Abstract
A variety of Chlamydia species belonging to the Chlamydiaceae family have been reported in reptilian hosts but scarce data about their occurrence in turtles and tortoises are available. In this study, research was conducted to acquire information on invasive alien species (IAS) of turtles and indigenous turtles and tortoises, living both free and in captivity, as possible reservoirs of Chlamydiaceae. Analysis of specimens (pharyngeal and cloacal swabs and tissues) from 204 turtles and tortoises revealed an overall Chlamydiaceae prevalence of 18.3% and 28.6% among free-living and captive animals respectively, with variable levels of shedding. Further testing conducted with a species-specific real-time PCR and microarray test was unsuccessful. Subsequently sequencing was applied to genotype the Chlamydiaceae-positive samples. Almost the full lengths of the 16S rRNA and ompA genes as well as the 16S-23S intergenic spacer (IGS) and 23S rRNA domain I were obtained for 14, 20 and 8 specimens respectively. Phylogenetic analysis of 16S rRNA amplicons revealed two distinct branches. Group 1 (10 specimens), specific to freshwater turtles and reported here for the first time, was most closely related to Chlamydia (C.) pneumoniae strains and the newly described Candidatus C. sanzinia. Group 2 (four specimens), detected in Testudo spp. samples, showed highest homology to C. pecorum strains but formed a separate sub-branch. Finally, molecular analysis conducted on positive samples together with their geographical distribution in places distant from each other strongly suggest that Group 1 specimens correspond to a new species in the Chlamydiaceae family. In-depth studies of Chlamydia spp. from turtles and tortoises are needed to further characterise these atypical strains and address arising questions about their pathogenicity and zoonotic potential.
Highlights
Chlamydiae are bacteria belonging to the Chlamydiaceae family
The presence of Chlamydiaceae was detected in 65.7% (23 out of 35) of Testudo spp. specimens based on the qPCR test
This study on Chlamydiaceae dissemination among free-living reptiles captured in lakes and rivers of eastern Poland was performed
Summary
Chlamydiae are bacteria belonging to the Chlamydiaceae family. Their host specificity varies: e.g. C_caviae is found in guinea pigs, whereas C. pneumoniae, C. pecorum or C. abortus can infect different animals as well as humans. Chlamydiaceae have been observed among more reptiles including the Burmese python [8, 9], emerald tree boa [10], western sand viper [11], chameleon [12], crocodiles (Nile, Indo-Pacific and Siamese) [5, 13, 14] and tortoises belonging to the Testudinidae family [15, 16]. Species determination in Chlamydiaceae-positive samples was not always possible but retrospective study and the newest research have shown C. pneumoniae presence in snakes and chameleon, and other yet unclassified Chlamydia spp. in the remaining cases [17]. Candidate species C. sanizinia was reported for the first time in a Madagascar tree boa (Sanzinia madagascariensis volontany) [3]
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