两个入侵烟粉虱隐种MEAM1与MED对番茄黄曲叶病毒水平传播能力的比较

【目的】烟粉虱 Bemisia tabaci 是番茄黄曲叶病毒（Tomato yellow leaf curl virus, TYLCV）在自然界的唯一传播媒介,除了可以直接取食获取TYLCV,烟粉虱还可以通过交配获取此植物病毒。虽然前人研究证明了烟粉虱Middle East-Asia Minor 1 （MEAM1）和Mediterranean （MED）隐种都可以通过交配在两性个体之间水平传播TYLCV,但有关MEAM1与MED隐种水平传播TYLCV的能力是否存在差异,不同研究的结果却并不一致。另外,目前尚无关于烟粉虱的水平传播行为是否有助于TYLCV在田间扩散的相关研究。【方法】从浙江、广东、云南和河南4个省份的田间采集MEAM1和MED隐种烟粉虱种群,在室内应用分子标记对各种群所属隐种鉴定后分别建立7个供试种群,然后观察每个种群内带毒成虫与不带毒成虫通过交配在不同性别个体之间水平传播TYLCV的能力,并选用采自广东的MEAM1隐种烟粉虱模拟群体交配实验探究通过交配获毒的烟粉虱个体是否具有传播TYLCV致健康番茄植株发病的能力。【结果】4个省内的MEAM1和MED隐种烟粉虱都可通过交配对TYLCV进行水平传播,但传播频率一般在10%以下。不同省份的MEAM1隐种种群之间在水平传播TYLCV的能力上无显著差异,不同省份的MED隐种种群之间也不存在显著差异。另外,同一省份的MEAM1隐种与MED隐种之间在水平传播TYLCV的能力上也不存在显著差异,而且在两个隐种中,带毒雄虫将病毒水平传播给不带毒雌虫与带毒雌虫将病毒水平传播给不带毒雄虫的频率没有显著差异。研究还表明,采自广东的MEAM1隐种烟粉虱个体通过交配水平获毒后不能致健康番茄植株发病。【结论】综合以上结果,我们推测TYLCV在中国境内两个入侵烟粉虱隐种MEAM1和MED各自种群内个体之间的水平传播概率较低,对该病毒在田间的扩散可能没有作用或作用不大。     

海南烟粉虱田间种群隐种鉴定及对溴氰虫酰胺和氟吡呋喃酮的敏感性检测

【目的】明确海南烟粉虱Bemisia tabaci田间种群对2种新型杀虫剂溴氰虫酰胺和氟吡呋喃酮的敏感水平,为杀虫剂的合理施用与烟粉虱的田间抗性监测提供参考。【方法】以2017年1月采自海南省6个地点烟粉虱田间种群为材料,利用线粒体细胞色素氧化酶Ⅰ基因(mtCOI)测序及mtCOI PCR-RFLP分子标记对采集种群隐种进行鉴定;采用浸叶法测定溴氰虫酰胺和氟吡呋喃酮2种药剂对各地区田间种群的致死中浓度(LC_(50))。【结果】在海南省6个烟粉虱田间种群中,三亚市吉阳区种群为MEAM1隐种,澄迈县永发镇种群和乐东黎族自治县冲坡镇种群以MED隐种为主的MEAM1(10%)与MED隐种(分别为94.6%和92.9%)混合种群,其他3个田间种群均为MED隐种。与溴氰虫酰胺敏感种群相比,除乐东黎族自治县十所种群已处于低水平抗性(RR:5~10倍),其他5个种群均处于敏感水平(RR5倍);所有6个田间种群对氟吡呋喃酮均处于敏感水平(RR3倍)。【结论】溴氰虫酰胺和氟吡呋喃酮可作为烟粉虱防控的新药剂,并可作为其抗性治理的替代药剂,但同时应密切关注烟粉虱对溴氰虫酰胺抗性水平变化情况。     

