Ticks and tick-borne diseases pose a significant threat to public health. The Tumen River Basin is located at the junction of China, North Korea and Russia, whose warm climate and favorable ecological environment are suitable for the growth and reproduction of ticks. At the same time, the cattle industry in this region is highly developed, with cattle serving as the primary economic source for the area. This study performed an epidemiological investigation and analysis of pathogens carried by ticks and cattle in the Tumen River basin. A total of 913 ticks and 247 bovine blood samples were collected from seven cities primarily focused on cattle farming in the Tumen River basin. Morphological and molecular biological identification of ticks was carried out to determine the distribution of ticks and their pathogens in the region. Through the detection of pathogens carried by cattle blood samples in the surrounding area, the correlation with tick distribution was confirmed. The species and distribution of ticks of different genders and in different collection environments, and the infection of pathogens in bovine blood samples were statistically analyzed. The results showed that the 913 ticks had 5 species, including Haemaphysalis longicornis, Haemaphysalis concinna, Haemaphysalis japonica, Dermacentor silvarum and Ixodes persulcatus. Three pathogens, Babesia ovata, Theileria orientalis and Theileria sinensis, were detected in the blood samples of vector ticks and cattle. These results provide a theoretical basis for the prevention and control of ticks and tick-borne diseases in the Tumen River basin.

African swine fever virus (ASFV), porcine circovirus type 2 (PCV2), and porcine circovirus type 3 (PCV3) represent important viral agents that threaten pig health. To date, no triplex real-time quantitative PCR (qPCR) method has been described for the concurrent identification of all three viruses. In this study, a triplex qPCR system was constructed using the ASFV B646L gene, PCV2 ORF1 gene, and PCV3 ORF1 gene as amplification targets, enabling simultaneous detection of ASFV, PCV2, and PCV3. The standard curves generated for the three viral targets all yielded R² values of 0.999. The detection thresholds for ASFV, PCV2, and PCV3 were 10 copies/μL. Specificity testing showed that the assay amplified only the three intended viral targets and did not cross-react with other prevalent swine pathogens. Intra-assay and inter-assay reproducibility analyses yielded coefficients of variation below 2%. Seventy-nine clinical samples were tested using the established triplex qPCR assay and a previously published reference method. Positive and negative results were identical between the two assays, giving a concordance rate of 100%. These findings indicate that the triplex real-time qPCR assay provides a sensitive, dependable, and specific method for detecting ASFV, PCV2, and PCV3 infections and may facilitate epidemiological surveillance, as well as pathogen prevention and control.