淡水紅樹林沼澤區螃蟹種類分佈之調查

[[volume]]19

[[alternative]]回顧、前瞻與借鏡

[[issue]]200

[[alternative]]科學、技學、社會(STS)之探討研究

[[issue]]190

淡水紅樹林沼澤螃蟹之生態研究

[[abstract]]紅樹林內之螃蟹經由一年調查所得，計有四科十屬、十五種，其中Uca formosensis 為台灣特有種，數量稀少而珍貴，分佈區土壤之分析，可分為二群Silt loam 及Sand y loam. Ocypodidae及Mictyridae較喜分佈在Silt loam.而Grapsidae 則喜歡在Silt loam. 所有之生態因子如溫度、有機質含量、含水量、導電度、植被之有無皆影響Cr abs 之分佈，由胃合物分析，知Portunidae是肉食性，Grapsidae 是雜食性，而Ocyp odidae及Mictyridae則是草食性。 Activity cycle因Crabs 之種類而異，一般而言，Grapsidae 比Ocypodidae早出現， 且活動時間較長Grabs population，經由統計分析，知名區Crabs ，大多呈Clumped distribution彼此間有重大差異，種與種之間，排斥性強，在同種之間，其♀♂混合 分佈，彼此數目成正相關。

Plane-Stress Model for Piezoelectric Phononic Crystals

聲子晶體的頻帶間隙特性造就許多有效的新裝置。本文研究具平面應力特性的二維壓電聲子晶體的波傳頻譜，基於平面應力忽略厚度方向上位移在動態行為下慣性造成的影響，所以提出了一修正平面應力模式，以期能適用於更大的頻率範圍。 首先利用平面應力條件配合上下表面電性的邊界條件，找出厚度方向上因為泊松效應造成的應變及相對應的厚度方向上的位移，使用漢彌爾頓原理找出此狀況下的統御方程式。再利用平面波展開法推導其波傳理論，使用倒晶格向量配合布洛赫理論對相關的材料常數與位移作傅立葉展開，代入統御方程式，推導出一廣義特徵方程式，利用數值方法可解出特徵值和特徵向量，由此求得二維壓電週期結構之波傳頻譜與位移場形。 使用前述理論推導，針對不同材料組成、厚度以及電性邊界下，探討其對頻帶間隙的影響。發現未修正模式與修正模式下低頻的色散曲線並無太大差異，但在高頻時修正平面應力模式下的色散曲線則會隨著厚度的增加而往低頻移動；並探討各個頻帶的位移場形圖，進一步瞭解各材料的運動情形，提供更多可資應用的特性。Possessing the properties of band gaps, phononic crystals have led the invention of many new devices. In order to handle this characteristic potential, the spectrum of a plane-stress piezoelectric phononic crystal is studied in this thesis. Base on plane-stress assumption neglect the inertia effect of the displacement in the direction of thickness in dynamic situation. Modified plane-stress model is suitable for use in a wider frequency range. First of all, the plane-stress and electric boundary conditions are employed to derive the strain and relative displacement because of Poisson effect, then we derive the governing equation by Hamilton principle. The plane wave expansion method and the Bloch theorem are used to modify the governing equation into the one fit for periodic structures. The material parameters and displacement fields are expanded with Fourier series with respect to reciprocal lattice vectors. Finally, a generalized eigenvalue problem is formed that is solved with numerical method to obtain the frequency spectrum and the displacement fields. The band gaps are found from the frequency spectrum. A study based on changing the materials, thickness, and electric boundary conditions is performed. We find that there is no difference between the dispersion curve of plane-stress model and modified plane-stress model in low frequency range. But the dispersion curve of modified plane-stress model will move down in high frequency range. Finally, by observing the phase of the displacement field, the dynamic situation of materials in the periodic structure is further understood that also provide useful information.