Bacterial community associated with Pyrodinium bahamense var. compressum during late exponential growth phase based on 16S rRNA gene sequence analysis

Aims@#Pyrodinium bahamense var. compressum is one of the principal causal agents of harmful algal blooms (HABs) in the coastal waters of Sabah, Malaysia. Seafood and aquaculture products tainted with lethal concentrations of the principal neurotoxin, saxitoxin, have been implicated in mortality and morbidity. The bacteria-algae association may play a key role in paralytic shellfish toxin (PST) production during a toxic bloom event. The production of PST during a harmful bloom is unclear and research on the bacterial diversity associated with Sabah P. bahamense is scarce. The present study examined the cultivable bacteria diversity associated with P. bahamense through 16S ribosomal RNA (rRNA) gene sequence analysis.@*Methodology and results@#The V3-V4 region of the 16S rRNA gene sequence was amplified and used to identify bacterial populations associated with P. bahamense var. compressum. A total of 62 isolates were successfully isolated, belonging to three different phyla, which were Proteobacteria; 55 (89%), Bacteroidetes; 6 (10%) and Actinobacteria; 1 (1%). Out of 55 Proteobacteria, 27 isolates were gamma-Proteobacteria (Marinobacter salsuginis) and 28 of the isolates were alpha-Proteobacteria; Mameliella atlantica (13), Roseibium denhamense (10) and Roseibium hamelinense (5). The remaining bacteria isolates from the phyla Bacteroidetes and Actinobacteria were identified as Muricauda lutimaris (6) and Micrococcus luteus (1), respectively.@*Conclusion, significance and impact of study@#The analysis of the bacterial 16S rRNA gene revealed multiple bacterial taxa associated with the toxic P. bahamense var. compressum bloom. The findings of the present work will pave the way for further studies aimed at isolating and characterizing genes involved in the saxitoxin biosynthesis in the associated bacteria.

Bacterial diversity of the abandoned Mamut Copper Mine in Sabah, Malaysia and its correlation with copper contamination

Aims@#The former Mamut Copper Mine, acid mine drainage site represents an anthropogenic altered landscape characterized by its acidic topsoil which is contaminated primarily with copper. Even though the mining operation was ceased at 1999, the bacterial diversity in this area has never been investigated. This study was conducted to ascertain the bacterial diversity of this abandoned copper mine and correlate it to the copper concentration in the soil. @*Methodology and results@#Soil samples were collected from 7 sites near the mine pit and the vicinity. Soil samples were assessed for soil copper elemental concentration using inductively coupled plasma optical emission spectrometry and bacteria were isolated via serial dilution followed by culture on nutrient agar plates. Phylogenetic analysis was done based on the full-length sequences of 16S rRNA gene. Twenty-four phylotypes were obtained from the 7 locations which originated from the phyla Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria. The results of the study indicated that site 2 (6.030223°; 116.658030°), located in between the mine pit and the mine factory with a copper concentration of 88.96 ppm, possessed the most diverse bacterial community with a Shannon diversity index (H) of 1.68, evenness (EH) of 0.94 and richness (S) of 6. @*Conclusion, significance and impact of study@#Current study revealed that there was a positive correlation between the copper concentration and the H index and the richness, but this was not reflected in the evenness. This is the first report of bacterial diversity from the former Mamut Copper Mine site. The data provided a valuable insight for the future monitoring of the bacterial community in this ecologically important niche.

Recombinant LipL32 Protein Developed Using a Synthetic Gene Detects Leptospiraspecific Antibodies in Human Serum Samples

Background: Synthetic biology is emerging as a viable alternative for the production of recombinant antigens for diagnostic applications. It offers a safe alternative for the synthesis of antigenic principles derived from organisms that pose a high biological risk. Methods: Here, we describe an enzyme-linked immunosorbent assay (ELISA) using the synthetic recombinant LipL32 (rLipL32) protein expressed in Escherichia coli for the detection of Leptospira-specific antibodies in human serum samples. The rLipL32-based ELISA was compared with a microscopic agglutination test (MAT), which is currently used as the gold standard for the diagnosis of leptospirosis. Results: Our results showed that all the MAT-positive serum samples were positive for Leptospira-specific IgG in an ELISA, while 65% (n = 13) of these samples were also positive for Leptospira-specific IgM. In the MAT-negative serum samples, 80% and 55% of the samples were detected as negative by an ELISA for Leptospira-specific IgM and IgG, respectively. Conclusion: An ELISA using the synthetic rLipL32 antigen was able to distinguish Leptospira-specific IgM (sensitivity 65% and specificity 80%) and IgG (sensitivity 100% and specificity 55%) in human serum samples and has the potential to serve as a rapid diagnostic test for leptospirosis.

Leptospirosis: recent incidents and available diagnostics – a review

Objective: The aim of this article was to review published research articles on leptospirosis, in particular the recent incidence of leptospirosis in Malaysia and the currently available diagnostic methods for the detection of leptospirosis. Methods: PubMed, Google Scholar and Google Search databases were searched using the key words Leptospira and leptospirosis. A total of seventy-six references were reviewed including sixty-seven research articles, three annual reports from Ministry of Health and six online newspaper articles. This review includes the following five sub-headings: introduction, leptospirosis transmission, leptospirosis incidents, laboratory diagnosis of leptospirosis and treatment and prevention of leptospirosis. Results: An increase in incidents of leptospirosis cases has been seen in recent years in Malaysia. The recent floods have contributed to the rise in the number of reported cases. Current diagnostic approaches such as dark field microscopy, microscopic agglutination test (MAT), Polymerase chain reaction and serological tests are inadequate as the organism is a slow grower. Conclusion: There is an urgent need to develop newer techniques for rapid detection of leptospirosis. The combination of PCR and ELISA are suggested for rapid and accurate diagnosis of leptospirosis. Studies on the mechanism of pathogenesis of Leptospira are needed for the development of vaccines that are safe for human use.