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In this study, 263 tapeworms were collected from eight road-killed red foxes in Xinjiang Uygur Autonomous Region (XUAR, northwestern China). The tapeworms were analyzed based on morphological characters and mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequences. Eighty-three Taenia and 180 Mesocestoides tapeworms were identified according to the presence or absence of rostellum, and the number, the length and the shape of the large rostellar hooks. The morphological and molecular analyses revealed that i) 180 Mesocestoides tapeworms, here named as Mesocestoides sp. (Vulpes vulpes), showed 99.21% (378/381 bp) identity to Mesocestoides sp. reported from red fox in Mongolia; and ii) 83 Taenia tapeworms belonged to three species. The first Taenia species [n = 16, named as Taenia sp. (Vulpes vulpes)], based on the length of large rostellar hooks (337–342 μm) and its cox1 sequence, was identified as a potentially novel species, which is phylogenetically close to Taenia laticollis. The second species [n = 54, named as Taenia sp. (Vulpes vulpes & Rhombomys opimus)], was morphologically similar to Taenia endothoracicus according to the number (n = 52), the length (319–332 μm) and the shape of the large rostellar hooks. This species, infecting three red foxes, shared 100% cox1 sequence identity with Taenia sp. (Rhombomys opimus) genotype C found previously in great gerbils (Rhombomys opimus) in the same region. The third species (n = 13, named as Taenia polyacantha-like), had shorter large rostellar hooks (178–180 μm) and showed 96.27% (361/375 bp) sequence identity to Taenia polyacantha reported from red fox in Italy. The “great gerbil-red fox” life cycle of Taenia sp. (Vulpes vulpes & Rhombomys opimus), belonging to the mitochondrial lineage of T. endothoracicus, is confirmed. The T. polyacantha-like species was firstly found in red fox in China. Taenia sp. (Vulpes vulpes) is a potentially novel species, which is close to T. laticollis based on its phylogenetic properties.

Abstract Background Previously, twelve Rickettsia species were identified in ticks, fleas, sheep keds (Melophagus ovinus), bats (Pipistrellus pipistrellus) and a tick-bitten patient in the Xinjiang Uygur Autonomous Region (XUAR) in northwestern China. Here we aimed to molecularly detect rickettsial agents in red fox (Vulpes vulpes), marbled polecat (Vormela peregusna) and their ticks. Methods During 2018–2019, 12 red foxes, one marbled polecat and their ticks were sampled in two counties and a city of the XUAR. The heart, liver, spleen, lung and kidney of these 13 carnivores were dissected, followed by DNA extraction. Hard ticks were identified both morphologically and molecularly. All samples were examined for the presence of rickettsiae by amplifying four genetic markers (17-kDa, gltA, ompA, sca1). Results A total of 26 adult ticks and 28 nymphs (38 Ixodes canisuga, nine Ixodes kaiseri, six Haemaphysalis erinacei and one Dermacentor marginatus) were collected from red foxes, and four Ha. erinacei ticks were removed from the marbled polecat. Analysis of cytochrome c oxidase subunit I (COI) gene sequences indicated that 2–32 nucleotides differed between I. canisuga, I. kaiseri and Ha. erinacei from northwestern China and Europe. Rickettsia raoultii was detected in three red foxes, Candidatus Rickettsia barbariae in a red fox, Rickettsia sibirica in a red fox and a marbled polecat, and R. raoultii in two tick species (I. canisuga and D. marginatus). Conclusions To the best of our knowledge, I. canisuga and I. kaiseri have not been previously reported from red foxes in China. The DNA of R. sibirica and R. raoultii was detected for the first time in the organs of red foxes, and R. sibirica in the organs of a marbled polecat. This is also the first molecular evidence for the presence of R. raoultii in I. canisuga. Our findings expand the range of tick-borne pathogens in wildlife species and associated ticks in China.

Twenty-five dog fleas (Ctenocephalides canis) and five ticks identified as Hyalomma asiaticum were collected from a Eurasian lynx (Lynx lynx) pup in northwestern China. Molecular analyses of four genetic markers showed the presence of Rickettsia aeschlimannii DNA in 5 out of 20 fleas. Only rickettsial 17-kDa gene was detected in the blood sample of the lynx. In addition, 2 out of 20 fleas were positive to Wolbachia endosymbiont by targeting 16S rDNA while there was no Bartonella DNA found both in 5 ticks and 20 fleas by using gltA and 16S-23S ITS. Our findings suggest that i) C. canis parasitizing wild Eurasian lynx harbors R. aeschlimannii and Wolbachia endosymbiont in the China-Kazakhstan border, and ii) Wolbachia endosymbiont in present study is closer to that in C. canis infesting sheltered dogs in Turkey.

