Tropical Estuarine Waters Research Articles

Biosurfactant Production from the Biodegradation of n-Paraffins, Isoprenoids and Aromatic Hydrocarbons from Crude Petroleum by Yarrowia lipolytica IMUFRJ 50682

Yarrowia lipolytica is a unique, strictly aerobic yeast with the ability to degrade efficiently hydrophobic substrates. In the present work, we evaluated the degrading potential of Yarrowia lipolytica IMUFRJ 50682, isolated from tropical estuarine water in Rio de Janeiro (Brazil), and the possible biomolecules produced during this process. To investigate which crude oil compounds are degraded by Y. lipolytica IMUFRJ 50682, this microorganism was grown in a medium containing Marlim petroleum (19 °API, American Petroleum Institute gravity) at 28 °C and 160 rpm for 5 days. The residual petroleum was submitted to gas chromatograph-mass spectrometric analysis (GC-MS). The chromatographic fingerprints of the residual petroleum were compared with the abiotic control test incubated in the same conditions. Y. lipolytica assimilates high molecular weight hydrocarbons, such as n-alkanes (C11-C19), isoprenoids (pristane and phytane), aromatics with two or three aromatics rings (naphthalene, methylnaphthalenes, dimethylnaphthalenes, trimethylnaphthalenes, phenanthrene, methylphenanthrenes, dimethylphenanthrenes, anthracene). This strain was also capable of consuming more complex hydrocarbons, such as tricyclic terpanes. During this biodegradation, the emulsification index of the culture medium increased significantly, showing that biosurfactant molecules can be produced from this process. Therefore, Y. lipolytica IMUFRJ 50682 showed to be a potential crude oil degrading yeast, which can be used for bioremediation processes and simultaneously produce bioproducts of commercial interest.

Dinoflagellates in tropical estuarine waters from the Maraú River, Camamu Bay, northeastern Brazil

Dinoflagellates display great diversity in tropical regions and play an important role in the complex microbial food webs of marine and brackish environments. The goal of this study is to identify planktonic dinoflagellates and their distribution in the estuary of the Maraú River, Camamu Bay, state of Bahia, in a region with increasing use of shellfish farming. Samples were carried out monthly from August 2006 to July 2007 at four stations along the estuary. Plankton was sampled with a 20 μm mesh net. We identified 20 dinoflagellate species. The greatest species richness was observed in the genera Protoperidinium (five spp.), Tripos (four spp.), and Prorocentrum (three spp.). Based on literature, six species were classified as potentially harmful: Akashiwo sanguinea, Dinophysis caudata, Gonyaulax spinifera, Prorocentrum micans, Scrippsiella cf. acuminata, and Tripos furca. Protoperidinium venustum was recorded for the first time in coastal waters of Bahia.

Using DNA Barcodes to Aid the Identification of Larval Fishes in Tropical Estuarine Waters (Malacca Straits, Malaysia).

Larval descriptions of tropical marine and coastal fishes are very few, and this taxonomic problem is further exacerbated by the high diversity of fish species in these waters. Nonetheless, accurate larval identification in ecological and early life history studies of larval fishes is crucial for fishery management and habitat protection. The present study aimed to evaluate the usefulness of DNA barcodes to support larval fish identification since conventional dichotomous keys based on morphological traits are not efficient due to the lack of larval traits and the rapid morphological changes during ontogeny. Our molecular analysis uncovered a total of 48 taxa (21 families) from the larval samples collected from the Klang Strait waters encompassing both spawning and nursery grounds of marine and estuarine fishes. Thirty-two (67%) of the larval taxa were identified at the species level, two taxa (4%) at the genus level, and 14 taxa (29%) at family level. The relatively low rate of species-level identification is not necessarily due to the DNA barcoding method per se, but a general lack of reference sequences for speciose and non- commercial fish families such as Gobiidae, Blenniidae, and Callionymidae. Larval morphology remains important in species diagnoses when molecular matches are ambiguous. A lower ethanol percentage (50%) for larva preservation is also useful to keep the body of larvae intact for morphological identification, and to preserve DNA for subsequent molecular analyses. The 10% Chelex resin used to extract DNA is also cost- effective for long term monitoring of larval fishes. Hence, the DNA barcoding method is an effective and easy way to aid the identification of estuarine larval fishes at the species level.

Screening of tropical estuarine water in south-west coast of India reveals emergence of ARGs-harboring hypervirulent Escherichia coli of global significance.

