Proteomic interactions between the parasitoid Diachasmimorpha longicaudata and the oriental fruit fly,Bactrocera dorsalis during host parasitism

The oriental fruit fly, Bactrocera dorsalis, is an important agricultural pest and biological control is one of the most effective control methodologies. We conducted an investigation on the molecular response of the fruit fly to parasitism by the larval parasitoid, Diachasmimorpha longicaudata using two-dimensional gel electrophoresis and mass spectroscopy. We identified 285 differentially expressed protein spots (109 proteins) during parasitism. The molecular processes affected by parasitism varied at different time point during development. Transferrin and muscle specific protein 20 are the only two proteins differentially expressed that play a role in host immunity 24 h after parasitism. Developmental and metabolic proteins from parasitoids (transferrin and enolase) were up-regulated to ensure establishment and early development of parasitoids 48 h post parasitism. 72 h after parasitism, larval cuticle proteins, transferrin and CREG1 were overexpressed to support the survival of parasitoids while host metabolism proteins and parasitoid regulatory proteins were down-regulated. Host development slowed down while parasitoid development went up at 96 h after parasitism. All developmental, regulatory, structural, and metabolic proteins were expressed at their optimum at 120 h post parasitism. Host development was reduced, metabolism and regulatory proteins were strongly involved in the activities. The development deteriorated further at 144 h after parasitism. Enolase and CREG1 were indicators of parasitoid survival. Hexamerin and transferrin from the parasitoid was peaked at 168–216 h after parasitism, strongly indicating that parasitoid would survive. This study represents the first report that reveals the molecular players involved in the interaction between the host and parasitoid.     

Additional measurements of distance-dependent capture probabilities for released males of Bactrocera cucurbitae and B. dorsalis (Diptera: Tephritidae) in Honolulu

This paper describes a mark-release-recapture study involving males of two economically important fruit flies (Diptera: Tephritidae), Bactrocera cucurbitae (Coquillett) and B. dorsalis (Hendel), conducted in Honolulu, Hawaii. In each of three residential neighborhoods in Honolulu, we placed two traps, one baited with cue lure and the other with methyl eugenol (male attractants for B. cucurbitae and B. dorsalis, respectively), in a single tree. For both species, dyed, mature males from recently established laboratory colonies were released 25, 50, 100, and 200 m from the traps along the four compass directions, and for B. dorsalis releases were also made 300 m from the traps. For both B. cucurbitae and B. dorsalis, the proportion of males captured varied significantly among study areas and among release distances. Averaged over the 3 sites, the percentage of released males captured varied from 14% for releases at 25 m to 0.5% for releases at 200 m for B. cucurbitae and from 21% for releases at 25 m to 3% for releases at 300 m for B. dorsalis. In general, the capture rates of B. cucurbitae males were similar between California and Hawaii, whereas the captures rates of B. dorsalis males were significantly higher in California than Hawaii for all releases distances tested. Distance-dependent capture rates were used to estimate detection sensitivities for the two species under a trapping regime utilizing 5 cue lure- and 5 methyl eugenol-baited traps per 2.59 km² (as currently employed in California) and then compare these sensitivities between California and Hawaii.     

Generation and analysis of expressed sequence tags from adductor muscle of Japanese scallop Mizuhopecten yessoensis

A normalized cDNA library was constructed from the adductor muscle of M. yessoensis and acquired 4595 high quality expressed sequence tags (ESTs). After clustering and assembly of the ESTs, 3061 unigenes containing 654 contigs and 2407 singletons were identified. The contig length ranged from 266 bp to 2364 bp and the average length of these contigs was 544 bp. Blastx nonredundant protein database analysis showed that 1522 unigenes had significant homology to known genes (E value ≤ 10^? 5). By comparing to Clusters of Orthologous Groups (COG) categories, 460 unigenes were annotated (E value ≤ 10^? 10). Using Kyoto Encyclopedia of Genes and Genomes (KEGG), 345 of 3061 unigenes were assigned into 103 pathways (E value ≤ 10^? 5). For InterProScan searches, 1237 unigenes were annotated containing 727 different types of protein domains. 941 of the 1237 unigenes were annotated for Gene Ontology (GO) classification using Uniprot2GO associations in any category (biological, cellular, and molecular). By sequences comparability and analysis of Blastx NCBI nonredundant protein database and KEGG, 66 unigenes were identified that may be involved in genetic information processing based on the known knowledge. The study provides a material basis as useful information for the genomic analysis of shellfish.     

