African swine fever virus (ASFV), the sole known DNA arbovirus, causes acute hemorrhagic fever in domestic pigs with case fatality rates approaching case fatality 100%. While less contagiously transmitted than viral agents like foot-and-mouth disease virus, ASFV’s epidemiological significance stems from its complex transmission ecology involving soft ticks. Ornithodoros ticks are established biological vectors, with eight species confirmed globally. However, potential vectors in China, where Ornithodoros lahorensis (O. lahorensis) is endemic and remain uncharacterized. This study tested the hypothesis that O. lahorensis serves as a competent biological vector for ASFV under laboratory conditions. We hypothesized that vector competence, defined as the ability to acquire, maintain, and transmit infectious African swine fever virus, differs between soft (O. lahorensis, Argas persicus) and hard (H. longicornis, R. sanguineus, D. silvarum) tick species, and tested this using experimental transmission assays. Results demonstrated that O. lahorensis efficiently supported transstadial persistence and transovarial transmission of ASFV. Naïve pigs exposed to infected O. lahorensis developed acute ASF with case fatality 100%. Viral replication in salivary glands, midgut, and Malpighian tubules was confirmed by qPCR and transmission electron microscopy, with no significant fitness costs to ticks. In contrast, A. persicus and all ixodid species failed to transmit. Here identifies O. lahorensis as a critical ASFV vector in China, informing targeted acaricidal control in endemic regions.

Investigating the prevalence and molecular genetic characteristics of Anaplasma ovis, Theileria spp., Anaplasma phagocytophilum, and hemotropic Mycoplasma infections in sheep populations across different regions of Gansu Province is of significant importance for the prevention and control of these pathogens. A total of 1523 sheep blood samples were collected from 19 counties (districts) in Gansu Province. Pathogen screening was conducted using PCR-based molecular detection techniques, followed by sequencing and phylogenetic analysis of specific genes (e.g., Msp4, 18S rRNA) from selected positive samples. Blood-borne pathogens infections in Gansu Province were widespread but unevenly distributed geographically. Theileria spp. and Anaplasma ovis were the dominant pathogens, with overall infection rates of approximately 16.7% and 9.6%, respectively. The highest Anaplasma ovis infection rate (82.5%) was observed in the Gannan region, where co-infections were common (24/97). An exceptionally high Theileria spp. infection rate (87.5%) was detected in the Zagana area. No pathogens were detected in Wuwei, Jingyuan, Huining, Jingtai, Qinghuan, or Maqu. Phylogenetic analysis revealed that the Msp4 gene sequences of Anaplasma ovis isolates from Gansu shared 99.48% homology with strains from Europe, Asia, and Africa. Anaplasma phagocytophilum isolates also showed high homology (99.53-99.84%) with multiple global strains. Seasonal data indicated significantly higher Theileria spp. infection rates in spring (23-34%) compared to other seasons (approximately 12%). Gansu Province is an endemic area for multiple blood-borne pathogens, with distinct regional clustering and seasonality in prevalence. The high conservation of pathogen gene sequences suggests genetic stability. This study provides essential epidemiological baseline data and a scientific foundation for targeted prevention and control of blood-borne pathogen diseases in sheep in Gansu Province.

Ticks, which are globally distributed, transmit a wide range of human and animal pathogens. Soft and hard ticks are two major tick families exhibiting distinct vector capacities and blood-feeding abilities. However, understanding the genetic basis of these differences is hindered by the scarcity of high-quality tick genomes, particularly for soft ticks. Here, we present high-quality genomes of two soft and two hard tick species of both evolutionary and medical importance, and compare the genomic characteristics of the two tick families. Compared with soft ticks, hard ticks exhibited faster genome evolution, characterized by larger genome sizes, higher rates of gene gain and loss, and a greater number of positively selected genes. Both tick families exhibited frequent positive selection across metabolism and organismal systems, sharing a core set of adaptive genes but undergoing refinement at different sites. Gene families underpinning various vector capacities, including those involved in cuticle structure, heme biosynthesis, egg waxing, and immune pathways, were identified. Additionally, we assembled the genomes of two novel Rickettsia endosymbionts from soft ticks, revealing high genomic divergence and lineage-specific adaptations to their hosts. These genomic resources provide fundamental insights into the genetic basis of tick biology and vector capacity, highlighting the divergent evolutionary strategies of blood-feeding arthropods shaped by their ecological niches.