The goal of this study was to investigate the involvement of a tropical Indian estuary in the emergence of antibiotic resistance genes (ARGs)-harboring hypervirulent E. coli of global significance. A total of 300 E. coli isolates was tested for antibiotic susceptibility to β-lactams, aminoglycosides, chloramphenicol, quinolones, sulphonamides, tetracyclines, and trimethoprim. The E. coli isolates were screened for the presence of antibiotic resistance genes (blaTEM, blaCTX-M, tetA, tetB, sul1, sul2, strA, aphA2, catI, dhfr1, and dhfr7), integrase (int1, int2, and int3), Shiga toxin genes (stx1 and stx2) and extraintestinal virulence genes (papAH, papC, sfa/focDE, kpsMT II, and iutA). The highest prevalence of antibiotic resistance was observed for ampicillin, followed by tetracycline, and nalidixic acid. Among E. coli isolates, 64% were resistant to at least one of the 15 antibiotics tested, and approximately 40% were multiple antibiotic-resistant (MAR). More than 40% (n = 122) of E. coli isolates had ARGs. Integrase 1 (int1) was found in 7.6% of E. coli isolates. Among E. coli isolates, 16.3% (n = 49) were extraintestinal pathogenic E. coli (ExPEC), and approximately 34.6% (n = 17) of ExPEC had ARGs. A hypervirulent ARGs-harboring STEC was isolated. The prevalence of Shiga toxin-producing E. coli (STEC) was low (n = 1). The prevalence of ARGs-harboring pathogenic E. coli isolates was higher in stations close to the City (urban area), than that of other stations. ERIC-PCR (enterobacterial repetitive intergenic consensus sequence polymerase chain reaction) analysis revealed a high degree of genetic diversity among the ARGs-harboring E. coli isolates. The results demonstrate a high prevalence of ARGs-harboring E. coli in estuarine water and confirm the need for a better wastewater treatment facility and proper control measures to reduce the discharge of sewage and wastewater into the aquatic environments.

High porewater exchange in a mangrove-dominated estuary revealed from short-lived radium isotopes

We hypothesise that mangroves play an important role in groundwater exchange processes in sub-tropical and tropical estuarine waters. To investigate this, multiple high resolution time series measurements of radium across a tidal estuary (Coffs Creek, NSW, Australia) were performed as well as a spatial survey in both bottom and surface layers. Results from the spatial survey revealed increasing radium concentrations in parts of the estuary surrounded by mangroves. The average radium concentration in estuary areas lined with mangroves was 2.5 times higher than the average concentration at the mouth of the estuary and 6.5-fold higher than upstream freshwater areas. Additionally, the area enriched in radium coincided with low dissolved oxygen concentrations, implying that porewater exchange may drive anoxia. A radium mass balance model based on 223Ra and 224Ra isotopes at different sections of the estuary confirmed higher porewater exchange rates from areas fringed with mangrove vegetation. Estimated porewater exchange rates were 27.8±5.3 and 13.6±2.1cmd−1 (0.8±0.1 and 0.4±0.1m3s−1) based on 223Ra and 224Ra isotopes, respectively. The average saline porewater exchange was∼10-fold larger than the upstream surface freshwater inputs to the estuary. We suggest that mangrove environments within subtropical estuaries are hotspots for porewater exchange due to the complex belowground structure of crab burrows and the effect of tidal pumping. Because porewater exchange releases carbon and nitrogen from coastal sediments, development and modification of mangrove areas in subtropical estuaries have a significant effect on coastal biogeochemical cycles.

OCCURRENCE OF PSEUDO-NITZSCHIA SPECIES (BACILLARIOPHYCEAE) AT KUCHING ESTUARIES FROM YEAR 2007 TO 2010

Amnesic shellfish poisoning (ASP) is a type of seafood poisoning due to human consumption on shellfish mollusks contaminated with algal toxin, domoic acid (DA) derived from the diatom Pseudo-nitzschia. Bloom dynamics of this diatom in tropical waters were poorly understood due to lack of long term study on the organism. Occurrence of Pseudo-nitzschia species was investigated at Santubong and Samariang estuarine waters, Kuching, Sarawak from 2007 to 2010. The estuaries were characterized by shallow water with semi-diurnal tidal cycle. Cell abundance was determined by microscopic enumeration. Temperature, pH, salinity and macronutrients at the sampling sites were determined at each sampling occasions. Highest cell density of Pseudo-nitzschia spp. was recorded 8.0 × 103 cells L in Santubong in April 2009 and P. brasiliana cells were found highest in October 2007 (2 × 104 cells L-1). Increase in cell abundance was coincided with high salinity and low precipitation rate. Water temperature and pH showed insignificant influence on the abundance of Pseudo-nitzschia spp. The result obtained in this study provided fundamental understanding on bloom dynamic of potential harmful Pseudo-nitzschia species in the tropical estuarine waters.

