Aquaculture is the fastest growing animal production sector. However, the production of marine fish is still hampered by the high mortality rate in the first few weeks after hatching. Mortality in larvae is often caused by microbial infections. Today, the incorporation of immunostimulants into microparticles provides us new tools to enhance disease resistance in marine larviculture. In this study, we prepared alginate microparticles loaded with the model antigen fluorescein isothiocyanate conjugated-bovine serum albumin. Optimum concentrations of alginate and CaCl2, the correct alginate viscosity and the appropriate preparatory conditions led to the creation of desirable microparticles with the correct size for oral feeding in gnotobiotic European sea bass larvae. The prepared alginate microparticles were stable in sea water and were successfully ingested by gnotobiotic sea bass larvae at day after hatching 7 without causing any negative effects. Results suggest the suitability of this drug delivery system for targeting the innate immune system of fish larvae in order to enhance disease resistance and thus reduce mortality in larviculture.
Burkholderia pseudomallei is an intracellular Gram-negative bacterial pathogen intrinsically resistant to a variety of antibiotics. Phages have been developed for use as an alternative treatment therapy, particularly for bacterial infections that do not respond to conventional antibiotics. In this study, we investigated the use of phages to treat cells infected with B. pseudomallei. Phage C34 isolated from seawater was purified and characterised on the basis of its host range and morphology using transmission electron microscopy (TEM). Phage C34 was able to lyse 39.5% of B. pseudomallei clinical strains. Due to the presence of contractile tail, phage C34 is classified as a member of the family Myoviridae, a tailed double-stranded DNA virus. When 2 × 105 A549 cells were exposed to 2 × 107 PFU of phage C34, 24 hours prior to infection with 2 × 106 CFU of B. pseudomallei, it was found that the survivability of the cells increased to 41.6 ± 6.8% as compared to 22.8 ± 6.0% in untreated control. Additionally, application of phage successfully rescued 33.3% of mice infected with B. pseudomallei and significantly reduced the bacterial load in the spleen of the phage-treated mice. These findings indicate that phage can be a potential antimicrobial agent for B. pseudomallei infections.
The study presents a baseline variability and climatology study of measured hydrodynamic, water properties and some water quality parameters of West Johor Strait, Singapore at hourly-to-seasonal scales to uncover their dependency and correlation to one or more drivers. The considered parameters include, but not limited by sea surface elevation, current magnitude and direction, solar radiation and air temperature, water temperature, salinity, chlorophyll-a and turbidity. FFT (Fast Fourier Transform) analysis is carried out for the parameters to delineate relative effect of tidal and weather drivers. The group and individual correlations between the parameters are obtained by principal component analysis (PCA) and cross-correlation (CC) technique, respectively. The CC technique also identifies the dependency and time lag between driving natural forces and dependent water property and water quality parameters. The temporal variability and climatology of the driving forces and the dependent parameters are established at the hourly, daily, fortnightly and seasonal scales.
Nineteen pairs of gaseous and surface seawater samples were collected along the cruise from Malaysia to the south of Bay of Bengal passing by Sri Lanka between April 12 and May 4, 2011 on the Chinese research vessel Shiyan I to investigate the latest OCP pollution status over the equatorial Indian Ocean. Significant decrease of α-HCH and γ-HCH was found in the air and dissolved water phase owing to global restriction for decades. Substantially high levels of p,p'-DDT, o,p'-DDT, trans-chlordane (TC), and cis-chlordane (CC) were observed in the water samples collected near Sri Lanka, indicating fresh continental riverine input of these compounds. Fugacity fractions suggest equilibrium of α-HCH at most sampling sites, while net volatilization for DDT isomers, TC and CC in most cases. Enantiomer fractions (EFs) of α-HCH and o,p'-DDT in the air and water samples were determined to trace the source of these compounds in the air. Racemic or close to racemic composition was found for atmospheric α-HCH and o,p'-DDT, while significant depletion of (+) enantiomer was found in the water phase, especially for o,p'-DDT (EFs = 0.310 ± 0.178). 24% of α-HCH in the lower air over the open sea of the equatorial Indian Ocean is estimated to be volatilized from local seawater, indicating that long-range transport is the main source.
