Differences in carbohydrate profiles in batch culture grown planktonic and biofilm cells of Amphora rostrata Wm. Sm.

Diatoms are abundant in biofilms developed on surfaces immersed in sunlit waters. In both the planktonic and the biofilm mode of growth, diatoms produce carbohydrate polymers which perform several functions including motility, protection, production of macro-aggregates and detoxification. However, little is known about the differences, if any, in the production and characterization at the molecular level of carbohydrates in planktonic and biofilm cells. In order to identify the differences in these two modes of growth, the concentration of total carbohydrates, carbohydrate fractions, neutral carbohydrates, uronic acids and amino sugars in planktonic and biofilm cells of Amphora rostrata were measured. The results showed that the distribution of carbohydrate fractions, uronic acids and amino sugars was different in biofilm and planktonic cells. Cell normalized concentrations of these components were two to five times greater in planktonic cells compared with biofilm cells. The concentrations of glucose and glucosamine decreased, whereas fucose increased in planktonic cells over the period of cultivation. Conversely, the concentrations of glucose and glucosamine increased while that of fucose decreased in attached cells. The study suggests that marked differences exist between the carbohydrates of the planktonic and the biofilm cells of A. rostrata.

Distribution of butyltins in waters and sediments of the Mandovi and Zuari estuaries, west coast of India.

Butyltins (TBT, DBT, MBT) were measured in water and sediment samples collected from various locations in the Mandovi (ten stations) and Zuari (nine stations) estuaries during November 2007. The total butyltin (TB) in water samples varied between approximately 12 and 73 and from 0.5 to 77 ng Sn l(-1), whereas for the sediments, it varied from approximately 15 to 118 and 6 to 119 ng Sn g(-1) dry weight for the Mandovi and the Zuari estuaries, respectively. The average TB concentration in the sediments from the Zuari estuary was higher as compared to the Mandovi estuary, whereas for the water samples, it was vice versa. For both the water and sediment samples, MBT was generally the most predominant butyltin in these two estuaries. This suggests that the butyltin contamination was not freshly derived. Fairly good relationships between the concentrations of TB and organic carbon in water and sediment samples of these estuaries implicate the importance of adsorption/desorption processes in controlling the levels of TB in these estuaries.

Biochemical composition of the marine conditioning film: implications for bacterial adhesion.

The conditioning film formed on glass panels was analysed for total carbohydrates (CFCHO), total proteins (CFP) and total uronic acids (CFURA). The influence of these compounds on the adhesion of three marine bacterial cultures, Pseudomonas sp. CE-2, Pseudomonas sp. CE-10 and Bacillus sp. SS-10 was also evaluated. One-way analysis of variance suggested a significant increase in the attachment of all three cultures to conditioned glass panels. Moreover, CE-2 (r = 0.874) and CE-10 (r = 0.879) showed a significant positive correlation with CFCHO. Conversely, SS-10 (r = -0.69) showed a significant negative correlation with CFCHO. Backward multiple linear regression analysis indicated that CFCHO were the most predictive component of the conditioning film in explaining bacterial adhesion to the conditioned glass panels.

Sources of hydrocarbons in sediments of the Mandovi estuary and the Marmugoa harbour, west coast of India.

Surface sediments were collected from various locations of the Mandovi estuary and the Marmugoa harbour. Sediments were analysed for organic carbon (OC), total lipids, n-alkanes concentration and composition. Concentrations of OC, total lipids and n-alkanes varied spatially and ranged from 1 to 2.5%, 176 to 1413 microg/g dry weight (dw) sediments, and 0.8 to 3.2 microg/g dw sediments of the Mandovi estuary, respectively; and from 0.6 to 2.9%, 233 to 1448 microg/g dw sediments, and 1.6 to 10.7 microg/g dw sediments in the Marmugoa harbour, respectively. Long chain, odd carbon n-alkanes (C23-C33) in the Mandovi estuary, whereas short chain, even carbon n-alkanes (C11-C21) in the Marmugoa harbour sediments were more abundant. The total HC concentrations, n-alkane composition, CPI, UCM and other evaluation indices suggest the dominance of terrestrial hydrocarbons in the estuarine while petroleum derived hydrocarbons in the harbour sediments. This conclusion was further supported by the abundance of hopanes with C29 to C34 alpha, beta compounds and steranes with C27, C28 and C29 compounds in the harbour sediments.

