Food safety is a matter of great public health concern worldwide and particularly crucial if the environment in which the food is handled is heavily contaminated. Most fresh foods particularly that of animal origin like beef is highly susceptible to microbial invasion and food poisoning. In poorly managed market environment particularly in Ghana, unhygienic practice is the major cause for food contamination. This study observed the hygienic practices and microbiological food safety standards of butchers who specifically sold beef in the Ashaiman market in Accra, Ghana. Hygienic practices of sixteen (16) butchers were randomly selected in a cross sectional study using an eight point scale checklist weekly over a period of four weeks. The microbial quality of one hundred and twenty-eight (128) fresh beef samples were aseptically collected and analysed using standard microbiological techniques. It was observed that majority of the butchers did not practice safe hygiene standards as recommended by the Ghana Food and Drugs Board and the Ghana Standards Board. The beef samples were contaminated with Aerobic mesophiles (189-23000 cfu/g), Staphylococcus aureus (22-59 cfu/g), Bacillus cereus (17-41 cfu/g), Clostridium perfringens (21-48 cfu/g) and Escherichia coli (31-2200 cfu/g). The pH of the beef samples were between 6.50 and 6.90. The butchers in Ashaiman market supplied fairly contaminated beef to the general public. Escherichia coli , which is a sign of faecal contamination, was the predominant microbial contaminant in the samples examined. The result of unhygienic practices and poor handling of beef by butchers in the Ashaiman market is the major cause of contaminated beef. There are chances that other meat sold by virtually the same group of persons could equally or even more be contaminated by food borne pathogens. Hence food industry and consumers should be made aware of the potential risk of food borne pathogens in beef sold by butchers in Ashaiman market.