Ticks and tick-borne diseases (TTBDs) are responsible for significant losses in terms of treatment costs, decreased productivity (such as milk and meat), reduced reproductive ability, and financial crisis to livestock owners. In Pakistan, it is crucial to periodically assess the risk of TTBDs and ecological factors, potential causes of acaricidal resistance in tick fauna, and the intensive increase in the spread of TTBDs. Participatory epidemiological approaches are key to assessing the livestock owners’ and stakeholders’ knowledge, attitude, and practices (KAP) about TTBDs. The current study determined the KAP about ticks and tick-borne diseases of respondents from Sindh, Pakistan. A total of 240 respondents were interviewed from different ecological zones: among them, 42.5% (n = 102) of the respondents practiced the manual removal of ticks from animals, while acaricide usage was indicated by 137 respondents (57.0%) as occurring sometimes, 50 (20.8%) monthly, 41 (17.0%) fortnightly, and 12 (5%) weekly, during the peak infestation season. Ticks were 2.6 times [OR = 2.5 (95% Cl = 1.47–4.06)] and viruses were 1.89 times [OR = 188 (95% Cl = 1.09–2.9)] more likely to cause the development of disease in animals than any other pathogen. Despite the appropriate usage of acaricides, the knowledge of participants was inadequate. The findings of this study emphasize the need to take into account identified knowledge gaps and to take the initiative in carrying out appropriate education activities and extension programs to enhance the adoption of effective tick prevention and control strategies.