Arnaout Youssef, Picard-Meyer Evelyne, Robardet Emmanuelle, Cappelle Julien, Cliquet Florence, Touzalin Frédéric, Jimenez Giacomo, Djelouadji Zouheira
Lyssavirus Unit, Nancy Laboratory for Rabies and Wildlife, ANSES, Malzéville, France.
USC 1233-INRAE Rongeurs Sauvages, Risque Sanitaire et Gestion des Populations, VetAgro Sup, Marcy l'Etoile, France.
PLoS One. 2023 Oct 20;18(10):e0292840. doi: 10.1371/journal.pone.0292840. eCollection 2023.
With over 1,400 species worldwide, bats represent the second largest order of mammals after rodents, and are known to host major zoonotic pathogens. Here, we estimate the presence of pathogens in autochthonous bat populations. First, we set out to check our samples for PCR amplification efficiency by assessing the occurrence of inhibited PCR reactions from different types of bat samples with amplifying the housekeeping gene β-actin. Second, we investigated the presence of five targeted pathogens in a French bat population using PCR. We targeted viral RNA of Canine distemper virus, Alphacoronavirus, Lyssavirus, Rotavirus and bacterial Leptospira DNA. To do so, we screened for these viruses in bat faecal samples as well as in oropharyngeal swab samples. The presence of Leptospira was assessed in urine, kidney, lung and faecal samples. Results showed a frequency of inhibited reactions ranging from 5 to 60% of samples, varying according to the sample itself and also suspected to vary according to sampling method and the storage buffer solution used, demonstrating the importance of the sampling and storage on the probability of obtaining negative PCR results. For pathogen assessment, rotavirus and alphacoronavirus RNA were detected in Myotis myotis, Myotis daubentonii, Myotis emarginatus and Rhinolophus ferrumequinum bats. Rotaviruses were also detected in Barbastella barbastellus. The presence of alphacoronavirus also varied seasonally, with higher frequencies in late summer and October, suggesting that juveniles potentially play an important role in the dynamics of these viruses. Leptospira DNA was detected in M. myotis and M. daubentonii colonies. The 16S rRNA sequences obtained from Leptospira positive samples showed 100% genetic identity with L. borgpetersenii. Neither canine distemper virus nor lyssavirus RNA were detected in any of the tested samples. This study is the first to show the presence of Leptospira in autochthonous French bats in addition to coronavirus and rotavirus RNA previously reported in European autochthonous bats.
蝙蝠是世界上种类超过1400种的哺乳动物，是仅次于啮齿动物的第二大目，并且已知携带着主要的人畜共患病原体。在此，我们估计本地蝙蝠种群中病原体的存在情况。首先，我们通过评估不同类型蝙蝠样本在扩增管家基因β-肌动蛋白时PCR反应受抑制的发生率，来检查我们样本的PCR扩增效率。其次，我们使用PCR调查了法国蝙蝠种群中五种目标病原体的存在情况。我们的目标是犬瘟热病毒、甲型冠状病毒、狂犬病病毒、轮状病毒的病毒RNA以及细菌钩端螺旋体的DNA。为此，我们在蝙蝠粪便样本以及口咽拭子样本中筛查这些病毒。在尿液、肾脏、肺和粪便样本中评估钩端螺旋体的存在情况。结果显示，受抑制反应的频率在样本的5%至60%之间，因样本本身而异，并且怀疑也因采样方法和所使用的储存缓冲溶液而异，这表明采样和储存对于获得阴性PCR结果的可能性至关重要。对于病原体评估，在鼠耳蝠、达氏鼠耳蝠、边缘鼠耳蝠和马铁菊头蝠中检测到轮状病毒和甲型冠状病毒RNA。在北棕蝠中也检测到了轮状病毒。甲型冠状病毒的存在也随季节变化，在夏末和10月频率较高，这表明幼体可能在这些病毒的动态变化中发挥重要作用。在鼠耳蝠和达氏鼠耳蝠群落中检测到钩端螺旋体DNA。从钩端螺旋体阳性样本中获得的16S rRNA序列与博氏钩端螺旋体显示出100%的基因同一性。在任何测试样本中均未检测到犬瘟热病毒和狂犬病病毒RNA。这项研究首次表明，除了先前在欧洲本地蝙蝠中报道的冠状病毒和轮状病毒RNA外，法国本地蝙蝠中还存在钩端螺旋体。