目 錄 中文摘要 i 英文摘要 ii 目錄 iii 表目錄 vi 圖目錄 vii 符號表 xi 第一章 緒論 1 1.1研究動機 1 1.2 文獻回顧 1 1.3本文目的與內容簡介 4 第二章 壓電變分原理與修正平面應力模式 6 2.1壓電變分原理 6 2.2平面應力 9 2.2.1修正的平面應力模式 10 2.3壓電材料 12 2.3.1氮化鋁、PZT壓電材料簡介 12 2.3.2平面應力 13 2.3.3電極效應 14 2.3.3.1上下表面有電極 14 2.3.3.2上下表面無電極 15 2.3.4修正平面應力模式 16 2.3.4.1上下表面有電極 16 2.3.4.2上下表面無電極 18 第三章 均質材料結構與週期結構之波傳分析 20 3.1均質材料結構 20 3.2 複合材料結構 21 3.2.1週期結構 21 3.2.1.1倒置晶格向量 21 3.2.1.2布里瓦區 23 3.2.1.3布洛赫理論 23 3.2.2平面波展開法 24 3.2.3材料性質的傅立葉級數係數 27 3.2.3.1兩種材料組成的週期結構 27 3.2.3.2圓形內含物之結構函數 29 3.2.3.3三種材料組成的週期結構 29 3.2.3.4圓形內含物之結構函數 31 第四章 平面應力結構之頻譜 32 4.1 數值方法 32 4.2 均質材料頻譜 33 4.3 平面波展開法之收斂性 34 4.4兩種材料與三種材料組成之週期結構比較 35 4.5 三種材料組成的週期結構之平面應力頻譜 36 4.5.1 不同厚度比較 36 4.5.2 上下表面電極的效應 36 4.5.3 不同壓電基材比較 37 4.6 二維壓電結構模態 38 第五章 結論與建議 40 參考文獻 42 附表 45 附圖 46 附錄 73 表 目 錄 表4.1 前15條色散曲線在n=10和n=9的差異 45 表4.2 上下表面有無電極第五條跟第十條色散曲線的差異 45 圖 目 錄 圖2.1 材料結構在 、 方向延伸， 方向上為有限厚度 46 圖3.1 （a）正方晶格向量（b）倒置晶格向量（c）第一布里瓦區 47 圖3.2 (a）兩種材料組成的二維週期結構示意圖 （b）兩種材料組成週期結構的單位晶胞示意圖 48 圖3.3 （a）三種材料組成的二維週期結構示意圖 （b）三種材料組成週期結構的單位晶胞示意圖 48 圖4.1 修正的平面應力模式下金的色散曲線 49 圖4.2 平面應力模式下金的色散曲線 49 圖4.3 平面應力與修正平面應力模式在不同厚度下金的頻譜圖 50 圖4.4 第一布里瓦區內最小不可重複區域 50 圖4.5 X點上前十個特徵頻率的收斂性質圖 51 圖4.6 （a）Au-PZT上下表面有電極週期結構之波傳頻譜圖 （b）Au-PMMA-PZT上下表面有電極週期結構之波傳頻譜圖 52 圖4.7 （a）Au-PZT上下表面無電極週期結構之波傳頻譜圖 （b）Au-PMMA-PZT上下表面無電極週期結構之波傳頻譜圖 53 圖4.8 （a）上下表面有電極三種材料組成週期結構圖 （b）上下表面有電極平面應力模式頻譜圖 54 (c）上下表面有電極修正平面應力模式 頻譜圖 （d）上下表面有電極修正平面應力模式 頻譜圖 55 圖4.9 （a）上下表面有電極平面應力模式及修正模式 頻譜比較圖 （b）上下表面有電極平面應力模式及修正模式 頻譜比較圖 56 圖4.10 (a）上下表面無電極三種材料組成週期結構圖 （b）上下表面無電極平面應力模式頻譜圖 57 （c）上下表面無電極修正平面應力模式 頻譜圖 （d）上下表面無電極修正平面應力模式 頻譜圖 58 圖4.11 （a）上下表面無電極平面應力模式及修正模式 頻譜比較圖 （b）上下表面無電極平面應力模式及修正模式 頻譜比較圖 59 圖4.12 上下表面有無電極的頻譜結構比較 60 圖4.13 （a）Au-PMMA-PZT上下表面無電極正方晶格之波傳頻譜圖 （b）Au-PMMA-AlN上下表面無電極正方晶格之波傳頻譜圖 61 圖4.14 Au-PMMA-AlN週期結構部分俯視圖 62 圖4.15 Au-PMMA-AlN週期結構第二條色散曲線 時 （a）位移 的實部、虛部 （b）位移 的實部、虛部 63 （c）位移 的振幅、相位 （d）位移 的振幅、相位 64 圖4.16 Au-PMMA-AlN週期結構第二條色散曲線 時 （a）位移 的實部、虛部 （b）位移 的實部、虛部 65 （c）位移 的振幅、相位 （d）位移 的振幅、相位 66 圖4.17 Au-PMMA-AlN週期結構第三條色散曲線 時 （a）位移 的實部、虛部 （b）位移 的實部、虛部 67 （c）位移 的振幅、相位 （d）位移 的振幅、相位 68 圖4.18 Au-PMMA-AlN週期結構第三條色散曲線 時 （a）位移 的實部、虛部 （b）位移 的實部、虛部 69 （c）位移 的振幅、相位 （d）位移 的振幅、相位 70 圖4.19 Au-PMMA-AlN週期結構 時 （a）第二條色散曲線中位移 的實部 （b）第三條色散曲線中位移 的實部 71 圖4.20 Au-PMMA-AlN週期結構第 時 （a）第二條色散曲線中位移 的實部 （b）第三條色散曲線中位移 的實部 7

