Rhizobacterial population density and nitrogen fixation in seagrass community of Gulf of Mannar, India.

Seagrass rhizosphere generally supports high bacterial population density which plays a major role in determining the nutrient cycles of the sea. Higher densities of total heterotrophic bacteria (26.3 ?06 CFU g-1), nitrogen fixing (27.3 ?03 CFUg-1), ammonifying (44.66 X 106 MPN g-1) and nitrifying bacteria (42.33 X 106 MPN g-1 ) have been registered in the seagrass areas than the non seagrass area. In particular, all these rhizosphere microbial population was higher in Thalassia hemprichii. The rates of nitrogen fixation was recorded in the different species of seagrasses such as Enhalus acoroides (1.166 n mol g-1d-1), Halophila ovalis (0.166 n mol g-1d-1), Thalassia hemprichii (18.5 n mol g-1d-1), Cymodocea serrulata (10.5 n mol g-1d-1), Halodule uninervis (5.375 n mol g-1d-1) and Syringodium isoetifolium (0.666 n mol g-1d-1) using gas chromatography. The average nitrogen fixation by the seagrasses of Gulf of Mannar alone was estimated to be 7640.58 n mol m-2 d-1 and the contributions from the rhizosphere microbes will increase the quantity to many fold.

Expression of heat shock protein70 in oral submucous fibrosis and oral squamous cell carcinoma: An immunohistochemical study.

Background: Heat shock proteins are a highly conserved group of protective cellular proteins whose synthesis is increased in response to a variety of environmental or pathophysiological stresses. Heat shock proteins are useful biomarkers for carcinogenesis in tissues and signal the degree of differentiation and the aggressiveness of cancers. Regulation of heat shock protein 70 (HSP70) expression in oral submucous fibrosis is not known much, and the aim of this study was to evaluate HSP70 expression in oral submucous fibrosis and oral squamous cell carcinoma by immunohistochemical method and to understand the role of HSP70 in tumorigenesis. Materials and Methods: Immunohistochemical method was used to detect HSP70 expression in normal oral mucosa, oral submucous fibrosis (n=30) and oral squamous cell carcinoma (n=20). HSP70 immunoreactivity was correlated with histological and clinicopathological features. Results: A significant increase in expression of HSP70 was observed (P<0.000) as the tissue progressed from oral submucous fibrosis towards oral squamous cell carcinoma. Conclusion: HSP70 is synthesized upon stress situations arising in cells of all living organisms. Expression of HSP70 indicates that stress plays an important role as a predisposing factor in oral submucous fibrosis and its subsequent progression to oral squamous cell carcinoma.

Biological activity of sea anemone proteins: II. Cytolysis and cell line toxicity.

Potent cytolytic activity was exhibited by proteins extracted from three sea anemones viz. Heteractis magnifica, Stichodactyla haddoni and Paracodylactis sinensis by affecting the red blood corpuscles (RBC) and the mouse fibroblast cell line (L929) and leukemia cell line (P388). Crude toxin of all the three anemone species induced spontaneous hemolysis of chicken, goat and human erythrocytes. The crude toxin of H. magnifica (0.98 mg/ml) elicited hemolysis at levels of 4096, 512 and 4096 HU (hemolytic unit) in chicken, goat and human erythrocytes respectively. Subsequently, the crude toxin of S.haddoni (0.82 mg/ml) exhibited a hemolytic activity of 256, 128 and 512 HU and that of P. sinensis (0.60 mg/ml) had a hemolytic activity of 128, 4096 and 512 HU. Most of the partially purified proteins of these anemones also exhibited the activity against the three different erythrocytes. The viability of L929 and P388 was adversely affected on adding the crude toxins. The symptoms of toxicity shown by the cells were rounding, lysis and detachment from the substratum. These effects were the least in S. haddoni, as compared to those the crude toxins of the other two species. Inhibition of growth of L929 exhibited by the toxin of the three species ranged between 61.08 and 93.38%. Similarly, inhibition of the growth of P388 ranged between 51.32 and 86.16%. The present investigation reveal the cytotoxic nature of anemone toxins.

Biological activity of sea anemone proteins: I. Toxicity and histopathology.

