Characterization of an entomopoxvirus of the lesser cornstalk borer (Elasmopalpus lignosellus)

A biochemical and limited morphological characterization of an entomopoxvirus infecting the lesser cornstalk borer, Elasmopalpus lignosellus, was made. The oval virions measure 270 × 200 nm and the spheroids average 1.5 μm in diameter. Sodium dodecyl sulfate polyacrylamide gel electrophoresis elucidated 32 structural polypeptides with molecular weights ranging from 13,000 to 145,000. The viral genome was examined with the restriction endonuclease EcoRI. Gel electrophoresis of the digested DNA yielded 26 bands and a total molecular weight of 140.8 × 10⁶.     

Effect of crowding, food deprivation, and diet on flight initiation and lipid reserves of the lesser grain borer, Rhyzopertha dominica

Effects of crowding, food deprivation, and type of cereal diet upon flight initiation, development, body weight, lipid content, and fatty acid composition of the lesser grain borer, Rhyzopertha dominica (F.), were studied in two field strains and one laboratory strain. Beetles of all strains reared under crowded conditions had significantly higher flight initiation than beetles reared on isolated kernels (uncrowded). Regardless of degree of crowding, flight initiation increased with the period of food deprivation up to a maximum at 24 h, after which flight initiation declined. Body weight and lipid content decreased as the food deprivation period increased, whereas fatty acid composition was not significantly affected by food deprivation. Beetles from a field strain collected in 1995 had higher flight initiation and increased lipid content compared with beetles from the laboratory strain. However, beetles from the laboratory strain were larger, developed faster, and were more fecund than beetles from this field strain. The cereal diet on which beetles were reared also had a significant effect on flight initiation, lipid content, and fatty acid composition. Beetles reared on whole rice and wheat produced adults with higher flight initiation, higher lipid content, and higher oleic acid concentration than beetles reared on whole corn and sorghum.     

Effects of harvest residue and tillage on lesser cornstalk borer (Lepidoptera: Pyralidae) damage to sugarcane

Lesser cornstalk borer, Elasmopalpus lignosellus (Zeller) (Lepidoptera: Pyralidae) is an important pest of sugarcane (a complex hybrid of Saccharum spp.) in southern Florida. Cultural controls for E. lignosellus were evaluated in preparation for the potential loss of effective insecticides. Field studies conducted in 2006 compared the effects of harvest residues from green-harvested sugarcane (no preharvest burning to remove leaf matter) on E. lignosellus stalk damage and yield. Damage by E. lignosellus was significantly lower in plant cane plots that were covered with harvest residues collected from a green-harvested field before shoot emergence compared with plots without harvest residue. There were no yield differences between plots with and without harvest residues in plant or ratoon sugarcane fields in the 2006 study. The effects of three postharvest tillage levels (conventional, intermediate, and no tillage) were evaluated in preharvest burned and green-harvested fields in 2008 and 2009. Significantly less E. lignosellus damage was observed in the green- versus preharvest burned fields in both years. Intermediate and no-tillage plots had very little damage in green-harvested field. Conventional tillage plots had the greatest damage in the green-harvested field and the lowest damage in the preharvest burned field. In 2008, biomass yield was greater in the intermediate than conventional tillage in the green-harvested field, but it was greater in the conventional than in other tillage levels in the preharvest burned field. These studies demonstrated that cultural controls could greatly reduce E. lignosellus damage in sugarcane without the use of insecticides.     

Transient gut retention and persistence of Salmonella through metamorphosis in the lesser mealworm, Alphitobius diaperinus (Coleoptera: Tenebrionidae)

