东方蜜蜂微孢子虫对密林熊蜂的致病机理

doi:10.3864/j.issn.0578-1752.2012.22.016

摘要/Abstract

摘要： 【目的】明确东方蜜蜂微孢子虫（Nosema ceranae）对密林熊蜂（Bombus patagiatus）的侵染性及致病机理。【方法】采用传统生物学和电镜超微结构观察的方法，结合qPCR定量分析对东方蜜蜂微孢子虫在密林熊蜂上的致病机理进行探讨。【结果】感染初期工蜂除取食减少和行动迟缓外无明显外观染病特征，感染后期工蜂萎靡、衰弱、飞翔无力。解剖后镜检发现中肠仅存少量孢子，但充满大量细菌；熊蜂肠道组织切片发现东方蜜蜂微孢子虫侵染中肠上皮细胞后导致核膨大并变形、线粒体体积变小甚至解体，内质网紊乱，但孢子只侵染寄主细胞质而不侵入细胞核，最终导致线粒体解体，细胞破裂；qPCR定量分析得出接种后3—4 d中肠和脂肪体中微孢子虫的感染量达到最高值，其它组织则基本未检测到。【结论】东方蜜蜂微孢子虫可侵染异源寄主熊蜂，致病机理为微孢子虫引起寄主中肠上皮细胞内溶物发生病变，并导致整个中肠上皮细胞的破裂和凋亡。

关键词: 密林熊蜂 , 东方蜜蜂微孢子虫 , 超微结构 , qPCR

Abstract: 【Objective】The objective of this study is to define the infectivity and pathogenic mechanism of Nosema ceranae to Bombus patagiatus. 【Method】 Traditional biology and ultrastructure under electronic microscope methods were used, and combined with the quantitative real-time PCR (qPCR) to explore the pathogenesis of N. ceranae to B. patagiatus. 【Result】 In the initial infection, the infected bees did not exhibit obvious external disease signs except decreased feeding and motor retardation. In the late period, the infected bees exhibited dispirited, weak and unable to fly. Through observation with light microscope, few N. ceranae but a lot of bacteria were found in the midgut of bees. N. ceranae mainly infected midgut of epithelial cells, the nuclear enlarged and out of shape, mitochondria become smaller even disintegrating and endoplasmic reticulum become disorders. However, the spores only infected the cytoplasm of the host instead of invading the nuclear, which lead to the disintegration of the mitochondria and cytoclasis. Quantitative analysis on RT-PCR showed that N. ceranae reached the highest level in the midgut and fat body 3-4 d after being infected and other organizations were barely detected.【Conclusion】N. ceranae can cross-species infect B. patagiatus and the pathological process starts with the pathological changes of intestinal cells, which lead to the cytoclasis and apoptosis.

Key words: Bombus patagiatus , Nosema ceranae , ultrastructure , qPCR

秦浩然, 和绍禹, 吴杰, 李继莲. 东方蜜蜂微孢子虫对密林熊蜂的致病机理[J]. 中国农业科学, 2012, 45(22): 4697-4704.

QIN Hao-Ran, HE Shao-Yu, WU Jie, LI Ji-Lian. Pathogenic Mechanism of Bombus patagiatus Infected by Nosema ceranae[J]. Scientia Agricultura Sinica, 2012, 45(22): 4697-4704.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2012.22.016

https://www.chinaagrisci.com/CN/Y2012/V45/I22/4697

参考文献

[1]Losey J E, Vaughan M. The economic value of ecological services provided by insects. BioScience, 2006, 56(4): 311-323.

[2]Breeze T D, Bailey A P, Balcombe K G, Potts S G. Pollination services in the UK: How important are honeybees? Agriculture, Ecosystems and Environment, 2011, 142(3/4): 137-143.

[3]Morse R, Calderone N W. The value of honey bees as pollinators of U.S. crops in 2000. Bee Culture, 2000, 128: 1-15.

[4]Higes M, Martín-Hernández R, Botías C, Bailón E G, González-Porto A V, Barrios L, Nozal M J D, Bernal J L, Jiménez J J, Palencia P G, Meana A. How natural infection by Nosema ceranae causes honeybee colony collapse. Environmental Microbiology, 2008, 10(10): 2659-2669.

[5]Chen Y P, Evans J D. Historical presence of Israeli acute paralysis virus in the United States. American Bee Journal, 2007, 147: 1027-1028.