Role of beta 1-4 linked polymers in the biofilm structure of marine Pseudomonas sp. CE-2 on 304 stainless steel coupons.

Pseudomonas sp CE-2 cells attach and form biofilms on 304-stainless steel (SS) coupons. A series of experiments were carried out in order to understand the role of exopolysaccharides (EPS) in the formation and maintenance of CE-2 biofilms on SS coupons. The biofilm density and EPS concentration increased over the period of incubation and the highest values for both were recorded after 72 h. Calcofluor and the lectin concanavalin A (Con A) showed a positive interaction with 72-h old biofilms, indicating the presence of beta 1-4 linked polymers, and alpha-d-glucose and alpha-d-mannose in the biofilm matrix of CE-2. When the CE-2 cells were grown in the presence of calcofluor (200 microg ml(-1)), biofilm formation was significantly reduced (approximately 85%). Conversely, the lectins Con A or WGA did not influence the CE-2 biofilms on the SS coupons. Furthermore, treatment with cellulase, an enzyme specific for the degradation of beta 1-4 linked polymers, removed substantial amounts of CE-2 biofilm from SS coupons. These results strongly suggest the involvement of beta 1-4 linked polymers in the formation and maintenance of Pseudomonas sp. CE-2 biofilms on SS coupons.

Application of thermoalkalophilic xylanase from Arthrobacter sp. MTCC 5214 in biobleaching of kraft pulp.

A thermoalkalophilic and cellulase-free xylanase produced from Arthrobacter sp. MTCC 5214 by solid-state fermentation using wheat bran as a carbon source was evaluated for prebleaching of kraft pulp. The UV absorption spectrum of the compounds released by enzyme treatment showed a characteristic peak at 280 nm, indicating the presence of lignin in the released colouring matter. Enzymatic prebleaching of kraft pulp showed 20% reduction in kappa number of the pulp without much change in viscosity. Enzymatic treatment reduced the amount of chlorine by 29% without any decrease in brightness. The viscosity of xylanase treated pulp was 4.0 p, whereas the viscosity of the pulp treated exclusively with chlorine was 4.1 p.

Purification and characterization of thermoalkalophilic xylanase isolated from the Enterobacter sp. MTCC 5112.

Thermoalkalophilic Enterobacter sp. MTCC 5112 was isolated from a sediment sample collected from the Mandovi estuary on the west coast of India. This culture produced extracellular xylanase. The xylanase enzyme was isolated by ammonium sulfate (80%) fractionation and purified to homogeneity using size exclusion and ion exchange chromatography. The molecular mass of the xylanase was approximately 43 kDa. The optimal pH of the xylanase activity was 9, and at room temperature it showed 100% stability at pH 7, 8 and 9 for 3 h. The optimal temperature for the enzyme activity was 100 degrees C at pH 9.0. At 80 degrees C and pH 9, 90% of the enzyme activity was retained after 40 min. At 70 and 60 degrees C, the enzyme retained 64% and 85% of its activity after 18 h, respectively, while at 50 degrees C and pH 9 the enzyme remained stable for days. For xylan, the enzyme gave a K(m) value of 3.3 mg ml(-1) and a V(max) value of 5,000 micromol min(-1) mg(-1) when the reaction was carried out at 100 degrees C and pH 9. In the presence of metal ions such as Co(2+), Zn(2+), Fe(2+), Cu(2+), Mg(2+) and Ca(2+) the activity of the enzyme increased, whereas strong inhibition of enzyme activity was observed in the presence of Hg(2+) and EDTA. To the best of our knowledge, this is the first report on the production of xylanase by this bacterium.