Mark-release-recapture of males of Bactrocera cucurbitae and B. dorsalis (Diptera: Tephritidae) in two residential areas of Honolulu

This paper describes a mark-release-recapture study involving males of two economically important tephritid fruit flies (Diptera: Tephritidae), Bactrocera cucurbitae (Coquillett) and B. dorsalis (Hendel), conducted over a 2-year period in Honolulu, Hawaii. In each of two residential neighborhoods, we placed two traps, one baited with cue lure and the other with methyl eugenol (male attractants for B. cucurbitae and B. dorsalis, respectively), in a single tree. Dyed, mature males from recently established laboratory colonies were released 100 or 500 m from the traps along the four compass directions in both winter and summer seasons. In each neighborhood, a total of 5,600 flight-able males of each species were released 100 m from the traps (14 dates × 4 directions × 100 males/release) and 56,000 flight-able males of each species were released 500 m from the traps (14 dates × 4 directions × 1,000 males/release). Within each study area, the number of males trapped did not vary significantly with direction or season for either species for either the 100- or 500-m releases. Significantly higher numbers of B. dorsalis males were captured than B. cucurbitae males for both the 100-m (16 versus 8 males/release) and 500-m (7 versus 2 males/release) releases (average values computed over both study areas). Also, following their release, B. dorsalis males were, in general, trapped more quickly than B. cucurbitae males. Given the strong attractancy of methyl eugenol, trap captures for B. dorsalis were lower than expected, and possible explanations are discussed.     

Optimization of Candida sp. 99-125 lipase catalyzed esterification for synthesis of monoglyceride and diglyceride in solvent-free system

Esterification of glycerol and oleic acid catalyzed by lipase Candida sp. 99-125 was carried out to synthesize monoglyceride (MAG) and diglyceride (DAG) in solvent-free system. Beta-cyclodextrin as an assistant was mixed with the lipase powder. Six reaction variables, initial water content (0–14 wt% of the substrate mass), the glycerol/oleic acid molar ratio (1:1–6:1), catalyst load (3–15 wt% of the substrate mass), reaction temperature (30–60 °C), agitator speed (130–250 r/min) and beta-cyclodextrin/lipase mass ratio (0–2) were optimized. The optimal conditions to the synthesis of MAG and DAG were different: the optimal glycerol/oleic acid molar ratio, beta-cyclodextrin/lipase mass ratio, catalyst load and reaction temperature were 6:1, 0, 5%, 50 °C for MAG, and 5:1, 1.5, 10%, 40 °C for DAG, respectively. The optimal water content and agitator speed for both MAG and DAG were 10% and 190 r/min, respectively. Under the optimal conditions, 49.6% MAG and 54.3% DAG were obtained after 8 h and 4 h, respectively, and the maximum of 81.4% MAG plus DAG (28.1% MAG and 53.3% DAG) was obtained after 2 h under the DAG optimal condition. Above 90% purity of MAG and DAG can be obtained by silica column separation.     