温室粉虱和烟粉虱3个隐种中热激蛋白基因hsp70和hsp90含量的比较分析

【目的】昆虫适应新环境的能力与其对温度的耐受能力密切相关。热激蛋白HSP70和HSP90具有提高生物体温度耐受性的功能。烟粉虱Bemisia tabaci (Gennadius)不同隐种和不同种粉虱对温度的适应能力有差异,这与它们的热激蛋白基因拷贝数的差异可能相关。【方法】利用实时荧光定量PCR方法,检测入侵型烟粉虱MED隐种和MEAM1隐种、本地型烟粉虱AsiaⅡ1隐种以及温室粉虱Trialeurodes vaporariorum (Westwood)基因组DNA中热激蛋白基因hsp70和hsp90的拷贝数。【结果】不同种类的粉虱和烟粉虱不同隐种体内的hsp70和hsp90的含量均有较大差异,其中hsp70和hsp90拷贝数在入侵型烟粉虱MED和MEAM1隐种中含量较其他两种均高,而在土著种AsiaⅡ1隐种中含量最低,在温室粉虱中居中。此外,相同物种雌雄成虫hsp70和hsp90的拷贝数也不同,雌虫hsp70和hsp90拷贝数约为雄虫的2倍。【结论】不同种粉虱及烟粉虱不同隐种的hsp70和hsp90的拷贝数可能与其耐热性差异相关。本研究为解释不同种粉虱、烟粉虱不同隐种及其不同性别的耐热性差异机制提供了进一步的依据。     

烟粉虱MEAM1和MED成虫在辣椒上传播番茄褪绿病毒的特性

【目的】分析与比较烟粉虱Bemisia tabaci MEAM1和MED成虫在辣椒Capsicum annuum植株上对番茄褪绿病毒(tomato chlorosis virus, ToCV)的传播差异,解析媒介昆虫-病毒-寄主植物间的互作关系,为田间辣椒上ToCV防治提供参考。【方法】利用烟粉虱MEAM1和MED成虫及ToCV cDNA侵染性克隆,对烟粉虱MEAM1和MED成虫在辣椒植株上获取和传播ToCV的能力进行比较,同时比较烟粉虱MEAM1和MED成虫在健康和携带ToCV辣椒植株上的取食偏好。【结果】在感染ToCV的辣椒植株上接虫后96 h内,烟粉虱MEAM1和MED成虫体内病毒积累量逐渐增加,MED成虫获毒速度和病毒积累量均大于MEAM1成虫的,MED成虫获毒量是MEAM1成虫的1.74倍;MED成虫在辣椒植株上的传毒量明显高于MEAM1成虫,传毒量平均相差9倍,且MED成虫传毒率比MEAM1成虫高29%。MEAM1和MED成虫对健康和感染ToCV的辣椒植株的取食偏好相似,都明显倾向于取食被ToCV侵染的辣椒植株。【结论】烟粉虱MED成虫在辣椒植株上获取、传播ToCV的能力均...     

利用mtCOI PCR-RFLP技术鉴定中国境内九个烟粉虱隐种

烟粉虱Bemisia tabaci (Gennadius)是一个物种复合体, 包括31个以上形态上无法区分的隐种, 其中少数隐种是世界性入侵害虫。目前, 在中国境内分布有2个入侵隐种和13个土著隐种。快速、 高效的鉴别方法对掌握烟粉虱田间发生规律及制定相关防控策略具有重要意义。然而到目前为止, 除了mtCOI基因测序比对外, 尚未有一种简便的方法可以有效地区分多个烟粉虱隐种。本研究采用mtCOI PCR-RFLP技术, 单独或组合使用TaqI, VspI, Van91I, NcoI和FokI这5种限制性内切酶, 酶切烟粉虱mtCOI的PCR扩增片段, 鉴别分布在中国境内的9个烟粉虱隐种。结果表明, 单独使用TaqI酶切, 可鉴别出MEAM1和China 1两个隐种, 使用TaqI+NcoI分步酶切可鉴定出MED和Asia 1两个隐种, 使用TaqI+Van91I分步酶切可鉴定出Asia II 3和Asia II 9两个隐种, 使用TaqI+VspI+FokI分步酶切可鉴定出Asia II 1, Asia II 6和Asia II 7 3个隐种。本研究为高效鉴别中国境内的多个烟粉虱隐种提供了方法。     