Vibrio vulnificus is a highly invasive human pathogen that exists naturally in estuarine environment and coastal waters. In this study, we used different PCR assays to detect V. vulnificus in 260 seafood and 80 seawater samples. V. vulnificus was present in about 34 (13%) of the 260 seafood samples and 18 (23%) of the 80 seawater samples. Repetitive extragenic palindromic PCR (REP-PCR) and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) were applied to subtype the V. vulnificus isolates. Twenty-five REP profiles and 45 ERIC profiles were observed, and the isolates were categorized into 9 and 10 distinct clusters at the similarity of 80%, by REP-PCR and ERIC-PCR, respectively. ERIC-PCR is more discriminative than REP-PCR in subtyping V. vulnificus, demonstrating high genetic diversity among the isolates.
Marine spatial planning tends to prioritise biological conservation targets over socio-economic considerations, which may incur lower user compliance and ultimately compromise management success. We argue for more inclusion of human dimensions in spatial management, so that outcomes not only fulfill biodiversity and conservation objectives, but are also acceptable to resource users. We propose a fuzzy logic framework that will facilitate this task- The protected area suitability index (PASI) combines fishers' spatial preferences with biological criteria to assess site suitability for protection from fishing. We apply the PASI in a spatial evaluation of a small-scale reef fishery in Sabah, Malaysia. While our results pertain to fishers specifically, the PASI can also be customized to include the interests of other stakeholders and resource users, as well as incorporate varying levels of protection.
Selective separation and sensitive detection of dissolved silicon and boron (DSi and DB) in aqueous solution was achieved by combining an electrodialytic ion isolation device (EID) as a salt remover, an ion-exclusion chromatography (IEC) column, and a corona charged aerosol detector (CCAD) in sequence. DSi and DB were separated by IEC on the H(+)-form of a cation exchange resin column using pure water eluent. DSi and DB were detected after IEC separation by the CCAD with much greater sensitivity than by conductimetric detection. The five-channel EID, which consisted of anion and cation acceptors, cathode and anode isolators, and a sample channel, removed salt from the sample prior to the IEC-CCAD. DSi and DB were scarcely attracted to the anion accepter in the EID and passed almost quantitatively through the sample channel. Thus, the coupled EID-IEC-CCAD device can isolate DSi and DB from artificial seawater and hot spring water by efficiently removing high concentrations of Cl(-) and SO4(2-) (e.g., 98% and 80% at 0.10molL(-1) each, respectively). The detection limits at a signal-to-noise ratio of 3 were 0.52μmolL(-1) for DSi and 7.1μmolL(-1) for DB. The relative standard deviations (RSD, n=5) of peak areas were 0.12% for DSi and 4.3% for DB.
Butyltin compounds (BTs) including mono-, di-, and tributyltin and total tin (sigmaSn), were determined in green mussels (Perna viridis) from various Asian developing countries, such as Cambodia, China (Hong Kong and southern China), Malaysia, India, Indonesia, the Philippines, and Vietnam, to elucidate the contamination status, distribution, and possible sources and to assess the risks on aquatic organisms and humans. Butyltin compounds were detected in green mussels collected from all the sampling location investigated, suggesting widespread contamination of BTs along the coastal waters of Asian developing countries. Among butyltin derivatives, tributyltin (TBT) was the predominant compound, indicating its ongoing usage and recent exposures in Asian coastal waters. Higher concentrations of BTs were found in mussels collected at locations with intensive maritime activities, implying that the usage of TBT as a biocide in antifouling paints was a major source of BTs. In addition, relatively high concentrations of BTs were observed in mussels from aquaculture areas in Hong Kong and Malaysia, as it has been reported in Thailand. With the recent improvement in economic status in Asia, it is probable that an increase in TBT usage will occur in aquaculture. Although contamination levels were generally low in mussel samples from most of the Asian developing countries, some of those from polluted areas in Hong Kong, India, Malaysia, the Philippines, and Thailand revealed levels comparable to those in developed nations. Furthermore, the concentrations of TBT in some mussels from polluted areas exceeded the threshold for toxic effects on organisms and estimated tolerable average residue levels as seafoods for human consumption. A significant correlation was observed between the concentrations of sigmaBTs and sigmaSn in mussels, and sigmaBTs were made up mostly 100% of sigmaSn in mussels taken from locations having intensive maritime/human activities. This suggests that anthropogenic BTs represent the major source of tin accumulation in mussels. To our knowledge, this is a first comprehensive report on butyltin pollution monitoring in developing countries in the Asia-Pacific region.