Substance Flow Analysis of arsenic in Taiwan

砷被廣泛、多元地應用於玻璃製造、木材防腐業及半導體、光電業等多項產業製程、產品中，在經濟活動的使用上有其必要性。但是，砷及其化合物具有致癌及非致癌之毒害特性；因此，在無法有效禁止砷及其化合物的使用下，有必要針對砷進行流布及流向分析、探討。究以物質流分析方法分別對砷於人類經濟活動之應用以及因人類活動所造成砷之環境釋放進行探討，以建構出台灣砷物質流網路模型；再者，利用物質流分析特性探討砷之使用時間趨勢、地區分布以及相關產業結構。最後將物質流分析結果整合投入產出分析方法，以投入產出分析方法連結環境及產業間關係，說明台灣砷產業使用現況，進一步探討砷於產業間投入產出之關聯性。究顯示，經濟圈主要使用量為玻璃製造業、水泥業、鉛蓄電池、木材防腐業以及半導體、光電業等五項產業，在環境砷釋放上，則是以水泥業、煤礦使用、原油煉解以及金屬礦物冶煉為主要污染釋放熱點。將物質流分析結果進行多年趨勢探討，說明砷物質之產業結構變化。較投入產出分析結果，發現非金屬礦物製品、其他金屬、電力等部門為每單位產值所需砷的使用量及環境釋放量較高者。含砷產業之環境負荷放大效應部分，服務業相較於製造業及初級產業有著較高的環境境負荷的累積、放大現象。在產業關聯性分析，發現同樣是以非金屬礦物製品、其他金屬、電力及石油煉製品等產業之前後關聯性較高(包含向前關聯、向後關聯)，以及將產業消費內容回歸到家計消費，發現在住、行及服務等產業類別有著高環境砷釋放量及使用量。最後比較砷之PTB、區域投入產出及消費型態之地區分布，發現高生活消費地區(如台北市、台中市、高雄市)之低環境砷釋放量，而部分低消費地區(花蓮縣、雲林縣)卻負擔高砷相關產業，即承受高環境砷負荷量，呈現出區域間存在著環境不公的現象。Arsenic and its compounds are widely applied in multiple industrial processes and products such as glasses, semiconductors and optoelectronics production, and wood preservatives. However, arsenic and its compounds have been characterized with both carcinogenic and non-carcinogenic toxicities. Therefore, flows of arsenic should be nalyzed because of their indispensability in a wide range of industrial processes. his study applied material flow analysis to investigate both the application of arsenic in human economic activities and the environmental release of arsenic due to human activities to build up the material flow network model of arsenic. The study also investigated the temporal pattern of arsenic usage, spatial distribution, and related industrial structure by using material flow analysis. Furthermore, the esults of material flow analysis were integrated with Input-Output Analysis method to link environmental burden with industrial structure. hrough the integrated analysis, current status of arsenic usage in Taiwan can be interpreted, and the input-output correlation of arsenic among industries can be further identified. ased on the results of material flow analysis, arsenic and its compounds are mainly applied in glasses, cement, lead acid batteries, semiconductors and optoelectronics production and wood preservatives industries in the anthroposphere. As for the environmental release, major eleasing hotspots are cements production, coal burning, oil and metal mineral refining processes. Multiple-year trend of arsenic flow were further dipcted to examine the industrial structural change in arsenic use. ccording to the results of input-output analysis, the sectors of non-metal mineral products, other metals, and electricity have greater arsenic use and environmental release per output value. As for amplification effect of environmental burdens from arsenic-containing ndustries, service industry revealed greater accumulation and amplification effects of environmental burdens than manufacturing and primary industries. The results also showed that the sectors of non-metal mineral products, other metals, electricity, and petroleum refining products have greater industrial linkages, including forward and ackward linkages. When industrial consumption was allocated into household consumption, housing, transportation, and service industries generated greatest arsenic use and environmental releases. Finally, omparing data of physical trade balance (PTB) of arsenic, regional input-output analysis and distribution of consumption patterns in different regions in Taiwan, high consumption regions (e. g. Taipei City, Taichung ity, and Kaohsiung City) were found to bear lower arsenic nvironmental releases, whereas some low consumption regions (e. g. Hualien County and Yunlin County) bear high environmental burden of arsenic due to industries with high arsenic intensity within these regions. The results demonstrated the environmental inequity among regions in Taiwan.頁次 要 ................................................................................................................................. i bstract ........................................................................................................................ iii 目錄 ........................................................................................................................ viii 目錄 ............................................................................................................................ x 一章 緒論 .................................................................................................................. 1 .1 研究背景 ........................................................................................................ 1 .2 研究目的 ........................................................................................................ 3 .3 研究架構 ........................................................................................................ 4 二章 文獻回顧 .......................................................................................................... 6 .1 砷之基本特性 ................................................................................................ 6 .1.1 砷及其化合物的物理化學性質 .......................................................... 6 .1.2 砷的毒理特性 ...................................................................................... 8 .1.3 砷的應用與經濟圈釋放 .................................................................... 10 .1.4 環境中砷的流布 ................................................................................ 11 .2 物質流分析 .................................................................................................. 14 .2.1 物質流概念 ........................................................................................ 14 .2.2 物質流分析之應用 ............................................................................ 14 .2.3 物質流之類別及步驟 ........................................................................ 15 .3 投入產出分析方法 ...................................................................................... 24 三章 研究方法 ........................................................................................................ 27 .1 研究步驟 ...................................................................................................... 27 .2 經濟圈內之物質流分析 .............................................................................. 29 .2.1 砷物質之網路模型 ........................................................................... 31 .2.2 含砷資料盤查分析 ........................................................................... 34 .3 環境圈物質流及釋放資訊 .......................................................................... 39 .3.1 空氣 ................................................................................................... 39 .3.2 水體 ................................................................................................... 40 .3.3 廢棄物 ............................................................................................... 40 .3.2 資源回收 ........................................................................................... 41 .4 結合經濟圈及環境圈之砷物質流 .............................................................. 42 .4.1 砷之生命週期模型建立 ................................................................... 42 .4.2 砷物質流結果分析及質量平衡 ....................................................... 46 .5 物質流投入產出分析方法 .......................................................................... 47 .6 物質流及相關指標 ...................................................................................... 52 四章 結果與討論 .................................................................................................... 55 .1 砷物質流結果分析 ...................................................................................... 55 .1.1 砷之經濟體流布及流量 ................................................................... 55 .1.2 砷之環境釋放分析 ........................................................................... 63 .1.3 結合經濟圈及環境圈之砷物質流分析結果 ................................... 68 .2 砷物質流之時間因子分析 .......................................................................... 72 .3 含砷物質進出口量之地區分布 .................................................................. 78 .4 砷物質產業活動與需求之環境衝擊 .......................................................... 82 .4.1 單位最終需求環境排放潛勢 ........................................................... 82 .4.2 砷應用產業之環境釋放關聯分析 ................................................... 85 .4.3 產業之乘數效應 ............................................................................... 88 .4.4 最終需求與環境衝擊分析 ............................................................... 89 .4.5 家計需求之砷物質之使用及環境釋放分布 ................................... 90 .5 各縣市砷物質使用及環境釋放比較 .......................................................... 91 .5.1 砷物質之消費型態分布 ................................................................... 91 .5.2 砷物質之區域 IO環境釋放分布 ..................................................... 93 .6 誤差探討 ...................................................................................................... 95 .1 結論 .............................................................................................................. 96 .2 建議 .............................................................................................................. 98 考文獻 ...................................................................................................................... 99 錄一 含砷物質之國內產銷 .................................................................................. 104 錄二 含砷物質之進出口資料 .............................................................................. 106 錄三台灣砷物質貿易趨勢 .................................................................................... 111 錄四 產業關聯部門定義 ...................................................................................... 113 錄五 消費支出調查 .............................................................................................. 11

