Chui Wei Bong

Complete degradation of dimethyl phthalate by biochemical cooperation of the Bacillus thuringiensis strain isolated from cotton field soil

Dimethyl phthalate (DMP), a phthalate ester, is widely used in cosmetics, perfumes, and plasticizers. It has been classified as a suspected endocrine disruptor by many countries. The present study describes the biodegradation of DMP by a new aerobic bacterium, isolated from soil samples of a cotton field by an enrichment culture technique utilizing DMP as the sole source of carbon and energy. The isolate was identified as Bacillus thuringiensis based on the morphological and biochemical characteristics as well as gene sequence analysis. Bacillus thuringiensis grows best in a mineral salt medium of pH 7.0 at 30 °C incubation for 48 hours. The effects of temperature, inoculum size, substrate concentration and incubation time on DMP degradation were also studied. Bacillus thuringiensis is able to biodegrade 400 mg L−1 of DMP under aerobic conditions with 99% degradation potential. A combination of GC and GC-MS analysis revealed a complete DMP biodegradation pathway. The results indicate that Bacillus thuringiensis may prove a promising source for DMP bioremediation at a commercial scale.

Temporal Variation of Bacterial Respiration and Growth Efficiency in Tropical Coastal Waters

ABSTRACT We investigated the temporal variation of bacterial production, respiration, and growth efficiency in the tropical coastal waters of Peninsular Malaysia. We selected five stations including two estuaries and three coastal water stations. The temperature was relatively stable (averaging around 29.5°C), whereas salinity was more variable in the estuaries. We also measured dissolved organic carbon and nitrogen (DOC and DON, respectively) concentrations. DOC generally ranged from 100 to 900 μM, whereas DON ranged from 0 to 32 μM. Bacterial respiration ranged from 0.5 to 3.2 μM O2 h−1, whereas bacterial production ranged from 0.05 to 0.51 μM C h−1. Bacterial growth efficiency was calculated as bacterial production/(bacterial production + respiration), and ranged from 0.02 to 0.40. Multiple correlation analyses revealed that bacterial production was dependent upon primary production (r2 = 0.169, df = 31, and P < 0.02) whereas bacterial respiration was dependent upon both substrate quality (i.e., DOC/DON ratio) (r2 = 0.137, df = 32, and P = 0.03) and temperature (r2 = 0.113, df = 36, and P = 0.04). Substrate quality was the most important factor (r2 = 0.119, df = 33, and P = 0.04) for the regulation of bacterial growth efficiency. Using bacterial growth efficiency values, the average bacterial carbon demand calculated was from 5.30 to 11.28 μM C h−1. When the bacterial carbon demand was compared with primary productivity, we found that net heterotrophy was established at only two stations. The ratio of bacterial carbon demand to net primary production correlated significantly with bacterial growth efficiency (r2 = 0.341, df = 35, and P < 0.001). From nonlinear regression analysis, we found that net heterotrophy was established when bacterial growth efficiency was <0.08. Our study showed the extent of net heterotrophy in these waters and illustrated the importance of heterotrophic microbial processes in coastal aquatic food webs.

Bacterial abundance and production, and their relation to primary production in tropical coastal waters of Peninsular Malaysia

In the present study, the relationship between bacteria and phytoplankton in tropical coastal waters was investigated. The bacterial abundance, bacterial production, chlorophyll a concentration and net primary production were measured at several locations in the coastal waters of Peninsular Malaysia. Chlorophyll a concentration ranged from 0.40 to 32.81 μg L–1, whereas bacterial abundance ranged from 0.1 to 97.5 × 106 cells mL–1. Net primary production ranged from 8.49 to 55.95 μg C L–1 h–1, whereas bacterial production ranged from 0.17 to 70.66 μg C L–1 h–1. In the present study, the carbon conversion factor used to convert bacterial production (cells mL–1 h–1) into carbon units ranged from 10 to 32.8 fg C cell–1, and was estimated from the bacterial size distribution measured at each location. Both phototrophic and heterotrophic biomass (bacteria–chlorophyll a) and activity (bacterial production–net primary production) were significantly correlated, although their correlation coefficients (r2) were relatively low (r2 = 0.188 and r2 = 0.218 respectively). Linear regression analyses provided the following equations to represent the relationship between: bacteria and chlorophyll a (Chl a), log Bacteria = 0.413 log Chl a + 6.057 (P = 0.003); and between bacterial production (BP) and net primary production (NPP), log BP = 0.896 log NPP – 0.394 (P = 0.004), which fitted with published results well. Comparison of annual carbon fluxes confirmed the prevalence of net heterotrophy in these coastal waters, and together with the low correlation coefficients, suggested the role of allochthonous organic matter in supporting heterotrophic activity.

