云南省蛛蝇和蝠蝇感染沃尔巴克氏体初步调查研究
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摘要
目的了解云南省蛛蝇和蝠蝇中沃尔巴克氏体(Wolbachia)的感染状况及其基因型别。方法 2012-2017年,在云南省芒市、腾冲、畹町、孟连、勐腊、祥云、墨江、永德县和保山市隆阳区共9个采样点采集蝙蝠体表蛛蝇和蝠蝇,PCR扩增沃尔巴克氏体的核糖体小亚基16S(16S rRNA)和细菌表皮蛋白(wsp)基因并测序,对所有16S r RNA和wsp基因进行系统进化分析。结果 2012-2017年,在云南省9个采集点采集蝙蝠体表蛛蝇747只(48批)和蝠蝇19只(6批),检测54批标本获得沃尔巴克氏体16S rRNA和wsp基因序列各24条。云南省蝙蝠体表蛛蝇和蝠蝇中,蛛虱蝇属(Phthiridium)6批和Eucampsipoda属37批中检出沃尔巴克氏体阳性分别为2批(2/6)和19批(19/37),蛛虱蝇属(Nycteribia)(0/4)和笔虱蝇属(Penicillidia)(0/1)未检出;蝠蝇均属于蝠虱蝇属(Brachytarsina),6批中检出沃尔巴克氏体阳性的有3批(3/6)。16S rRNA基因系统进化结果提示,24株沃尔巴克氏体均位于一个大的分支中,与1株来源于白蚁(DQ837204)的同源性最高达98.80%～100%。wsp基因的系统进化分析提示,24株沃尔巴克氏体位于B组和F组基因型。结论沃尔巴克氏体在云南省蛛蝇和蝠蝇中广泛而长期存在,wsp基因系统进化分析属于B组和F组基因型。
Objective To investigate the infection status and genotypes of Wolbachia in Nycteribiidae and Streblidae in Yunnan province, China. Methods From 2012 to 2017, Nycteribiidae and Streblidae were collected from bats in 9 spots in Yunnan province, i.e., Mangshi, Tengchong, Wanding, Menglian, Mengla, Xiangyun, Mojiang, Yongde county, and Baoshan city. The 16 S rRNA and wsp genes of Wolbachia were amplified and sequenced by PCR. Phylogenetic analyses were performed on all the 16 S rRNA and wsp genes. Results From 2012 to 2017, a total of 747(48 batches) Nycteribiidae flies and 19(6 batches) Streblidae flies were collected from the fur of bats in the 9 collection spots in Yunnan province. A total of24 pairs of 16 S rRNA and wsp genes of Wolbachia were identified in the 54 batches of samples. For the Nycteribiidae from the fur of bats in Yunnan province, Wolbachia infection was positive in 2 out of 6 batches of Phthiridium and 19 out of 37 batches of Eucampsipoda, but negative in any batch of Nycteribia(0/4) and Penicillidia(0/1). All 6 batches of Streblidae belonged to Brachytarsina, 3 out of which were positive for Wolbachia infection. Phylogenetic analysis of the 16 S rRNA gene showed that the 24 Wolbachia strains shared the same evolutionary branch with one from termites(DQ837204), with a 98.80%-100% homology in nucleotide sequence. Phylogenetic analysis of the wsp gene revealed that the 24 Wolbachia strains belonged to the supergroups B and F. Conclusion Wolbachia has been widely distributed in Nycteribiidae and Streblidae in Yunnan province for a long time. Phylogenetic analysis of the wsp sequences suggests that all identified Wolbachia strains belong to the supergroups B and F.
Objective To investigate the infection status and genotypes of Wolbachia in Nycteribiidae and Streblidae in Yunnan province, China. Methods From 2012 to 2017, Nycteribiidae and Streblidae were collected from bats in 9 spots in Yunnan province, i.e., Mangshi, Tengchong, Wanding, Menglian, Mengla, Xiangyun, Mojiang, Yongde county, and Baoshan city. The 16 S rRNA and wsp genes of Wolbachia were amplified and sequenced by PCR. Phylogenetic analyses were performed on all the 16 S rRNA and wsp genes. Results From 2012 to 2017, a total of 747(48 batches) Nycteribiidae flies and 19(6 batches) Streblidae flies were collected from the fur of bats in the 9 collection spots in Yunnan province. A total of24 pairs of 16 S rRNA and wsp genes of Wolbachia were identified in the 54 batches of samples. For the Nycteribiidae from the fur of bats in Yunnan province, Wolbachia infection was positive in 2 out of 6 batches of Phthiridium and 19 out of 37 batches of Eucampsipoda, but negative in any batch of Nycteribia(0/4) and Penicillidia(0/1). All 6 batches of Streblidae belonged to Brachytarsina, 3 out of which were positive for Wolbachia infection. Phylogenetic analysis of the 16 S rRNA gene showed that the 24 Wolbachia strains shared the same evolutionary branch with one from termites(DQ837204), with a 98.80%-100% homology in nucleotide sequence. Phylogenetic analysis of the wsp gene revealed that the 24 Wolbachia strains belonged to the supergroups B and F. Conclusion Wolbachia has been widely distributed in Nycteribiidae and Streblidae in Yunnan province for a long time. Phylogenetic analysis of the wsp sequences suggests that all identified Wolbachia strains belong to the supergroups B and F.
