Proteomics/qPCR protocol to estimate physical ages of wild male oriental fruit flies, Bactrocera dorsalis (Hendel)

Chiou Ling Chang 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.2

Laboratory reared fruit flies may have different behavioral phenotypes as compared to the wild in the field. The domesticated lab setting and the absence of factors from the natural environment on lab reared insects may result in different behavioral patterns, such as the length of life cycle. This study was performed to conduct a regression analysis of the ratio of odorant binding protein 99b (OBP99b) expression from 4 to 12-d-old oriental fruit fly males, relative to 1-d-old males to estimate the physical ages of wild male fruit flies. We established two polynomial linear regression equations based on 4–12 days old lab-reared flies using proteomics and qPCR. The equations are Y = −0.7768 + 0.7205X, R2 = 0.89 (for proteomics) and Y = −0.6478 + 0.344X, R2 = 0.64 (for qPCR). We used these equations to estimate the physical ages of wild-caught male fruit flies. These results indicate that despite multiple behavioral differences between laboratory reared and field-caught flies, the physical ages of both groups are identical.We suggest proteomics and qPCR analysis of selected genes and the proteins they encode may be developed into reliable tools for determining the ages of wild-caught animals, including oriental fruit flies.

Effects of Aspergillus oryzae Fermentation Extract on Performance of Lactating Cows in the Summer and Winter in Taiwan

Chiou, Peter Wen-Shyg,Chen, Chao-Ren,Yu, Bi Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.3

The aims of this study is to evaluate the effect of Aspergillus oryzae Fermentation Extract (AFE) on the performance of lactating cows in summer (May to July) and winter (December to February). The experiment was a completely randomized design (CRD) and dietary treatments were 1) basal diet without AFE, 2) basal plus 3 g/d AFE into the basal total mixed ration (TMR), 3) basal plus 45.4 mg AFE/kg the ensiling corn silage and 4) AFE inclusion in silage and TMR. Twenty-eight cows from each trial were selected and randomly allocated into the four treatment groups, confined in individual pens, and fed ad libitum for 8 weeks in both seasons of feeding trials. Results showed that AFE inclusion in corn silage significantly improved DM intake by 4.4% and milk yield by 3.1% (p<0.05) during summer. In the winter season, AFE inclusion in the diet significantly improved milk yield by 10%. Direct addition of AFE to the TMR even further significantly improved milk yield over the addition through corn silage by 7.4% in winter (p<0.05). An additive effect of AFE inclusion into TMR and through corn silage was also demonstrated in the winter-feeding. AFE inclusion however, did not improve DM intake during the winter trial. In the summer trial, inclusion of AFE showed an adverse effect on the percentage of milk fat, but did not impact on the milk fat yield. Adding AFE through corn silage showed a trend towards alleviating the negative effects of milk fat from direct AFE inclusion in TMR. The similar trend occurred in the winter trial. The inclusion of AFE through corn silage significantly lowered the milk protein content over direct AFE addition, but did not significantly impacted the milk protein yield in summer. AFE supplementation during the winter season significantly increased milk protein content. Adding AFE to the corn silage significantly increased milk protein content over direct AFE addition in winter although inclusion of AFE significantly decreased total milk solid content in the summer (p<0.005). During the winter season, inclusion of AFE required less DM to produce a unit of milk. Inclusion of AFE into corn silage required less DM, energy and protein to produce a unit of milk. But inclusion of AFE did not alleviate heat stress on the lactating cows.