The crude as well as partially purified protein fractions from anemone species viz. Heteractis magnifica, Stichodactyla haddoni and Paracodylactis sinensis, collected from the Gulf of Mannar, south east coast of India were found to be toxic at different levels to mice. The mice showed behavioral changes such as loss of balance, opaque eyes, tonic convulsions, paralysis, micturiction, flexing of muscles, prodding (insensitive to stimulii), foaming from mouth and exophthalmia. The toxic proteins upon envenomation produced several chronic and lethal histopathological changes like formation of pycnotic nuclii and glial nodules in the brain; heamolysis, thrombosis and myocardial haemorrhage in the heart; granulomatous lesions, and damage to the hepatic cells in the liver and haemorrhage throughout the kidney parenchyma and shrinkage of glomerular tufts in the kidney. The toxins proved to be neurotoxic, cardiotoxic, nephrotoxic and hepatotoxic by their action on internal organ systems. The toxins were also thermostable till 60oC and had considerable shelf life.

Spatial and temporal variations in phytoplankton in coral reef and seagrass ecosystems of the Palk Bay, southeast coast of India.

Spatial and temporal behavior of distribution of phytoplankton of the coral reef and seagrass environment of the Palk Bay was studied during April 2002 to March 2003. A total of 133 species of phytoplankton was recorded during the study period, of which, 98 species belong to Bacillariophyceae, 15 species belong to Dinophyceae, 12 species belong to Cyanophyceae and 8 species belong to Chlorophyceae. Diatoms (57.14 to 94.10%) contributed more towards the percentage composition of different groups of phytoplankton at the two stations, followed by dinoflagellates (3.12 to 28.57%), blue-greens (2.43 to 12.5%) and greens (3.7 to 7.69%). Higher phytoplankton population density was recorded during the summer season at both stations (St.1. 62,000 cells l-1 and St.2. 55,000 cells l-1). Coral reef environment was two-fold more productive (2.10-130.21 mg C m-3 hr-1) than the seagrass environment (3.30 - 85.56 mg C m-3 hr -1). Chlorophyll ‘a’ concentration showed higher values at station 1, corresponding to the higher phytoplankton population density recorded at this station along with primary productivity.

Spectral reflectance properties of mangrove species of the Muthupettai mangrove environment, Tamil Nadu.

In the Muthupettai mangrove environment, spectral properties of six mangrove species viz. Avicennia marina, Aegiceras corniculatum, Excoecaria agallocha, Acanthus ilicifolius, Suaeda monoica and S. maritima was studied using Multi band Ground Truth Radiometer (Model-041). The study found that the chlorophyll concentration of different mangrove leaves varies between 0.05 and 0.36 mg g(-1), registering the minimum in S. maritima and maximum in E. agallocha. Interestingly species with higher chlorophyll concentration showed lower reflectance values alteast in the spectral bands 1 and 2. E. agallocha registered 0.36 mg g(-1) of chlorophyll while it recorded only 2.18 and 2.43% reflectance where as S. maritima recorded 3.16 and 3.27% of reflectance in bands 1 and 2. This indicates chlorophyll concentration is one of major factors responsible in determining the reflectance pattern of the pant communities. The spectral properties of mangroves were largely differed with that of the water and soil samples collected from the same locations, these results favourd the utilization of remotely sensed data for depicting various water and soil quality parameters from that of mangrove species in the mangrove environment. This study also found that the difference in reflectance of mangroves at canopy level is not only influenced by the chlorophyll content of species but also by the prevailing environmental condition and background reflectance of soil and water as well.

Physico-chemical and microbial characteristics of the coral reef environment of the Gulf of Mannar marine biosphere reserve, India.

Investigation on physico-chemical parameters and bacteial characteristics of the coral reef environs of the Gulf of Mannar biosphere reserve was studied. The study found the influence of different physico-chemical parameters on one another and also on the distribution of the total heterotrophic bacteria (THB) in the coral reef areas. Nutrients exhibited considerable seasonal and spatial variations with influence on the bacterial population. Coral reef areas recorded higher bacterial population density both in water (3.5 to 18 x 10(5) CFU ml(-1)) and sediment (1 to 14 x 10(7) CFU g(-1)) samples than the non coral reef areas (3.4 to 10.5 x 10(4) CFU ml(-1) in water and 0.9 to 7 x 10(6) CFU g(-1)). The study also found the dominance of gram negative groups at all the three stations (64.73, 63,5 and 72.59%) with Pseudomonas contributing maximum number of strains in all the samples. In addition Vibrio, Aeromonas, Flavobacterium, Cytophaga, Enterobacter and Alcaligenes were also recorded. The gram positive group was represented by Bacillus, Micrococcus, Arthrobacter and Corynebacterium. The genetic composition of THB isolated from the coral mucus revealed the presence of Vibrio and Micrococcus in all the coral mucus.