引文
[1]郭颂,凌锋,王金娜,等.浙江省中东部白纹伊蚊共生菌沃尔巴克氏体的调查研究[J].中国媒介生物学及控制杂志,2015,26(6):545-549.DOI:10.11853/j.issn.1003.4692.2015.06.002.
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[7]Stouthamer R,Breeuwer JAJ,Hurst GDD.Wolbachia pipientis:Microbial manipulator of arthropod reproduction[J].Annu Rev Microbiol,1999,53:71-102.DOI:10.1146/annurev.micro.53.1.71.
[8]Werren JH,Baldo L,Clark ME.Wolbachia:master manipulators of invertebrate biology[J].Nat Rev Microbiol,2008,6(10):741-751.DOI:10.1038/nrmicro1969.
[9]Hosokawa T,Nikoh N,Koga R,et al.Reductive genome evolution,host-symbiont co-speciation and uterine transmission of endosymbiotic bacteria in bat flies[J].ISME J,2012,6(3):577-587.DOI:10.1038/ismej.2011.125.
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[12]Witsenburg F,Salamin N,Christe P.The evolutionary host switches of Polychromophilus:a multi-gene phylogeny of the bat malaria genus suggests a second invasion of mammals by a haemosporidian parasite[J].Malar J,2012,11:53.DOI:10.1186/1475-2875-11-53.
[13]Olival KJ,Dick CW,Simmons NB,et al.Lack of population genetic structure and host specificity in the bat fly,Cyclopodia horsfieldi,across species of Pteropus bats in Southeast Asia[J].Parasit Vectors,2013,6:231.DOI:10.1186/1756-3305-6-231.
[14]Poinar GO Jr.Vetufebrus ovatus n.gen.,n.sp.(Haemospororida:Plasmodiidae)vectored by a streblid bat fly(Diptera:Streblidae)in Dominican amber[J].Parasit Vectors,2011,4:229.DOI:10.1186/1756-3305-4-229.
[15]Morse SF,Olival KJ,Kosoy M,et al.Global distribution and genetic diversity of Bartonella in bat flies(Hippoboscoidea,Streblidae,Nycteribiidae)[J].Infect Genet Evol,2012,12(8):1717-1723.DOI:10.1016/j.meegid.2012.06.009.
[16]Kamani J,Baneth G,Mitchell M,et al.Bartonella species in bats(Chiroptera)and bat flies(Nycteribiidae)from Nigeria,West Africa[J].Vector Borne Zoonotic Dis,2014,14(9):625-632.DOI:10.1089/vbz.2013.1541.
[17]Brook CE,Bai Y,Dobson AP,et al.Bartonella spp.in fruit bats and blood-feeding ectoparasites in Madagascar[J].PLoS Negl Trop Dis,2015,9(2):e0003532.DOI:10.1371/journal.pntd.0003532.
[18]Aznar-Lopez C,Vasquez-Moron S,Marston DA,et al.Detection of rhabdovirus viral RNA in oropharyngeal swabs and ectoparasites of Spanish bats[J].J Gen Virol,2013,94:69-75.DOI:10.1099/vir.0.046490-0.
[19]Van Vuren PJ,Wiley M,Palacios G,et al.Isolation of a novel fusogenic orthoreovirus from Eucampsipoda africana bat flies in South Africa[J].Viruses,2016,8(3):65.DOI:10.3390/v8030065.
[20]Feng Y,Li YY,Fu SH,et al.Isolation of Kaeng Khoi virus(KKV)from Eucampsipoda sundaica bat flies in China[J].Virus Res,2017,238(15):94-100.DOI:10.1016/j.virusres.2017.06.007.
[21]Wilkinson DA,Duron O,Cordonin C,et al.The Bacteriome of bat flies(Nycteribiidae)from the Malagasy region:a community shaped by host ecology,bacterial transmission mode,and hostvector specificity[J].Appl Environ Microbiol,2016,82(6):1778-1788.DOI:10.1128/AEM.03505-15.