Carbon Flux Through Bacteria in a Eutrophic Tropical Environment: Port Klang Waters

Port Klang waters is eutrophic and long term data showed that its water quality is deteriorating. We observed the occurrence of hypoxia due to very low GPP that was limited by NO3. Although primary production is the basis of aquatic food web, and supported both CR and BCD, episodes of uncoupling were observed. Uncoupling occurred especially when GPP was very low, and generally BCD and GPP were balanced. Our results showed that only 2% of C consumed by bacteria were passed onto protists. This suggested that the microbial loop was not an efficient pathway to recycle organic matter as a substantial amount of carbon was lost.

Investigating the decay rates of Escherichia coli relative to Vibrio parahemolyticus and Salmonella Typhi in tropical coastal waters.

Using the size fractionation method, we measured the decay rates of Escherichia coli, Salmonella Typhi and Vibrio parahaemolyticus in the coastal waters of Peninsular Malaysia. The size fractions were total or unfiltered, <250 μm, <20 μm, <2 μm, <0.7 μm, <0.2 μm and <0.02 μm. We also carried out abiotic (inorganic nutrients) and biotic (bacterial abundance, production and protistan bacterivory) measurements at Port Dickson, Klang and Kuantan. Klang had highest nutrient concentrations whereas both bacterial production and protistan bacterivory rates were highest at Kuantan. We observed signs of protist-bacteria coupling via the following correlations: Protistan bacterivory-Bacterial Production: r = 0.773, df = 11, p < 0.01; Protist-Bacteria: r = 0.586, df = 12, p < 0.05. However none of the bacterial decay rates were correlated with the biotic variables measured. E. coli and Salmonella decay rates were generally higher in the larger fraction (>0.7 μm) than in the smaller fraction (<0.7 μm) suggesting the more important role played by protists. E. coli and Salmonella also decreased in the <0.02 μm fraction and suggested that these non-halophilic bacteria did not survive well in seawater. In contrast, Vibrio grew well in seawater. There was usually an increase in Vibrio after one day incubation. Our results confirmed that decay or loss rates of E. coli did not match that of Vibrio, and also did not correlate with Salmonella decay rates. However E. coli showed persistence where its decay rates were generally lower than Salmonella.

The contribution of heterotrophic nanoflagellate grazing towards bacterial mortality in tropical waters: comparing estuaries and coastal ecosystems

Heterotrophic nanoflagellate (HNF) grazing depends on both temperature and trophic status of an ecosystem. As most microbes already function at their temperature optimum in tropical waters, we hypothesised that HNF grazing rates would be higher in more productive sites such as estuaries than in less productive areas such as coastal waters. We sampled two estuaries and three coastal sites along the Straits of Malacca and the South China Sea near the Malaysia Peninsula. Bacterial abundance ranged 0.9–6.3 × 106 cells mL–1, whereas HNF abundance ranged 1.8–10.1 ×103 cells mL–1. Bacterial production ranged 1.1–12.7 × 105 cells mL–1 h–1, whereas HNF grazing rates were an order of magnitude lower at 1.0–78.5 × 104 cells mL–1 h–1. Bacterial abundance, net bacterial production and HNF grazing rates were higher in estuaries than coastal waters but HNF abundance did not differ between the two areas. Across all stations, HNF grazing rates increased with bacterial production, and accounted for ~33% of bacterial production. Our results suggest that in the tropical waters studied, there was a bacterial production–grazing imbalance. Other loss factors such as viral lysis, sedimentation or the presence of benthic filter feeders could account for this imbalance.

