High prevalence and genetic variation of Bartonella species inhabiting the bats in southwestern Yunnan

doi:10.17520/biods.2021028

Abstract

Abstract:

Aims: Bats are natural reservoir hosts of many zoonotic viruses and non-viral pathogens. Several lines of evidence indicate that bats harbor diverse groups of Bartonella species and genotypes. Little is still known about how prevalence varies and the genetic diversity of Bartonella spp. in bats in China.
Methods: Between 2015 and 2017, bats were captured in four areas in southwestern Yunnan by the net trapping method. Liver and spleen tissues of bats were collected, and nucleic acids were extracted after mixing and grinding. The ssrA gene of Bartonella spp. was detected by using the TaqMan real-time fluorescent quantitative PCR assay (qPCR). The positive products of qPCR were amplified and sequenced to identify the species of Bartonella. The phylogenetic relationships were constructed by binary and network methods.
Results: We present the results of the investigation of bat-borne Bartonella in four areas of Yunnan Province, which have shown a high prevalence of Bartonella and high genetic diversity in the local bat population. Bartonella gene ssrA was detected by using qPCR in 56.4% (172/305) bats captured from four sampling sites, including 22 Rhinolophus affinis, 18 Rhinolophus blythi, and 132 Rousettus leschenaultii. There was no statistically significant difference (χ² = 1.135, P = 0.567) in the infection rates among species. The infection rates of Lincang, Xishuangbanna, Baoshan, and Ruili were 50.0% (22/44), 61.7% (29/47), 62.1% (18/29), and 55.7% (103/185), respectively. Thirty-seven ssrA sequences of Bartonella were obtained by second amplification from qPCR and sequencing. The binary tree revealed 10 phylogroups, one of which was related to B. elizabethae, B. tribocorum and B. krasnovii, the other sequences were far from the known Bartonella spp. and were closely related to bat-borne Bartonella from bats in Asia, Europe, and America. The genetic diversity analysis showed that the nucleotide diversity index (π) was 0.11381 ± 0.00928, and the genotypic diversity index (Hd) was 0.985 ± 0.010, which brought about 29 genotypes. The genetic networks constructed by 115 ssrA sequences exhibited that Bartonella spp. from global bats had an incomplete geographical distribution and host specificity.
Conclusion: We found that the bat-borne Bartonella in Yunnan was genetically diverse, geographically widespread, and endemic and that bats play a part in transmitting Bartonella species locally. The analysis of phylogenetic networks based on the global dataset of ssrA sequences of bat-borne Bartonella points to an evolutionary association of Bartonella spp. with their bat hosts and the effect of geographical isolation.

Key words: Bartonella, bat, genetic diversity, phylogenetic analysis, reservoir

Dongmei Li, Weihong Yang, Qingduo Li, Xi Han, Xiuping Song, Hong Pan, Yun Feng. High prevalence and genetic variation of Bartonella species inhabiting the bats in southwestern Yunnan[J]. Biodiv Sci, 2021, 29(9): 1245-1255.

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URL: https://www.biodiversity-science.net/EN/10.17520/biods.2021028

https://www.biodiversity-science.net/EN/Y2021/V29/I9/1245

Figures/Tables 4

Table 1 Number of species and geographical origins of the bats infected with Bartonella spp.

蝙蝠种类 Species of bats	感染率(阳性数/检测数) Infection rate (positive/detected)				合计 Total
蝙蝠种类 Species of bats	临沧市 Lincang City	西双版纳州 Xishuangbanna Prefecture	保山市 Baoshan City	德宏州 Dehong Prefecture
中菊头蝠 Rhinolophus affinis	50.0% (22/44)	0	0	0	50.0% (22/44)
小菊头蝠 Rhinolophus blythi	0	0	62.1% (18/29)	0	62.1% (18/29)
棕果蝠 Rousettus leschenaultii	0	61.7% (29/47)	0	55.7% (103/185)	56.9% (132/232)
合计 Total	50.0% (22/44)	61.7% (29/47)	62.1% (18/29)	55.7% (103/185)	56.4% (172/305)

Fig. 1 Phylogenetic tree highlighting the position of ssrA sequences of Bartonella from bats relative to reference species of Bartonella spp. The bat species in the inner circle of the phylogenetic tree, represented by red, blue, and brown lines; bat sequence names in this study are marked with red font, the same as specimen no. in Appendix 2. BSLY, Lujiang Town, Longyang district, Baoshan City; LCZK, Nansan Town, Zhenkang County, Lincang City; BNML, Yaoqu Town, Mengla County, Xishuangbanna Prefecture; DHRL, Wanding Town, Ruili City, Dehong Prefecture; I-X in the center represent the phylogroups of ssrA sequence from bats in this study, while Brucella melitensis CP044985 and Rhizobuim sp. NXC24 CP024311 are outgroups.

Fig. 2 Median-joining network of ssrA genotypes for Bartonella spp. from bat specimens in this study. Genotypes are denoted as pies, in which the numbers are number of individuals, with a size proportional to genotypic frequency.

Fig. 3 Phylogenetic network diagram generated from ssrA sequences of Bartonella spp. from bat specimens in this study and GenBank using NeighborNet algorithm. There are 152 ssrA sequences of Bartonella spp. in the network diagram, including 37 sequences in this study and 115 sequences downloaded from the GenBank database. A and B are the networks of bat-borne Bartonella from the globe according to the geographic distributions and the host taxonomy associations.

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