[22]Saurav GK,Daimei G,Rana VS,et al.Detection and localization of Wolbachia in Thrips palmi Karny(Thysanoptera:Thripidae)[J].India J Microbiol,2016,56(2):167-171.DOI:10.1007/s12088-016-0567-7.
[23]Zhou WG,Rousset F,O’Neill S.Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences[J].P Proc Roy Soc B:Biol Sci,1998,265(1395):509-515.DOI:10.1098/rspb.1998.0324.
[24]Lei WW,Guo XY,Fu SH,et al.Isolation of Tibet orbivirus,TIBOV,from Culicoides collected in Yunnan,China[J].PLoSOne,2015,10(8):e0139646.DOI:10.1371/journal.pone.0136257.
[25]Werren JH,Windsor D,Guo LR.Distribution of Wolbachia among Neotropical arthropods[J].Pros Roy Soc B:Biol Sci,1995,262(1364):197-204.DOI:10.1098/rspb.1995.0196.
[26]Van Meer MMM,Witteveldt J,Stouthamer R.Phylogeny of the arthropod endosymbiont Wolbachia based on the wsp gene[J].Insect Mol Biol,1999,8(3):399-408.DOI:10.1046/j.1365-2583.1999.83129.x.
[27]Sakamoto H,Ishikawa Y,Sasaki T,et al.Transinfection reveals the crucial importance of Wolbachia genotypes in determining the type of reproductive alteration in the host[J].Genet Res,2005,85(3):205-210.DOI:10.1017/S0016672305007573.
[28]Cordaux R,Michel-Salzat A,Frelon-Raimond M,et al.Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare:evolutionary implications[J].Heredity,2004,93(1):78-84.DOI:10.1038/sj.hdy.6800482.
[29]宋社吾,赵彤言,董言德,等.我国蚊虫中昆虫共生微生物Wolbachia感染的研究[J].中国媒介生物学及控制杂志,2002,13(1):19-21.DOI:10.3969/j.issn.1003-4692.2002.01.006.
[30]Iturbe-Ormaetxe I,Walker T,O’Neill SL.Wolbachia and the biological control of mosquito-borne disease[J].EMBO Rep,2011,12(6):508-518.DOI 10.1038/embor.2011.84.
[31]Calisher CH,Childs JE,Field HE,et al.Bats:important reservoir hosts of emerging viruses[J].Clin Microbiol Rev,2006,19(3):531-545.DOI:10.1128/CMR.00017-06.
[32]O’Shea TJ,Cryan PM,Cunningham AA,et al.Bat flight and zoonotic viruses[J].Emerg Infect Dis,2014,20(5):741-745.DOI:10.3201/eid2005.130539.
[33]Wynne JW,Wang LF.Bats and viruses:friend or for?[J].PLoSPathog,2013,9(10):e1003651.DOI:10.1371/journal.ppat.1003651.
[34]徐保海.中国虱蝇总科记述(昆虫纲:双翅目)[J].中国人兽共患病学报,2011,27(1):67-75.DOI:10.3969/j.issn.1002-2694.2011.01.016.
[35]Glowska E,Dragun-Damian A,Dabert M,et al.New Wolbachia supergroups detected in quill mites(Acari:Syringophilidae)[J].Infect Genet Evol,2015,30:140-146.DOI:10.1016/j.meegid.2014.12.019.
[36]Jiggins FM,Von Der Schulenburg JHG,Hurst GDD,et al.Recombination confounds interpretations of Wolbachia evolution[J].Proc Biol Sci,2001,268(1474):1423-1427.DOI:10.1098/rspb.2001.1656.
[37]Baldo L,Hotopp JCD,Jolley KA,et al.Multilocus sequence typing system for the endosymbiont Wolbachia pipientis[J].Appl Environ Microbiol,2006,72(11):7098-7110.DOI:10.1128/AEM.00731-06.
[38]胡亚萍,刘小波,桑少伟,等.多位点序列分型技术在沃尔巴克氏体基因分型中的研究进展[J].中国媒介生物学及控制杂志,2016,27(2):197-201.DOI:10.11853/j.issn.1003.8280.2016.02.028.
[2]Herting M,Wolbach SB.Studies on rickettsia-like microorganisms in insect[J].J Med Res,1924,44(3):329-374.
[3]Hertig M.The rickettsia,Wolbachia pipientis(gen.et sp.n.)and associated inclusions of the mosquito,Culex pipiens[J].Parasitology,1936,28(4):453-486.DOI:10.1017/S0031182000022666.
