Melophagus ovinus (sheep ked) is a hematophagous ectoparasite that mainly parasitizes sheep. In addition to causing inflammation, wool loss, and skin damage to the animal hosts, M. ovinus also serves as a vector for a variety of pathogens and is highly likely to participate in the life and transmission cycle of pathogenic organisms. Herein, we investigated the presence and molecular characterization of vector-borne pathogens in M. ovinus from Qinghai-Tibet Plateau, China. A total of 92 M. ovinus pools collected from the Qinghai province of China were screened for the presence of selected vector-borne pathogens. The overall positive rate of A. ovis, A. bovis, A. phagocytophilum, and T. ovis in M. ovinus was 39.1%, 17.4%, 9.8%, and 89.1%, respectively. All of the samples were negative for Border disease virus (BDV), other Anaplasma species, Babesia spp., Rickettsia spp., and Borrelia spp. Co-infection of different Anaplasma species and T. ovis occurred in 51.2% of all samples with T. ovis. The positive rates of A. ovis, A. bovis, and A. phagocytophilum in different regions and altitudes of the sampling sites were significantly different. Sequence and phylogenetic analysis of target genes confirmed their identity with corresponding pathogens. Our results elucidate the occurrence and molecular characterization of Anaplasma spp. and Theileria spp. in M. ovinus, which could act as potential zoonotic reservoirs. To the best of our knowledge, this is the first report of the detection of A. bovis and A. phagocytophilum DNA in M. ovinus. This study gives the first extensive molecular survey of vector-borne pathogens with veterinary and public health significance in M. ovinus from the Qinghai-Tibet Plateau, China.

Background: Anaplasma, Babesia and Theileria are tick-borne pathogens (TBPs) that affect livestock worldwide. However, information on these pathogens in yaks (Bos grunniens) and Tibetan sheep (Ovis aries) on the Qinghai-Tibet Plateau (QTP), China, is limited. In this study, Anaplasma spp., Babesia spp. and Theileria spp. infections were assessed in yaks and Tibetan sheep from Qinghai Province. Methods: A total of 734 blood samples were collected from 425 yaks and 309 Tibetan sheep at nine sampling sites. Standard or nested polymerase chain reaction was employed to screen all the blood samples using species- or genus-specific primers. Results: The results showed that 14.1% (60/425) of yaks and 79.9% (247/309) of Tibetan sheep were infected with at least one pathogen. Anaplasma ovis, Anaplasma bovis, Anaplasma capra, Anaplasma phagocytophilum, Babesia bovis and Theileria spp. were detected in this study, with total infection rates for all the assessed animals of 22.1% (162/734), 16.3% (120/734), 23.6% (173/734), 8.2% (60/734), 2.7% (20/734) and 19.3% (142/734), respectively. For yaks, the infection rate of A. bovis was 6.4% (27/425), that of B. bovis was 4.7% (20/425) and that of Theileria spp. was 3.3% (14/425). Moreover, 52.4% (162/309) of the Tibetan sheep samples were infected with A. ovis, 30.1% (93/309) with A. bovis, 56.0% (173/309) with A. capra, 19.4% (60/309) with A. phagocytophilum and 41.4% (128/309) with Theileria spp. Conclusions: This study revealed the prevalence of Anaplasma spp., Babesia spp. and Theileria spp. in yaks and Tibetan sheep in Qinghai Province, China, and provides new data for a better understanding of the epidemiology of TBPs in these animals in this area of the QTP, China. Graphical