Aims: This study was undertaken to determine the retention of Salmonella through Alphitobius diaperinus metamorphosis and its contribution, through defecation, to external contamination. Methods and Results: Insects were exposed to a tagged Salmonella enterica and evaluated for external elimination. (i) Each day for 3 weeks, a filter collected frass from a restrained insect for analysis. (ii) Exposed larvae in a closed container were followed through pupation, and newly emerged adults were examined for their retention of marker bacteria. Conclusions: Exposed adults and larvae produced Salmonella‐positive frass for an average of 8 days, ranging from 6 to 11 days and 6 to 12 days, respectively. Nineteen per cent of the larvae carried Salmonella through metamorphosis and eclosion, with 5% of the pupal exuviae being positive as well. Significance and Impact of the Study: Many sources of foodborne pathogens within the poultry production facilities, including reservoir populations, currently go unrecognized. This diminishes the ability of producers to mitigate the transfer of pathogens between animals, humans and the environment. Poultry management standards accept the reutilization of litter. Alphitobius diaperinus survive between flock rotations on the reutilized litter, and it was demonstrated in this study that the Salmonella they carry can survive with them.     

A newly recorded entomopoxvirus B in Anacridium aegyptium (Orthoptera: Acrididae)

An entomopoxvirus infecting fat body cells of Anacridium aegyptium is reported. Spheroid virus occlusion bodies, ranging in size from 3 to 8 μm, contain virions 336–356 nm long and 210–217 nm wide. The virion ultrastructure indicates that the newly discovered virus belongs to the genus Entomopoxvirus B known to infect Lepidoptera and Orthoptera.     

Expression of a Bacillus thuringiensis cryIA(c) gene in transgenic peanut plants and its efficacy against lesser cornstalk borer

The invasion of peanut (Arachis hypogaea L.) pods and seeds by aflatoxin-forming species of Aspergillus is linked to injury by the lesser cornstalk borer and frequently causes a severe reduction in crop quality. The lesser cornstalk borer is susceptible to the lepidopteran-active Bacillus thuringiensis insecticidal crystal protein. We have introduced a codon-modified Bacillus thuringiensis cryIA(c) gene into peanut using microprojectile bombardment. The toxin-coding region of a Bt cryIA(c) gene was reconstructed for expression in plants and the resulting 3.4 kb gene cassette (promoter: 1.8 kb coding: 3) was directly cloned into the BglII site of plant transformation vectors. The vectors contained the hph gene, conferring resistance to the antibiotic hygromycin. Somatic embryos initiated from immature peanut cotyledons of two cultivars were used as the target for bombardment. DNA from hygromycin-resistant embryogenic cell lines, regenerated plants, and a progeny plant showed the presence and integration of hph and Bt genes by PCR and/or Southern blot analyses. ELISA immunoassay of the CryIA(c) protein from the hygromycin-selected plants showed the expression of CryIA(c) protein up to 0.18% of total soluble protein. Insect feeding bioassay of transformed plants indicated various levels of resistance to the lesser cornstalk borer, from complete larval mortality to a 66% reduction in larval weight. A negative correlation between percent survival or larval weight and the amount of Bt CryIA(c) protein was recorded indicating in general that the higher the protein level the lower the survival or larval weight of the insect. Based on leaf bioassay, transformation of peanut with vectors containing the Bt cryIA(c) gene may be effective in protecting the peanut plants from damage by lepidopteran insect larvae of lesser cornstalk borer     

Assessing toxicity of Lake Diefenbaker (Saskatchewan,Canada) sediments using algal and nematode bioassays

Lake Diefenbaker, on the South Saskatchewan River, Saskatchewan, Canada, receives, on average, 90% of its inflow from snowmelt and rainfall in the Rocky Mountains. The inflowing rivers also receive irrigation return flows and municipal and industrial effluents which may result in the contamination of lake sediments. The sediments were assessed by nematode and algal bioassays.The toxicity of five chemical fractions of the sediment was determined using the nematode Panagrellus redivivus as the test organisms. The results suggest that the sediment chemical fractions frequently inhibit growth and maturation, while lethality was observed at 4 of 12 sites.Samples from 3 of these sites were further evaluated using conventional elutriate Algal Fractionation Bioassays (AFB) with both natural Lake Diefenbaker phytoplankton and a mixed laboratory grown algal culture. The natural phytoplankton showed inhibition at sediment: water ratios of 10: 1; whereas the algal cultures showed both enhancement and inhibition. Evidently, the sediments are frequently toxic to the species tested except for the algal culture. The AFB assesses the mitigative and synergistic effects of contaminants and nutrients and being a conventional elutriate, is more realistic and potentially more acceptable than the chemical fractionation/nematode bioassay technique which essentially considers potential trace organic contaminant effects.     

