Pathogenic Escherichia coli (E. coli) is a significant foodborne pathogen that poses a serious global public health risk. These pathogenic strains of E. coli can be found in various environments such as soil, water, livestock, livestock products, and humans. This review aims to evaluate the role of livestock and the environment as potential sources and reservoirs of pathogenic E. coli in Malaysia over the past two decades. A systematic search was conducted on PubMed, Scopus, Web of Science, and Google Scholar databases using standardized keywords (Escherichia coli, Livestock, Abattoir workers, Farm workers, and Malaysia) to identify relevant original research articles published between 2001 and 2023 and our systematic review protocol was published in Prospero (Registration number: CRD42023433199). A total of 20 eligible articles were included in this review. The prevalence of multi-drug resistant E. coli among livestock production system and the environment, ranged from 4% to 100%. E. coli strains isolated from livestock production systems and environment in Malaysia exhibited high potential for pathogenicity, with diverse enterotoxins, genes associated with antimicrobial resistance, and virulence factors. Phylogroup B and D were noted among the E. coli isolates indicating that the strains in Malaysia are highly virulent and extra-intestinal. High levels of resistance to antimicrobials particularly ESBL and colistin which are last resort antimicrobials were noted. Butchering or processing of livestock, contamination of plants via contaminated waterways or livestock manure are the main sources and reservoir for E. coli to spread in Malaysia. Continued surveillance, monitoring and a comprehensive 'One Health' approach are recommended to effectively prevent and control livestock drug-resistant E.coli strains throughout the country.