Diverse and abundant multi-drug resistant E. coli in Matang mangrove estuaries, Malaysia

E.coli, an important vector distributing antimicrobial resistance in the environment, was found to be multi-drug resistant, abundant, and genetically diverse in the Matang mangrove estuaries, Malaysia. One-third (34%) of the estuarine E. coli was multi-drug resistant. The highest antibiotic resistance prevalence was observed for aminoglycosides (83%) and beta-lactams (37%). Phylogenetic groups A and B1, being the most predominant E. coli, demonstrated the highest antibiotic resistant level and prevalence of integrons (integron I, 21%; integron II, 3%). Detection of phylogenetic group B23 downstream of fishing villages indicates human fecal contamination as a source of E. coli pollution. Enteroaggregative E. coli (1%) were also detected immediately downstream of the fishing village. The results indicated multi-drug resistance among E. coli circulating in Matang estuaries, which could be reflective of anthropogenic activities and aggravated by bacterial and antibiotic discharges from village lack of a sewerage system, aquaculture farms and upstream animal husbandry.

Mangrove Oyster (Crassostrea belcheri) as a Biomonitor Species for Bioavailability of Polycyclic Aromatic Hydrocarbons (PAHs) from Sediment of the West Coast of Peninsular Malaysia

ABSTRACT Rapid industrialization and urbanization in the west coast of Peninsular Malaysia has caused increasing pollution particularly of petroleum and petroleum by-products. Surface sediment and mangrove oyster (Crassostrea belcheri) were collected from five mangrove ecosystems in the west coast of Peninsular Malaysia and investigated for bioavailability of polycyclic aromatic hydrocarbons (PAHs). Sampling locations were selected from both remote areas with few or no previous records of petroleum pollution such as Pulau Merambong and polluted areas that are under international attention such as Klang mangrove ecosystem. PAH fractions were obtained through soxhlet extraction and two-step column chromatography and the fractions were injected to gas chromatography-mass spectrometry (GC-MS) for analysis. The concentrations of PAHs ranged from 151 to 4973 ng g−1 dw in the sediments, while from 309 to 2225 ng g−1 dw in the oysters. When tested for diagnostic ratios, a predominance of pyrogenic source PAHs was detected in the sediments, whereas PAHs in the oysters had mixed petrogenic and pyrogenic sources. A significant correlation (p < 0.05) was found between high molecular weight (HMW) PAHs in the sediments and oysters and biota accumulation factors (BAFs) of PAHs were approaching or exceeding unity indicating the ability of mangrove oyster in bioaccumulation of PAHs. Overall, this study indicates that mangrove oyster (C. belcheri) can be used as a biomonitor species for PAHs in an aquatic environment.

Assessment of heavy metal pollution in surficial sediments from a tropical river-estuary-shelf system: A case study of Kelantan River, Malaysia

To understand the source-to-sink of pollutants in the Kelantan River estuary and the adjacent shelf area in Malaysia, a total of 42 surface sediment samples were collected in the Kelantan River-estuary-shelf system to analyze for grain size, total organic carbon (TOC) content, Al and heavy metals (Cr, Ni, Cu, Zn, Cd and Pb). The surficial sediments were mainly composed of clayey silt and the TOC content in sediments decreased from the river to the shelf. The surficial sediments experienced Pb pollution; Cr only showed a certain level of pollution in the coastal area of the estuary but not in other areas, and Ni, Cu, Zn, and Cd showed no pollution. The heavy metals mainly originated from natural weathering and erosion of rocks and soils in the catchment and enriched near the river mouth. Total organic carbon can promote the enrichment of heavy metals in sediments.

Aliphatic hydrocarbons and triterpane biomarkers in mangrove oyster (Crassostrea belcheri) from the west coast of Peninsular Malaysia

The Straits of Malacca is one of the worlds busiest shipping routes where frequent oil spills occur. Rapid development in the west coast of Peninsular Malaysia is the other major source of petroleum pollution in this narrow waterway. In order to identify occurrence and origin of hydrocarbons in the Straits, mangrove oysters (Crassostrea belcheri) were collected from five sampling locations and analysed for n-alkanes and biomarkers. Soxhlet apparatus and two step column chromatography were used for extraction, purification and fractionation of the oysters. Petroleum origin n-alkanes were detected in majority of the sampling locations which is indicative of anthropogenic activities in this region. Using source and maturity diagnostic ratios for hopanes revealed used crankcase oil as the main source of petroleum hydrocarbons in oysters from all sampling locations except for the Pulau Merambong where signature of South East Asia crude oil (SEACO) was detected.