Characterization of midgut trypsin-like enzymes and three trypsinogen cDNAs from the lesser grain borer, Rhyzopertha dominica (Coleoptera: Bostrichidae)

Protein digestion in the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), results from the action of a complex of serine proteinases present in the midgut. In this study we partially characterized trypsin-like enzyme activity against N-alpha-benzoyl-L-arginine p-nitroanilide (BApNA) in midgut preparations and cloned and sequenced three cDNAs for trypsinogen-like proteins. BApNAase activity in R. dominica midgut was significantly reduced by serine proteinase inhibitors and specific inhibitors of trypsin, whereas BApNAase activity was not sensitive to specific inhibitors of chymotrypsin or aspartic proteinases. However, trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) inhibited BApNAase activity by about 30%. BApNAase was most active in a broad pH range from about pH 7 to 9.5. The gut of R. dominica is a tubular tract approximately 2.5 mm in length. BApNAase activity was primarily located in the midgut region with about 1.5-fold more BApNAase activity in the anterior region compared to that in the posterior region. Proteinases with apparent molecular masses of 23-24 kDa that were visualized on casein zymograms following electrophoresis were inhibited by TLCK. Three cDNAs for trypsinogen-like proteins were cloned and sequenced from mRNA of R. dominica midgut. The full cDNA sequences consisted of open reading frames encoding 249, 293, and 255 amino acid residues for RdoT1, RdoT2, and RdoT3, respectively. cDNAs RdoT1, RdoT2, and RdoT3 shared 77-81% sequence identity. The three encoded trypsinogens shared 54-62% identity in their amino acid sequences and had 16-18 residues of signal peptides and 12-15 residues of activation peptides. The three predicted mature trypsin-like enzymes had molecular masses of 23.1, 28, and 23.8 kDa for RdoT1, RdoT2, and RdoT3, respectively. Typical features of these trypsin-like enzymes included the conserved N-terminal residues IVGG62-65, the catalytic amino acid triad of serine proteinase active sites (His109, Asp156, Ser257), three pairs of conserved cysteine residues for disulfide bridges, and the three residues (Asp251, Gly274, Gly284) that determine specificity in trypsin-like enzymes. In addition, RdoT2 has both a PEST-like sequence at the C-terminus and a free Cys158 near the active site, suggesting instability of this enzyme and/or sensitivity to thiol reagents. The sequences have been deposited in GenBank database (accession numbers AF130840 for RdoT1, AF130841 for RdoT2, and AF130842 for RdoT3).     

Production and partial characterization of monoclonal antibodies for detection of entomopoxvirus from Melanoplus sanguinipes

Entomopoxviruses (EPV) are currently being considered as candidate grasshopper (Orthoptera: Acrididae) microbial control agents. Classical techniques for diagnosing infections in grasshoppers are laborious, time consuming, and sometimes inaccurate. Specific murine monoclonal antibodies were developed against an EPV from Melanoplus sanguinipes (Fab) for use in a nitrocellulose-based enzyme-linked immunoassay (dot-blot). An IgG_2b monoclonal antibody was used to diagnose infections in grasshoppers 13 days following injection with virions. Of 25 grasshoppers that had patent infections microscopically, 22 produced positive results on the dot-blot. In a second test, 39 patently infected grasshoppers all produced positive results. Seven additional grasshoppers in the first test and 2 in the second test gave positive reactions in the dot-blot method but virus was not detected upon microscopic examination. The monoclonal antibody did not cross-react with other commonly occurring grasshopper pathogens. The dot-blot method detected as few as 2.5×10⁶ purified EPV virions. The improvement over existing detection techniques should facilitate evaluation of EPV for field use.

