本文關(guān)鍵詞：小睪并殖吸蟲(chóng)的形態(tài)學(xué)及DNA序列分析　出處：《昆明醫(yī)學(xué)院》2009年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 小睪并殖吸蟲(chóng) 形態(tài)學(xué) DNA序列分析 ITS2 CO1

【摘要】： [目的]應(yīng)用形態(tài)學(xué)分類及DNA序列分析相結(jié)合的方法深入研究小睪并殖吸蟲(chóng)(Paragonimus microrchis Hsia Zhou Chang,1978)的蟲(chóng)種獨(dú)立性,在并殖科吸蟲(chóng)中的分類學(xué)地位,以及與其它并殖吸蟲(chóng)蟲(chóng)種的親緣關(guān)系,逐步澄清并殖科吸蟲(chóng)的蟲(chóng)種爭(zhēng)議,探討更加客觀和易于鑒定的分類學(xué)方法,并為進(jìn)一步研究各蟲(chóng)種的生物學(xué)特性、地理分布、對(duì)人體的致病性,以及開(kāi)展相關(guān)的防治工作奠定重要基礎(chǔ)。 [方法]從小睪并殖吸蟲(chóng)流行區(qū)采集第二中間宿主,分離囊蚴、脫囊蚴、感染實(shí)驗(yàn)動(dòng)物獲得成蟲(chóng)和蟲(chóng)卵。通過(guò)生物體視鏡、顯微鏡觀察囊蚴、后尾蚴、成蟲(chóng)及蟲(chóng)卵形態(tài),并測(cè)量其大小;掃描電鏡觀察成蟲(chóng)的皮棘形態(tài)特征。結(jié)合DNA序列分析,Blast軟件同源性比較,及MEGA4.0軟件構(gòu)建基因進(jìn)化樹(shù)。 [結(jié)果]小睪并殖吸蟲(chóng)成蟲(chóng)活體呈肉紅色,固定后呈磚灰色,封片標(biāo)本測(cè)量:蟲(chóng)體大小為8.5-12.9×3.6-5.0mm,平均10.47±1.94×4.35±0.74mm。寬長(zhǎng)之比1:2.1-2.8,平均1:2.41�？谖P橫徑為0.511-0.778mm,平均0.657±0.103mm。腹吸盤位于體前1/3與中1/3橫線處或稍后緣,大小為0.648-0.821×0.648-0.828mm,平均0.748±0.107×0.745±0.102mm。兩吸盤橫徑之比為1:1.13。卵巢位于腹吸盤的左或右下方,大小為0.720-1.598×0.662-1.397mm,平均1.238±0.475×1.032±0.376mm,自中心體發(fā)出6-8葉,每葉又可發(fā)出3-4小葉。睪丸位于體中1/3與后1/3橫線處或前緣,無(wú)分支或具1-5個(gè)芽狀突起,左睪大小為0.662-1.390×0.504-0.979mm,平均0.974±0.334×0.732±0.250mm;右睪為0.526-1.570×0.504-1.210mm,平均0.989±0.437×0.701±0.344mm。睪丸明顯地小于卵巢。掃描電鏡觀察成蟲(chóng)的皮棘,除口、腹吸盤之外均披有單生皮棘,蟲(chóng)體前中部的皮棘密集,排列規(guī)則,體后部的稀疏,排列不規(guī)則,形狀似尖刀狀或三角型�？谖P內(nèi)壁為縱向皺襞,腹吸盤內(nèi)壁為環(huán)形皺襞。 小睪并殖吸蟲(chóng)后尾蚴呈長(zhǎng)橢圓形,排泄囊巨大,位于兩腸支間。封片標(biāo)本測(cè)量其大小0.878-1.210×0.432-0.634mm,平均1.034±0.167×0.509±0.108mm�？谖P橫徑為0.115-0.180mm,平均為0.140±0.025;腹吸盤大小為0.130-0.202×0.130-0.209mm,平均為0.170±0.027×0.173±0.031mm,口腹吸盤之比平均為1:1.236。 小睪并殖吸蟲(chóng)囊蚴呈球形,個(gè)體巨大,具有雙層囊壁,外壁薄而透明,易破裂,內(nèi)壁較厚。蚴體常蜷曲充滿于內(nèi)層囊腔內(nèi),可見(jiàn)蚴體盤曲而折光的腸支,排泄囊呈袋狀位于兩腸支間。未加蓋玻片測(cè)量其大小(有外壁)為0.760-1.406×0.689-1.399mm,平均1.006×0.951mm,外壁厚度為0.003-0.014mm,平均為0.008mm。未加蓋玻片測(cè)量只有內(nèi)壁的囊蚴的大小為0.632-0.852×0.596-0.831mm,平均為0.728×0.703mm,內(nèi)壁厚為0.007-0.031mm,平均為0.011mm。 小睪并殖吸蟲(chóng)蟲(chóng)卵一般呈寬橢圓形,金黃色,大多數(shù)卵蓋明顯,卵殼薄厚不均,且末端增厚。加蓋玻片測(cè)量其大小為73.1-95.9×42.3-55.3μm,平均為81.37±9.90×47.67±5.58μtm。 通過(guò)Blast同源性比較,小睪并殖吸蟲(chóng)與老撾哈氏并殖吸蟲(chóng)的ITS2和CO1同源性分別為100%和99%,與泰國(guó)曼谷并殖吸蟲(chóng)同源性分別為98%和96%,與中國(guó)浙江的哈氏并殖吸蟲(chóng)同源性分別為96%和88%。ITS2和CO1基因進(jìn)化樹(shù)顯示小睪并殖吸蟲(chóng)與老撾的哈氏并殖吸蟲(chóng)聚為一支、此支再與泰國(guó)曼谷并殖吸蟲(chóng)聚為一支,此大支再與中國(guó)浙江的哈氏并殖吸蟲(chóng)聚為一支。 [結(jié)論]小睪并殖吸蟲(chóng)成蟲(chóng)、后尾蚴、囊蚴、蟲(chóng)卵的形態(tài)與夏代光教授等(1978年)、周本江教授(1988年)描述的各期形態(tài)一致,符合小睪并殖吸蟲(chóng)的形態(tài)特征。首次應(yīng)用掃描電鏡觀察小睪并殖吸蟲(chóng)成蟲(chóng)皮棘,為單生型,形狀似尖刀狀或三角型,體前中部的皮棘密集,排列規(guī)則,體后部稀疏,排列不規(guī)則�？谖P內(nèi)壁為縱向皺襞,腹吸盤內(nèi)壁為環(huán)形皺襞,其形態(tài)結(jié)構(gòu)的特點(diǎn)與并殖吸蟲(chóng)的攝食、附著及運(yùn)動(dòng)功能有關(guān)。就基因序列而言,小睪并殖吸蟲(chóng)與老撾的哈氏并殖吸蟲(chóng)為同一蟲(chóng)種,老撾的哈氏并殖吸蟲(chóng)的真實(shí)性有待進(jìn)一步澄清。小睪并殖吸蟲(chóng)與泰國(guó)曼谷并殖吸蟲(chóng)同源性較高,親緣關(guān)系較近,按照Blair.D的觀點(diǎn)小睪并殖吸蟲(chóng)與泰國(guó)的曼谷并殖吸蟲(chóng)應(yīng)為同一蟲(chóng)種的復(fù)合型(Complex)。
[Abstract]:[Objective] the application of analysis method of morphological classification and DNA sequence combination research of Paragonimus microrchis (Paragonimus microrchis Hsia Zhou Chang, 1978) of the species independence, the taxonomic status of Paragonimus trematodes of Paragonimus and other kinds of relationship, species gradually clarify the dispute Paragonimus trematodes, taxonomic studies method is more objective and easy to identify, and for the further study of the biological characteristics, species geographical distribution, the pathogenicity of the human body, and carry out the relevant prevention and control work to lay an important foundation.
[Methods] from the testis of the intermediate host of Paragonimus epidemic area were collected second, separated metacercaria, removal of metacercariae, adult worms and eggs for animal experimental infection. Through the observation of organisms mirror, metacercariae, microscope after cercariae, egg and adult form, and measure its size; observe the morphological characteristic spines of adult worms were combined with scanning electron microscopy. Sequence analysis of DNA Blast software, homology, phylogenetic tree construction and MEGA4.0 software.