[4]Hilgenboecker K,Hammerstein P,Schlattmann P,et al.How many species are infected with Wolbachia?-a statistical analysis of current data[J].FEMS Microbiol Lett,2008,281(2):215-220.DOI:10.1111/j.1574-6968.2008.01110.x.
[5]Zug R,Hammerstein P.Still a host of hosts for Wolbachia:analysis of recent data suggests that 40%of terrestrial arthropod species are infected[J].PLoS One,2012,7(6):e38544.DOI:10.1371/journal.pone.0038544.
[6]Werren JH.Biology of Wolbachia[J].Annu Rev Entomol,1997,42(1):587-609.DOI:10.1146/annurev.ento.42.1.587.
[7]Stouthamer R,Breeuwer JAJ,Hurst GDD.Wolbachia pipientis:Microbial manipulator of arthropod reproduction[J].Annu Rev Microbiol,1999,53:71-102.DOI:10.1146/annurev.micro.53.1.71.
[8]Werren JH,Baldo L,Clark ME.Wolbachia:master manipulators of invertebrate biology[J].Nat Rev Microbiol,2008,6(10):741-751.DOI:10.1038/nrmicro1969.
[9]Hosokawa T,Nikoh N,Koga R,et al.Reductive genome evolution,host-symbiont co-speciation and uterine transmission of endosymbiotic bacteria in bat flies[J].ISME J,2012,6(3):577-587.DOI:10.1038/ismej.2011.125.
[10]Morse SF,Bush SE,Patterson BD,et al.Evolution,multiple acquisition,and localization of endosymbionts in bat flies(Diptera:Hippoboscoidea:Streblidae and Nycteribiidae)[J].Appl Environ Microbiol,2013,79(9):2952-2961.DOI:10.1128/AEM.03814-12.
[11]Morse SF,Dick CW,Patterson BD,et al.Some like it hot:evolution and ecology of novel endosymbionts in bat files of caveroosting bats(Hippoboscoidea,Nycterophiliinae)[J].Appl Environ Microbiol,2012,78(24):8639-8649.DOI:10.1128/AEM.02455-12.
[12]Witsenburg F,Salamin N,Christe P.The evolutionary host switches of Polychromophilus:a multi-gene phylogeny of the bat malaria genus suggests a second invasion of mammals by a haemosporidian parasite[J].Malar J,2012,11:53.DOI:10.1186/1475-2875-11-53.
[13]Olival KJ,Dick CW,Simmons NB,et al.Lack of population genetic structure and host specificity in the bat fly,Cyclopodia horsfieldi,across species of Pteropus bats in Southeast Asia[J].Parasit Vectors,2013,6:231.DOI:10.1186/1756-3305-6-231.
[14]Poinar GO Jr.Vetufebrus ovatus n.gen.,n.sp.(Haemospororida:Plasmodiidae)vectored by a streblid bat fly(Diptera:Streblidae)in Dominican amber[J].Parasit Vectors,2011,4:229.DOI:10.1186/1756-3305-4-229.
[15]Morse SF,Olival KJ,Kosoy M,et al.Global distribution and genetic diversity of Bartonella in bat flies(Hippoboscoidea,Streblidae,Nycteribiidae)[J].Infect Genet Evol,2012,12(8):1717-1723.DOI:10.1016/j.meegid.2012.06.009.
[16]Kamani J,Baneth G,Mitchell M,et al.Bartonella species in bats(Chiroptera)and bat flies(Nycteribiidae)from Nigeria,West Africa[J].Vector Borne Zoonotic Dis,2014,14(9):625-632.DOI:10.1089/vbz.2013.1541.
[17]Brook CE,Bai Y,Dobson AP,et al.Bartonella spp.in fruit bats and blood-feeding ectoparasites in Madagascar[J].PLoS Negl Trop Dis,2015,9(2):e0003532.DOI:10.1371/journal.pntd.0003532.
[18]Aznar-Lopez C,Vasquez-Moron S,Marston DA,et al.Detection of rhabdovirus viral RNA in oropharyngeal swabs and ectoparasites of Spanish bats[J].J Gen Virol,2013,94:69-75.DOI:10.1099/vir.0.046490-0.
[19]Van Vuren PJ,Wiley M,Palacios G,et al.Isolation of a novel fusogenic orthoreovirus from Eucampsipoda africana bat flies in South Africa[J].Viruses,2016,8(3):65.DOI:10.3390/v8030065.