Phosphate solubilizing actinomycetes in the estuarine environment: an inventory.

Sediment samples were collected from different stations of the Vellar estuary for isolation of total actinomycetes and phosphate solubilizing actinomycetes. Phosphatase activity in the sediments was also investigated Consistently a higher number of actinomycetes, phosphate solubilizing actinomycetes and phosphatase activity were recorded from the clay sediments than the sandy sediments at all the stations. In all, 7 strains showed positive phosphatase activity. Among them, one strain PS-3 exhibited good activity and was further investigated for optimum phosphorus solubilization at different pH (6, 6.5, 7, 7.5 and 8) and incubation (1st day to 20th day) periods. The solubilizing activity was maximum at the pH 7 and an incubation period of 13 days was required for an appreciable quantity of phosphorus to be leached into the medium. Based on the chemotaxonomical and conventional methods of identification, the strain PS-3 has been tentatively identified as Streptomyces galbus. The present study indicates that phosphatase enzyme and S. galbus along with other actinomycetes species would play a major role in solubilizing the phosphate in the estuarine ecosystem and increasing the soluble phosphate concentration thereby enhancing the productivity

Partial purification and anti-leukemic activity of L-asparaginase enzyme of the actinomycete strain LA-29 isolated from the estuarine fish, Mugil cephalus (Linn.).

The actinomycete strain LA-29 isolated from the gut contents of the fish, Mugil cephalus of the Vellar estuary showed excellent L-asparaginase activity The enzyme was purified 18-fold and the final recovery of protein was 1.9%, which exhibited an activity of 13.57 IU/mg protein. The partially purified L-asparaginase inhibited the growth of leukemia cells in male wistar rats. Average survival period of the rats was more in an optimum enzyme dose of 100 units and the survival period was less when the dosages were increased and at the same time the enzyme became less effective when the dosages were decreased. Higher survival of 17.2 days was recorded when 100 units of the enzyme was given in three intermittent doses (50/25/25 units) at the interval of 24 hr. Analysis of cell components of the strain LA-29 has revealed the wall type-I which is the characteristic of the genus Streptomyces. Further the morphological, physiological and biochemical features along with the micromorphological results obtained for the strain LA-29 were compared with that of the Streptomyces species found in Bergey's Manual of Determinative Bacteriology and the strain LA-29 has been tentatively identified as Streptomyces canus.

Studies on L-asparaginase enzyme of actinomycetes isolated from estuarine fishes.

Actinomycetes were isolated from different organs viz. skin, gills and gut contents of three species of fishes viz. Mugil cephalus (Linnaeus, 1758), Chanos chanos (Forskal, 1775) and Etroplus suratensis (Bloch, 1780) using three different media from the Vellar estuary, situated along the southeast coast of India. Among the three fishes, M. cephalus harboured highest number of actinomycetes population in all the three body parts examined followed by C. chanos and E. suratensis. Out of the three body parts of all fishes, gut contents had highest actinomycetes population followed by gills and skin. Among the three media used for isolation of actinomycetes, Kuster's agar medium was found to be suitable than the starch casein agar and glucose asparagine agar media. Out of the 40 strains isolated, only six strains (LA-2, LA-8, LA-15, LA-20, LA-29 and LA-35) showed significant L-asparagianse activity and were taken up for further studies. Impact of various physical and chemical factors such as pH, temperature, sodium chloride concentration, carbon sources and amino acids on the growth of actinomycetes and L-asparaginase activity was also studied. Optimum growth and enzyme activity was noticed under pH 7 to 8, temperature 37 degrees C, 1-2% sodium chloride concentration, sucrose as carbon source and without any amino acids. Analysis of the cell components of the isolated strains has revealed the wall type-I (the wall type-I is typical for the genus Streptomyces) and the strains were micromorphologically similar to the genus Streptomyces. Hence, the morphological, physiological and biochemical along with the micromorphological results obtained for the L-asparaginase producing strains were compared and the strains were tentatively identified as Streptomyces aureofasciculus (LA-2), S. chattanoogenesis (LA-8), S. hawaiiensis (LA-15), S. orientalis (LA-20), S. canus (LA-29) and S. olivoviridis (LA-35).