[results] Paragonimus microrchis adult living donor was red meat, after a fixed gray brick, mounting were measured: body size is 8.5-12.9 * 3.6-5.0mm, average 10.47 + 1.94 * 4.35 + 0.74mm. width to length ratio of 1:2.1-2.8, the average 1:2.41. oral sucker 0.511-0.778mm diameter, average 0.657 + 0.103mm. ventral sucker in the front 1/3 and 1/3 in the horizontal line or later edge, the size of 0.648-0.821 * 0.648-0.828mm, average 0.748 + 0.107 * 0.745 + two 0.102mm. sucker 1:1.13. diameter of ovarian in the ventral sucker left or right below the ratio of the size of 0.720-1.598 * 0.662-1.397mm, average 1.238 + 0.475 * 1.032 + 0.376mm, since the centrosome sends 6-8 leaves, each can send 3-4 leaflets. Leaves testicles in the 1/3 and 1/3 lines in the body or at the front, no branch or 1-5 bud like protrusions, left testis size is 0.662-1.390 * 0.504-0.979mm, average 0.974 + 0.334 * 0.732 + 0.250mm; right testis 0.526-1.570 * 0.504-1.210mm, an average of 0.989 + 0.437 * 0.701 + 0.344mm. testis significantly less than ovary. The spines were observed with scanning electron microscope, in addition, with a single ventral sucker beyond rawhide spines, the worm before the central spines arranged regularly, dense, sparse body back, irregular arrangement, shaped like a knife. Oral sucker or triangle. The inner wall is longitudinal folds, ventral sucker wall ring folds.
After cercariae of Paragonimus microrchis elongatedoval, excretory bladder is huge, in two. The intestinal branch mounting samples to measure the size of 0.878-1.210 * 0.432-0.634mm, average 1.034 + 0.167 * 0.509 + 0.108mm. oral sucker transverse diameter was 0.115-0.180mm, the average was 0.140 + 0.025; ventral sucker size is 0.130-0.202 * 0.130-0.209mm, an average of 0.170 + 0.027 * 0.173 + 0.031mm, the ratio of the average for 1:1.236. sucker
Metacercaria of Paragonimus microrchis is spherical, great individual, with a double wall, the outer wall is thin and transparent, easily broken, the inner wall is thicker. Larvae body often curled up on the inner layer with cystic cavity, visible body known as the refractive index of intestinal larvae, excretory sac forms a bag shape in two. The intestinal branch without cover measuring the size (wall) is 0.760-1.406 * 0.689-1.399mm * 0.951mm, an average of 1.006, the outer wall thickness is 0.003-0.014mm, the average is 0.008mm. not only the inner wall of the metacercariae coverslip measuring the size of 0.632-0.852 * 0.596-0.831mm, the average is 0.728 * 0.703mm, the wall thickness is 0.007-0.031mm, the average is 0.011mm.
Paragonimus microrchis eggs are generally wide oval, golden yellow, egg cover most obviously, the unevenness of the thickness, and the end cover slip thickening. Measure the size of 73.1-95.9 * 42.3-55.3 m, average 81.37 + 9.90 * 47.67 + 5.58 tm.
Through the Blast homology comparison of Paragonimus microrchis and Laos Paragonimus harinasutai ITS2 and CO1 homology were 100% and 99%, Thailand and Bangkok Paragonimus homology were 98% and 96%, and China Zhejiang Paragonimus harinasutai homology were 96% and 88%.ITS2 and CO1 gene evolution the tree shows of Paragonimus microrchis and Laos Paragonimus harinasutai were clustered into one clade, this branch and Thailand of Paragonimus in Bangkok were clustered into one clade, the branch and Chinese Zhejiang Paragonimus harinasutai clustered into one clade.
[Conclusion] of Paragonimus microrchis adults after cercariae, metacercariae, egg shape with Professor Xia Daiguang (1978), Professor Zhou Benjiang (1988) the consistent morphological description of the morphological features of Paragonimus microrchis. The first application of scanning electron microscope observation of Paragonimus microrchis adult spines, as solitary type, shaped like a sharp knife shaped or triangular, before the middle of the body of the spines arranged regularly, is dense, sparse, irregular arrangement. The oral sucker wall longitudinal folds, ventral sucker wall ring folds, the morphological structure and characteristics of Paragonimus in food intake, attachment and motor function. The gene sequence, Paragonimus microrchis and Laos Paragonimus harinasutai for the same species, the authenticity of the Lao Paragonimus harinasutai needs further clarification. Paragonimus microrchis and Thailand Bangkok Paragonimus homology, close relationship, according to Blair.D The point of view of Paragonimus Paragonimus and Paragonimus Bangkok in Thailand should be the compound type of the same insect species (Complex).