烟粉虱MEAM1隐种卵黄原蛋白受体基因cDNA的克隆、 序列分析及在不同发育时期的表达

卵黄原蛋白受体（vitellogenin receptor, VgR）是卵黄原蛋白被卵母细胞摄取的关键因子, 在卵黄发生和卵母细胞发育等生理过程中发挥着重要作用。为探讨烟粉虱Bemisia tabaci VgR的功能, 我们采用RT-PCR和RACE等技术扩增了烟粉虱MEAM1隐种B. tabaci Middle East-Asia Minor 1 (MAEM1) 的VgR基因cDNA 全长序列。生物信息学分析表明, 烟粉虱MEAM1隐种的VgR基因cDNA全长5 774 bp, 编码1 919个氨基酸, 推测分子量约201 kDa, N-端前31个氨基酸为信号肽。烟粉虱MEAM1隐种的VgR属于低密度脂蛋白受体（low density lipoprotein receptor, LDLR）家族, 蛋白质三维结构预测分析表明, 该受体具有LDLR家族基因典型的保守功能结构域。通过实时荧光定量PCR技术研究了烟粉虱MEAM1隐种VgR基因不同发育时期的表达, 结果表明VgR基因在伪蛹期开始表达, 并在羽化后1 d达到高峰, 此后逐渐降低, 3 d后又逐渐升高, 直至羽化后7 d达到峰值。研究结果丰富了卵黄原蛋白受体家族基因的数据库, 为今后深入研究并揭示烟粉虱卵黄发生的调控机制奠定了基础。     

烟粉虱MEAM1隐种磷脂氢谷胱甘肽过氧化物酶基因克隆与表达分析

【目的】本研究旨在从烟粉虱Bemisia tabaci中东-小亚细亚1隐种(Middle East-Asia Minor 1, MEAM1)中克隆磷脂氢谷胱甘肽过氧化物酶(phospholipid hydroperoxide glutathione peroxidase, PHGPX)基因,鉴定其在烟粉虱不同发育阶段及吡虫啉处理不同时间后雌成虫体内的表达情况,明确其在烟粉虱应对外界环境压力中的功能。【方法】利用3′RACE克隆和测定烟粉虱MEAM1隐种内PHGPX基因的cDNA全长序列,并对其编码的氨基酸序列进行生物信息学分析;利用定量RT-PCR技术对该基因在烟粉虱MEAM1隐种不同发育阶段及吡虫啉处理不同时间后雌成虫体内的表达量进行分析。【结果】获得了烟粉虱MEAM1隐种两个磷脂氢谷胱甘肽过氧化物酶基因的全长cDNA序列,分别命名为BtB-PHGPX1(GenBank登录号:KY312116)和BtB-PHGPX2(GenBank登录号:KY312117)。序列分析表明,BtB-PHGPX1基因开放阅读框全长732 bp,编码243个氨基酸;BtB-PHGPX2基因开放阅读框全长567 bp,编码188个氨基酸。序列比对结果表明两基因的编码蛋白内均具有谷胱甘肽过氧化物酶保守的半胱氨酸、谷氨酰胺和色氨酸残基位点。BtB-PHGPX1在烟粉虱MEAM1隐种卵内表达量显著高于其在若虫、伪蛹、雌成虫和雄成虫内的表达量,BtB-PHGPX2在烟粉虱MEAM1隐种卵内的表达量显著低于其在若虫、伪蛹和雌成虫内的表达量(P<0.05)。BtB-PHGPX1和BtB-PHGPX2在雌成虫内的表达量均显著高于雄成虫内。吡虫啉处理雌成虫2 h时两基因的表达量均较对照显著提高(P<0.05),处理后5, 10和24 h时其表达量均较对照显著下降(P<0.01)。【结论】本研究克隆了烟粉虱MEAM1隐种两个PHGPX基因的序列全长,明确了其在不同发育阶段及吡虫啉处理不同时间后雌成虫体内的差异表达,推测PHGPX在烟粉虱抵御环境压力及杀虫剂胁迫时可能发挥着重要的防御作用。     