New record of Akashiwo sanguinea (Dinophyta) in the tropical estuarine waters of Northeastern Brazil (Western Atlantic).

This study reports the occurrence and the effect of the environmental factors on the spatial and temporal distribution of the dinoflagellate Akashiwo sanguinea (Hirasaka) Hansen & Moestrup in estuarine waters of northeastern Brazil. Samples were collected at seven stations from March 2007 to February 2008 during high tide and low tide, using Van Dorn bottles. The samples were immediately fixed with Lugol and analyzed with the Utermöhl method. Water samples were also collected for the identification of the hydrological characteristics of the area. Akashiwo sanguinea occurred throughout the annual cycle and at all sampling sites with densities ranging between 5 and 410 x 103 cells.L-1. The highest densities were recorded at low tide, especially during the months of the rainy season (July: 210 x 103 cells.L-1; August: 410 x 103 cells.L-1). Density values were within the normal range and blooms were not detected. Despite being common in the area, the species showed preference for sites with high concentrations of orthophosphate and total dissolved phosphorus and with salinity in the mesohaline regime.

Factors affecting biosurfactant production by oil degrading Aeromonas spp. isolated from a tropical environment

An Aeromonas spp. was isolated from tropical estuarine water. The organism grew on crude oil and produced biosurfactant that could emulsify hydrocarbons. The peak growth and biosurfactant production was on the 8th day. The organism grew on a range of hydrocarbons that include crude oil and hexadecane while no growth was recorded on some hydrocarbons that include benzene. The biosurfactant produced by the organism emulsified a range of hydrocarbons with diesel ( E 24 = 65) as the best substrate and hexane ( E 24 = 22) as the poorest. After purification, the biosurfactant was found to contain about 38% carbohydrate and an unidentified lipid. No protein was present in the purified biosurfactant. Production of biosurfactant was highest in medium with glucose and lowest in the medium with diesel + acetate. Soybean was the best nitrogen source for biosurfactant production. The activity of the biosurfactant was enhanced optimally at NaCl concentration of 5%, pH of 8.0 and temperature of 40 °C. The biosurfactant retained 77% of its original activity after 120 min of exposure to heat at a temperature of 100 °C. Biosurfactant may be produced with this organism using non-hydrocarbon substrates such as glucose and soybean that are readily available and would not require extensive purification for use in food and pharmaceutical industries.

Survival of Escherichia coli in a tropical estuary

The survival of Escherichia coli in tropical estuarine water has been studied under controlled laboratory conditions using microcosms. The survival has been assessed in terms of various self purifying factors of the natural waters such as biological, chemical and physical factors. The biological factors considered included competition from other microorganisms, predation by protozoa and coliphages. The suitability of the chemical composition of estuarine water has been studied under chemical factors and negative impact of sunlight has been studied under physical factors. The results revealed that sunlight exerted maximum negative impact, followed by biotic factors contained in the estuarine water. However, the chemical composition of the estuarine water is found to be suitable for the growth and survival of E. coli. The injury exerted by each of the above factors was also evaluated by using a selective and non-selective medium in conjunction. It was found that sunlight resulted in 100% injury of the cells as the cells failed to develop in a selective medium. While, sunlight resulted in the extinction of 90% of the E. coli cells within the first two hours of exposure, biotic factors took nearly 24 hours to remove the same amount of population.

Photochemical oxygen consumption in marine waters: A major sink for colored dissolved organic matter?

Quantification of photochemical O2 uptake provides a measure of total chemical photooxidation of dissolved organic matter (DOM). Here we study this process and present estimates suggesting that photooxidation has the potential to significantly modify marine DOM pools, complementing or exceeding oxidation via coupled chemical‐biological pathways. We measured apparent quantum yields (AQYs) of photobleaching, O2 uptake, and H2O2 production in several coastal marine samples and in dilutions of a tropical estuarine water with oligotrophic seawater. O2‐loss AQYs varied little among samples or with dilution but decreased linearly from 1.2×10–3 at 300 nm to 0.3×10–3 at 400 nm and dropped about threefold to near‐constant values with increasing absorbed light dose. H2O2 production, about 45% of O2 uptake, showed similar dependencies, whereas singlet oxygen (O2 (1Δg)) reactions contributed less than 1% of O2 uptake for typical coastal water. Implications of these findings for photochemical O2, H2O2, and DOM cycling are discussed.Modeling the dose‐dependence of O2 loss and photobleaching at 310 nm required three DOM pools. In the simplest case, about 90% is a weakly absorbing, low‐AQY pool of DOM admixed with two similar‐sized pools of more photochemically reactive DOM. This result suggests that rigorously extrapolating laboratory data to the environment requires detailed mapping of dose‐wavelength‐photobleaching AQY surfaces.Action spectra and DOM flux estimates for coastal photooxidative chemistry were derived. Site‐specific potential rates are comparable to available in situ data. Globally, the DOM photolysis capacity appears to be larger than estimated coastal DOM inputs, especially in tropical and temperate areas, including areas with maximal DOM inputs.

