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.

Salmonella species (spp.) and Escherichia coli (E. coli) are the most common infectious pathogens in poultry. Antimicrobials are given either as growth promoters or as treatment, thereby increasing the possibility of the emergence of antimicrobial resistance (AMR). We determined the prevalence of AMR for both pathogens isolated from broiler farms in the East Coast of Peninsular Malaysia from 2018–2019. A total of 384 cloacal swabs were collected, followed by bacterial isolation, confirmation, and antimicrobial susceptibility tests. The overall prevalence of Salmonella spp. and E. coli were 6.5% and 51.8%, respectively. Salmonella spp. and E. coli displayed resistance towards the following antimicrobials: erythromycin (100% for both pathogens), chloramphenicol (76.2% and 84.5%, respectively), tetracycline (62% and 94.6%, respectively), ampicillin (47.7% and 87%, respectively), sulfamethoxazole/trimethoprim (42.9% and 83.3%, respectively), ciprofloxacin (4.8% and 23.8%, respectively), nalidixic acid (9.6% and 60.7%, respectively), streptomycin (19% and 66%, respectively), kanamycin (28.6% and 57%, respectively), cephalothin (0% and 11%, respectively), and gentamicin (0% and 20.2%, respectively). Multidrug resistance (MDR) was recorded in 82% of Salmonella spp. and 100% of E. coli. These findings demonstrate the high prevalence of AMR in both pathogens in broiler farms on the East Coast of Peninsular Malaysia. These findings could be attributed to the excessive use of antimicrobial agents by poultry farm owners. Enhanced control measures and a strong monitoring system should be urgently implemented in order to reduce the emergence of antimicrobial resistance.

Antimicrobial therapy is the most applied method for treating and preventing bacterial infection in livestock. However, it becomes less effective due to the development of antimicrobial resistance (AMR). Therefore, there is an urgent need to find new antimicrobials to reduce the rising rate of AMR. Recently, antimicrobial peptides (AMPs) have been receiving increasing attention due to their broad-spectrum antimicrobial activity, rapid killing activities, less toxicity, and cell selectivity. These features make them potent and potential alternative antimicrobials to be used in animals. Here, we discuss and summarize the AMPs in animals, classification, structures, mechanisms of action, and their potential use as novel therapeutic alternative antimicrobials to tackle the growing AMR threat.

Background: S. aureus is one of the causative agents of bovine mastitis. The treatment using conventional antimicrobials has been hampered due to the development of antimicrobial resistance and the ability of the bacteria to form biofilms and localize inside the host cells. Objectives: Here, the efficacy of graphene oxide (GO), a carbon-based nanomaterial, was tested against the biofilms and intracellular S. aureus invitro. Following that, the mechanism for the intracellular antimicrobial activities and GO toxicities was elucidated. Methods: GO antibiofilm properties were evaluated based on the disruption of biofilm structure, and the intracellular antimicrobial activities were determined by the survival of S. aureus in infected bovine mammary cells following GO exposure. The mechanism for GO intracellular antimicrobial activities was investigated using endocytosis inhibitors. GO toxicity towards the host cells was assessed using a resazurin assay. Results: At 100 ug/mL, GO reduced between 30 and 70% of S. aureus biofilm mass, suggesting GO’s ability to disrupt the biofilm structure. At 200 ug/mL, GO killed almost 80% of intracellular S. aureus, and the antimicrobial activities were inhibited when cells were pre-treated with cytochalasin D, suggesting GO intracellular antimicrobial activities were dependent on the actin-polymerization of the cell membrane. At < 250 ug/mL, GO enhanced the viability of the Mac-T cell, and cells were only affected at higher dosages. Conclusion: The in vitro efficacy of GO against S. aureus in vitro suggested the compound could be further tested in Vivo to zrecognize its potential as one of the components of bovine mastitis therapy.

The escalating threat of infectious diseases, exacerbated by antimicrobial resistance (AMR) and biofilm formation, necessitates innovative therapeutic strategies. This review presents a comprehensive exploration of the potential of nanoparticles synthesized from natural sources, including plant extracts, microbial products, and marine compounds, as antimicrobial agents. These naturally derived nanoparticles demonstrated significant antibiofilm and antivirulence effects, with specific examples revealing their capacity to reduce biofilm mass by up to 78% and inhibit bacterial quorum sensing by 65%. The integration of bioactive compounds, such as polyphenols and chitosan, facilitates nanoparticle stability and enhances antimicrobial efficacy, while green synthesis protocols reduce environmental risks. Notably, the review identifies the potential of silver nanoparticles synthesized using green tea extracts, achieving 85% inhibition of polymicrobial growth in vitro. Despite these promising results, challenges such as standardization of synthesis protocols and scalability persist. This study underscores the transformative potential of leveraging naturally sourced nanoparticles as sustainable alternatives to conventional antimicrobials, offering quantitative insights for their future application in combating mono‐ and polymicrobial 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.