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.     

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.     

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.     

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.     

Entomopathogenic fungi (Hypocreales) for control of potato psyllid, Bactericera cockerelli (Šulc) (Hemiptera: Triozidae) in an area endemic for zebra chip disease of potato

Potato psyllid, Bactericera cockerelli, is a serious pest of potato and other solanaceous vegetables in the United States, Mexico, Central America, and New Zealand and is responsible for transmission of Candidatus Liberibacter solanacearum which causes a disease known as “zebra chip” (ZC). Entomopathogenic fungi could provide a viable component for an integrated pest management strategy for control of B. cockerelli and other potato pest insects. Three field trials of commercial formulations of Metarhizium anisopliae (F 52®, Novozymes Biologicals) and Isaria fumosorosea (Pfr 97®, Certis USA) and abamectin (Agri-Mek®, Syngenta, USA) were conducted in Weslaco, Texas. Rates are expressed in quantity of product delivered in 375–470 l of water/ha. F 52 applied at 0.51, 1.1, and 2.2 l/ha and Agri-Mek applied at 584 ml/ha produced reductions of B. cockerelli eggs and nymphs of 45%, 59%, 67%, and 63%, respectively. Only Agri-Mek significantly reduced plant damage. Pfr 97 at 1.1 kg/ha with and without 1% Trilogy® (neem oil, Certis, USA), and Agri-Mek at 584 ml/ha resulted in psyllid reductions of 78%, 76%, and 84%, respectively. Significantly decreased plant damage and ZC symptoms were observed for all treatments. Tuber yields for Pfr plus Trilogy and Agri-Mek were significantly higher than the control. F 52 applied at 1.1 and 2.2 l/ha and Pfr 97 at 1.1 and 2.2 kg/ha produced 62%, 62%, 66%, and 65% reduction, respectively. Tuber yield for both rates of Pfr and the high rate of F 52 were significantly higher than the control. All fungal treatments significantly reduced plant damage and ZC symptoms.     

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.     

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.     

Proteome analysis of male accessory gland secretions in Leucinodes orbonalis Guenee (Lepidoptera: Crambidae), a Solanum melongena L. pest

Male accessory gland (MAG) proteins are transferred along with the sperm to females at the time of mating and have diverse effects on female reproductive physiology in a wide range of insects. In this study, we sought to identify the MAG proteins in Leucinodes orbonalis Guenee, a Solanum melongena L. pest, by analyzing the MAG proteins of virgin and mated male moths by nano-LC-ESI-MS/MS techniques. A total of 142 and 131 proteins in virgin and mated males were identified, respectively, among which 17 (12.0%) and 10 (7.6%) proteins were found to show secretory signals in virgin and mated males, respectively. These secretory proteins were shown to be involved in several biological processes in insects, including egg development, sperm-related functions/capacitation, defense, metabolism, and protein chaperoning. To the best of our knowledge, this is the first study to perform a proteome analysis of the MAG proteins of L. orbonalis, and offers an opportunity for further investigation of the functions of these proteins. In insects, certain MAG proteins are known to inhibit mating whereas others accelerate egg-laying. Therefore, the identification of these proteins in L. orbonalis may be useful for pest control.     

Construction and characterization of a cDNA library from shark regenerated hepatic tissue

Sharks are a type of fish with a full cartilaginous skeleton and have big livers. To better understand liver regeneration in sharks and screening for the important genes participated in disease-defense, in this study, a cDNA library of regenerated liver tissues of shark, Chiloscyllium plagiosum, was constructed. A total of 2103 expressed sequence tags (ESTs), which represents 997 unique genes, were sequenced. Among these genes, 434 (43.53%) of them showed significant similarity (E-values < 10^?5) to the sequences in NCBI Nt database, 685 (68.71%) of these unique genes showed significant similarity (E-values < 10^?5) to the sequences in NCBI Nr database, and 662 (66.40%) of these unique genes showed significant similarity (E-values < 10^?5) to the Swiss-Prot database. Preliminary analysis of unique genes according to COG database showed that unigenes were further grouped into 21 functional categories including inorganic ion transport and metabolism, energy production and conversion, posttranslational modification, protein turnover and chaperones, general function prediction only, translation, and ribosomal structure and biogenesis. Several possible candidate genes involved in liver regeneration were selected to analyze their expression with relative quantification real-time PCR. This study may contribute to our better understanding of the molecular mechanism of regeneration in shark liver. Furthermore, the EST cataloguing and profiling of shark will be also benefited to the functional genomic research in this marine species.     

