Soft ticks are important hematophagous ectoparasites that are found in all zoogeographical regions of the world. In addition to causing direct economic losses, they are also important as disease causing agents. However, in many countries, including Pakistan, their diversity and associated pathogens remained poorly studied. This study aimed to minimize this knowledge gap by conducting morpho-molecular confirmation of soft ticks and molecular assessment of associated Borrelia in this country. A total of 62 ticks were collected from nesting environment of pigeons in Khyber Pakhtunkhwa, Pakistan. Of these, 24.19% (15/62) were identified as Argas hermanni (males = 9, 60%, females = 6, 40%), 62.90% (39/62) as Argas persicus (males = 17, 43.58%, females = 14, 35.89%, nymphs = 8, 20.51%), and 12.90% (8/62) (all nymphs) as an undetermined Argas species. Twenty-nine ticks, representing all three species, were randomly selected for DNA extraction, and subjected to polymerase chain reaction (PCR) to amplify partial fragments of 16S rDNA and CO1 for ticks, and flaB for the associated Borrelia species. Among these, Borrelia DNA was detected in 4/18 (22.2%) A. persicus ticks (2/8 males and 2/7 females). The 16S rDNA and CO1 sequences of A. hermanni showed maximum 98.17% and 99.30% identities, respectively, with conspecifics from Iran, while the CO1 sequence of A. persicus showed 100% identity with conspecifics from Kenya and Kazakhstan. The 16S rDNA and CO1 sequences of Argas sp. showed 96.48% and 97.33% identities, respectively, with A. persicus from Pakistan and Kenya. The flaB sequence associated with A. persicus showed 100% identity with Borrelia anserina from Brazil and Pakistan. In the phylogenetic trees of ticks, A. hermanni (16S rDNA and CO1) and A. persicus (CO1) from the current study clustered with conspecifics, while Argas sp. formed a sister clade with A. persicus group (16S rDNA and CO1). In the flaB phylogenetic tree, B. anserina from the current study clustered with corresponding species in the relapsing fever group. This study provides the first molecular record of A. hermanni, the first report of A. persicus associated with pigeons, with detection of B. anserina, and also documents a novel lineage closely related to A. persicus, all from Pakistan.

First molecular record of Argas hermanni and Argas sp. closely related to Argas persicus, with detection of Borrelia anserina in Argas persicus from pigeons in Pakistan

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发表日期：

2026年5月26日

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Zia Ur Rahman,
Mehran Khan,
Zaibullah Khan,
Wali Khan,
Ahmad Khan,
Iram Liaqat,
Abid Ali,
Mashal M. Almutairi

摘要

Soft ticks are important hematophagous ectoparasites that are found in all zoogeographical regions of the world. In addition to causing direct economic losses, they are also important as disease causing agents. However, in many countries, including Pakistan, their diversity and associated pathogens remained poorly studied. This study aimed to minimize this knowledge gap by conducting morpho-molecular confirmation of soft ticks and molecular assessment of associated Borrelia in this country. A total of 62 ticks were collected from nesting environment of pigeons in Khyber Pakhtunkhwa, Pakistan. Of these, 24.19% (15/62) were identified as Argas hermanni (males = 9, 60%, females = 6, 40%), 62.90% (39/62) as Argas persicus (males = 17, 43.58%, females = 14, 35.89%, nymphs = 8, 20.51%), and 12.90% (8/62) (all nymphs) as an undetermined Argas species. Twenty-nine ticks, representing all three species, were randomly selected for DNA extraction, and subjected to polymerase chain reaction (PCR) to amplify partial fragments of 16S rDNA and CO1 for ticks, and flaB for the associated Borrelia species. Among these, Borrelia DNA was detected in 4/18 (22.2%) A. persicus ticks (2/8 males and 2/7 females). The 16S rDNA and CO1 sequences of A. hermanni showed maximum 98.17% and 99.30% identities, respectively, with conspecifics from Iran, while the CO1 sequence of A. persicus showed 100% identity with conspecifics from Kenya and Kazakhstan. The 16S rDNA and CO1 sequences of Argas sp. showed 96.48% and 97.33% identities, respectively, with A. persicus from Pakistan and Kenya. The flaB sequence associated with A. persicus showed 100% identity with Borrelia anserina from Brazil and Pakistan. In the phylogenetic trees of ticks, A. hermanni (16S rDNA and CO1) and A. persicus (CO1) from the current study clustered with conspecifics, while Argas sp. formed a sister clade with A. persicus group (16S rDNA and CO1). In the flaB phylogenetic tree, B. anserina from the current study clustered with corresponding species in the relapsing fever group. This study provides the first molecular record of A. hermanni, the first report of A. persicus associated with pigeons, with detection of B. anserina, and also documents a novel lineage closely related to A. persicus, all from Pakistan.