Utilization of cyclohexanol by bacteria in a tropical estuarine water.

The Lagos lagoon is a wide expanse of estuarine water. The range of temperature, pH and conductivity of samples collected from 3 sources were 27-30 degrees C, 7.8-8.3 and 2.34-2.85 mS respectively. Consistent increase and decrease in the population of indigenous bacteria occurred in the test and control experiments respectively. Organisms in samples from Apapa and Iddo utilized cyclohexanol better than those in the sample from the University of Lagos. The cyclohexanol degrading bacteria were identified as species of Pseudomonas. Acinetobacter, Vibrio, Micrococcus and Flavobacterium. Pseudomonas sp. had the highest growth potential on cyclohexanol. These organisms play important roles in reducing the pollutant loads of the Lagos lagoon.

Variations in condition and body constitution in a tropical estuarine fish with year-round recruitment

Fish condition (Fulton's K) and the abundance of biochemical constituents (water, protein, lipid and carbohydrate) of juveniles of the tropical estuarine fish, Ambassis vachelli, were monitored over a 15 month period. Fulton's K and biochemical constituents of juveniles recruiting into the adult habitat varied significantly throughout the study. Correlations between Fulton's K and all biochemical measures were weak, with a maximum correlation of 0.1379 (with protein abundance). Significant differences in condition and body constitution of individuals suggest that juveniles recruiting into the adult habitat are not in equal condition and therefore survivorship and success may vary within and among cohorts. Temperature (and salinity) display greater variation in tropical estuarine waters than in other tropical waters and are likely to play a role in determining recruit condition and body constitution. However, the exact role of physical parameters in determining condition and body constitution of fishes is still unknown.

The distribution and abundance of urea in tropical estuarine waters in relation to hydrochemical parameters

A field study was conducted to determine the role of hydrochemical influence on the abundance and distribution of urea‐nitrogen in a tropical estuary. The dissolved urea exhibited prominent features of seasonal estuarine build‐up during low flow conditions. At times of high biological production, this parameter was correspondingly lower. The paper further studies the influence of physical factors like temperature and salinity (estuarine hydrography) as well as chemical factors like pH, DO, DOC and Chlorophyll‐a to bring out the associated features. The uptake of urea as a preferred nutrient during biological activity is also reported. Industrial or urban waste discharges did not contribute significantly to the abundance of urea for this part of the waterbody. The tropicality of nutrient cycling as revealed from urea‐nitrogen distribution signatures the phenomenal regional diversity.

Early diagenesis of mangrove leaves in a tropical estuary: Bulk chemical characterization using solid-state 13C NMR and elemental analyses

Changes in the chemical composition of mangrove ( Rhizophora mangle) leaves during decomposition in tropical estuarine waters were characterized using solid-state 13C nuclear magnetic resonance (NMR) and elemental (CHNO) analysis. Carbohydrates were the most abundant components of the leaves accounting for about 50 wt% of senescent tissues. Tannins were estimated to account for about 20 wt% of leaf tissues, and lipid components, cutin, and possibly other aliphatic biopolymers in leaf cuticles accounted for about 15 wt%. Carbohydrates were generally less resistant to decomposition than the other constituents and decreased in relative concentration during decomposition. Tannins were of intermediate resistance to decomposition and remained in fairly constant proportion during decomposition. Paraffinic components were very resistant to decomposition and increased in relative concentration as decomposition progressed. Lignin was a minor component of all leaf tissues. Standard methods for the colorimetric determination of tannins (Folin-Dennis reagent) and the gravimetric determination of lignin (Klason lignin) were highly inaccurate when applied to mangrove leaves. The N content of the leaves was particularly dynamic with values ranging from 1.27 wt% in green leaves to 0.65 wt% in senescent yellow leaves attached to trees. During decomposition in the water the N content initially decreased to 0.51 wt% due to leaching, but values steadily increased thereafter to 1.07 wt% in the most degraded leaf samples. The absolute mass of N in the leaves increased during decomposition indicating that N immobilization was occurring as decomposition progressed.