Abstract In this study, two road-killed marbled polecats (Vormela peregusna) were molecularly analysed for tick-borne pathogens. Rickettsia raoultii, “Candidatus Rickettsia barbariae” and a novel Babesia genotype have been identified, for the first time in marbled polecat. DNA of this Babesia sp. genotype was also present in four out of 15 Haemaphysalis erinacei ticks collected from the Babesia PCR-positive marbled polecat. Results of this study suggest that marbled polecats may serve as reservoirs for these bacteria and protozoans.

Background Ye Mule Aries sheep is one of the most important sheep breeds in Xinjiang, China. This breed is well adapted to harsh environmental conditions and displays strong disease resistance, fast growth, and high cold tolerance. To analyze the clonal expression and immunogenicity of the Ye Mule Aries sheep inhibin gene, total RNA was extracted from sheep ovarian tissue and used as a template to generate a eukaryotic expression vector and study inhibin immunogenicity. Methods Primers were designed to amplify the inhibin A gene via polymerase chain reaction and the amplified product was cloned between the ScalI and EcoRI restriction sites of the expression vector pEGFP-N1 to construct a recombinant plasmid, pEGFP-INHα. Following the validation of successful cloning, the pEGFP-INHα plasmid was transfected into BHK cells to verify expression in eukaryotes and subsequently utilized as an antigen in rabbits. Rabbits were tested for anti-inhibin antibodies and serum follicle-stimulating hormone (FSH) concentrations. Results The analysis of the INHα gene sequence revealed that INHα is 1109 bp long and is translated to an approximately 40 KDa protein. Bioinformatics approach indicated that the INHα gene is highly conserved between organisms. Immunization with the eukaryotic expression vector, pEGFP-INHα, which expresses the INHα gene elicited immune response and generatigeneration on of anti-INHα antibody. The antibody had a significant regulatory effect on the serum concentration of FSH in rabbits and led to higher levels of FSH, indicating increased ovary function. Conclusions The present work resulted in a successful construction of eukaryotic expression plasmid pEGFP-INHα and verified the immunogenicity of this highly conserved protein. Further, the expression of pEGFP-INHα was shown to have a significant impact on the secretion of FSH, indicating a potential regulatory role in ovarian function. In conclusion, our current findings can serve as a working model for studying the effect of INHα on the breeding performance of Ye Mule Aries sheep, providing a novel strategy to improve their reproduction rates.

The Ye Mule Aries (YM) Sheep is a valuable local sheep breed from Xinjiang, China. This breed is well adapted to harsh environmental conditions and displays strong disease resistance, fast growth, and high cold tolerance. The effect of small interfering RNA (siRNA) on the expression of follistatin inhibin alpha gene (INHa) in YM sheep was analyzed, and the INHa gene was amplified from total RNA of the ovary tissue of YM sheep and was successfully cloned. The eukaryotic siRNA expression vector was constructed with the cloned gene as the target and validated at the cellular level. Three RNA interference fragments at different sites were designed based on obtained cDNA of the target gene and the siRNA expression vector pGenesil 10-3p-siRNA was constructed. The siRNA expression vector was digested, identified and sequenced, and then transfected into YM sheep granulosa cells. The sequence of YM sheep INHa gene was obtained and uploaded on the NCBI database (accession number KP-113678.1). The INHa gene from YM sheep is 1109 bp long and translates to approximately 40 KDa protein. Q-PCR analysis showed that the plasmid pGenesil 10-3p-siRNA could interfere with the expression of INHa in granulosa cells to an efficiency of 83%, which was also confirmed through western Blot assay. This study successfully constructed the eukaryotic expression plasmid pGenesil 10-3p-siRNA, and confirmed that the plasmid interfered significantly with the expression of INHa gene in YM sheep. Our current findings can serve as a model for studying the effect of INHa on the breeding performance of YM sheep, providing a novel strategy to improve their reproductive capacities. [ABSTRACT FROM AUTHOR]