Introduction: The Maximum Entropy models (MaxEnt) is commonly employed for early detection of disease transmission, particularly effective in assessing the risk zones and intensity of tick-borne disease transmission based on climatic factors.Methods: Utilizing the diversity data of common ticks in Qinghai, along with surveillance statistics tick-borne pathogens of ticks and epidemiological information, we have charted and predicted the prevalence of tick and tick-borne pathogens across Qinghai province.Results and Discussion: The findings indicate that the pivotal environmental factors influencing the distribution of ticks and tick-borne pathogens include land use and land cover (Lucc), elevation (Elev), annual precipitation (Bio12) and annual mean temperature (Bio1). Notably, for Dermacentor nuttalli, the significant environmental factors accounted for 35.2% for Lucc, 24.7% for Elev and 18.6% for Bio12. In the case of Dermacentor silvaru, the pivotal factors contributed 33.2% for Lucc, 23.7% for Elev and 22.6% for Bio1. For Haemaphysalis qinghaiensis, the key environmental variables were 45% for Elev, 30.9% for Lucc and 18.4% for Bio12. Regarding the pathogens, the environmental factors influencing Borrelia burgdorferi contributed 61.5% for Lucc, 13.3% for Elev and 11.9% for Bio1. For Piroplasmida, the contribution was 62.1% for Lucc, 16.7% for Bio1 and 9.9% for Bio12. Rickettsia was influenced by factors accounting for 34.2% for Lucc, 29.7% for Elev and 17.3% for Bio12, while Anaplasma variables contributed 38.2% for Bio1, 26.6% for Lucc and 18.9% for Bio12. The finding indicated that the three predominantly ticks species (D. nuttalli, D. silvaru, and H. qinghaiensis) and the primary tick-borne pathogens (B. burgdorferi, Piroplasmida, Rickettsia, and Anaplasma) are predominantly concentrated in the source regions of the Yellow River, the Yangtze River, the Lancang River, and the eastern part of the Haixi Autonomous Prefecture. Projected under the ssp245 scenario, there is a notable expansion trend in the risk areas for ticks and tick-borne pathogens These findings are consistent with previous epidemiological studies on major ticks and tick-borne diseases in Qinghai conducted by our laboratory, this suggests the feasibility of using the MaxEnt model to predict the distribution of tick-borne disease transmission and compensates for the paucity of research on the Maxent model in the Qinghai Province.

The Qinghai–Tibet Plateau Area (QTPA) has a complex natural ecosystem, causing a greatly increased risk of spreading various tick-borne diseases including rickettsial infections, which are regarded as one of the oldest known vector-borne zoonoses. However, the information of one of its pathogen, spotted fever group Rickettsia (SFG Rickettsia), is limited in tick vectors and animals in this area. Therefore, this study focused on the investigation of SFG Rickettsia in tick vectors, yaks (Bos grunniens), and Tibetan sheep (Ovis aries) in the QTPA. A total of 1,000 samples were collected from nine sampling sites, including 425 of yaks, 309 of Tibetan sheep, 266 of ticks. By morphological examination, PCR, and sequencing, we confirmed the species of all collected ticks. All tick samples, all yak and Tibetan sheep blood samples were detected based on SFG Rickettsia ompA and sca4 gene. The results showed that all tick samples were identified to be Haemaphysalis qinghaiensis, and the positive rates of SFG Rickettsia were 5.9% (25/425), 0.3% (1/309), and 54.1% (144/266) in yaks, Tibetan sheep, and ticks, respectively. All positive samples were sequenced, and BLASTn analysis of the ompA gene sequences of SFG Rickettsia showed that all positive samples from animals and ticks had 99.04–100% identity with yak and horse isolates from Qinghai Province, China. BLASTn analysis of the sca4 gene sequences of SFG Rickettsia showed that all positive samples had 97.60–98.72% identity with tick isolates from Ukraine. In addition, the phylogenetic analysis showed that all the SFG Rickettsia ompA and sca4 sequences obtained from this study belong to the same clade as Rickettsia raoultii isolated from livestock and ticks from China and other countries. Molecularly, this study detected and characterized SFG Rickettsia both in the tick vectors and animals, suggesting that the relationship between SFG Rickettsia, tick species and animal hosts should be explored to understand their interrelationships, which provide a theoretical basis for preventing control of this pathogen.