Effect of natural preservatives on the growth of histamine producing bacteria.

Present study deals with the hampering of the growth of histamine producing bacteria (HPB), by using NaCl and spices which are easily available and cheaper cost wise. For this experiment, four strains of HPB viz. Vibrio parahaemolyticus, Bacillus cereus, Pseudomonas aeruginosa and Proteus mirabilis were tested against 1 to 10% concentrations of NaCl and 1 to 5% concentrations of natural preservatives (turmeric, ginger and garlic) in a basal medium. HPB showed different growth rates at different concentrations of NaCl and natural preservatives. V. parahaemolyticus, B. cereus and Ps. aeruginosa showed no growth at 10% concentration. When the HPB growth was tested with garlic, turmeric and ginger extracts, growth of all the bacteria was inhibited by garlic and turmeric extracts at 5% concentration. In ginger, V. parahaemolyticus, B. cereus and P. mirabilis were totally inhibited at 5% concentration. But Ps. aeruginosa showed very less growth at this concentration.

Nutrient characteristics and sediment texture of the seagrass beds of the Gulf of Mannar.

Changes in nutrient concentrations and sediment texture of the seagrass beds were recorded for two years from July 1996 to June 1998 at monthly intervals at two stations (Station 1--Manoli island; Station 2--Chinnapalam) of the Gulf of Mannar Marine Biosphere Reserve. Different nutrients in water showed varied concentrations in different seasons. Phosphate concentration varied between 2.32 and 7.32 microM, nitrate concentration ranged from 5.2 to 18.78 microM and nitrite concentration from 1.92 to 732 microM. Sand was a major component of the sediments, followed by silt and clay at both the stations. At station 1, sand composition varied between 85.9 and 95.3% while at station 2, it varied between 81.9 and 93.6%. Silt composition varied between 3.1 and 9.4% at station land at station 2, it varied between 4.0 and 12.4%. At station 1, clay composition varied from 1.6 to 4.6% and at station 2, it varied between 2.1 and 5.6%. The present study revealed that the nutrients and sediment characteristics of the seagrass beds of the Gulf of Mannar has clear spatial and temporal variations. They are largely interlinked and influenced by the human activities and climatic changes and are individually or jointly governing the distribution, diversity, biomass and productivity of the seagrasses of the Gulf of Mannar.

Water quality and phytoplankton characteristics in the Palk Bay, southeast coast of India.

The present study was carried out to determine the water quality in terms of physicochemical characteristics and plankton distribution in the coastal waters of Kattumavadi, Palk Bay for a period of one year from April 2002 to March 2003. Air and surface water temperatures varied from 28 degrees C to 32.50 degrees C and from 27.5 to 32.0 degrees C while light extinction coefficient (LEC) varied between 0.95 and 1.85. Salinity ranged from 26.0 to 34.5 per thousand and the pH ranged between 7.95 and 8.35. Variation in dissolved oxygen content was from 4.15 to 7.18 ml(-1), and the particulate organic carbon (POC) content varied between 0.49 and 2.28 mgCl(-1). Concentrations of nutrients viz. nitrate (2.15 to 8.28 microM), nitrite (0.12 to 0.62 microM), inorganic phosphate (1.28 to 2.15 microM) and reactive silicate (5.15 to 12.52 microM) also varied independently. Chlorophyll a content ranged from 0.28 to 1.48 mg m(-3) and the primary productivity, from 4.19 to 16.08 mgCm(-3) hr(-1). The present study recorded a total of 43 species of planktonic diatoms and two species of blue-green algae. Population density of phytoplankton varied from 18,000 to 34,000 cells l(-1). Percentage composition, of the diatoms showed minimum values during the monsoon season and the maximum values during the premonsoon season.

Optimum conditions for L-glutaminase production by actinomycete strain isolated from estuarine fish, Chanos chanos (Forskal, 1775).

Actinomycetes were isolated from skin, gills and gut contents of estuarine fish. Chanos chanos using Kuster's agar medium. Out of 20 strains tested, the strain LG-10 which was tentatively identified as Streptomyces rimosus showed L-glutaminase activity. Optimum production of L-glutaminase enzyme (17.51 IU/ml) was observed after 96 h of incubation at 27 degrees C, pH 9 and glucose and malt extract as carbon and nitrogen sources, respectively. The present study indicated scope for the use of S. rimosus as an ideal organism for the industrial production of extracellular L-glutaminase.