Infectious disease caused by pathogenic bacteria continues to be the primary challenge to humanity. Antimicrobial resistance and microbial biofilm formation in part, lead to treatment failures. The formation of biofilms by nosocomial pathogens such as Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), and Klebsiella pneumoniae (K. pneumoniae) on medical devices and on the surfaces of infected sites bring additional hurdles to existing therapies. In this review, we discuss the challenges encountered by conventional treatment strategies in the clinic. We also provide updates on current on-going research related to the development of novel anti-biofilm technologies. We intend for this review to provide understanding to readers on the current problem in health-care settings and propose new ideas for new intervention strategies to reduce the burden related to microbial infections.

S. aureus is the pathogen that is commonly associated with subclinical mastitis, causing significant economic losses to dairy farms. This infection responds poorly to antimicrobial treatment, which could be due to the development of AMR, biofilm formation, and the intracellular invasion of S. aureus into bovine mammary cells leading to treatment failure. Thus, it is important to understand the challenge of this problem. Therefore, the present study aims to determine the prevalence, antimicrobial resistance, and characterization of S. aureus that was isolated from subclinical bovine mastitis in East Coast Malaysia. A total of 235 milk samples from dairy cows were collected from selected farms in Kelantan and Pahang. The samples were subjected to a somatic cell analysis to identify subclinical mastitis, followed by bacteria isolation and antimicrobial susceptibility testing. The isolated S. aureus were further analyzed for their ability to form biofilms and invade the bovine mammary epithelial cells (MAC-T cells) in in vitro infections modeling using a gentamicin protection assay. The overall total of 74/235 (31.4%; 95%CI = 0.31; 0.32) of the milk samples demonstrated >200,000 somatic cells/mL, suggesting the presence of subclinical mastitis in the animals. A total of 39/235 (16.5%; 95% CI = 0.16, 0.17) of the milk samples harbored S. aureus which demonstrated resistance towards the following antimicrobials: penicillin (18/39, 46%), ampicillin (17/39, 43.6%), oxacillin (12/39, 31%), tetracycline (10/39, 26%), and erythromycin (7/39, 18%). AMR was recorded for a total of (17/39, 43.6%) of S. aureus isolates. All isolates formed biofilms, with (8/30, 27%) strongly biofilm-forming, (18/30, 60%) moderately biofilm-forming, and the remaining (4/30, 13%) of isolates weakly biofilm-forming. Interestingly, the AMR isolates appear to produce weak and moderate biofilm. Moreover, (6/20, 30%) of the S. aureus isolates were invasive towards MAC-T cells, as indicated by their ability to evade gentamicin treatment. The study demonstrated the presence of AMR, invasiveness, and biofilm formation in S. aureus that was isolated from subclinical mastitis. This characteristic presents additional challenges to existing antimicrobial therapy.