漆酶-1基因在烟粉虱MEAM1隐种和温室粉虱中的表达模式比较

【目的】漆酶-1(LAC1)在粉虱-植物互作中发挥重要作用,本研究旨在探讨烟粉虱Bemisia tabaci MEAM1隐种和温室粉虱Trialeurodes vaporariorum两种存在竞争关系的粉虱中LAC1基因表达模式是否存在差异。【方法】测定烟粉虱MEAM1隐种和温室粉虱唾液漆酶-1活力;对温室粉虱LAC1基因进行了克隆和测序;通过荧光定量PCR测定和比较了LAC1在两种粉虱不同发育阶段和成虫不同组织中的表达谱以及取食外施水杨酸(0. 5 mmol/L)的番茄植株后两种粉虱不同发育阶段和成虫不同组织中LAC1的表达水平,同时检测了成虫存活率和产卵量。【结果】酶活力测定结果表明两种粉虱的唾液中漆酶-1活力差异不显著。温室粉虱PCR扩增后得到大小639 bp的LAC1基因中间片段。发育表达谱结果表明,LAC1基因在两种粉虱各个发育阶段均有表达,在烟粉虱MEAM1隐种成虫期表达量逐渐升高,而在温室粉虱各发育时期的表达量无显著差异;烟粉虱MEAM1隐种成虫期LAC1的表达量高于温室粉虱。组织表达结果表明,LAC1在两种粉虱成虫的头胸部、中肠和腹部残骸中均有表达,但在中肠中丰度最高。取食外施水杨酸番茄植株的烟粉虱MEAM1隐种,随取食时间的延长成虫存活率下降,与对照(取食喷施清水的植株)相比差异显著。羽化后14 d时,取食外施水杨酸植株的烟粉虱MEAM1隐种存活率仅为对照的一半,而取食外施水杨酸植株的温室粉虱的存活率与对照相比差异不显著。无论是取食外施水杨酸的植株还是取食对照植株,烟粉虱MEAM1隐种体内LAC1表达量和产卵量均高于温室粉虱。【结论】LAC1在两种粉虱体内表达模式存在差异,这可能在两种粉虱的竞争适应中具有一定作用。     

烟粉虱MEAM1和AsiaⅡ7隐种对不同寄主的选择性和生态适应性

广东省感染木尔坦棉花曲叶病毒(CLCuMuV)的朱槿上烟粉虱隐种为MEAM1和AsiaⅡ7混合种群,并且AsiaⅡ7是该病毒有效传毒介体,明确这两个隐种的寄主选择性差异,能够为介体烟粉虱的防治提供依据。本研究选择锦葵科的棉花、黄秋葵和茄科的茄子、番茄4种寄主植物为供试寄主,通过笼内自由扩散、实验室嗅觉测定、发育观察,比较了MEAM1和AsiaⅡ7隐种对不同寄主的选择性和生态适应性。结果表明:烟粉虱MEAM1和AsiaⅡ7对4种寄主存在明显的寄主选择性,MEAM1成虫和产卵选择趋性为棉花>茄子>番茄>黄秋葵;AsiaⅡ7成虫和产卵选择趋性为棉花>黄秋葵>茄子>番茄。MEAM1和AsiaⅡ7在4种寄主上的发育历期、存活率有显著差异,MEAM1的存活率(卵到成虫)在棉花、茄子、黄秋葵上均较高(80%以上),番茄上最低(59%);AsiaⅡ7的存活率(卵到成虫)为棉花(90%)>黄秋葵(71%)>茄子(64%),在番茄上发育至1龄全部死亡。上述结果表明,4种作物均适宜MEAM1生长发育,锦葵科的2种作物适宜AsiaⅡ7隐种生长发育,茄科的2种作物不适宜。应避免棉花与黄秋葵近距离种植,防止传毒介体烟粉虱将黄秋葵作为桥梁寄主将病毒传播至棉花。     

一种含特异性侵染内共生菌Rickettisa massilia的烟粉虱种群建立方法

【目的】烟粉虱Bemisia tabaci(Gennadaius)广泛分布于全球除南极洲外各大洲的90多个国家和地区,其寄主范围广,危害严重,属于农业上的重要害虫。烟粉虱体内的内共生菌有助于其扩散传播,提高其寄主适合度,但其体内的内共生菌大都为次生内共生菌,很难进行体外培养,因此,加大了其内共生菌研究的难度。【方法】采集我国部分地区的烟粉虱种群,然后利用基于mt DNA COⅠ基因的种特异性引物进行种群生物型鉴定,再用内共生菌的特异性引物来鉴定含有特异性侵染内共生菌种群,确定个体含内生菌类型后,采取继代饲养方法建立种群。【结果】经鉴定烟粉虱的生物型为Q型(MED隐种),经过5代种群筛选后,可以建立稳定遗传的含特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。【结论】通过本方法可以获得稳定的含有特异性侵染内共生菌Rickettisa massilia菌的烟粉虱种群。该方法对研究烟粉虱与体内内共生菌、植物三者之间以及烟粉虱体内某几种内共生菌之间的互作有重要意义,同时也为明确烟粉虱体内Rickettisa菌的作用研究打下基础。     