Bacterial extracellular polymeric substance (EPS): a carrier of heavy metals in the marine food-chain.

The ecological implications of metal binding properties of bacterial EPS and its possible role in the bioaccumulation of pollutants in the marine food-chain was investigated using a partially purified and chemically characterized microbial EPS isolated from a species of Marinobacter. Various factors influencing metal sorption by the EPS including the influence of initial metal concentrations, incubation time, pH and sodium chloride concentrations on binding of lead (Pb2+) and copper (Cu2+) were evaluated. The bacterial EPS selectively bound more amount of Cu2+ per mg of EPS than Pb2+. Both copper and lead were sorbed more at near neutral pH than acidic pH. The sorption of Cu2+ increased with increasing copper concentration. The estimated maximum binding ability (MBA) of the EPS was 182 nmol copper and 13 nmol lead mg(-1) EPS. However, the sorption of these metals decreased with the increase in sodium chloride concentration. Furthermore, up to 35% of 14C-labeled Marinobacter was ingested by a benthic polychaete Hediste diversicolor. On an average, 29% of the ingested EPS was absorbed into tissues and 49% of the EPS was respired. It was apparent that the animals used the EPS as a source of energy and nutrition. The labile nature of the bacterial EPS and its ability to bind heavy metals might route the bound metals through the marine food chain, thereby transferring and aiding bioaccumulation of metal pollutants in the higher trophic animals.

Butyltins in the sediments of Kochi and Mumbai harbours, west coast of India.

Surface sediment samples were collected from various locations in the Mumbai and Kochi harbours, west coast of India, to assess the presence of butyltin compounds. Tributyltin (TBT) and dibutyltin (DBT) varied from 16 to 16,816 ng/g dry wt. and from undetected to 469 ng/g dry wt., respectively, of the sediment in Kochi harbour. In Mumbai harbour, the values of TBT and DBT ranged between 4.5 and 1193 ng/g dry wt. and from undetected to 131 ng/g dry wt. of the sediments, respectively. The concentrations of both TBT and DBT showed strong seasonal variation probably due to the effect of tides and currents. Nevertheless, the levels of butyltin compounds were generally higher at sites influenced by shipping activities such as navigation, dry dock and ship-building activities. The presence of DBT indicates the abiotic or microbiological degradation of TBT. Compared to TBT, DBT was relatively less abundant, suggesting either fresh inputs of TBT and/or less degradation of TBT. The concentrations of TBT showed significant positive relationships with organic carbon and lipid, implying that both lipophilic and ionic interactions were probably involved in controlling the abundance of TBT at these sediments. The observed levels of butyltin compounds are much higher than those required to induce toxic effects on marine organisms, suggesting that these sediments were contaminated with butyltin compounds.

Butyltins in water, biofilm, animals and sediments of the west coast of India.