Theileria annulata causes tropical theileriosis in cattle, yet the molecular basis of host-parasite crosstalk across intracellular stages remains incompletely defined. We combined RNA sequencing and untargeted metabolomics to profile paired uninfected and infected bovine leukocytes (schizont stage) and erythrocytes (piroplasm stage), together with purified schizonts and piroplasms. Integrated analyses revealed pronounced, cell type-specific reprogramming. Infected leukocytes showed activation of immune signalling, amino acid metabolism and energy-producing pathways, consistent with leukocyte transformation, whereas infected erythrocytes preferentially engaged glutathione metabolism and redox homeostasis. Parasite stage comparisons uncovered extensive transcriptional and metabolic rewiring, including stage-biased expression of mitochondrial components, antioxidant systems and putative stage-regulated transcription factors. These coherent host-parasite adaptations likely facilitate parasite survival and persistence within distinct cellular niches. This work delineates a stage-resolved multi-omics landscape of T. annulata infection spanning host and parasite compartments and identifies signalling and metabolic pathways that merit functional validation as candidates for improved diagnostics and targeted interventions against bovine tropical theileriosis.

African swine fever virus (ASFV) causes an incurable swine disease with nearly 100% mortality, posing a catastrophic threat to global pig production. The soft tick Ornithodoros lahorensis acts as a critical biological vector that sustains persistent ASFV replication and mediates long-distance viral transmission, yet the molecular mechanisms governing ASFV-tick interplay remain poorly understood. Here, we integrated transcriptomics and metabolomics to systematically dissect molecular changes in O. lahorensis across three infection stages: Uninfected control, early infection (7 days post-infection, dpi), and late persistent infection (21 dpi). Multi-omics integration revealed that ASFV extensively remodels tick host metabolism, predominantly activating purine/pyrimidine metabolism, lipid biosynthesis, and energy metabolism. We further characterized a conserved regulatory module consisting of 12 core genes and 8 signature metabolites that collectively support ASFV genome replication and virion assembly. Three hub metabolic genes (TK1, ATP5F1B, and IMPDH) were selected for functional validation via siRNA silencing in ticks; individual gene silencing suppressed ASFV loads by 89.2%, 91.5%, and 87.8%, respectively (p < 0.001***). This work represents the first comprehensive multi-omics investigation of ASFV infection in O. lahorensis. We identified tick-specific molecular targets to block vector-mediated ASFV spread and established a standardized multi-omics analytical pipeline for tick-virus interaction research. Our findings elucidate the mechanistic basis of long-term ASFV persistence in soft ticks and deliver novel actionable clues for developing vector-targeted ASF intervention strategies.