Hexabromocyclododecanes (HBCDs), 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE), and decabromodiphenyl ethane (DBDPE) used as alternatives for polybrominated diphenyl ethers (PBDEs) are also persistent in the environment as PBDEs. Limited information on these non-PBDE brominated flame retardants (BFRs) is available; in particular, there are only few publications on environmental pollution by these contaminants in the coastal waters of Asia. In this regard, we investigated the contamination status of HBCDs, BTBPE, and DBDPE in the coastal waters of Asia using mussels as a bioindicator. Concentrations of HBCDs, BTBPE, and DBDPE were determined in green (Perna viridis) and blue mussels (Mytilus edulis) collected from the coastal areas in Cambodia, China (mainland), SAR China (Hong Kong), India, Indonesia, Japan, Malaysia, the Philippines, and Vietnam on 2003-2008. BTBPE and DBDPE were analyzed using GC-MS, whereas HBCDs were determined by LC-MS/MS. HBCDs, BTBPE, and DBDPE were found in mussels at levels ranging from <0.01 to 1,400, <0.1 to 13, and <0.3 to 22 ng/g lipid wt, respectively. Among the three HBCD diastereoisomers, α-HBCD was the dominant isomer followed by γ- and β-HBCDs. Concentrations of HBCDs and DBDPE in mussels from Japan and Korea were higher compared to those from the other Asian countries, indicating extensive usage of these non-PBDE BFRs in Japan and Korea. Higher levels of HBCDs and DBDPE than PBDEs were detected in some mussel samples from Japan. The results suggest that environmental pollution by non-PBDE BFRs, especially HBCDs in Japan, is ubiquitous. This study provides baseline information on the contamination status of these non-PBDE BFRs in the coastal waters of Asia.
The genus Pseudo-nitzschia has attracted attention because of production of the toxin, domoic acid (DA), causing Amnesic Shellfish Poisoning (ASP). Pseudo-nitzschia blooms occur frequently in Chinese coastal waters, and DA has been detected in several marine organisms, but so far no Pseudo-nitzschia strains from Chinese waters have been shown to produce DA. In this study, monoclonal Pseudo-nitzschia strains were established from Chinese coastal waters and examined using light microscopy, electron microscopy and molecular markers. Five strains, sharing distinct morphological and molecular features differentiating them from other Pseudo-nitzschia species, represent a new species, Pseudo-nitzschia simulans sp. nov. Morphologically, the taxon belongs to the P. pseudodelicatissima group, cells possessing a central nodule and each stria comprising one row of poroids. The new species is characterized by the poroid structure, which typically comprises two sectors, each sector located near opposite margins of the poroid. The production of DA was examined by liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses of cells in stationary growth phase. Domoic acid was detected in one of the five strains, with concentrations around 1.05-1.54 fg cell-1. This is the first toxigenic diatom species reported from Chinese waters.
The distributions of Cu, Zn, and Pb concentrations in the selected soft tissues (foot, cephalic tentacle, mantle, muscle, gill, digestive caecum, and remaining soft tissues) and shells of the mud-flat snail Telescopium telescopium were determined in snails from eight geographical sites in the south-western intertidal area of Peninsular Malaysia. Generally, the digestive caecum compared with other selected soft tissues, accumulated higher concentration of Zn (214.35+/-14.56 microg/g dry weight), indicating that the digestive caecum has higher affinity for the essential Zn to bind to metallothionein. The shell demonstrated higher concentrations of Pb (41.23+/-1.20 microg/g dry weight) when compared to the selected soft tissues except gill from Kuala Sg. Ayam (95.76+/-5.32 microg/g dry weight). The use of different soft tissues also can solve the problem of defecation to reduce error in interpreting the bioavailability of heavy metals in the intertidal area.