Biofilm, fish, oyster, mussel, clam, surface seawater, suspended particulate matter (SPM), and sediment samples were collected from marine and/or estuarine waters of the west coast of India. These samples were analysed for butyltin derivatives such as dibutyltin (DBT) and tributyltin (TBT). The concentrations DBT plus TBT varied between 2.4 and 8.3, 163 and 363 ng/l, 5 and 2853 ng/g dry wt in the SPM, seawater and sediment samples, respectively, of the Marmugao harbour. The values of DBT plus TBT ranged between 0.60 and 29, 123 and 242 ng/l and 1.4 and 65 ng/g dry wt in SPM, water and sediment samples, respectively, collected from the Mandovi estuary. In the Dona Paula Bay the DBT plus TBT varied from 10 to 89 ng/l in surface seawater, and TBT from 10 to 513 ng/g in biofilm samples. For the coastal sediment samples the concentration of DBT plus TBT ranged between 36 and 133 ng/g dry wt of sediment. For the animal samples the DBT plus TBT ranged between 58 and 825 ng/g dry wt of the tissue. Mussel tissues contained the highest amount of DBT plus TBT (825 ng/g dry wt tissue), whereas highest TBT concentration was recorded in the oyster (732 ng/g dry wt). TBT was generally the most abundant butyltin compound in most of the samples suggesting fresh inputs and/or less degradation of TBT. A wide range of the observed butyltin concentrations suggests the presence of localized areas of contamination. Leaching of tributyltin-containing antifouling paints from the ocean going ships, fishing and recreational boats, barges, and the inputs of TBT from the Goa shipyard and dry dock facility situated in the harbour are the probable sources of the DBT and TBT in the samples of the west coast of India. Higher levels of TBT were observed in biofilm relative to that in the surrounding seawater. When fed on TBT contaminated biofilm of the diatom Navicula subinflata, butyltin concentrations in the clam Paphia malabarica increased over the period of feeding suggesting the importance of biofilm in the transfer of butyltins to higher group of organisms.

Distribution of butyltins in the waters and sediments along the coast of India.

Water and surface sediment samples were analyzed for butyltins (TBT, DBT, MBT) from various ports along the east and west coast of India. The total butyltin (TB) in water samples varied between ~1.7 and 342 ng S nl⁻¹, whereas for sediments it varied between below detection limit to 14861 ng S ng⁻¹ dry weight of sediment. On an average Chennai port recorded the highest level of butyltins in the sediments while Paradip recorded the highest level of butylins in the waters. A fairly good relationship between the TB in the sediment and overlying water samples, as well as between organic carbon and TB, implicates the importance of adsorption/desorption process in controlling the levels of TBT in these port areas. In India the data on organotin pollution is very sparse; most of the port areas have been surveyed for butyltins for the first time during this study.

Organotins in the sediments of the Zuari estuary, west coast of India.

Industrial use of organotins such as butyltins and phenyltins has increased several folds during the last two decades. Butyltins and phenyltins are synthetic, multipurpose chemicals, which have been extensively used in marine antifouling paints. They have been known to be extremely poisonous to mollusc fishery resources (oysters, clams, etc.). Contamination of Zuari estuary sediments was assessed by quantitative determination of butyltins and phenyltins by using GC-MS using the electron ionization mode. Butyltins predominated in the whole area over phenyltins. Butyltins contributed about 70-90% of the organotins in general. The concentration of butyltins in sediments ranged from 20 to 7621 ng Sn/g. The concentration of phenyltins in sediments ranged from 0 to 46 ng Sn/g. Degradation indices for butyltins and phenyltins were calculated. The Butyltin degradation index (BDI) for the Zuari sediments ranged from 0 to 2.7 indicating a lot of fresh input of butyltins in the estuary and a lower degradation rate. The phenyltin degradation index (PhDI) ranged from 1 to 10 implying that different processes were prevalent at different stations. The observed organotin levels in the Zuari estuary indicate some highly localized areas of contamination which are severe enough to cause harmful effects on marine flora and fauna. Therefore, there is a need to regulate the use of butyltins and phenyltins as biocides in marine antifouling paints.

Effects of DNP on the cell surface properties of marine bacteria and its implication for adhesion to surfaces.

The effect of 2, 4-dinitrophenol (DNP) on the extracelluar polysaccharides (EPS), cell surface charge, and the hydrophobicity of six marine bacterial cultures was studied, and its influence on attachment of these bacteria to glass and polystyrene was evaluated. DNP treatment did not influence cell surface charge and EPS production, but had a significant effect on hydrophobicity of both hydrophilic (p = 0.05) and hydrophobic (p = 0.01) cultures. Significant reduction in the attachment of all the six cultures to glass (p = 0.02) and polystyrene (p = 0.03) was observed after DNP treatment. Moreover, hydrophobicity but not the cell surface charge or EPS production influenced bacterial cell attachment to glass and polystyrene. From this study, it was evident that DNP treatment influenced bacterial cell surface hydrophobicity, which in turn, reduced bacterial adhesion to surfaces.