烟粉虱的分类地位及在中国的分布

烟粉虱广泛分布于全球热带和亚热带地区。近20多年,烟粉虱的一些遗传群入侵世界各地,严重危害作物生产。烟粉虱遗传结构的多样性和复杂性早已被关注,但其分类地位,尤其是烟粉虱到底是一个包含多个生物型的种还是一个包含许多隐种的物种复合体,一直颇受争议。近几年,有关烟粉虱种系发生和系统学的研究取得长足进展,有证据推论其是一个包含至少31个隐种的物种复合体,但生殖隔离证据仍显不足,种系发生分析结果也因仅依据COI一个基因而受到质疑。因此,在大多数从事烟粉虱研究的同行接受其为一个物种复合体的概念的同时,仍有同行沿用生物型的概念。在我国境内已先后报道了包括13个本地种和2个全球入侵种在内的15个烟粉虱隐种。本地种主要分布在我国南部及包括海南岛和台湾岛的东南沿海地区,隐种的多样性由南向北逐渐降低。入侵种“中东一小亚细亚1”隐种（MEAMl）（即“B型”）和“地中海”隐种（MED）（即“Q型”）分别于20世纪90年代中后期和2003年前后入侵我国,并在许多地区迅速取代了本地种而占据优势地位。全国范围内的调查数据显示,这2个入侵种可在大部分区域共同存在,但自2005年以来,MED在许多地区陆续取代MEAMl,这很可能与MED对大量使用的新烟碱类杀虫剂有较强抗性有关。本文还讨论了烟粉虱隐种复合体分类所面临的命名等难题以及大范围抽样调查的数据偏差问题。     

A multiple PCR‐LDR assay for identification of two invasive cryptic species of whiteflies Bemisia tabaci

The MEAM1 and MED species of the cryptic species complex Bemisia tabaci are important invasive pests that cause tremendous crop losses worldwide. A rapid and highly reliable molecular technique is necessary to identify these species because they are morphologically indistinguishable. Therefore, a multiple polymerase chain reaction coupled with a ligase detection reaction (PCR‐LDR) that was based on polymorphisms in the mitochondrial cytochrome oxidase I (mtCOI) gene of B. tabaci was developed to distinguish the two cryptic species. An assessment of the method indicated that PCR‐LDR provided high specificity and sensitivity in discriminating MEAM1 (SHB) and MED (SHQ) whiteflies. In field tests, PCR‐LDR genotyping was performed in one 96‐well plate to identify 93 individuals collected from 8 districts in the suburbs of Shanghai. Complete concordance was observed between PCR‐LDR and sequencing methods. The method was used to confirm that MEAM1 and MED were found in two districts, but only the MED was found in the other six districts. PCR‐LDR, which is a transplantable platform, provides an alternative method for species identification of B. tabaci at low cost.     

Extensive settlement of the invasive MEAM1 population of Bemisia tabaci (Hemiptera: Aleyrodidae) in the Caribbean and rare detection of indigenous populations

Bemisia tabaci populations belonging to Middle East-Asia Minor one (MEAM1) and Mediterranean (MED) groups (formerly biotype B and Q, respectively) have spread throughout the world. Although the introduction of MEAM1 is documented from several Caribbean islands, it is generally not known whether MED has also been introduced; whether indigenous populations have survived; and if in the affirmative, to which group(s) they belonged. Whiteflies were collected from seven islands on various plant species. The prevalence of MEAM1 and non-MEAM1 individuals was assessed using a microsatellite approach validated with sequences of the mitochondrial cytochrome oxidase I (mtCOI) gene. Of the 262 samples tested, 247 exhibited the MEAM1 pattern, whereas none showed the MED pattern. The mtCOI gene was partially sequenced from a sample of individuals exhibiting MEAM1 (n = 15) and non-MEAM1 patterns (n = 8) and compared with type sequences. The 15 individuals exhibiting the MEAM1 pattern were confirmed to belong to MEAM1. Of the eight individuals representative of the six non-MEAM1 patterns, two belonged to the indigenous New World (NW) group of B. tabaci (NW), one belonged to a distinct species of Bemisia, and five belonged to MEAM1. One individual belonging to NW exhibited 99.9% nucleotide identity with a NW individual from Puerto Rico. The other was identified as the most divergent individual of the North and Central American genetic cluster. We conclude that a highly homogenous MEAM1 population has extensively settled in the Caribbean and that heterogeneous NW populations were still detectable although severely displaced.     