Arthropod including ticks, fleas, and lice have been found to be infected with Rickettsia massiliae, a pathogenic member of spotted fever group, causing rickettsiosis. This comprehensive scoping review summarize the known facts about its transmission, diagnostic-methods, phylogenetic position, human-case-reports, preventive-measures, and distribution in Palearctic and Oriental regions. Three main steps were followed to compile this study: explanation of objective(s), identification of relevant literature, and retrieval of data determined by inclusion and exclusion criteria. Various databases, including Science Direct, Web of Science, PubMed, Scopus, Cochrane, and Google Scholar were screened for relevant literature. Our objectives were to collect data regarding transmission method(s) of R. massiliae, identification-assay(s), phylogenetic position, clinical-reports globally, and distribution via possible vectors host animals surviving in Palearctic and Oriental regions. Descriptive analysis has been conducted to plot the frequency graphs of reported numbers in different countries and hosts. Findings presented that R. massiliae have been found across Palearctic and Oriental regions in 5 tick genera (Rhipicephalus, Hyalomma, Haemaphysalis, Ixodes, Amblyomma, and Dermacentor), 1 of louse (Haematopinus), 1 of sheep ked (Melophagus), and 1 of flea (Archaeopsylla), although their role as a vector(s) is still unknown. Dogs, sheep, cattle, and goats were recorded as epidemiologically important host animals for infected arthropods in 30, 23, 17, and 16 different studies, highlighting their role in its possible transmission. Additionally, Rh. sanguineus sensu lato (s.l) and Rh. turanicus were reported to infest the aforementioned animals, which is also being recognized as potential vector for its transmission. Highest number of R. massiliae reports (10) from different animals and vectors were recorded in China. Italy was recorded with highest (3) clinical cases of humans to-date. Moreover, transmission methods like transovarial via Rh. turanicus and horizontal/artificial-feeding via Rh. sanguineus s.l. and various diagnostic methods for R. massiliae have been documented. R. massiliae has been widely documented throughout Palearctic and Oriental regions, with human cases reported in about six countries. Rh. sanguineus and Rh. turanicus stands out the potential vectors, while further investigation into the implications of diverse range of arthropods in epidemiology of this bacterium.

Ticks are obligate blood-sucking ectoparasite and common vector of a wide variety of zoonotic pathogens. In this research, we sequenced and assembled the circular complete mitogenome information of Chinese isolates of Hyalomma anatolicum maintained at the Laboratory. The 14,714 bp mitogenome consists of 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes and 2 ribosomal RNA (rRNA). The nucleotide sequence of Hy. anatolicum mitogenome includes approximately A (35.05%), G (8.70%), T (41.20%) and C (11.26%), the proportion of A + T (80.04%) is substantially higher than G + C (19.96%). We compared mitogenome selection pressure of the Hy. anatolicum and found that positive selection signatures were detected in atp8, contrasting sharply with the strong purifying selection observed in cox1. Phylogenomic reconstruction using concatenated PCGs robustly placed Hy. anatolicum within a clade sister to Hy. excavatum. We further validated cox3, cytb, nad1, nad2, nad3, nad4, nad4L, nad5 and nad6 as lineage-specific markers mirroring full mitogenome topology, proposing their cost-effective use in Hyalomma systematics. The divergence time analysis suggests that Hy. anatolicum was the most recent species in Hyalomma, 1.95 Mya. As far as we know, this is the first study exploring the complete mitogenome for Hy. anatolicum. Here, we obtained the whole mitogenome sequence for the species Hy. anatolicum, which enriched the mitogenome data of ticks, provided new idea for future research on the molecular evolution, population heredity and systematics of ticks.

The prevalence of tick-borne bacterial and viral diseases, which pose a serious threat to human and livestock health, is increasing worldwide. At present, only a limited number of tick-borne pathogens have been reported, and no analysis of the microbial pathogen community in ticks has been carried out. We sequenced the viral metagenome of Ornithodoros lahorensis species of ticks from the Chinese mainland and identified 390 RNA viruses with unique microbial compositions. A total of 992 assembled viral transcriptomes revealed the breadth and diversity of the genome structure of tick-borne viruses, reflecting the importance of ticks as RNA viral pools. We analyzed the phylogeny of different virus families to investigate virus evolution and found that the most diverse tick-associated viruses belonged to the family Siphoviridae, which diverged earlier in evolutionary time than other arboviruses. There were only a few tick-specific viruses, whereas the number of vertebrate-infecting viruses in ticks was greater. We hope that our virus sequencing dataset will facilitate future important research on viruses carried by ticks that can infect vertebrates.