Alpha-L-arabinofuranosidases: the potential applications in biotechnology.

Recently, alpha-L-arabinofuranosidases (EC3.2.1.55) have received increased attention primarily due to their role in the degradation of lignocelluloses as well as their positive effect on the activity of other enzymes acting on lignocelluloses. As a result, these enzymes are used in many biotechnological applications including wine industry, clarification of fruit juices, digestion enhancement of animal feedstuffs and as a natural improver for bread. Moreover, these enzymes could be used to improve existing technologies and to develop new technologies. The production, mechanisms of action, classification, synergistic role, biochemical properties, substrate specificities, molecular biology and biotechnological applications of these enzymes have been reviewed in this article.

Dynamics of amino acids in the conditioning film developed on glass panels immersed in the surface seawaters of Dona Paula Bay.

The conditioning film developed on glass panels immersed in surface seawater over a period of 24 h was analysed for total organic carbon (OC), total organic nitrogen (ON), and total hydrolyzable amino acid (THAA) concentrations and composition. The concentrations of C and N and THAA increased, whereas the C/N ratio decreased over the period of immersion. The amino acid-C and N accounted for 3.7-6.7% and 10.3-65.3% of OC and ON, respectively. The relative contribution of glycine plus threonine and serine to the total amino acids decreased while that of valine, phenylalanine, isoleucine and leucine increased over the period of immersion. Principal component analysis (PCA) based on mole% amino acid composition showed that the degradation indices (DI) for the conditioning film organic matter increased over the period of immersion. A high C/N ratio, a low %THAA-C, % THAA-N and DI values and the abundance of glycine plus threonine and serine in the conditioning film organic matter during the first few hours following immersion imply that the adsorbed organic matter was mostly derived from degraded organic matter.

Analysis of microfouling products formed on metallic surfaces exposed in a marine environment.

Mild steel (MS), stainless steel (SS) and copper (Cu) test panels were immersed in the surface water of Dona Paula Bay over a period of 15 d. During the immersion period data on the hydrography, nutrients and suspended matter were also collected. The suspended matter and fouling products on the MS, SS and Cu panels were analysed for organic carbon (OC), organic nitrogen (ON), chlorophyll a (chl a), protein and carbohydrate concentration and composition, and the dry weight (DW) was recorded. Compared to suspended matter, the chemical and biochemical components of the fouling products showed strong temporal and substratum related differences. The microfouling biomass (as DW, OC, ON, chl a and protein) on all the test panels generally increased over the period of immersion. Carbohydrates were more abundant in the suspended matter whereas fouling products were enriched in proteins. The contribution of protein-carbon to the total carbon increased over the period of immersion for the microfouling products on MS and SS whilst it did not show a consistent trend on Cu. Whereas, the carbohydrate-carbon contribution to the total carbon increased for the fouling products on MS, it did not exhibit a particular pattern on SS or Cu over the period of immersion. Capillary gas chromatographic analysis showed the presence of glucose, galactose, mannose, arabinose, xylose fucose and ribose in both the fouling products and suspended matter. However, there were differences in the relative distribution of these monosaccharides in the suspended matter and the fouling products. Glucose was the most abundant monosaccharide, which showed strong temporal variations in suspended matter. In contrast, the wt % concentrations of individual monosaccharides showed large temporal differences for the fouling products, which were strongly influenced by the period of immersion and the type of test substratum. Glucose and fucose were relatively more abundant in the fouling products on SS and Cu, whilst glucose was the most abundant monosaccharide on MS. The monosaccharide and chemical composition data suggest strong temporal changes in the composition of the fouling products.