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Prodution et caractérisation partielle d'anticorps monoclonaux pour la détection d'entomopoxyvirus de Melanoplus sanguinipes

Résumé Les criquets sont très nuisibles aux pâturages de l'Ouest des USA et du Canada. Les méthodes classiques de protection sont basées sur les traitements chimiques lors des pullulations. Un entomopoxvirus (EPV) extrait de M. sanguinipes est généralement considéré comme un outil, pour le contrôle des populations sur une longue période, en vue de la suppression des criquets. Les méthodes actuelles d'isolement de EPV sont longues, pénibles et peu fiables. Les tests d'adsorption des antigènes sur l'anticorps fixé et le dosage per l'anticorps enzymatiquement marqué sont efficaces, sûrs et donnent à temps des résultats pour déceler des entomopathogènes. Nous avons produit des anticorps monoclonaux de souris contre EPV de M. sanguinipes, et les avons utilisés dans des dosages immunoenzymatiques d'extraits protéiques adsorbés sur nitrocellulose. Les EPV sont recherchés sur des criquets injectés de virions 13 jours avant. Sur 25 criquets qui présentaient des infections nettes lors d'examens microscopiques, 22 ont donné des résultats positifs par sérologie. Dans un second test, sur 38 criquets nettement contaminés, tous ont donné des résultats positifs; 7 criquets suppl'ementaires mentaires dans le premier test et 2 dans le second test ont donné des résultats positifs en sérologie alors que l'examen microsopique n]avait pas test ont donné des résultats positifs en sérologie alors que l'examen microsopique n'avait pas révélé de virus. Ceci montre que ce type de détection est plus sûr que la méthode ordinaire. Les anticorps monoclonaux ne donnent pas de réactions avec les autres pathogènes courants des criquets. La méthode sérologique a permis de détecter même une concentration de 2,5×10⁶ virions EPV.

     

Age-specific dose-mortality effects of Beauveria bassiana (Deuteromycotina: Hyphomycetes) on the European corn borer,Ostrinia nubilalis (Lepidoptera: Pyralidae)

Three isolates of Beauveria bassiana were evaluated under laboratory conditions for pathogenicity against the European corn borer, Ostrinia nubilalis. Dose-mortality regressions and nonparametric statistical analyses revealed significant differences in pathogenicity between isolates and across O. nubilalis instars. First instars were found to be the most susceptible of the five larval stages. Little difference was noted for instars 2, 3, and 5 within any of the three B. bassiana isolates. Fourth instar larvae were found to be the most tolerant in all cases.     

Morphological study of the reproductive system and ovarian growth of the univoltine bamboo borer,Omphisa fuscidentalis (Hampson, 1896) (Lepidoptera: Crambidae)

ABSTRACT

The female reproductive system and protein deposition in the ovaries during development have never been examined in the bamboo borer Omphisa fuscidentalis Hampson. The aim of this study was thus to study the morphology of the female reproductive system of each stage of development. The female reproductive system of the borer consists of a pair of ovary, oviduct and accessory glands. Each ovary is composed of four polytrophic ovarioles that connect to lateral oviducts, fused with a common oviduct. The size of the ovary in diapausing larvae for 9 months was determined. The length and width of the ovaries were the smallest in September larvae (0.343 ± 0.03 and 0.071 ± 0.01 mm, respectively). The ovaries were the largest during ovarian development in May (0.752 ± 0.08 mm long and 0.084 ± 0.01 wide). Additionally, ovarian size was significantly larger in adults than in pupae. The ovarian protein concentration of larvae in May was 0.59 ± 0.06 mg/ml and increased to 16.61 ± 7.5 and 37.42 ± 5.5 mg/ml in pupae (June) and adults (July), respectively. The results showed ovarian development in all life stages of this holometabolous insect, which has a longer life cycle than other lepidopterans.