Ticks are ectoparasites that feed on blood and pose a threat to both the livestock industry and public health due to their ability to transmit pathogens through biting. However, the impact of factors such as bloodmeal and geographic regions on the bacterial microbiota of Haemaphysalis qinghaiensis remains poorly understood. In this study, we used the v3-v4 region of the 16S rRNA gene to sequence the microbiota of Haemaphysalis qinghaiensis from eight groups (HY_M, YS_M, XH_M, LD_M, BM_M, LD_F_F, LD_F, and BM_F_F) in Qinghai Province. Significant differences in bacterial richness were observed between LD_F_F, BM_F_F, and LD_F (P < 0.01), and among the five groups (HY_M, YS_M, XH_M, BM_M, and LD_M) (P < 0.05). The bacterial diversity also differed significantly between LD_F_F, LD_F, and BM_F_F (P < 0.01), as well as among the five groups (HY_M, YS_M, XH_M, LD_M, and BM_M) (P < 0.01). The group with the highest number of operational taxonomic units (OTUs) was LD_F, accounting for 23.93 % (419/1751), while BM_F_F accounted for at least 0.80 % (14/1751). At the phylum level, Firmicutes was the most abundant, with relative abundance ranging from 7.44 % to 96.62 %. At the genus level, Staphylococcus had the highest abundance, ranging from 1.67 % to 97.53 %. The endosymbiotic bacteria Coxiella and Rickettsia were predominantly enriched in LD_F_F. Additionally, the 16S gene of Coxiella showed the highest identity of 99.07 % with Coxiella sp. isolated from Xinxiang hl9 (MG9066 71.1), while the 16S gene of Rickettsia had 100 % identity with Candidatus Rickettsia hongyuanensis strains (OK 662395.1). Functional predictions for the prokaryotic microbial community indicated that the main functional categories were Metabolic, Genetic information processing, and Environmental information processing across the eight groups. This study provides a theoretical basis for the prevention and treatment of tick-borne diseases, which is of great significance for public health.

The Qinghai Lake National Nature Reserve (QLNNR), renowned for its abundant natural resources and diverse ecological habitats, serves as an ideal environment for ticks, thereby increasing the risk of various tick-borne pathogens (TBPs) transmission. This study aimed to investigate the prevalence of TBPs in ticks collected from Przewalski's gazelle and Tibetan sheep within the QLNNR. A total of 313 tick samples were collected from the vicinity of Qinghai Lake. Tick species identification was conducted using both morphological and molecular biology techniques. Polymerase chain reaction (PCR) amplification was performed to detect the presence of spotted fever group (SFG) Rickettsia, Coxiella burnetii, Anaplasma phagocytophilum, Babesia microti, Theileria spp, Borrelia burgdorferi, Brucella spp, and Anaplasma ovis was performed using specific primers. Positive samples were sequenced and analyzed using BLASTn, followed by phylogenetic tree construction. The ticks collected from the Qinghai Lake area were identified as Dermacentor nuttalli. The overall prevalence rates of ticks carrying SFG Rickettsia and C. burnetii were 42.8 % (134/313) and 4.8 % (15/313), respectively. Three SFG Rickettsia species were detected, including R. raoultii 33.9 % (106/313), R. slovaca 3.8 % (12/113) and R. sibirica 7.7 % (24/113), with R. raoultii being the predominant species. The prevalence rates of SFG Rickettsia and C. burnetii in ticks from Tibetan sheep was 44.7 % (115/257) and 4.7 % (12/257), respectively,and in ticks from Przewalski's gazelle were 33.9 % (19/56) and 5.4 % (3/56). Furthermore, the study revealed a positive linear relationship between the abundance of Przewalski's gazelle and the number of ticks, as well as the prevalence of TBPs. The current study has identified Dermacentor nuttalli as the predominant tick vector species within the QLNNR region. The detection of SFG Rickettsia and C. burnetii has augmented our understanding of the epidemiological profile of ticks and TBPs in this area, thereby providing a robust theoretical foundation for the implementation of effective prevention and control strategies against TBPs.