Screening of vip genes from a Spanish Bacillus thuringiensis collection and characterization of two Vip3 proteins highly toxic to five lepidopteran crop pests

A Spanish Bacillus thuringiensis strain collection was screened for the presence of vip genes. One hundred strains from a Canary Island collection were screened for vip1 and vip2 genes and 7% contained potentially novel vip1 and vip2-like genes, as indicated by the low degree of similarity with previously known vip1 and vip2 genes. Four hundred strains from a collection originating from the Spanish mainland were screened for vip3 genes and 14.5% of them contained potentially novel vip3-like genes. Reconstruction of the full-length vip sequences could only be achieved for two vip3 gene variants encoding 789 and 787 amino acid proteins that were designated as Vip3Aa45 and Vip3Ag4, respectively. These proteins showed 82% pairwise identity between them and differed from Vip3Aa1 in the putative signal peptide, two specific proteolytic processing sites and the 66-kDa insecticidal fragment. The purified proteins were tested against nine lepidopteran pest species and displayed toxicity, expressed as mean lethal concentration, for five of them. The two toxins were highly toxic for Lobesia botrana (∼1–2 μg/ml) and Spodoptera littoralis (∼20 ng/cm²), moderately toxic for Spodoptera exigua (∼100–300 ng/cm²), and varied greatly in their toxicity for Mamestra brassicae or Chrysodeixis chalcites, with high toxicity for Vip3Aa45 in M. brassicae (∼40 ng/cm²) and for Vip3Ag4 in C. chalcites (∼45 ng/cm²).     

S-Arachidonoyl-2-thioglycerol synthesis and use for fluorimetric and colorimetric assays of monoacylglycerol lipase

We report the first synthesis of 2-thioglycerol and S-arachidonoyl-2-thioglycerol (the thioester analog of the endocannabinoid 2-arachidonoylglycerol) in an eight or nine step procedure with a yield of ～25% and establish the use of this substrate for maleimide-based fluorescent and dithiobis(2-nitrobenzoic acid)-based colorimetric assays of human recombinant monoacylglycerol (MAG) lipase (hMAGL) and human brain membrane MAG hydrolase activity. Inhibitor structure–activity relationships observed here for hMAGL and 2-ATG correlate well (r² = 0.93, n = 9) with earlier findings for mouse brain MAG hydrolase with non-thiol substrates.     

Sampling of Bemisia tabaci adults using a pre-programmed autonomous pest control robot

An autonomous robot (Cabbot) was built to control insect pests in a plastic greenhouse and utilized to sample adult whiteflies (Bemisia tabaci) on paprika plants (Capsicum annum var. angulosum). To accomplish this, a sampling device consisting of an air compressor for pest agitation, a sticky trap (100 mm × 150 mm) and an image processing system for pest identification were installed on the Cabbot. The sampling precision of the Cabbot (D = 0.16) was higher than that of the sticky trap (D = 0.19) when sampling of adult white flies was conducted in caged pots. The Cabbot could also collect a substantial number of individuals within a short duration (3 minutes). The collection efficiency (i.e., percent of samples collected at least n (1 in this case) individuals to the total sample number) of insects by the Cabbot was markedly high at low population size, showing approximately 30% when the population size was ≤ 16 individuals per plant. The sampling precision and collection efficiency suggested that the Cabbot is effective in local, short-term sampling and could be used for early warning of pest occurrences.     

The complete mitogenome of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) and comparison with other lepidopteran insects

The complete mitochondrial genome (mitogenome) of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) was determined to be 15,653 bp, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a A + T-rich region. It has the typical gene organization and order of mitogenomes from lepidopteran insects. The AT skew of this mitogenome was slightly positive and the nucleotide composition was also biased toward A + T nucleotides (81.31%). All PCGs were initiated by ATN codons, except for cytochrome c oxidase subunit 1 (cox1) gene which was initiated by CGA. The cox1 and cox2 genes had incomplete stop codons consisting of just a T. All the tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The A + T-rich region of the mitogenome was 495 bp in length and consisted of several features common to the lepidopteras. Phylogenetic analysis showed that the B. mori Dazao was close to Bombycidae.     