Abbreviation: TEM: transmission electron microscope     

Characterization of midgut trypsin-like enzymes and three trypsinogen cDNAs from the lesser grain borer, Rhyzopertha dominica (Coleoptera: Bostrichidae)

Protein digestion in the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), results from the action of a complex of serine proteinases present in the midgut. In this study we partially characterized trypsin-like enzyme activity against N-α-benzoyl--arginine p-nitroanilide (BApNA) in midgut preparations and cloned and sequenced three cDNAs for trypsinogen-like proteins. BApNAase activity in R. dominica midgut was significantly reduced by serine proteinase inhibitors and specific inhibitors of trypsin, whereas BApNAase activity was not sensitive to specific inhibitors of chymotrypsin or aspartic proteinases. However, trans-epoxysuccinyl--leucylamido-(4-guanidino) butane (E-64) inhibited BApNAase activity by about 30%. BApNAase was most active in a broad pH range from about pH 7 to 9.5. The gut of R. dominica is a tubular tract approximately 2.5 mm in length. BApNAase activity was primarily located in the midgut region with about 1.5-fold more BApNAase activity in the anterior region compared to that in the posterior region. Proteinases with apparent molecular masses of 23–24 kDa that were visualized on casein zymograms following electrophoresis were inhibited by TLCK.Three cDNAs for trypsinogen-like proteins were cloned and sequenced from mRNA of R. dominica midgut. The full cDNA sequences consisted of open reading frames encoding 249, 293, and 255 amino acid residues for RdoT1, RdoT2, and RdoT3, respectively. cDNAs RdoT1, RdoT2, and RdoT3 shared 77–81% sequence identity. The three encoded trypsinogens shared 54–62% identity in their amino acid sequences and had 16–18 residues of signal peptides and 12–15 residues of activation peptides. The three predicted mature trypsin-like enzymes had molecular masses of 23.1, 28, and 23.8 kDa for RdoT1, RdoT2, and RdoT3, respectively. Typical features of these trypsin-like enzymes included the conserved N-terminal residues IVGG^62–65, the catalytic amino acid triad of serine proteinase active sites (His¹⁰⁹, Asp¹⁵⁶, Ser²⁵⁷), three pairs of conserved cysteine residues for disulfide bridges, and the three residues (Asp²⁵¹, Gly²⁷⁴, Gly²⁸⁴) that determine specificity in trypsin-like enzymes. In addition, RdoT2 has both a PEST-like sequence at the C-terminus and a free Cys¹⁵⁸ near the active site, suggesting instability of this enzyme and/or sensitivity to thiol reagents. The sequences have been deposited in GenBank database (accession numbers AF130840 for RdoT1, AF130841 for RdoT2, and AF130842 for RdoT3).     

A study of host specificity in the entomopathogenic fungus Beauveria bassiana (Hypocreales,Clavicipitaceae)

Beauveria bassiana (Balsamo – Crivelli) Vuillemin based mycoinsecticides are used against agricultural, veterinary and medical insect pests. The fungus has a very diverse and extensive host range. Variation in virulence among isolates of B. bassiana to different insect species has been abundantly documented. Given the effect of multiple factors on virulence, it is not certain whether the observed difference in virulence can be labelled as host specificity. Environmental conditions and susceptibility of the insect population are two main factors that affect successful fungal infection. Keeping the environmental factors constant, if virulence of an isolate to different insect species and different populations within an insect species is compared, the scale of difference between the two responses can be estimated. If differences in virulence of an isolate to different insect species are greater than the difference in virulence to different insect populations within an insect species, then, the isolate can be considered as exhibiting specific preference to those insect species towards which it exhibits high virulence. To examine this feature, a worldwide sample of B. bassiana was bioassayed on nine insect species and two different populations within two insect species. Laboratory bioassays were done on: Bombyx mori (Lepidoptera), Spodoptera litura (Lepidoptera), Chilo partellus (Lepidoptera), Helicoverpa armigera (Lepidoptera), Epilachna vigintioctopunctata (Coleoptera), Mylabris pustulata (Coleoptera), Aphis craccivora (Homoptera), Maconellicoccus hirsutus (Hemiptera) and Oecophylla smaragdina (Hymenoptera). The range of variation in virulence of a B. bassiana isolate to different insect species was not more than that observed with different populations within a single insect species. B. bassiana is thus a generalist with no strict host preference. B. bassiana based biopesticide can be used as a broad spectrum insecticide against a myriad of insect pests.     

