International Journal of Molecular Veterinary Research, 2024, Vol.14, No.5, 194-201 http://animalscipublisher.com/index.php/ijmvr 196 disease (FMD) can lead to high morbidity and mortality rates, further exacerbating economic losses and necessitating stringent control measures (Damaty et al., 2021). Effective disease management and prevention strategies are essential to mitigate these impacts and ensure the sustainability of water buffalo farming. In summary, both infectious and non-infectious diseases pose significant challenges to water buffalo health, with far-reaching economic and public health implications. Addressing these challenges requires comprehensive disease surveillance, effective management practices, and robust prevention strategies. 3 Molecular Diagnostic Tools 3.1 PCR-based techniques in disease diagnosis Polymerase chain reaction (PCR) is a cornerstone in the molecular diagnosis of diseases in water buffaloes. It is widely used for detecting various pathogens, including Anaplasma marginale and Babesia species. For instance, PCR assays have been employed to identify tick-borne pathogens in water buffaloes in the Philippines, revealing a significant prevalence of Anaplasma marginale and Babesia bovis (Galon et al., 2019). Similarly, nested PCR techniques have been used to detect Cryptosporidium spp. in buffaloes, showing a higher sensitivity compared to traditional microscopic methods (Al-Amery et al., 2022). PCR-based methods are also crucial in diagnosing Mycobacterium avium subspecies paratuberculosis, the causative agent of Johne's disease, with qPCR assays providing a higher detection rate than serological tests (Uy et al., 2018). 3.2 High-throughput sequencing and its applications High-throughput sequencing technologies, such as Next Generation Sequencing (NGS), have advanced the molecular diagnostics of infectious diseases in water buffaloes. These technologies allow for comprehensive profiling of pathogen genomes and host responses. For example, NGS has been utilized to characterize miRNA signatures in water buffaloes infected with Brucella abortus, identifying potential biomarkers for non-invasive diagnosis (Lecchi et al., 2019). This approach provides insights into the host-pathogen interactions and can aid in the development of targeted diagnostic and therapeutic strategies. 3.3 Serological and immunological diagnostics in molecular testing Serological and immunological diagnostics complement molecular techniques by providing additional layers of information about disease status and immune response. Enzyme-linked immunosorbent assays (ELISA) are commonly used to detect antibodies against pathogens such as Mycobacterium aviumsubspecies paratuberculosis, although they may have lower sensitivity compared to molecular methods like qPCR (Uy et al., 2018). In the context of schistosomiasis, multi-epitope recombinant antigens have been developed to improve the sensitivity and specificity of serological tests, demonstrating potential for enhanced diagnostic accuracy (Lv et al., 2018). Additionally, immunophenotyping using flow cytometry has been applied to assess lymphocyte alterations in buffaloes with brucellosis, providing valuable data for disease diagnosis and management (Grandoni et al., 2023). In summary, the integration of PCR-based techniques, high-throughput sequencing, and serological diagnostics offers a comprehensive toolkit for the molecular diagnosis of diseases in water buffaloes. These methods enhance the detection, characterization, and understanding of various pathogens, contributing to improved disease management and control strategies. 4 Comparative Analysis of Molecular Diagnostics for Specific Diseases 4.1 Diagnostic approaches for bovine tuberculosis Bovine tuberculosis (bTB) in water buffalo is primarily caused by Mycobacterium bovis. Traditional diagnostic methods, such as the intradermal tuberculin test, have limitations in sensitivity and specificity. Recent studies have explored alternative diagnostic methods, including histopathological analysis, microbiological culture, and molecular techniques like PCR-HRM (High-Resolution Melting) analysis. In a study conducted in Colombia, histopathological tests revealed that 52% of tuberculin-positive buffaloes had lesions compatible with bTB, while PCR-HRM confirmed Mycobacterium bovis in 34% of cases, even in the absence of histological lesions (Jaiswal
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