Structure and ultrastructure of spermatozoon in six species of Drosophilidae (Diptera)

The Drosophilidae family is formed by Brachycera Diptera distributed widely across different regions of the planet. It is composed of about 4000 species, 304 of which are found in Brazil. The objective of this work was to characterize morphologically the structure of the male internal reproductive apparatus and the ultrastructure of the spermatozoon in four Neotropical (Drosophila cardini, D. mercatorum, D. nebulosa and D. sturtevanti) and two invasive (D. simulans and Zaprionus indianus) species of drosophilids. The structural aspect of the internal reproductive apparatus corresponds with that described for other drosophilids; however, there are differences in the size and coloration of the structures, such as the testes, in each species analyzed. The spermatozoon of these species was seen to be long and fine, presenting morphological variation. The ultrastructure of the spermatozoon revealed that the morphological pattern is similar to that found in the majority of insects. The head region presents a nucleus with condensed chromatin and the acrosome positioned laterally to the nucleus. In the tail region, the axoneme presents the 9 + 9 + 2 pattern commonly described for other species of Diptera. The species presented differences regarding the shape and size of the mitochondrial derivatives. Cytochemical analysis using EPTA also revealed differences in terms of the location of the basic proteins in the mitochondrial derivates. The results obtained contribute to expanding the database for the Drosophilidae family, providing information that may contribute to intra- and inter-specific identification and supplying phylogenetic analyses.     

Biology and demography of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) on mulberry

Lesser mulberry pyralid, Glyphodes pyloalis Walker, is a monophagous pest of mulberry and has recently been reported in northern Iran. The biology and life table of this pest were studied in controlled conditions (24 ± 1 °C, 75 ± 5% RH and 16:8 LD). Mortality rate, metamorphosis, appearance of adult insects and adult sex ratio were recorded daily. Data were analyzed based on an age-stage, two-sex life table. Developmental rate among individuals and between sexes were also considered. The developmental periods for the egg, first through fifth instar larvae, prepupae and pupae were 4.06 ± 0.03, 2.93 ± 0.03, 2.03 ± 0.02, 2.01 ± 0.01, 2.10 ± 0.03, 4.09 ± 0.03, 2.04 ± 0.02 and 9.7 ± 0.09 days, respectively. The mean total developmental period from egg to adult was 35.40 ± 0.37 days. The maximum adult longevity was 7 and 11 days for males and females, respectively. The pre-oviposition period lasted 2.14 ± 0.04 days. Laboratory observation showed that adults are nocturnal and female lay eggs at night. The intrinsic rate of increase (r_m) and the finite rate of increase (λ) were 0.14 ± 0.005 d^? 1 and 1.15 ± 0.01 d^? 1. The net reproductive rate (R₀), gross reproductive rate (GRR) and mean generation time (T) were 134.67 ± 20.6 female offspring, 294.71 ± 4.07 eggs/female and 34.44 ± 0.2 days, respectively. Life expectancy of freshly laid egg was 32.15 days.     

Multi-chaperone-peptide-rich mixture from colo-carcinoma cells elicits potent anticancer immunity

Background: Chaperones play an important role in inducing anti-cancer immunity. To explore the probability of using chaperone-peptide-rich complexes extracted from colo-carcinoma cells as anti-cancer vaccine, we extracted and prepared chaperone-peptide-rich complexes from CT26 cells, which were subsequently investigated on anti-cancer efficacy. Methods: The crude extracts of the CT26 cells treated with heat and Trichosanthin were precipitated with salt and dialyzed to remove proteins below 50 kDa and above 300 kDa in molecular weight; the proteins with the molecular weights in 70 kDa, 90 kDa, 95 kDa, 110 kDa and 170 kDa were collected through gel filtration and SDS-PAGE. After confirmation, the purified proteins were used to determine their effects on lymphocyte proliferation, the activities of NK and CTL, tumor suppression and the tumor-bearing mouse survival. Results: The majority of the chaperone-peptides of anti-cancer immunity in CT26 cells, including HSP70-antigen peptide, HSP90-antigen peptide, gp96-antigen peptide, HSP-110 antigen peptide, HSP170-antigen peptide, was satisfactorily extracted that the multi-chaperone-peptide-rich mixtures were obtained. All the mixtures prepared could elicit lymphocyte proliferation, enhance the activities of CTL and NK, reinforce the tumor suppression and prolong the mouse survival. Conclusions: The multi-chaperone-peptide-rich mixtures could be prepared via dialysis and gel filtration combining with SDS-PAGE. Both the heat stress and Trichosanthin could induce and increase the mixtures, of which that treated by 42 °C heat and Trichosanthin was found to possess the strongest anti-cancer efficacy.     