Biology and damage traits of emerald ash borer planipennis Fairmaire）in China

Emerald ash borer （Agrilus planipennis Fairmaire） （Coleoptera： Buprestidae） is a major stem borer of ash （Fraxinus spp.）. It is univoltine in Tianjin, while it is semivoltine in Heilongjiang Province, and both univoltine and semivoltine in Changchun, Jilin Province, where the majority is univoltine. The longevity of emerald ash borer adults is 17.2 ± 4.6 days （n = 45）, eggs 9.0 5：1.1 days （n = 103）, univoltine larvae 308 days, semivoltine larvae 673 days, and pupae 61.2 ± 1.6 days （n = 45）. It takes about 100 days from the time larvae bore into the phloem to when they complete the pupal cell. In a 10-year-old velvet ash （Fraxinus velutina Tort.） plantation in Tianjin, emerald ash borer preferred to oviposit on the regions of boles from 50-150 cm above ground, accounting for 76.7% of the total girdling. Girdling on the south side of the tree boles accounted for 43.40% of the total girdling. The emerald ash borer population density is higher at the edge of the plantation compared with the center.     

Male copulatory patterns in the lesser bushbaby (Galago moholi) in captivity

I observed seven pairs of sexually mature, captiveGalago moholi during the females’ ovarian cycles to determine the pattern of male copulatory behavior and its relationship to female receptivity. In this species, a copulatory lock was not apparent, a variety of thrusting patterns was displayed, and the male was capable of ejaculating following a single mount and intromission and may have ejaculated more than once during the 30-min test. Male mating indices did not differ significantly on consecutive days of the mating period, though declines in certain mating latencies on the second and third days of mating suggested increased female receptivity. Mating indices did not differ notably between the first and the second copulation within a test, though both showed some differences compared to the third copulation. InG. moholi, male copulatory behavior is typical of a prosimian species with a dispersed — nongregarious — mating system, in which copulation has a single prolonged intromission that extends into the postejaculatory period. This is characteristic of species in which more than one male has access to an estrous female.     

Diapause termination in the spotted stem borer, Chilo partellus (Lepidoptera: Pyralidae) in the laboratory

The effect of temperature, photoperiod, artificial diet and water on the termination of diapause by larvae of the stem borer, Chilo partellus (Lepidoptera: Pyralidae), was studied in the laboratory. Termination of diapause as indicated by pupation was affected mainly by a combination of high temperature and a long day photoperiod. Total darkness did not prevent termination of diapause and pupation occurred also in larvae which were never exposed to water. Long days accelerated pupation, but, under 16 h daylength, termination of diapause was faster than under constant illumination. Provision of artificial diet had no effect or slowed down pupation but water decreased the time to pupation. Under 28°C, 16 h daylength and availability of water, C. partellus diapausing larvae terminated diapause and pupated in about 9 days.     

Laboratory evaluation of the virulence of Beauveria bassiana isolates to the sorghum shoot borer Chilo partellus Swinhoe (Lepidoptera: Pyralidae) and their characterization by RAPD-PCR

Beauveria bassiana has long been used as a mycopesticide. It has a wide host range; isolates have been reported to differ in host range and virulence to a given insect species. Identification of a molecular marker linked to a virulent phenotype to a target pest would be useful in screening for isolates effective against it. Twenty B. bassiana isolates were tested for their virulence to the second instar larvae of Chilo partellus Swinhoe in laboratory bioassays and their DNA fingerprints were generated by RAPD-PCR. Three arbitrary categories of aggressiveness were chosen; isolates that caused >70%, between 70 and 40% and <40% larval mortality were grouped as highly, medium and less aggressive types, respectively. In the random amplified polymorphic DNA (RAPD) analysis a 30% variability was observed among the isolates; which clustered into three major groups. The groups based on virulence rating did not match with the RAPD clusters. One of the highly aggressive isolates clustered with less aggressive isolates in one cluster and the other grouped along with the medium aggressive isolates in a different cluster. The B. bassiana isolates were classified phenotypically based on the taxonomic order of the original insect host and the climatic zone (tropical/temperate) from which they were isolated. No correlation between the aggressiveness of the isolate and the relatedness of the original insect host to the tested insect was observed; both the highly aggressive isolates were from coleopteran insects. A correlation was found between the RAPD grouping and the phenotypic classification of the isolates. All the lepidopteran isolates grouped into one major cluster, most sub clusters were constituted by isolates from the same climatic zone.     