A study was conducted to investigate the influence of Asian monsoon on chlorophyll-a (Chl-a) content in Sabah waters and to identify the related oceanographic conditions that caused phytoplankton blooms at the eastern and western coasts of Sabah, Malaysia. A series of remote sensing measurements including surface Chl-a, sea surface temperature, sea surface height anomaly, wind speed, wind stress curl, and Ekman pumping were analyzed to study the oceanographic conditions that lead to large-scale nutrients enrichment in the surface layer. The results showed that the Chl-a content increased at the northwest coast from December to April due to strong northeasterly wind and coastal upwelling in Kota Kinabalu water. The southwest coast (Labuan water) maintained high concentrations throughout the year due to the effect of Padas River discharge during the rainy season and the changing direction of Baram River plume during the northeast monsoon (NEM). However, with the continuous supply of nutrients from the upwelling area, the high Chl-a batches were maintained at the offshore water off Labuan for a longer time during NEM. On the other side, the northeast coast illustrated a high Chl-a in Sandakan water during NEM, whereas the northern tip off Kudat did not show a pronounced change throughout the year. The southeast coast (Tawau water) was highly influenced by the direction of the surface water transport between the Sulu and Sulawesi Seas and the prevailing surface currents. The study demonstrates the presence of seasonal phytoplankton blooms in Sabah waters which will aid in forecasting the possible biological response and could further assist in marine resource managements.
In Malaysia, harmful algal blooms often occur along the coastal waters of west Sabah, where one of the causative organisms is the toxin-producing dinoflagellate, Pyrodinium bahamense var. compressum. A total of five P. bahamense var. compressum isolates were obtained from four locations and were cultured in f/2 medium. A Polymerase Chain Reaction (PCR) based technique was developed and used to screen for the presence of the dinoflagellate, P. bahamense var. compressum. A dinoflagellate-specific primer pair was designed based on sequences of P. bahamense var. compressum to amplify the 18S small subunit ribosomal DNA (rDNA) sequences. The rDNA of the P. bahamense var. compressum isolates were obtained. A species-specific primer pair was designed to target a 600 bp rDNA sequence of the target dinoflagellate. The primer pair targeting P. bahamense var. compressum did not yield any product with the fifteen algae cultures used as negative controls, but only amplified the rDNA of P. bahamense var. compressum cultures. The PCR method for identification of P. bahamense var. compressum was also applied on twenty field samples collected with plankton net. P. bahamense var. compressum cells were detected by PCR in five field samples and were confirmed by direct sequencing. From this study, a species-specific primer pair was obtained to identify the target species, P. bahamense var. compressum, among the natural complex communities of seawater.
Near-shore surface sediment was collected from five stations off Redang Island located on the eastern coast of Peninsular Malaysia. Freeze-dried sediments were Soxhlet extracted and then fractionated using column chromatography into aliphatic and polar fractions. Determination of these fractions was carried out using gas chromatography mass spectrometry. The concentration of total resolved aliphatic hydrocarbons in sediments ranged from 157 to 308 ng/g. The distribution of aliphatic fraction showed the presence of n-alkanes ranging from nC15 to nC33 with a minor odd-to-even predominance exhibiting carbon maximum, depending on station, at nC17, nC26, nC29 or nC31. Calculation of Carbon Preference Index (CPI) for CPI(15-33) gave values ranging from 1.09 to 1.46. n-Alkanol in all sediment exhibits even-to-odd carbon predominance ranging from nC16 to nC28 and maximising at nC22. n-Fatty acids distribution ranged from nC14 to nC24 with a dominant maximum at nC16 and exhibiting high values of short chain fatty acids (≤nC20) to long chain fatty acids (>nC20) ratios. Unsaturated fatty acids, particularly nC16:1 and nC18:1 is also ubiquitous in all samples. Cholesterol is the most abundant compound amongst the sterol group ranging from 42.8 to 62.6% of the total sterols. β-Sitosterol, brassicasterol and stigmasterol, are also present but of relatively lower amount. These observations suggest that the aliphatic lipids and sterols in the study area originate, mainly, from biogenic sources of marine microbial with minor contribution from epiticular waxes of terrestrial plants.
This study aims to determine the concentrations of surfactants in the surface microlayer (SML), subsurface water (SSW) and fine mode aerosol (diameter size
The microbial composition in coastal water of the Port Dickson beach in Negeri Sembilan, Malaysia was analyzed using several microbial indicators for the purpose of selecting the best indicator for marine water pollution. The indicators studied were total coliform (TC), fecal coliform (FC), fecal streptococci (FS) and coliphage. Five locations were selected along the Port Dickson beaches and samplings were carried out in 1998 and 2001. The results showed an increase in the number of total coliform (TC), fecal coliform (FC) and fecal streptococci (FS) between these two sampling by 98.12%, 86.12% and 99%, respectively. The numbers of TC, FC and FS exceeded the recommended limit for recreational seawater based on U.S. EPA 1986 standard. There was a positive correlation between TC, FC and FS and negative to coliphages.