Technical Development and Research Improvement in Biological Control of Plant Pests (II)

環保意識的抬頭與人類對食品安全的關注,使得農作物病蟲害的生物防治工作受到社會大眾的重視,因此,本計畫目的在於研發作物病蟲害生物防治策略,以便能有效在農業生態體系中推展應用.本計畫涵蓋有九個子計畫,它們個別的工作目標如下:即 (1) 由夏威夷引進銀葉粉蝨的天敵Encarsia pergandiella, E. hispida及E. mineoi,探討它們防蟲之功效與飼養、檢疫等工作; (2) 探討斑潛蠅發生生態,調查本土現有斑潛蠅之天敵種類及引進有效的寄生性天敵,作為生物防治之用; (3) 評估黑殭菌各菌株致病力與抗環境逆境;利用黑殭菌DNA多元性特定條帶作出DNA-probe,供田間生態及施用存活調查; (4) 探討大量飼育捕植蹣類天敵,建立捕植對卵孢子發芽、感染與傳播的影響;施用增產菌FG-1菌株與滅達樂,以期延緩病害發生與促進薑株生長; (6) 利用本地產蟲生線蟲(Steinernema abbasi),研發出適合台灣蔬菜害蟲之安全有效的生物防治技術; (7) 觀察飼育紅點唇瓢蟲之基本需求,評估其生物防治梨圓介殼蟲之潛力; (8) 利用生物分析法篩選具有抑制病原菌的作物殘渣與植體,並輔以HPLC或GC分析它們所含有的硫配糖體種類及分解產物,藉以瞭解各分解產物的殺菌與抑菌功效; (9) 針對本地種蟲生線蟲Steinernema abbasi對蔬菜主要害蟲如小菜蛾、白粉蝶、大菜螟及菜心螟蟲的致病力進行生物檢定與施用技術.Biological control of plant pests has received considerable attention throughout the world, drawing momentum from the growing public concern regarding the widespread use of hazardous chemicals in pest control. In order to develop and apply biocontrol strategies in agriecosystem, the objectives of this project including nine sub-projects are (1) to import a whitefly parasitoids, Encarsia pergandiella, E. hispida and E. mineoi from Hawaii and study their efficacy on controlling the silver leaf whitefly; (2) to study the distribution, occurrence, seasonal abundance, population dynamics and damage of leafminers in southern area and test the basic biology, parasitic rates of introduced parasitoids; (3) to screen Metarhizium anisopliae strains having high entomopathogenicity, more resistant against UV light and the fungicides; to analyze the genetic diversity and relationship of the tested green mascardine fungi by using electrophoretic fingerprinting; (4) to develop biocontrol technologies of the spider mites with their predaceous mites, which depends upon the availble mass rearing skills and their lowering costs determined by the design of artificial rearing arena and supplemental foods; (5) to study the ecology of ginger soft rotdisease including the transmission of the disease through seed ginger; the temperature effect on the oospore germination, infection and spreading. Also, to study the use of both metalaxyl and YIB (yield increasing bacteria) isolate Fg-1 to delay the disease development and increase the plant growth; (6) to investigate biological control of vegetable pests using the indigenous species, Steinernema abbasi, in safe and effective control measures; (7) to study the basic rearing requirement for Chilocorus kuwanae and to provide a preliminary assessment of its potential as a biological agent of the San Jose Scale, Quadraspidiotus perniciosus; (8) to determine the effect of volatile compound released from leaf and seed of Brassica species on the natural isothiocynate fumigant for integrated control of crop soilborne diseases; (9) to develop a vermiculite, formulation of Steinernema abbasi to control four vegetable pests, such as diamondback moth, imported cabbageworm, cabbage pyralid and cabbage webworm