Phylogeographic pattern and regional evolutionary history of the maize stalk borer Busseola fusca (Fuller) (Lepidoptera: Noctuidae) in sub-Saharan Africa

Busseola fusca (Fuller) (Lepidoptera: Noctuidae) is one of the major cereal pests in sub-Saharan Africa. Previous phylogeographic investigations on samples collected in Kenya, Cameroon and West-Africa showed the presence of three main clades (W, KI, KII) originated from populations isolated in West and East Africa around one million years ago. Demographic and phylogenetic analyses suggested that this event was followed by local demographic expansion and isolation by distance. These hypotheses were tested by a more comprehensive sampling across B. fusca’s geographic range in Africa. Comparisons of sequences of partial mitochondrial DNA gene (cytochrome b) from 489 individuals of 98 localities in southern, central, eastern and western African countries confirmed the presence of the three main clades. Phylogenetic, F-statistics, demographic parameters and nested clade phylogeographic analyses confirmed that the clades experienced geographic and demographic expansion with isolation by distance after their isolation in three refuge areas. The geographic range of clade KII, already known from East to Central sub-Saharan Africa was extended to Southern Africa. Mismatch distribution analysis and the negative values of Tajima’s D index are consistent with a demographic expansion hypothesis for these three clades. Significant genetic differentiations were revealed at various hierarchical levels by analysis of molecular variance (AMOVA). Hypotheses about the geographic origin of the three main clades are detailed.     

Field evaluation of spinosad as a grain protectant for stored wheat in Australia: efficacy against Rhyzopertha dominica (F.) and fate of residues in whole wheat and milling fractions

Abstract  The potential of spinosad as a grain protectant for the lesser grain borer, Rhyzopertha dominica , was investigated in a silo-scale trial on wheat stored in Victoria, Australia. Rhyzopertha dominica is a serious pest of stored grain, and its resistance to protectants and the fumigant phosphine is becoming more common. This trial follows earlier laboratory research showing that spinosad may be a useful pest management option for this species. Wheat (300 t) from the 2005 harvest was treated with spinosad 0.96 mg/kg plus chlorpyrifos-methyl 10 mg/kg in March 2006, and samples were collected at intervals during 7.5 month storage to determine efficacy and residues in wheat and milling fractions. Chlorpyrifos-methyl is already registered in Australia for control of several other pest species, and its low potency against R. dominica was confirmed in laboratory-treated wheat. Grain moisture content was stable at about 10%, but grain temperature ranged from 29.3°C in March to 14.0°C in August. Bioassays of all treated wheat samples over 7.5 months resulted in 100% adult mortality after 2 weeks exposure and no live progeny were produced. In addition, no live grain insects were detected during outload sampling after a 9 month storage. Spinosad and chlorpyrifos-methyl residues tended to decline during storage, and residues were higher in the bran layer than in either wholemeal or white flour. This field trial confirmed that spinosad was effective as a grain protectant targeting R. dominica .     

Free amino acids of the haemolymph of the southwestern corn borer and the European corn borer in relation to their diapause

The titres of free amino acids present in the haemolymph of diapausing larvae of the southwestern corn borer, Diatraea grandiosella and the European corn borer, Ostrinia nubilalis, were examined. High titres of serine were found in the haemolymph of both species. Serine may serve as a storage form of compounds that are required for the synthesis of uric acid and other purines. The high titres of proline found in the haemolymph of O. nubilalis during the fall and winter may contribute to the freezing tolerance of this species. Alanine accumulated in the haemolymph of both species during the winter.