[20]Feng Y,Li YY,Fu SH,et al.Isolation of Kaeng Khoi virus(KKV)from Eucampsipoda sundaica bat flies in China[J].Virus Res,2017,238(15):94-100.DOI:10.1016/j.virusres.2017.06.007.
[21]Wilkinson DA,Duron O,Cordonin C,et al.The Bacteriome of bat flies(Nycteribiidae)from the Malagasy region:a community shaped by host ecology,bacterial transmission mode,and hostvector specificity[J].Appl Environ Microbiol,2016,82(6):1778-1788.DOI:10.1128/AEM.03505-15.
[22]Saurav GK,Daimei G,Rana VS,et al.Detection and localization of Wolbachia in Thrips palmi Karny(Thysanoptera:Thripidae)[J].India J Microbiol,2016,56(2):167-171.DOI:10.1007/s12088-016-0567-7.
[23]Zhou WG,Rousset F,O’Neill S.Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences[J].P Proc Roy Soc B:Biol Sci,1998,265(1395):509-515.DOI:10.1098/rspb.1998.0324.
[24]Lei WW,Guo XY,Fu SH,et al.Isolation of Tibet orbivirus,TIBOV,from Culicoides collected in Yunnan,China[J].PLoSOne,2015,10(8):e0139646.DOI:10.1371/journal.pone.0136257.
[25]Werren JH,Windsor D,Guo LR.Distribution of Wolbachia among Neotropical arthropods[J].Pros Roy Soc B:Biol Sci,1995,262(1364):197-204.DOI:10.1098/rspb.1995.0196.
[26]Van Meer MMM,Witteveldt J,Stouthamer R.Phylogeny of the arthropod endosymbiont Wolbachia based on the wsp gene[J].Insect Mol Biol,1999,8(3):399-408.DOI:10.1046/j.1365-2583.1999.83129.x.
[27]Sakamoto H,Ishikawa Y,Sasaki T,et al.Transinfection reveals the crucial importance of Wolbachia genotypes in determining the type of reproductive alteration in the host[J].Genet Res,2005,85(3):205-210.DOI:10.1017/S0016672305007573.
[28]Cordaux R,Michel-Salzat A,Frelon-Raimond M,et al.Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare:evolutionary implications[J].Heredity,2004,93(1):78-84.DOI:10.1038/sj.hdy.6800482.
[29]宋社吾,赵彤言,董言德,等.我国蚊虫中昆虫共生微生物Wolbachia感染的研究[J].中国媒介生物学及控制杂志,2002,13(1):19-21.DOI:10.3969/j.issn.1003-4692.2002.01.006.
[30]Iturbe-Ormaetxe I,Walker T,O’Neill SL.Wolbachia and the biological control of mosquito-borne disease[J].EMBO Rep,2011,12(6):508-518.DOI 10.1038/embor.2011.84.
[31]Calisher CH,Childs JE,Field HE,et al.Bats:important reservoir hosts of emerging viruses[J].Clin Microbiol Rev,2006,19(3):531-545.DOI:10.1128/CMR.00017-06.
[32]O’Shea TJ,Cryan PM,Cunningham AA,et al.Bat flight and zoonotic viruses[J].Emerg Infect Dis,2014,20(5):741-745.DOI:10.3201/eid2005.130539.
[33]Wynne JW,Wang LF.Bats and viruses:friend or for?[J].PLoSPathog,2013,9(10):e1003651.DOI:10.1371/journal.ppat.1003651.
[34]徐保海.中国虱蝇总科记述(昆虫纲:双翅目)[J].中国人兽共患病学报,2011,27(1):67-75.DOI:10.3969/j.issn.1002-2694.2011.01.016.
[35]Glowska E,Dragun-Damian A,Dabert M,et al.New Wolbachia supergroups detected in quill mites(Acari:Syringophilidae)[J].Infect Genet Evol,2015,30:140-146.DOI:10.1016/j.meegid.2014.12.019.
[36]Jiggins FM,Von Der Schulenburg JHG,Hurst GDD,et al.Recombination confounds interpretations of Wolbachia evolution[J].Proc Biol Sci,2001,268(1474):1423-1427.DOI:10.1098/rspb.2001.1656.
[37]Baldo L,Hotopp JCD,Jolley KA,et al.Multilocus sequence typing system for the endosymbiont Wolbachia pipientis[J].Appl Environ Microbiol,2006,72(11):7098-7110.DOI:10.1128/AEM.00731-06.
[38]胡亚萍,刘小波,桑少伟,等.多位点序列分型技术在沃尔巴克氏体基因分型中的研究进展[J].中国媒介生物学及控制杂志,2016,27(2):197-201.DOI:10.11853/j.issn.1003.8280.2016.02.028.