Development of Techniques for Rapid Identification of Pest Organisms on Animals and Plants

1. Identification techniques and information compilation of arthropod vectors transmitting important infectious diseases of animals and man: The objective of this project is to collect, compile and analyze all the domestic and international research information regarding vector-borne infectious diseases (including zoonoses) and their transmission arthropod vectors. A book with a title of “Introduction to arthropod vectors and the infectious diseases they transmitted” will be published by the Bureau of Animal and Plant Health Inspection and Quarantine timely. This book will be beneficial to the professional of all related field. 2. Development and application of the integrated diagnostic kit for citrus systemic diseases: The systemic pathogens such as the bacteria causing citrus huanglongbing, Citrus tristeza virus (CTV), Citrus tatter leaf virus (CTLV) and Citrus exocortis viroid (CEVd) are predominant and destructive to citrus industry in Taiwan. These pathogens are mainly transmitted by vegetative propagation. Establishment of pathogen-free nursery systems and development of rapid and sensitive diagnostic probes are considered to be the most important methods for the pest control. This research project is dedicated to develop new diagnostic probes and improve several devised diagnostic methods using immunological and molecular techniques. An integrated diagnostic kit will be produced through combination of all our devised detection methods for these citrus systemic pathogens. This kit will be helpful to the inspection and management of the citrus pests, and it is worthy of international patent for world-wide extension. 3. Development nano-particle-sensitized QCM sensor for the real-time diagnosis and identification of pathogens in animals and plants: Utilizing gold nanoparticle binding with virus to increase virus weight, overcome detection limit and enhance detection response. This project will carry out the following important works this year: 1. Preparation of Gold nanoparticle; 2. To obtain and prepare sample; 3. To combine Gold nanoparticle and virus; 4. To develop mechanisms of sample's verifications and tests. 4. Development of techniques for rapid discrimination of various insect pests in imported grain: The goal of this project is to develop species-specific PCR primers for rapid discrimination of insect pests in imported grain. The approach will be founded on using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Firstly, to compare the RAPD-PCR profiles of 3 major lepidopterous pests of grains, i.e. Plodia interpunctella H., Sitotroga cerealella O., and Cadra cautella; secondly, to isolate the species-specific amplicons from the gel and to further clone and sequence; thirdly, to design specific PCR primers based on the sequence; finally, to test the specificity and stability of PCR reactions with the designed primers. By the end of the research, we expect to develop 2-3 pairs of species-specific PCR primers for rapid species discrimination of each of these 3 pests. Furthermore, we also expect that this method is able to expend to be used on quarantine inspection of imported grain and to avoid the possible invasion of these pests. 