Halophilic micro-organisms have received much interest because of their potential biotechnological applications, among which is the capability of some strains to synthesize polyhydroxyalkanoates (PHA). Halomonas sp. SK5, which was isolated from hypersaline microbial mats, accumulated intracellular granules of poly(3-hydroxybutyrate) [P(3HB)] in modified accumulation medium supplemented with 10% (w/v) salinity and 3% (w/v) glucose.
Widespread accumulation and distribution of microplastics at the sea surface raise concerns as the habitat is a feeding ground for zooplankton. As primary consumers, these organisms are closely connected to microplastic input in the marine food chain. Little comparative information currently exists about this problem in estuary and offshore systems. This study investigates microplastic distribution in the surface water and the potential ingestion of microplastics in selected taxonomic groups of zooplankton from the Terengganu Estuary to offshore waters, Malaysia. In the surface water, three types of microplastics were found (fibres, fragments and pellets). Fibres made up the highest percentage, comprising 80.8% and 73.8% of microplastics in offshore waters and estuaries, respectively. The highest total density of microplastics was found in the Terengganu Estuary (545.8 particles m-3). Microplastics sampled from the offshore waters were identified as polyamide, polyethylene, and polypropylene, which possibly originated from secondary microplastic sources. Two types of microplastics were detected in zooplankton: fibres and fragments. Fibres were the most commonly ingested microplastic type in zooplankton collected from offshore waters (94%) and estuaries (77.7%). The average sizes of ingested fibres and fragments were 361.7 ± 226.8 μm and 96.8 ± 28.1 μm, respectively, with a wider range of sizes ingested observed in offshore waters than in estuaries. The concentration of microplastics in seven zooplankton groups varied from 0.01 ± 0.002 particles ind.-1 (Harpacticoida) to 0.2 ± 0.14 particles ind.-1 (Aphragmophora). Notwithstanding the conformity of our results (increased anthropogenic activities led to greater plastic pollution within the estuary), no significant correlation was observed between the levels of microplastic ingestion and microplastic concentration in the surface water within both areas. Our results provide an important baseline reference on microplastic pollution from estuary to offshore waters, as well as proving that zooplankton act as a repository for microplastic in the marine ecosystem.
The establishment of next-generation technology sequencing has deepened our knowledge of marine sponge-associated microbiota with the identification of at least 32 phyla of Bacteria and Archaea from a large number of sponge species. In this study, we assessed the diversity of the microbial communities hosted by three sympatric sponges living in a semi-enclosed North Sea environment using pyrosequencing of bacterial and archaeal 16S ribosomal RNA gene fragments. The three sponges harbor species-specific communities each dominated by a different class of Proteobacteria. An α-proteobacterial Rhodobacter-like phylotype was confirmed as the predominant symbiont of Halichondria panicea. The microbial communities of Haliclona xena and H. oculata are described for the first time in this study and are dominated by Gammaproteobacteria and Betaproteobacteria, respectively. Several common phylotypes belonging to Chlamydiae, TM6, Actinobacteria, and Betaproteobacteria were detected in all sponge samples. A number of phylotypes of the phylum Chlamydiae were present at an unprecedentedly high relative abundance of up to 14.4 ± 1.4% of the total reads, which suggests an important ecological role in North Sea sponges. These Chlamydiae-affiliated operational taxonomic units may represent novel lineages at least at the genus level as they are only 86-92% similar to known sequences.
Fungi and other eukaryotes represent one of the last frontiers of microbial diversity in the sponge holobiont. In this study we employed pyrosequencing of 18S ribosomal RNA gene amplicons containing the V7 and V8 hypervariable regions to explore the fungal diversity of seven sponge species from the North Sea and the Mediterranean Sea. For most sponges, fungi were present at a low relative abundance averaging 0.75% of the 18S rRNA gene reads. In total, 44 fungal OTUs (operational taxonomic units) were detected in sponges, and 28 of these OTUs were also found in seawater. Twenty-two of the sponge-associated OTUs were identified as yeasts (mainly Malasseziales), representing 84% of the fungal reads. Several OTUs were related to fungal sequences previously retrieved from other sponges, but all OTUs were also related to fungi from other biological sources, such as seawater, sediments, lakes and anaerobic digesters. Therefore our data, supported by currently available data, point in the direction of mostly accidental presence of fungi in sponges and do not support the existence of a sponge-specific fungal community.