Proteomic identification of a potential sex biomarker for 2 fruit fly species at pupal stage

Chiou Ling Chang 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.1

Fruit flies are one of the most destructive pests worldwide. The sterile insect technique (SIT) has been one of the most effective fruit fly control techniques. Genetic sexing strains have been developed to facilitate the separation ofmales and females based on color at the pupal stage for the oriental fruit fly, Bactrocera dorsalis, andthemedfly, Ceratitis capitate, known as dorsalis translocation white pupae (DTWP) and temperature sensitive lethal mutation strain (TSL). In both cases, the females are white and the males are brown. By releasing only sterile males, the control efficiency was increased, and fruit damage due to sterile females reduced. There remains, however, a gap in our understanding of this phenomenon because there is no information on the physiological basis of the sex differences. Here, we report the outcome of a study designed to identify pupal proteins related to the sex differences using these two strains. We collected whole pupae of both species each day for first 10 days for protein analysis. Protein expression was analyzed by 2D electrophoresis, gel densitometry and mass spectrometry. The general odorant binding protein, OBP56d, was differentially expressed in white pupae versus brown pupae for both species. We also confirmed this result with phenotypical characteristics to prove these proteins were not derived from puparia. These proteins had molecular weights between 10 and 15 kDa and a pI of 6.73 for DTWP and 5.71 for TSL.

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

Chiou Ling Chang,Guang Hong Liang,Scott M. Geib 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.1

The oriental fruit fly, Bactrocera dorsalis, is an important agricultural pest and biological control is one of themost effective control methodologies. We conducted an investigation on the molecular response of the fruit fly toparasitism by the larval parasitoid, Diachasmimorpha longicaudata using two-dimensional gel electrophoresis andmass 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. Transferrinand muscle specific protein 20 are the only two proteins differentially expressed that play a role in host immunity24 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 afterparasitism, larval cuticle proteins, transferrin and CREG1 were overexpressed to support the survival of parasitoidswhile host metabolism proteins and parasitoid regulatory proteins were down-regulated. Host developmentslowed 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. Hostdevelopment was reduced, metabolism and regulatory proteins were strongly involved in the activities. Thedevelopment deteriorated further at 144 h after parasitism. Enolase and CREG1 were indicators of parasitoidsurvival. Hexamerin and transferrin from the parasitoid was peaked at 168–216 h after parasitism, stronglyindicating that parasitoid would survive. This study represents the first report that reveals the molecular playersinvolved in the interaction between the host and parasitoid.

The current status of three-dimensional ultrasonography in gynaecology

Chiou Li Ong 대한초음파의학회 2016 ULTRASONOGRAPHY Vol.35 No.1

Ultrasonography (US) is the most recent cross-sectional imaging modality to acquire threedimensional (3D) capabilities. The reconstruction of volumetric US data for multiplanar display took a significantly longer time to develop in comparison with computed tomography and magnetic resonance imaging. The current equipment for 3D-US is capable of producing highresolution images in three different planes, including real-time surface-rendered images. The use of 3D-US in gynaecology was accelerated through the development of the endovaginal volume transducer, which allows the automated acquisition of volumetric US data. Although initially considered an adjunct to two-dimensional US, 3D-US is now the imaging modality of choice for the assessment of Müllerian duct anomalies and the location of intrauterine devices.

Comparative proteomic profiling within each developmental stage of the solanum fruit fly, Bactrocera latifrons Hendel

Chiou Ling Chang,Scott M. Geib 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.4

Fruit fly development has been widely studied in details, understanding of “what”, “when”, “where”, “why”, and “how” many hundred thousand proteins exist in an insect cell interact and express during development at molecular level largely remained to be clarified. We conducted proteome mapping in all developmental stages of the solanum fruit fly, Bactrocera latifrons (Hendel), by comparing all ages within a stage to their 1-d-old, using 2-D gel electrophoresis and mass spectrometry. Samples of designated ages of each stage of B. latifrons were collected, analyzed, and described. A custom peptide database, derived from a publically available de novo B. latifrons transcriptome assembly was adopted for peptide identification. Identified differentially expressed proteins (DEPs) and their putative protein functions were annotated in representative SDS gel images, charts, and tables. Based on our proteomic data, we constructed a preliminary and descriptive reference proteome maps which not only provide valuable information toward a comprehensive understanding of fruit fly development, but also build a foundation for development of novel advanced fruit fly control techniques or further studies related to sterilization insect technique (SIT) and genome deletion. Any epigenetic impacts due to abiotic or biotic environmental factors will be easier to be identified, manipulated, and further led to gene editing research.