5. Development of Molecular Diagnostic method for the rapid differentiation of Baylisascaris procyonis and Toxocara canis eggs: Baylisascaris procyonis is commonly found in raccoon and has been recognized as an emerging helmintic zoonosis in recent years. When B. procyonis embryonated eggs are ingested by a host other than the raccoon, larva migrans ensues. In human, this larval can invade the brain and eye, causing severe disease and sometimes death. Clinical findings include eosinophilic meningoencephalitis, eosinophilic cardiac pseudotumor and chorioretinitis. B. procyonis can also infect dog, using it as an intermediate host as well as final host. The infected dog can also shed B. procyonis eggs, thus posing a diagnostic problem for those dogs that are infected with Toxocara canis. Apart from the size difference, B. procyonis eggs and T. canis eggs are quite morphologically similar. Thus, it is important to develop a molecular biological technique to differentiate the two types of eggs. We propose to develop a molecular diagnostic method, using PCR, for the rapid differentiation of B. procyonis and T. canis eggs. 6. Application of bio-Nanotechnology for diagnosis and detection of Lily mottled virus in lilium: In recent years, the development of molecular biology is fast. Now many kinds of nucleic acid probes and polymerase chain reaction (PCR) methods have been applied to identify and detect the plant pathogens because of the fast, uniqueness and sensitive characteristic of biotechnology. Although the sensitivity of PCR is up to 50-100 fg, but it still needs a professional technician for operation and takes approximately 6.5 hours, and its cost is expensive. So it is difficult for widely application on detection of pathogens. Therefore, a rapid, simple, and high sensitive technique with low expenses for detection of plant pathogens is necessary. The aim of the present study was to develop an ultra-sensitive and excellent photostability probe for detection of Lily mottle virus in Lilium by using bio-nanotechnology. 7. Development of Ehrlichiosis Serodiagnosis: Ehrlichiosis was Zoonosis in dog and human. E. canis and E. chaffeensis reside and replicates within the mononuclear cells, the clinic signs were included: weight loss, anemia, lymphadenomegaly, and anorexia. This study will utilize bioinformatics to predict the hydrophilic region (HR) of E. canis outer membrane protein Omp30, and then expressed the Omp30HR and NucA-Omp30HR in E. coli expression system. They will be purified and quantities with Ni+ column and ELISA, separately. These results enable us to utilize Omp30HR or NucA-Omp30HR for Dot-ELISA to clinic serodiagnosis of dog or human ehrlichiosis. We will develop the good, convenient, sensitive, fast and cheap Dot-ELISA kit, in order to remote the quarantine epidemic prevention plan of Zoonosis. 