Many chemical acaricides have been used to mitigate the detrimental effects of Haemaphysalis longicornis. However, the excessive use of chemical acaricides may facilitate the development of resistance in H. longicornis. Larval packet tests (LPTs) were conducted on larvae aged 14–21 days collected from 10 counties in China to evaluate the effectiveness of different acaricides, including cyhalothrin, cypermethrin, deltamethrin, amitraz, and fipronil. Additionally, genomic DNA was extracted from tick samples collected from 14 counties, and partial fragments of the voltage-gated sodium channel (VGSC), beta-amino-oxidoreductase (β AOR), and gamma-aminobutyric acid (GABA)-gated chloride channel genes were amplified via PCR. According to the LPT findings, it can be inferred that ticks from all localities exhibited resistance to at least three different acaricides. Multidrug resistance to five acaricides was detected in 40

Global distribution of ticks and their associated tick-borne pathogens (TBPs) presents substantial health concerns for both humans and animals. The present study aimed to investigate the distribution, morpho-molecular identification, and associated TBPs of diverse tick species collected from Khyber Pakhtunkhwa and Punjab provinces of Pakistan. Morphological identifed ticks were molecularly confirmed via cytochrome oxidase I (COI) and 16S rRNA genes, which showed 15 different tick species. Among them, we found the highest prevalence rate of Rhipicephalus (Rh) microplus (404/1803; 22.41%), while the lowest prevalent tick species were Haemaphysalis (Hae) montgomeryi (44/1803; 2.44%). Similarly, the highest tick load was found on cattle (544/186; 2.92), while least was found on goats (272/164; 1.66). Various TBPs, including Rickettsia (R) spp. (via gltA, sca4, ompA, ompB genes), Anaplasma (A) sp. (via 16S rRNA gene), Coxiella burnetii (via IS1111 gene), Piroplasm and Hepatozoon (H) spp. (via 18S rRNA gene) were screened. The Rh. microplus tick species showed highest positivity rate (11.63%) for various TBPs, whereas Hae. sulcata ticks were recorded as TBPs free in current study. Among the detected TBPs, Coxiella burnetii was most prevalent (1.72%), followed by A. phagocytophilum & R. slovaca (1.44% each), R. sibirica (0.78%), R. raoultii (0.61%), H. canis (0.55%), R. conorii subsp. raoultii (0.50%), Hepatozoon sp., (0.44%), Theileria (Th) uilenbergi (0.39%), Babesia (B) microti (0.33%), and Th. luwenshuni (0.28%). This study provides the first report of TBPs, including R. slovaca, R. sibirica, and B. microti, in ticks from Pakistan. Phylogenetic analysis was performed based on the aforementioned genetic markers, in which ticks and their associated TBPs formed distinct clades with their corresponding isolates. This research work provides a deep insight regarding the characterization of different ticks and their associated TBPs in Pakistan and, significantly, it presents evidence of potential zoonotic threats to both animal and public health due to these newly detected pathogens.

Haemaphysalis longicornis is associated with the transmission of 30 various types of pathogens, having medical and veterinary importance. Hence, this study was designed to investigate the prevailing pathogen in Zhangjiachuan county, Gansu province, China. For the said purpose, we performed a molecular investigation of Rickettsia and Piroplasms pathogens in morpho-molecularly confirmed H. longicornis tick species, collected from cattle in the study area. Furthermore, the efficacy of deltamethrin was evaluated using larval packet tests (LPTs) against this tick species, followed by molecular and in silico analysis of voltage-gated sodium channels (VGSCs) gene. Moreover, their amplified VGSCs genes (Domain II and a part of its linker region with Domain III) were comparatively aligned with insecticide-susceptible Rhipicephalus (Rh) microplus (AF134216.2, OQ349192.1, KM073932.1), Rh. sanguineus (KU886031.1) and resistant Rh. microplus (MH986341.1, KM073928.1, KM073930.1, KM073931.1, KM073933.1), Rh. annulatus (MN535171.1), and Rh. appendiculatus (MH053429.1). Our findings revealed 149/469 (31.77%) and 92/469 (19.62%) prevalence rates of single infection for Candidatus Rickettsia jingxinensis (via 17-KDa and ompB genes) and Theileria orientalis (via 18S rRNA gene), respectively. Additionally, LPTs results expressed diverse mortality rates of 8.17%, 17.63%, 22.26%, 43.18%, 73.46%, and 87.51% via the application of 0.00006, 0.0006, 0.006, 0.06, 0.6, and 6 mg/mL deltamethrin concentrations, respectively, indicating the presence of reduced susceptible larvae in the targeted region. Comparative alignment of VGSCs gene (domain-II) showed 29 single nucleotide polymorphisms (SNPs) including 5 nonsynonymous (NS) SNPs. All five NS-SNPs were in the linker regions, playing a crucial role in protein channels (VGSCs) formation. Whereas comparison of the obtained partial domain-III of VGSCs gene with Rh. microplus and Rh. sanguineus tick species showed 100% conservativeness as no SNP was identified in its domain-III. In silico analysis regarding the 3D structure of VGSCs protein revealed suitability for attachment of deltamethrin and recorded binding affinity was -524.58 kcal/mol. This study presented the first ever molecular report of VGSC gene of H. longicornis tick species which will be helpful in their management as well as to mitigate their negative impacts.