【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R383.2;R532.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王文林,雷霖,周本江,劉愛(ài)華,王蕊芳,林世英;豐宮并殖吸蟲(chóng)染色體核型的研究[J];地方病通報(bào);2001年03期

2 胡文慶，周世祜;異盤并殖吸蟲(chóng)第二中間宿主感染情況的調(diào)查研究[J];地方病通報(bào);1995年01期

3 樊培方,陳黛霞,安耕九;三種并殖吸蟲(chóng)光學(xué)與掃描電鏡觀察[J];動(dòng)物學(xué)報(bào);1979年04期

4 何毅勛,林宇光,鐘文怡;斯氏并殖吸蟲(chóng)的掃描電鏡觀察[J];動(dòng)物學(xué)報(bào);1982年02期

5 陳心陶;巨睪并殖(吸蟲(chóng))成蟲(chóng)的形態(tài)研究和并殖科分類的探討[J];動(dòng)物學(xué)報(bào);1964年03期

6 劉德廣,姚其芳,黃紹芬;異盤并殖吸蟲(chóng)囊蚴的觀察[J];動(dòng)物學(xué)雜志;1982年03期

7 胡文慶;異盤并殖吸蟲(chóng)母雷蚴及子雷蚴的掃描電鏡觀察[J];廣西預(yù)防醫(yī)學(xué);1998年02期

8 胡文慶;異盤并殖吸蟲(chóng)尾蚴體被結(jié)構(gòu)的掃描電鏡觀察[J];廣西醫(yī)科大學(xué)學(xué)報(bào);1999年01期

9 劉娟,李雍龍;DNA序列分析在吸蟲(chóng)種株基因差異研究方面的應(yīng)用[J];國(guó)外醫(yī)學(xué).寄生蟲(chóng)病分冊(cè);2004年03期

10 李友松;;中國(guó)并殖吸蟲(chóng)蟲(chóng)種及分組的探討[J];福建醫(yī)藥雜志;1989年03期

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