Aims: Vibrio harveyi is a serious pathogen for marine organisms particularly in hatcheries and grow-out ponds that attack their immune system. The rapid detection of V. harveyi is urgently needed to prevent bacterial spread. Here we described a rapid and specific visual detection method based on the Loop-Mediated Isothermal Amplification in combination with Lateral Flow Dipstick (LAMP-LFD). Methodology and results: A set of six novel primers were designed to target the toxR gene. These include the biotin-labelled inner primer that complements specifically to the target sequences. The resulting biotinylated LAMP amplicons were hybridised to the FAM-labelled probe resulting in lateral flow detection on the dipstick. The addition of loop primers improved the reaction time of LAMP by more than half and rapid detection was observed within 10-15 min. In comparison, the sensitivity of PCR-UV analysis was only at 104 copies while LAMP-LFD was able to detect lower amounts at 103 copies. The LFD provided higher specificity and selectivity since hybridization with specific probes to the LAMP amplicons was employed. In addition, detection of V. harveyi infected grouper was successful using the LAMP-LFD method described here. Conclusion, significance and impact of study LAMP-LFD is specific to V. harveyi. Our method provides a useful tool to rapidly detect and monitor the outbreaks of the pathogen.

Aims: Enterococcus bacteria, including some strains that are resistant to antibiotics like vancomycin can pose a threat to public health. The purpose of this study is to identify the species, antibiotic susceptibility profile and VanA/VanB gene frequencies in Enterococci isolated from patients at a tertiary hospital in Kota Kinabalu, Sabah. Methodology and results : Various bodily fluid specimens were collected from 162 patients between July 2019 and June 2021. Species confirmation and susceptibility testing were performed using an automated system. Subsequently, PCR was used to determine the presence of VanA and VanB genes. Species identification revealed the presence of five enterococcal species, namely E. faecalis (91), E. faecium (64), E. gallinarum (3) E. casseliflavus (2), along with one isolate each of E. hirae and E. avium. Overall, resistance to antibiotics like ampicillin, quinolones, tetracycline, gentamicin-syn, nitrofurantoin, glycopeptides and linezolid was generally low (<50%). However, a significant number of isolates displayed high resistance to erythromycin (>50% of samples), while resistance to tetracycline was more moderate. The frequencies of VanA and VanB genes were low (0.6 and 0%, respectively) and they were only detected in E. faecium. Conclusion, significance and impact of study The results indicate that while the prevalence of vancomycin-resistant enterococci (VRE) may be low, there is an increasing incidence of multidrug-resistant enterococci, particularly with regards to erythromycin.