Ticks are globally important vectors of human and animal pathogens. This study characterized microbial communities in Haemaphysalis longicornis (from humid Zhangjiachuan County) and Hyalomma dromedarii (from arid Minqin County, Gansu Province, China) using metagenomic sequencing. We identified diverse bacteria and viruses including known pathogens (Anaplasma phagocytophilum, Totivirus spp., Escherichia spp.) and potentially novel agents (Totivirus-like and Trachysalambria curvirostris-associated sequences). These results highlight the role of these tick species in pathogen transmission across different environments and emphasize the need for region-specific surveillance. Our findings contribute to the understanding of tick-borne disease risks and inform strategies for targeted control.

Ovine theileriosis is an important tick-borne protozoan disease. It has been reported that three Theileria species are responsible for ovine theileriosis in China, which are T. luwenshuni, T. uilenbergi, and T. ovis. Here, we established three detection techniques based on loop-mediated isothermal amplification (LAMP) and nanoparticle-based lateral flow biosensor (LFB) for the infection of the three Theileria species. Three LAMP primer sets were designed targeting the nucleotide sequences of the 28S rRNA gene of T. luwenshuni, T. uilenbergi, and T. ovis. We used LAMP coupled with real-time fluorescence detection to optimize the concentrations of dNTP Mix, MgSO4, and Bst 2.0 DNA polymerase, as well as the reaction temperature of the LAMP assay, and then combined LAMP with LFB (LAMP-LFB). The entire detection assay process, including genomic DNA extraction (40 min), LAMP reaction (40 min), and LFB readout (<5 min), can be completed within 85 min. The established assays can specifically detect species of T. luwenshuni, T. uilenbergi, and T. ovis infection without cross-reaction with other Theileria, Babesia, and Anaplasma species. The detection limits of the LAMP-LFB assays for T. luwenshuni, T. uilenbergi, and T. ovis plasmid templates were 2.72 × 102 copies/μL, 2.96 × 103 copies/μL, and 3.05 × 101 copies/μL, respectively. Finally, we compared the established LAMP-LFB assay with the traditional PCR assay. The results showed that the total coincidence rates were 96.67 % (T. luwenshuni), 96.67 % (T. uilenbergi), and 93.33 % (T. ovis), respectively. In general, we developed a rapid, simple, sensitive, and specific technique for differential detection of T. luwenshuni, T. uilenbergi, and T. ovis infection in small ruminants.