Evaluation of tomato cultivars to Helicoverpa armigera using two-sex life table parameters in laboratory

Helicoverpa armigera (Hübner) is one of the most important pests of a wide range of agricultural crops worldwide. Resistance of 10 tomato cultivars (‘Primoearly,’ ‘Riogrand,’ ‘CaljN3,’ ‘Kingstone,’ ‘Earlyurbana,’ ‘Petomech,’ ‘EarlyurbanaY,’ ‘Mobil,’ ‘Imprial’ and ‘Petoearly’) to H. armigera was evaluated under laboratory conditions at 25 ± 1 °C, 60 ± 5% RH and a photoperiod of 16:8 (L:D) hours using age-stage, two-sex life table parameters. The larval period ranged from 19.96 on ‘Riogrand’ to 24.58 days on ‘Imprial.’ The insects reared on ‘Imprial’ had the longest total preoviposition period (44.80 days) and those reared on ‘Riogrand’ had the shortest one (35.44 days). The longest adult longevity for female and male was observed on ‘EarlyurbanaY’ (14.40 days) and ‘Kingstone’ (15.00 days), respectively. Using age-stage, two-sex life table, the value of the net reproductive rate (R₀) varied from 7.8 on ‘Imprial’ to 186.9 offspring per individual on ‘Petomech.’ The lowest value of the intrinsic rate of increase (r) and finite rate of increase (λ) was on ‘Imprial’ (0.0410 and 1.0423 day^− 1, respectively) and the highest was on ‘Petomech’ (0.1274 and 1.1359 day^− 1, respectively). The mean generation time (T) on different cultivars varied from 39.9 to 48.2 days. The results revealed that ‘Petomech’ was the most susceptible (suitable) and ‘Imprial’ was the most resistant (unsuitable) cultivar to this pest among the tomato cultivars tested.     

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.     

Higher Growth Hormone Levels are Associated with Erectile Dysfunction in Male Patients With Acromegaly

Objective: To investigate in vivo correlates of erectile dysfunction (ED) in male patients with acromegaly.Methods: Fifty-one male patients with acromegaly were assessed by the International Index of Erectile Function-5 and Acromegaly Quality of Life (Acro-QoL) questionnaires. The measurement of serum nitric oxide (NO) were performed in patients and age-matched nonacromegalic controls.Results: Among 51 patients analyzed, 32 (62.7%) had ED. Patients with ED showed lower Acro-QoL scores regarding global (69.8 ± 17.7 versus 79.4 ± 11.2; P = .035) and personal relationship dimensions (59.6 ± 22.1 versus 76.8 ± 17.6; P = .012) than non-ED patients. ED patients were older (44.5 ± 11.2 years versus 33.2 ± 8.5 years; P = .04) and showed higher growth hormone (GH) levels (15.5 μg/L &lsqb;interquartile range of 9.5 to 34.5 μg/L] versus 5.9 μg/L &lsqb;interquartile range of 3.4 to 13.9 μg/L]; P = .001) compared to non-ED patients. The cutoff values for identifying ED were 7.9 μg/L for random GH and 5.3 μg/L for GH nadir after oral administration of 75 g of glucose. There was no significant difference in total testosterone levels between the two groups (6.36 ± 4.24 nmol/L versus 9.54 ± 5.50 nmol/L; P = .299). The NO levels in patients with acromegaly were significantly lower than those in nonacromegalic controls (8.77 ± 1.78 μmol/L versus 19.19 ± 5.02 μmol/L, respectively; P = .049). Furthermore, the NO levels were even lower in ED patients than those in non-ED patients (5.14 ± 0.98 μmol/L versus 12.09 ± 3.44 μmol/L; P = .027).Conclusion: Our study showed that ED is prevalent in male acromegalic patients and may be associated with systemic endothelial dysfunction induced by excessive GH. Further studies investigating the mechanism of GH and ED are required.Abbreviations: Acro-QoL = Acromegaly Quality of Life; ED = erectile dysfunction; FSH = follicle-stimulating hormone; GH = growth hormone; IGF-1 = insulin-like growth factor 1; IIEF-5 = international index of erection function-5; LH = luteinizing hormone; MRI = magnetic resonance imaging; NO = nitric oxide; OGTT = oral glucose tolerance test; QoL = quality of life; ROC = receiver operating characteristic     

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).     

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.