Testosterone and immune-reproductive tradeoffs in healthy women

Although testosterone (T) has been characterized as universally immunosuppressive across species and sexes, recent ecoimmunology research suggests that T's immunomodulatory effects (enhancing/suppressing) depend on the organism's reproductive context. Very little is known about the immune effects of T in healthy females, and even less about how reproductive effort modulates the immune effects of T in humans. We investigated how the interaction between endogenous T and sexual activity predicted menstrual cycle-related changes in several measures of immunity: inflammation (indexed by interleukin-6, IL-6), adaptive immunity (indexed by immunoglobulin A, IgA), and functional immunity (indexed by bactericidal assay). Thirty-two healthy women (sexually abstinent, N = 17; sexually active with one male partner, N = 15) provided saliva samples at four points in the menstrual cycle: menses, follicular, ovulation, and luteal phases. Among sexually abstinent women, T was positively associated with IL-6 across the cycle; for sexually active women, however, T was positively associated with IL-6 in the luteal phase only, and negatively associated with IL-6 at ovulation. High T predicted higher IgA among women who reported infrequent intercourse, but lower IgA among women who reported very frequent intercourse. Finally, across groups, T was positively associated with greater bacterial killing at menses, but negatively associated in the luteal phase. Overall, rather than being universally immunosuppressive, T appeared to signal immunomodulation relevant to reproduction (e.g., lowering inflammation at ovulation, potentially preventing immune interference with conception). Our findings support the hypothesis that the immunomodulatory effects of endogenous T in healthy females depend on sexual and reproductive context.     

Ghrelin and peptide YY increase with weight loss during a 12-month intervention to reduce dietary energy density in obese women

Reducing dietary energy density (ED) promotes weight loss; however, underlying mechanisms are not well understood. The purpose of this study was to determine if low-ED diets facilitate weight loss through actions on ghrelin and peptide YY (PYY), independent of influences of psychosocial measures. Seventy-one obese women (BMI 30–40 kg/m²) ages 22–60 years received counseling to reduce ED. Fasting blood samples were analyzed for total ghrelin and total PYY by radioimmunoassay at months 0, 3, 6, and 12. Restraint, disinhibition, and hunger were assessed by the Eating Inventory. Body weight (−7.8 ± 0.5 kg), BMI (−2.9 ± 0.2 kg/m²), body fat (−3.0 ± 0.3%), and ED (−0.47 ± 0.05 kcal/g or −1.97 ± 0.21 kJ/g) decreased from months 0 to 6 (p < 0.05) after which no change occurred from months 6 to 12. Ghrelin increased in a curvilinear fashion (month 0: 973 ± 39, month 3: 1024 ± 37, month 6: 1109 ± 44, and month 12: 1063 ± 45 pg/ml, p < 0.001) and PYY increased linearly (month 0: 74.2 ± 3.1, month 3: 76.4 ± 3.2, month 6: 77.2 ± 3.0, month 12: 82.8 ± 3.2 pg/ml, p < 0.001). ED, body weight, and hunger predicted ghrelin, with ED being the strongest predictor (ghrelin = 2674.8 + 291.6 × ED − 19.2 × BW − 15 × H; p < 0.05). There was a trend toward a significant association between ED and PYY (PYY = 115.0 − 43.1 × ED; p = 0.05). Reductions in ED may promote weight loss and weight loss maintenance by opposing increases in ghrelin and promoting increases in PYY.