The protozoan pathogen, Theileria annulata, is transmitted by ticks which is very detrimental to various breeds of cattle, leading to the development of theileriosis. Early diagnosis and control of the disease is a crucial aspect of preventing and treating any infection. In order to detect T. annulata on-site, we selected sporozoite surface antigen gene (SPAG) as the target gene to design Recombinase Polymerase Amplification (RPA) primers and crRNA to build a one-pot SHERLOCK method. The results showed the assay could specifically detect T. annulata infection, but have no-cross reaction with the genomic DNA of T. annulata, Theileria sinensis, Theileria orientalis, Babesia bigemina and Babesia bovis. For its sensitivity, the assay could detect 1 × 103 copies/μL at least. In comparison with the published PCR assay, the relative specificity and sensitivity of the one-pot SHERLOCK assay was 100 % and 100 % respectively, and the coincidence rate was 100 %. Meanwhile, to improve the practicality, we used the lysis buffer to handle the blood of the infected animal to detect the T. annulata. The findings presented that T. annulata can be detected by using the one-pot SHERLOCK assay, which had preliminarily solved the problem of genome extraction in SHERLOCK field applications. Although this assay has a decrease in sensitivity but it reduced the number of operational steps and the probability of aerosol contamination. This assay has the potential to be developed into a commercial detection method and provide good support for the prevention and control of theileriosis.

Emerging infectious diseases caused by various tick-borne microorganisms (TBMs) pose public and animal health concerns, including camels, with no defined global distribution. In this study, 150 blood samples and 288 ticks were collected from symptomatic two-humped camels (Camelus bactrianus) in Gaotai County, Gansu Province, China. Morphologically identified ticks were confirmed using cytochrome oxidase I (COI), and the findings revealed two species, Hyalomma asiaticum and Haemaphysalis longicornis (prevalence: 245/288 [88.19 %] and 34/288 [11.81 %], respectively). The extracted Genomic DNA from blood and ticks was processed by conventional PCR to investigate the existing TBMs based on 16S rRNA, 18S rRNA, and 17-kDa genes. Different TBMs, including Anaplasma bovis, Colpodella sp., Rickettsia rickettsii, and Candidatus Rickettsia jingxinensis, have been documented as single infections at different rates. High single infection rates (198/218; 90.83 % and 117/150; 78.00 %) of A. bovis in Hy. asiaticum and camel blood were recorded, whereas the lowest single infection rate (3/22; 13.64 %) of R. rickettsii was noted in Hae. longicornis. Co-infection with Rickettsia spp. + A. bovis (20/288; 6.94 %), Colpodella sp. + A. bovis (14/288; 4.86 %), Colpodella sp. + Rickettsia spp. (1/288; 0.35 %), and Colpodella sp. + Rickettsia spp. + A. bovis (1/288; 0.35 %) were recorded as concurrent infection. Phylogenetic analysis revealed that the representative TBMs have close similarities and clustered together with their corresponding isolates from China, South Korea, India, the USA, Mexico, Bangladesh, Malawi, Japan, Pakistan, Cyprus, Nigeria, Poland, and Brazil. These findings present a preliminary baseline regarding TBMs infection in camel blood and ticks and provide a framework for further studies on the prevalence and effective control measures for ticks and tick-associated diseases.