8. The identification and detection of brown rot pathogens on peach: Brown rot disease is one of the most disruptive diseases on stone fruits worldwide and has been reported in Taiwan. This disease can be caused by Monilinia fructicola, M. laxa, M. fructigena and Monilia polystroma. M. fructicola and M. laxa attacks mainly stone fruit, while M. fructigena attacks both stone and pome fruits but is found more frequently in the latter. Each brown rot pathogen of stone fruits has a different regulatory status. For example, M. fructigena is listed as a quarantine pathogen in the USA and in Australia. To understand the species of brown rot pathogens exist in peach orchards in Taiwan and to monitor the species of brown rot pathogens on imported fruits, a fast and accurate identification method to identify the species of brown rot pathogen needs to be established. In this study, we will isolate brown rot pathogens of peach from different peach growing areas in Taiwan and from imported stone fruits including cherry, peach and nectarine. The preliminary identification of those collected cultures will be performed based on the colony morphology, conidial size and sporulation type. A PCR based identification method with specific molecular markers will be developed to identify the species of collected brown rot pathogens. The genetic variation of brown rot pathogens within those collected from orchards in Taiwan and those from other countries will also be studied. 9. Application of liquid array for detection of avian Mycobacteriosis in zoo birds: Mycobacterium avium is pathogenic to most avian including domestic birds, pet birds, cage birds, wild birds, and zoo birds but most frequently infects domestic birds. It has also been reported in swine, rabbit, deer, cattle, lamb, and marten and is recognized as an important zoonosis. Histopathologically, multifocal nodules were observed in many organs. The goal of this study is to establish liquid array in combination with pathological examination, Ziehl-Neelsen staining, xMAP, and nucleotide sequencing to detect avian tuberculosis. 10. Development of immunoproteomics platforms to identify candidate antigens of classical swine fever virus for discriminating diagnosis: Classical swine fever (hog cholera) is a highly contagious disease of pigs and caused by classical swine fever virus (CSFV). The commonly used vaccine in Taiwan against hog cholera is an attenuated LPC strain of CSFV that made from type 820 CSFV. At least 141 CSFV strain have been isolated in the wild field of Taiwan. Base on their genomic sequences, all the CSFV can be divided into three genotypes; LPC strain belongs to genotype 1 and wild field CSFV belongs to genotype 2. Because LPC strain is different from wild field CSFV in the genotype and vaccination or infection pathways, and that may induce different antibody responses. In this plan, we would like to identify the differences of candidate antigens between two genotypes of CSFV by immunoproteomic technique, and then use to distinguish between LPC-vaccinated and wild CSFV infected pigs. Sera from pigs with LPC vaccine or wild field CSFV infections and control group of sera from health individuals will be analyzed for reactivity by western blot against LPC virus or wild field CSFV total proteins separated by 2-dimemional electrophoresis. We predict several immunogenic protein spot will be identified by immunoproteomic analysis, MALTI-TOF and LC-MS-MS. In the future, the candidate antigens will be developed to a rapid and easy diagnosis kit and use to discriminate between LPC vaccinated and wild CSFV infected pigs.1. 