法国食虫虻（半翅目：异翅亚目）及其共生体的分子和 MALDI-TOF MS 鉴定。
Molecular and MALDI-TOF MS identification of swallow bugs Cimex hirundinis (Heteroptera: Cimicidae) and endosymbionts in France.
机构信息
IHU-Méditerranée Infection, Marseille, France.
Aix Marseille Univ, IRD, AP-HM, SSA, VITROME, Marseille, France.
出版信息
Parasit Vectors. 2021 Nov 27;14(1):587. doi: 10.1186/s13071-021-05073-x.
BACKGROUND
The Cimicidae are obligatory blood-feeding ectoparasites of medical and veterinary importance. We aim in the current study to assess the ability of MALDI-TOF MS to identify Cimex hirundinis swallow bugs collected in house martin nests.
METHODS
Swallow bugs were picked out from abandoned nests of house martin swallows and identified morphologically to the species level. The bugs were randomly selected, dissected and then subjected to MALDI-TOF MS and molecular analyses.
RESULTS
A total of 65 adults and 50 nymphs were used in the attempt to determine whether this tool could identify the bug species and discriminate their developmental stages. Five adults and four nymphs of C. hirundinis specimens were molecularly identified to update our MS homemade arthropod database. BLAST analysis of COI gene sequences from these C. hirundinis revealed 98.66-99.12% identity with the corresponding sequences of C. hirundinis of the GenBank. The blind test against the database supplemented with MS reference spectra showed 100% (57/57) C. hirundinis adults and 100% (46/46) C. hirundinis nymphs were reliably identified and in agreement with morphological identification with logarithmic score values between 1.922 and 2.665. Ninety-nine percent of C. hirundinis specimens tested were positive for Wolbachia spp. The sequencing results revealed that they were identical to Wolbachia massiliensis, belonging to the new T-supergroup strain and previously isolated from C. hemipterus.
CONCLUSIONS
We report for the first time to our knowledge a case of human infestation by swallow bugs (C. hirundinis) in France. We also show the usefulness of MALDI-TOF MS in the rapid identification of C. hirundinis specimens and nymphs with minimal sample requirements. We phylogenetically characterized the novel Wolbachia strain (W. massiliensis) infecting C. hirundinis and compared it to other recognized Wolbachia clades.
背景
蚰蜒科是具有医学和兽医重要性的强制性吸血外寄生虫。我们旨在当前的研究中评估 MALDI-TOF MS 识别在燕子巢中收集的食虫虻的能力。
方法
从燕子窝中取出食虫虻，并对其进行形态学鉴定到种水平。随机选择虻，解剖后进行 MALDI-TOF MS 和分子分析。
结果
共使用 65 只成虫和 50 只若虫来确定该工具是否可以识别虻种并区分其发育阶段。从 C. hirundinis 标本中分子鉴定了 5 只成虫和 4 只若虫，以更新我们的 MS 自制节肢动物数据库。这些 C. hirundinis 的 COI 基因序列的 BLAST 分析显示与 GenBank 中 C. hirundinis 的相应序列具有 98.66-99.12%的同一性。用数据库补充 MS 参考光谱进行的盲测试显示，100%（57/57）的 C. hirundinis 成虫和 100%（46/46）的 C. hirundinis 若虫得到可靠识别，与形态学鉴定一致，对数评分值在 1.922 和 2.665 之间。测试的 99%的 C. hirundinis 标本对 Wolbachia spp.呈阳性。测序结果表明，它们与 Wolbachia massiliensis 相同，属于新的 T-超级群菌株，以前从 C. hemipterus 中分离出来。
结论
我们首次报告了在法国发生的人类被食虫虻（C. hirundinis）叮咬的病例。我们还展示了 MALDI-TOF MS 在快速识别 C. hirundinis 标本和若虫方面的有用性，所需样本量最小。我们对感染 C. hirundinis 的新型 Wolbachia 菌株（W. massiliensis）进行了系统发育特征分析，并将其与其他公认的 Wolbachia 进化枝进行了比较。
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