Southern cattle tick, Rhipicephalus (R) microplus, is a major challenge in transmitting Babesia spp., Theileria spp., and Anaplasma spp., worldwide. Mitigation of this tick and associated pathogens is crucial for animal and human- health. In present study, laboratory-based larval packet tests (LPTs) were performed to assess fipronil efficacy against R. microplus ticks collected from four Chinese localities, Nanning, Guangshui, Macheng, and Tengchong. Additionally, we reported first molecular characterization of the full-length gamma-aminobutyric acid (GABA)gated chloride channel gene of R. microplus ticks from China and Pakistan (Khyber Pakhtunkhwa and Punjab). For LPTs bioassay, fipronil's concentrations (5-ppm, 25-ppm, 50-ppm, 75-ppm, 100-ppm) in acetone and DNasefree water were prepared. The highest average mortality rate of larvae was observed in Nanning (4.34-98.13 %) while lowest in Tengchong (5.70-82.29 %) via triplicate LPTs. The recorded LC50 values were 33.2 ppm, 35.7 ppm, 49.7 ppm, and 55.4 ppm, respectively, for the four localities (Guangshui, Macheng, Nanning, and Tengchong) while the LC99 values were 722 ppm, 827 ppm, 949 ppm, and 1342.3 ppm, respectively. The resistance factors (RR50) were 13.83, 14.88, 20.71, and 23.08, indicating the development of level-II resistance in the tick populations from these regions respectively. Molecularly, consensus sequences of R. microplus GABAgene from China and Pakistan resulted 99.24-99.52 % and 99.10-99.39 % identities with dieldrin-susceptible NRFS (GQ398111.1) and dieldrin-resistant (GQ398112.1) R. microplus ticks, respectively. Findings revealed 21 single nucleotide polymorphisms (SNPs) [8-non-synonymous (NS) and 13-synonymous (S)], including specifically 10-SNPs from Nanning, 9-SNPs from Macheng, 7-SNPs from Guangshui, 10-SNPs from Tengchong, and 9SNPs from Pakistani R. microplus full-length GABA gene. Explicitly, we obtained one NS-SNP in TM3 (T-871-G; F291-V) and two NS-SNPs in TM4 (A-1438-G; N-480-D and A-1439-G; N-480-G) regions of Chinese and Pakistani, while one NS-SNP in TM2 (A-763-G; T-255-A) region of Pakistani R. microplus ticks was documented. Coexistence among SNPs in TM3 (T-871-G; F-291-V), TM4 (A-1439-G; N-480-G) and one linker SNP (A-1378-G; S-460-G) were also recorded. Presence of SNPs and their coexistence suggest the possible tri-dimensional structural modification in GABA-Cl channels that might interfere with binding of fipronil, resulting in development of resistance. This comprehensive research will provide a basis for governmental and pharmaceutical industries for development of effective tick control-strategies as well as to manage the effectiveness and functionality of the available acaricides.

Owing to the lack of research on pesticide-sensitive strains of Chinese Haemaphysalis longicornis, monitoring the resistance status of H. longicornis field populations is impossible, thus posing a risk to the control of drug resistance. For this purpose, the diagnostic doses (DDs) of 8 neurotoxic chemical acaricides in the Chinese pesticide-sensitive strain H. longicornis were detected via the larval packet test recommended by the Food and Agriculture Organization of the United Nations. The DDs were 0.080% w/v for cyhalothrin, 0.526%w/v for cypermethrin, 0.064% w/v for deltamethrin, 0.164%w/v for diazinon, 1.675%w/v for dichloro-diphenyl-trichloroethane (DDT), 0.880%w/v for ivermectin, 0.242%w/v for amitraz and 0.036%w/v for fipronil. This work represents the first record of a standard sensitive strain of Chinese H. longicornis. The obtained median lethal concentration (LC50) and DD values can be used as reference data in field populations for the detection of resistance in H. longicornis in different regions of China. These results can be used to evaluate the effectiveness of neurotoxic chemical pesticides on the market, optimize the usage strategy of pesticides, and provide a theoretical basis for improving the monitoring system of drug resistance. Additionally, we provide new genomic data for this species of tick, which allows for the development of a new method to screen for mutations associated with pesticide resistance, especially pyrethroids.

African swine fever is an acute, highly contagious infectious disease that affects both domestic and wild pigs. ASFV is the only known DNA virus transmitted by arthropod vectors, with acute infection in pigs leading to morbidity and mortality rates as high as 100%. The virus can persist in a transmission cycle among wild boars, soft ticks, and domestic pigs. To date, nine Ornithodoros spp. have been confirmed to be capable of transmitting ASFV worldwide. However, the potential soft tick species capable of transmitting ASFV in China remain unclear. In this study, we compared the ability of Ornithodoros lahorensis and Argas persicus, as well as Ixodidae spp., to transmit ASFV via animal transmission experiments. The results revealed that O. lahorensis soft ticks, but not A. persicus or Ixodes, could act as competent vectors through transstadial and transovarial transmission of ASFV. These viruses were detected in the nymphs developed from larvae, adults developed from nymphs, and larvae hatched from eggs by adults. Thus, O. lahorensis are likely an important natural vectors of ASFV.