[6]Chen Y P, Evans J D, Smith I B, Pettis J S. Nosema ceranae is a long-present and wide-spread microsporidian infection of the European honey bee (Apis mellifera) in the United States. Journal of Invertebrate Pathology, 2008, 97(2): 86-88.

[7]Goulson D. Effects of introduced bees on native ecosystems. Annual Review of Ecology, Evolution and Systematics, 2003, 34: 1-26.

[8]Fries I, Feng F, da Silva A, Slemenda S B, Pieniazek N J. Nosema ceranae n. sp. (Microspora, Nosematidae), morphological and molecular characterization of a microsporidian parasite of the Asian honey bee Apis cerana (Hymenoptera, Apidae). European Journal of Protistology, 1996, 32(3): 356-365.

[9]梁正之. 对中蜂孢子虫病的初步探讨. 中国蜂业, 1980(4): 15-16

Liang Z Z. Nosema disease in honeybee (Apis cerana cerana). Apiculture in China, 1980(4): 15-16. (in Chinese)

[10]Singh Y. Nosema in Indian honey bee (Apis cerana indica). American Bee Journal, 1975, 115: 59.

[11]Yakobson B, Pothichot S, Wongsiri S. Possible transfer of Nosema apis from Apis mellifera to Apis cerana//Asian Honey Bees and Bee Mites. Bangkok, 1992.

[12]Fries I, Martín R, Meana A, García-Palencia P, Higes M. Natural infections of Nosema ceranae in European honey bees. Journal of Apicultural Research, 2006, 45(4): 230-233.

[13]Morse R A, Flottum K. Honey Bee Pests, Predators and Diseases, 3rd ed. Ohio: A. I Root Company, 1997: 59-76.

[14]Huang W F, Bocquet M, Lee K C, Sung I H, Jiang J H, Chen Y W, Wang C H. The comparison of rDNA spacer regions of Nosema ceranae isolates from different hosts and locations. Journal of Invertebrate Pathology, 2008, 97: 9-13.

[15]Plischuk S, Marthín-Hernández R, Prieto L, Lucía M, Botías C, Meana A, Abrahamovich A H, Lange C, Higes M. South American native bumblebees (Hymenoptera: Apidae) infected by Nosema ceranae (Microsporidia), an emerging pathogen of honeybees (Apis mellifera). Environmental Microbiology Reports, 2009, 1(2): 131-135.

[16]陈文锋, 李继莲, Schmid-Hempel P, 吴杰, 彭文君, 安建东. 我国四省区熊蜂中微孢子虫的自然感染率. 福建农林大学学报: 自然科学版, 2010, 39(3): 295-300.

Chen W F, Li J L, Schmid-Hempel P, Wu J, Peng W J, An J D. Natural infection rate of microsporidia in bumblebees from four different locations of China. Journal of Fujian Agriculture and Forestry University: Natural Science Edition, 2010, 39(3): 295-300. (in Chinese)

[17]Li J L, Chen W F, Wu J, Peng W J, An J D, Schmid-Hempel P, Schmid-Hempel R. Diversity of Nosema associated with bumblebees (Bombus spp.) from China. International Journal for Parasitology, 2012, 42: 49-61.

[18]Deredec A, Courchamp F. Extinction thresholds in host-parasite dynamics. Annales Zoologici Fennici, 2003, 40: 115-130.

[19]Higes M, Garcia-Palencia P, Martin-Hernandez R, Meana A. Experimental infection of Apis mellifera honeybees with Nosema ceranae (Microsporidia). Journal of Invertebrate Pathology, 2007, 94: 211-217.

[20]黄少康, 杨守深, 王丽华, 付中民. 中蜂来源的微孢子虫对意蜂工蜂的侵染性研究. 中国蜂业, 2007, 58(1): 7-8, 12.

Huang S K, Yang S S, Wang L H, Fu Z M. Infectivity of microsporidium from Apis cerana cerana to Intalian honey bee worker. Apiculture of China, 2007, 58(1): 7-8, 12. (in Chinese)

[21]Chen Y P, Evans J D, Zhou L, Boncristiani H, Kimura K, Xiao T, Litkowski A M, Pettis J S. Asymmetrical coexistence of Nosema ceranae and Nosema apis in honey bees. Journal of Invertebrate Pathology, 2009, 101(3): 204-209.