Effects of Aspergillus oryzae Inclusion on Corn Silage Fermentation

Chiou, Peter Wen-Shyg,Ku, Hsiao-Che,Chen, Chao-Ren,Yu, Bi Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.11

This study is aimed at evaluating the effect of Aspergillus oryzae fermentation extract (AFE) on corn silage fermentation characteristics. Trial included two groups of treatments, with or without AFE inclusion in corn ensilage. Sixty corn silage containers, including two treatments with thirty replicates each, were processed in a laboratory scale mini-silo of 21 cm radius by 45 cm height. Three replicate containers were opened and sampled for analysis at 0, 0.5, 1, 2, 3, 4, 6, 10, 18 and 34 days after being ensiled. One silage container from each treatment was installed with a remote controlled electronic thermometer to record the temperature changes. Analysis included silage temperature, pH, fermentation acids, the water-soluble carbohydrates and chemical compositions and the silage protein fractions. Results showed that on the first day, the temperature of the ensiled corn was slightly higher than room temperature, but returned to room temperature on the second day. The pH and concentrations of WSC, ADF, lignin and acetic acid in the AFE treated silage were significantly lower than the control groups (p<0.05). The lactic acid and crude protein on the other hand were significantly higher in the AFE treated silage as compared to the control (p<0.05) at the end of the ensilage period. The DM content was significantly higher (p<0.05) whereas the butyric acid content of the AFE treated silage was significantly lower (p<0.05) than the control at the end of the 34 day ensilage period. Titratable acid and buffering capacity in the corn silage were not significantly different between treatment groups (p>0.05). Ammonia N concentration in the AFE treated silage showed a trend of decrease (p>0.05). NPN and the protein fraction A in both groups increased during the conservation period, but fraction A in the AFE treated corn silage was significantly higher than the control silage (p<0.05). During the conservation period, the AFE treated corn silage showed a trend toward a decrease in fractions $B_1$, $B_3$ and C (p<0.05). The protein fraction B2 showed a trend toward increase in the control group and an inconsistent trend in the AFE treated silage during the ensiling period. The AFE treated silage showed a better Flieg score over the control silage (97 vs. 75) as calculated from the concentrations of lactic acid, acetic acid and butyric acid.

Effect of High Dietary Copper on the Morphology of Gastro-Intestinal Tract in Broiler Chickens

Chiou, P.W.S.,Chen, C.L.,Chen, K.L.,Wu, C.P. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.4

An experiment was conducted to study the effects of high dietary copper supplementation on the gastrointestinal tract morphology of broiler chickens. Eighty 3-week-old broiler chicks were divided randomly into eight groups of four dietary treatments and over three week were fed isoenergetic and isonitrogenous diets that contained 0, 100, 250, or 500 mg/kg of supplemental copper from cupric sulfate. The copper supplementation in the broiler diet up to 250 mg/kg did not significantly influence broilers' performance. A high dietary copper supplementation of 500 mg/kg did significantly depress growth and feed conversion in the broilers (p<0.05). Copper supplementation more than 250 mg/kg in the broiler diet significantly influenced the morphology of the GI tract, as shown by severe oral lesions and gizzard erosion. It also significantly depressed the villi height and significantly thickened the muscular layer in the duodenum (p<0.05). The severely damaged villi were observed by scanning electronic microscope from the duodenum samples of broilers fed a 500 mg/kg copper supplemented diet. The 500 mg/kg copper supplemented diet also significantly influenced the plasma constituents. Plasma glucose concentration was significantly depressed (p<0.05).