Phenolics,rather than glucosinolates,mediate host choice of Bemisia tabaci MEAM1 and MED on five cabbage genotypes

Worldwide, the most two important cryptic species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are MEAM1 (Middle East–Asia Minor 1, “B” biotype) and MED (Mediterranean, “Q” biotype). Although both B. tabaci MEAM1 and MED are polyphagous, they differ greatly in host choice and performance on various host plants. MEAM1 prefer to settle and perform better than MED on cabbage (Brassica oleracea), for example, but the underlying mechanism is largely unexplored. In the present study, we first measured the contents of the main secondary insect-resistant substances (glucosinolates and phenolics) and main nutrients (soluble proteins, total amino acids and total nitrogen) in five cabbage genotypes. We then examined the settling and oviposition choices of MEAM1 and MED on the five cabbage genotypes, respectively. The settling and oviposition preferences of both MEAM1 and MED were negatively related to the content of total phenolics rather than to the content of glucosinolates or main nutrients. Furthermore, our results showed that MEAM1 ranked the host quality of the cabbage genotypes more accurately than MED. The results at least in part indicate that total phenolics rather than glucosinolates mediate the host choice of B. tabaci MEAM1 and MED on the five cabbage genotypes.     

A new record for Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) species complex of Turkey

In this study, species complex of Turkish Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations was determined by PCR‐based DNA analysis. According to phylogenetic analyses, the B. tabaci samples have been identified within three generic groups. A major part of the samples belonged to two invasive species, either Middle East–Asia Minor 1 (MEAM1) or Mediterranean (MED). In addition to these two invasive species, several samples collected from greenhouses and cotton fields have been found to be related to Middle East–Asia Minor 2 (MEAM2), which is the first record of Turkish B. tabaci species complex.     

Genetic diversity and geographic distribution of Bemisia tabaci and Trialeurodes vaporariorum in Costa Rica

The tobacco whitefly Bemisia tabaci (Gennadius) cryptic species complex and of the greenhouse whitefly Trialeurodes vaporariorum (Westwood) are extensively reported as destructive pests in vegetable crops worldwide. A survey was conducted in 2011 and 2012 to determine the occurrence and genetic diversity present in the populations of these whiteflies in the major vegetable production areas of Costa Rica. Insect samples were collected from sweet pepper (Capsicum annuum L.), tomato (Solanum lycopersicum L.), common bean (Phaseolus vulgaris L.) and weeds present in commercial crops either in open field or greenhouse conditions. PCR‐RFLP analysis of mitochondrial cytochrome c oxidase subunit 1 gene (mtCOI) sequences of 621 whitefly individuals confirmed the presence of the Mediterranean (MED) type of the B. tabaci and of T. vaporariorum in most sampled regions. Also, individuals of the Middle East‐Asia Minor 1 (MEAM1) type of the B. tabaci were observed in low numbers. Contingency analyses based on type of crop, geographical region, whitefly species, year of collection and production system confirmed that T. vaporariorum was the most frequent species in vegetable production areas in Costa Rica, both in greenhouses and in open fields. B. tabaci MED is likely spreading to new areas of the country, whereas B. tabaci MEAM1 was mostly absent or rarely found. Comparisons of mtCOI sequences from B. tabaci individuals revealed the presence of four B. tabaci sequence haplotypes (named MED‐i, MED‐ii, MEAM1‐i, MEAM1‐xviii) in Costa Rica, three of them identical to B. tabaci haplotypes previously reported in the Western Hemisphere and other parts of the world. Analysis of sequences of T. vaporariorum individuals revealed a more complex population with the presence of 11 haplotypes, two of which were identical to T. vaporariorum sequences reported from other countries.