, toadflax patches were found to both decrease in density and become more fragmented over time, with 15% of patches disappearing completely. These findings show that M. janthinus has had a significant negative impact on both the density and distribution of Dalmatian toadflax throughout a large part of its range in British Columbia.     

Resistance of Maize Landraces to the Maize Weevil Sitophilus zeamais Motsch. (Coleoptera: Curculionidae)

The maize weevil Sitophilus zeamais Motsch. is an important pest of maize that attacks the grain both in the field and during storage. The damage caused by the maize weevil S. zeamais on maize landraces, Amarelo Antigo, Asteca, Caiano, Carioca, and Ferrinho, was evaluated by no-choice tests under laboratory conditions. The commercial varieties Sol da Manh?, BR 106, BR 451, and the synthetics PC 0203 and PC 9903 were evaluated for comparisons with the maize landraces. The parameters evaluated were susceptibility index, number of weevil progeny, development time, weevil progeny dry weight, and grain dry weight loss. The landraces were more susceptible to the maize weevil as compared to the commercial varieties. Based on the cluster analysis, two groups of susceptibility to the maize weevil were observed: one of more susceptible populations formed by local landraces and BR 451, and another less susceptible, with commercial varieties, synthetics, and the landrace Amarelo.