Application of Cornell Net Carbohydrate and Protein System to Lactating Cows in Taiwan

Chiou, Peter Wen-Shyg,Chuang, Chi-Hao,Yu, Bi,Hwang, Sen-Yuan,Chen, Chao-Ren Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.6

The aim of this study was to apply the Cornell net carbohydrate and protein system (CNCPS) in subtropical Taiwan. This was done by means of 3 trials, viz, in situ, lactation and metabolic trials, the latter using the urinary purine derivatives (UPD) to estimate the ruminal microbial yield. Dietary treatments were formulated according to different nutrient requirement systems including, (1) a control NRC78 group on NRC (1978), (2) a NRC88 group on NRC (1988), and (3) a CNCPS group on Cornell Net carbohydrate and protein system model. Results from the lactation trial showed that DM intake (DMI) was higher (p<0.05) in the NRC78 than the other treatment groups. The treatments did not significantly influence milk yield, but milk yield after covariance adjustment for DMI was higher in the CNCPS group (p<0.05). The FCM, milk fat content and yield were greater in both the NRC78 and the NRC88 group over the CNCPS group (p<0.05). The treatments did not significantly influence the DMI adjusted FCM. The solid-non-fat and milk protein contents were higher in the CNCPS group (p<0.05) with or without DMI covariance adjustment. Lactating efficiency was higher in the CNCPS group (p<0.05) compared to the other groups. The significantly lowest milk urea-N (MUN) with better protein utilization efficiency in the CNCPS group (p<0.05) suggested that less N would be excreted into the environment. Cows in the CNCPS group excreted significantly more and the NRC88 group significantly less urinary purine derivatives (UPD) implying that more ruminal microbial protein was synthesized in the CNCPS over the NRC88 group. The CNCPS could become the most useful tool in predicting the trends in milk yield, microbial yield and MUN.

Fruit fly nutrition and proteomics

Chiou Ling Chang 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

Fruit fly is one of the most important pests for vegetables and crops worldwide. Since 1895, four species of fruit flies has invaded into Hawaii. In 2000, a group of scientists from Hawaii has initiated and implemented an area wide pest management program to suppress fruit fly population in Hawaii. Six techniques developed within the program has been transferred to many countries that have the fruit fly problem. Four techniques (monitoring, sanitation, bait spray, and male annihilation) are readily done by farmers. The other two techniques (sterile insect release and augmentative parasitoid release) involve mass fruit fly stock. Sterile insect technique (SIT) used in sterile insect release requires continuous mass rearing. Current mass rearing system has been satisfactory for rearing need. However, there are problems such as pesticide contamination of supporting material, spent diet management, labor intensive, and space issue. USDA-Agricultural Research Service looked for alternatives. In 2004, a novel fruit fly liquid diet has been developed. The core of this diet is using an inert substance (sponge cloth) to replace biological supporting material for mill feed (wheat product). During this diet development process, we have observed that fruit fly performance changes associate with the change of diet components. One of the most significant components is wheat germ oil. Larval diet supplemented with wheat germ oil (WGO) causes physiological reactions, such as increased fecundity and fertility, in some insects. Although the impact of WGO on insect physiology is important, the mechanisms of these actions are poorly understood. In this presentation, we will confirm our hypothesis that the addition of WGO to medium developed for larval oriental fruit flies modulates gene expression in the corresponding adults and further to identify when and how these gene expressed during different life cycle stages. We separately reared larvae of Bactrocera dorsalis on diets lacking or supplemented with WGO, and analyzed for expressed proteins in the resulting adult males and females by 2D-electrophoresis. Analysis of the gels revealed significant changes in expression levels of >70 proteins, 64 of which were identified by mass spectrometric analysis on MALDI-TOF/TOF. Apparent changes in expression levels for 6 of these proteins were confirmed by quantitative real-time PCR, showing that the changes in mRNA expression were reflected in changes in protein expression. These findings support the hypothesis that one mechanism of WGO actions in insect nutrition is the modulation of gene expression. Our goal is to identify molecular markers that serve as early indicators of the quality of insect culture media. Markers of deficient culture media will increase the efficiency of developing optimal systems for mass rearing beneficial insects and some pest species because decisions on culture media quality can be made without waiting through one or several life cycles.