[22]Fries I, Feng F. Cross infectivity of Nosema apis in Apis mellifera and Apis cerana//Proceedings of the Apimondia 34th International Apicultural Congress. Bucharest, Romania, 1995: 151-155.

[23]McIvor C A, Malone L A. Nosema bombi, a microsporidian pathogen of the bumble bee Bombus terrestris (L.). New Zealand Journal of Zoology, 1995, 22: 25-31.

[24]Van den Eijnde J, Van den Vette N. Nosema infection in honeybees (Apis mellifera L) and bumble bees (Bombus terrestris L). Proceedings of Section Experimental and Applied Entomology of the Netherlands Entomological Society, 1993, 4: 205-208.

[25]Shafer A B A, Williams G R, Shutler D, Rogers R E L, Stewart D T. Cophylogeny of Nosema (Microsporidia: Nosematidae) and bees (Hymenoptera: Apidae) suggests both cospeciation and host-switch. The Journal of Parasitology, 2009, 95(1): 198-203.

[26]刘锋, 周婷, 王强, 代平礼. 东方蜜蜂微孢子虫 (N. cernane)提纯方法优化//中国养蜂学会中蜂协作委员会2009年学术交流会论文集. 南宁, 2009: 28-31.

Liu F, Zhou T, Wang Q, Dai P L. The optimization of N. cernane purification method//Proceedings of China Beekeeping Society Bee Partnership Committee in 2009 Symposium. Nanning, 2009: 28-31. (in Chinese)

[27]李继莲, 吴杰, 蒋皖, 彭文君, 安建东, 黄家兴. 熊蜂微孢子虫对熊蜂的危害. 蜜蜂杂志, 2007(9): 5-8.

Li J L, Wu J, Jiang W, Peng W J, An J D, Huang J X. The harm of Nosema bombi to bumblebees. Journal of Bee, 2007(9): 5-8. (in Chinese)

[28]Antúnez K, Martín-Hernández R, Prieto L, Meana A, Zunino P, Higes M. Immune suppression in the honey bee (Apis mellifera) following infection by Nosema ceranae (Microsporidia). Environmental Microbiology, 2009, 11(9): 2284-2290.

[29]Li J L, Wu J, Peng W J, An J D, Guo Z B, Tong Y M. Nosema bombi, a microsporidian pathogen of the bumble bee Bombus lucorum. Journal of Apicultural Science, 2005, 49(1): 53-57.

[30]Huang W F, Jiang J H, Chen Y W, Wang C H. A Nosema ceranae isolate from the honeybee Apis mellifera. Apidologie, 2007, 38: 30-37.

[31]史景泉, 陈意生, 卞修武. 超微病理学. 北京: 化学工业出版社, 2005.

Shi J Q, Chen Y S, Bian X W. Ultrastructural Pathology. Beijing: Chemical Industry Press, 2005. (in Chinese)

[32]Klee J, Besana A M, Genersch E, Gisder S, Nanetti A, Tam D Q, Chinh T X, Puerta F, Ruz J M, Kryger P, Message D, Hatjina F, Korpela S, Fries I, Paxton R J. Widespread dispersal of the microsporidium Nosema ceranae, an emergent pathogen of the western honey bee, Apis mellifera. Journal of Invertebrate Pathology, 2007, 96: 1-10.

[33]Giersch T, Berg T, Galea F, Hornitzky M. Nosema ceranae infects honey bees (Apis mellifera) and contaminates honey in Australia. Apidologie, 2009, 40: 117-123.

[34]Paxton R J, Klee J, Korpela S, Fries I. Nosema ceranae has infected Apis mellifera in Europe since at least 1998 and may be more virulent than Nosema apis. Apidologie, 2007, 38: 558-565.

[35]Chen Y P, Evans J D, Smith I B, Pettis J S. Nosema ceranae is a long-presentand wide-spread microsporidean infection of the European honey bee (Apis mellifera) in the United States. Journal of Invertebrate Pathology, 2008, 97: 186-188.

[36]Invernizzia C, Abuda C, Tomascoa I H, Harriet J, Ramalloc G, Campá J, Katz H, Gardiol G, Mendoza Y. Presence of Nosema ceranae in honeybees (Apis mellifera) in Uruguay. Journal of Invertebrate Pathology, 2009, 101: 150-153.