媒介重要人畜傳染病之有害生物鑑定資訊與技術建立：本計畫主要收集並分析有關國內外媒介重要人、家畜及人畜共通傳染病節肢動物的研究資料，再加上這些傳染病基本資訊，邀集國內學術單位等專家撰稿審查，依進度出版「媒介重要人畜傳染病的有害生物-節肢動物篇」一書，以提供動植物防檢疫局和相關單位人員參考。使我國動物疫病防疫檢疫單位有完整的參考資料，對有害生物尤其是媒介傳播人畜傳染病生物媒介之鑑定編輯成冊以提供國內執行防檢疫人員參考。 2. 柑橘系統性病害整合性診斷試劑套組之開發與應用：台灣重要柑橘系統性病原包括黃龍病細菌、萎縮病毒(CTV)、破葉病毒(CTLV)與鱗砧次病毒(CEVd)等，這些病原經由帶病種苗普遍傳播而危害嚴重，生產無病種苗搭配敏感且正確的檢疫技術乃是防治這些病害的首要之道。本計畫以原本初步研發成功的柑橘病原檢疫技術為基礎，進一步再研發與改善，目標在於建立一套方便且快速的整合性偵測套組。具體工作項目包括： (1) 製備出柑橘破葉病毒之單元抗體； (2) 研發CEVd快速RT-PCR偵測技術； (3) 改良黃龍病細菌、CTV、CTLV之分子偵測法等。所得研究成果不但可供國內防檢疫之應用，更因這些病原皆為國際性重要病原，未來可申請專利而推展於國際。 3. 開發奈米粒子敏感化石英晶片感測器以即時診斷鑑定動植物有害生物：利用奈米金粒子(gold nanoparticle, GNP)和病毒結合，以增加病毒本身重量，以突破QCM的感測極限及增加偵測反應。本計畫今年將執行下列重要工作： (1) 製備金奈米粒子(Gold nanoparticle, GNP)； (2) 樣本取得及製備； (3) 奈米金粒子與病毒結合； (4) 樣本檢測驗證機制。 4. 進口大宗穀物害蟲之快速鑑定技術開發：本研究的主要目的在於利用聚合酶連鎖反應-逢機增幅多態型DNA (random amplified polymorphic DNA-polymerase chain reaction, RAPD-PCR)的方法，研發及尋求特定的DNA標誌，以供快速檢測與鑑定積穀害蟲之用。本計畫主要將先利用RAPD技術比對三種主要鱗翅目積穀害蟲，包括印度穀蛾(Plodia interpunctella H.)、麥蛾(Sitotroga cerealella O.)與粉斑螟蛾(Cadra cautella)等，尋求具種特異性的DNA片段，再進而依此片段之序列設計具有蟲種專一性的引子，並更進一步檢測其專一性與穩定性，以提高鑑定的準確度。預計本研究結束時，可建立利用PCR快速檢測上述三種積穀害蟲的方法，將可作為日後應用此技術以監測大宗進口穀物是否夾帶外來種之鱗翅目害蟲，進而有效地防堵外來害蟲種入侵的機會。 5. 開發分子生物診斷法來快速鑑定及識別浣熊蛔蟲與犬蛔蟲：浣熊蛔蟲主要寄生於浣熊，近幾年來被視為一種的新興疾病。當含有仔蟲的蟲卵被浣熊以外的宿主攝入時，會造成幼蟲移行症。當人被感染時，幼蟲可能移行至腦部及眼球而造成嚴重的臨床症狀甚至死亡。臨床症狀包括嗜酸性球性腦膜腦炎、嗜酸性球性心臟偽腫瘤及脈絡膜視網膜炎等。犬隻可擔任浣熊蛔蟲的中間宿主及終宿主。犬隻感染浣熊蛔蟲或犬蛔蟲時皆會排出蟲卵，因而造成鑑定上的困難。兩種蛔蟲之蟲卵的外觀除大小之差異外，在型態學上非常相似。因此開發分子生物診斷法來快速鑑定及識別有其重要性。本計畫將利用PCR法來建立快速診斷法鑑別浣熊蛔蟲及犬蛔蟲。 6. 生物奈米技術在百合斑駁病毒病害診斷與偵測之運用：近年來，分子生物學的發展迅速，生物技術因其具有快速、專一性及靈敏的特性，目前已有多種核酸探針及聚合酵素連鎖反應(PCR)技術被發展應用於鑑定及偵測植物病原菌的研究。然而PCR偵測DNA的靈敏度雖高達50-100 fg，但需要專業技術人才的操作、耗時6.5小時及費用昂貴，所以不易推廣。因此，發展一快速、簡易、高靈敏度且費用低的作物疫病檢測技術，亦有其必要性。本研究擬利用生物奈米技術(bio-nanotechnology)研發具高靈敏度與高光穩定度的百合斑駁病毒螢光奈米診斷探針，並能在短時間內迅速檢測出百合斑駁病毒，供為植物病害檢疫之參考。 7. 艾利希體症檢測技術之開發：艾利希體症是犬及人等之人畜共通傳染病，其病原體主要寄生於單核球，臨床症狀包括體重下降、貧血、淋巴腫大、厭食等症狀。故本試驗將延續先前之研究成果，將犬艾利希體之外膜蛋白Omp30利用生物資訊學進行抗原親水區(hydrophilic region; HR)之預測，再以大腸桿菌表現系統大量表現、純化及定量後，使我們可利用具親水區的Omp30HR或NucA-Omp30HR，作為艾利希體之感染檢測試劑組，並進行艾利希體症病犬之臨床篩檢，以開發具備良好、簡便、靈敏、快速且便宜的檢測試劑，而能有效推動人畜共通傳染病之檢疫防疫計畫。 8. 桃褐腐病菌之診斷偵測：桃褐腐病為對核果類植物最具破壞性之病害之一，此一世界性病害亦曾在台灣被報導過。此病害可由M. fructicola, M. laxa, M. fructigena及M. polystroma感染造成。Monilinia fructicola及M. laxa主要感染核果類植物，而M. fructigena主要感染蘋果及梨，但亦可侵入桃等核果類植物。每種菌各有其被管制之狀態，例如M. fructigena為美國及澳洲兩國禁止輸入之檢疫病菌。為了瞭解國內桃果園所存在及由國外進口水果所攜帶之褐腐病菌種類，必須建立一快速而準確的檢定方法。本研究將收集分離自國內桃果園及進口水果之褐腐病菌，初步菌種鑑定將以培養特性、孢子大小及產孢形態等為依據。以PCR反應為主之分子診斷方法將被建立以了解國內褐腐病菌之種類並可為進出口核果類水果、蘋果及梨等檢測用。本計畫亦將進一步進行國內外桃褐腐病菌株分子層次差異之比對以了解國外菌株與國內菌株之親緣性。 9. 禽類結核病(avian tuberculosis)由鳥型結核桿菌(Mycobacterium avium)所引起之慢性、接觸性、漸進性傳染病，以消化道病變為主，為人畜共通傳染病。本菌對鳥類、豬、貂、兔子、綿羊、鹿、牛等均具敏感性，病禽呈漸進性消瘦、羽毛蓬鬆、下痢等症狀，各臟器均可見多發白色結節，死亡率高。 本計畫擬建立微珠核酸探針技術。輔以病理學檢查、Ziehl-Neelsen氏抗酸染色法、xMAP (Flexible Multi-analyte Profiling)及核酸定序，檢測為禽類結核病。 10. 建立豬瘟疫苗免疫與野外感染區別診斷之技術平台：目前市面上豬瘟疫苗種毒820病毒株，都是由家畜衛生試驗所統一供應，再經動物疫苗廠製作成兔化豬瘟疫苗。台灣至今所收集到的野外豬瘟分離毒共有141株，經核酸部份定序後約可歸類為三種不同的基因型，兔化豬瘟疫苗屬於第一基因型；而現階段田間流行的豬瘟病毒則歸屬於第二基因型。由於接種疫苗及田間感染的病毒株，屬於不同基因型; 田野豬瘟病毒是自然接觸性感染，而兔化豬瘟疫苗是經肌肉注射感染。基於上述兩種不同點，我們認為兔化豬瘟疫苗和田間自然感染病毒，會引起動物產生不同的抗體反應。利用免疫蛋白體學的相關技術，可以將其相對的抗原找出來，抗原找出來後，可利用此抗原製成檢驗試劑，用以區分非感染、野病毒株感染或接種兔化豬瘟疫苗的豬隻，而本年度計畫目標就是企圖找出這樣有差異性的豬瘟抗原。我們預期若實驗順利將可得到非感染、野病毒株感染或接種兔化豬瘟疫苗豬隻的血清抗體可結合的抗原圖譜。經統計分析非感染、野病毒株感染或接種兔化豬瘟疫苗豬隻的血清反應圖譜，找到顯著差異的抗原或未曾發現的抗原。新發現的抗原，可利用MALTI-TOF及LC-MS-MS等技術，定其氨基酸序列，並反推其相對DNA序列，鑑定其身份。之後的延續計畫可利用基因工程法，構築質體在E. coli內大量生產抗原，純化抗原做為檢驗試劑用以鑑別分非感染、野病毒株感染或接種兔化豬瘟疫苗的豬隻