In vitro cytotoxicity MTT assay in Vero, HepG2 and MCF -7 cell lines study of Marine Yeast.

Marine yeasts isolated from coastal mangrove ecosystem namely Candida albicans, Kuraishia capsulate and Sacchromyces cerevisae were screened for the cytotoxicity against a non-transformed Vero (African green monkey kidney normal cell line) and two cancer cell lines human breast carcinoma cells (HepG2), human breast carcinoma cells (MCF -7) in different concentrations (1000 to 1.953 μg/ml). Lower doses enhanced the viability of the cultured cells, MTT assay .higher doses decreased viability of the cells by 50% or more. MTT assay was used to measure the cell proliferation and survival. Amongst three yeast strains, Sacchromyces cerevisae showed more than 80% cell viability in Vero cell lines and were studied for further cytotoxicity against HepG2, MCF -7 cell lines respectively.

Anatomical Changes during Rooting of Mangroves - Avicennia officinalis and Excoecaria agallocha.

Mangrove forests continue to disappear all over the world due to a number of reasons. This is the study made to screen the anatomical and biochemical changes during the rooting process. Two important mangrove species have been examined based on their salt relationship natures: Avicennia officinalis subsp. australasica (salt excreting plant) and Excoecaria agallocha L (salt accumulating plant). The plants were treated with different root promoting hormones like IBA, IAA and NAA at different concentrations for three minutes each. After 45 days of the growth period root growth and anatomical changes were observed. The roots originated from the deeper zone of the secondary xylem after several anatomical variations in the hormone treated plants. This is the first trial which may help to determine the mass propagation of these commercial and medicinally important mangroves.

Genetic diversity in three populations of Avicennia marina along the eastcoast of India by RAPD markers.

Genetic diversity was analysed in three populations of the mangrove species, Avicennia marina by using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Ten random decamer primers were used to score the diversity from three locations of eastcoast of India: Parangipettai (Tamil Nadu), Kakkinada (Andhra Pradesh) and Sundarbans (West Bengal). These primers produced 388 scorable DNA fragments, of which 252 (64.98%) were polymorphic, 182 (46.90%) were monomorphic, and 14 (3.61%) were unique. RAPD banding patterns displayed variations between and within the populations, while, there was no morphological variation.

Isolation of antibacterials from the mangrove, Avicennia marina and their activity against multi drug resistant Staphylococcus aureus

Objective: leaf extract of A. marina was tested on the growth of clinically isolated multi-drug resistant Staphylococcus aureus and its bioactive compounds were attempted. Method: Clinical strain of Staphylococcus aureus, were isolated from sputum, pus and blood of different patients and 22 strains were screened for antibiotic susceptibility. Avicennia marina was extracted in different solvents and antibacterial assay was carried out using Kirby-Bauer’s disk diffusion method. Crude methanol extract of the mangrove was loaded on a silica gel column and eluted with chloroform and methanol (9:1 to 1:9) followed by ethyl acetate and methanol (9:1 to 1:9). Based on in vitro assay, the 12th fraction was subjected for minimum inhibitory concentration (MIC). The active fraction was analysed by using a Clarus 500 Perkin Elmer gas chromatography. Result: Based on the antibiotic susceptibility test, six strains (RMSA 6, RMSA12, RMSA16, RMSA18, RMSA19 and RMSA21) were resistance against methicillin, vancomycin and ciprofloxacin. The results indicated that the methanolic leaf extract showed the highest antibacterial activity against all the tested strains RMSA 6 (16mm), RMSA12 (15 mm), RMSA16 (13 mm), RMSA18 (10 mm), RMSA19 (17 mm) and RMSA21 (16 mm). The MIC of the partially purified extract showed potential results against all the multidrug resistant strains however, the lowest concentration was recorded against RMSA 6, RMSA19 and RMSA21 strain. In the GC-MS results, 5 bioactive compounds were identified from the partially purified extract of A.marina. Conclusion:The methanolic extract of A. marina has the more potential candidate to inhibit against multidrug resistant S. aureus.

Effect of neem extract against the bacteria isolated from marine fish.

Marine ornamental fishes are exceedingly valuable due to their high demand in domestic and international markets. There is a growing global interest to rear the fishes in captivity. But problem due to bacteria and fungi are the major hitch in captive condition. Since, the use of antibiotics is banned, an attempt was made to ascertain in vitro assay of the neem leaves extract against the bacterial pathogens isolated from infected fishes. Bacterial strains isolated from infected regions of the clown fishes Amphiprion sebae and A. ocellaris were identified as Aeromonas hydrophila, Enterobacter sp., E. coli, Pseudomonas aeruginosa, Proteus sp., Streptococcus sp., Vibrio cholerae, V. alginolyticus, V. parahaemolyticus and Yersinia enterocolitica. Ethanol and methanol extracts were highly inhibitory to the bacterial isolates when compared to other solvents. Ethanol extracts exhibited low minimum inhibitory concentration (75 - 250 ,g ml-1) as compared to other extracts. The present finding revealed that the neem leaf extract significantly reduces the bacterial pathogens and their infection in marine ornamental fishes.

Biomedical and pharmacological potential of tetrodotoxin-producing bacteria isolated from marine pufferfish Arothron hispidus (Muller, 1841)

Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55%, in Cellulomonas fimi with 620, 300, 10 and 128.42%, and in Bacillus species with 40, 200, 849.98 and 158.69%, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43% whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16% at the lowest dilution.Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55 percent, in Cellulomonas fimi with 620, 300, 10 and 128.42 percent, and in Bacillus species with 40, 200, 849.98 and 158.69 percent, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43 percent whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16 percent at the lowest dilution.(AU)

Antimicrobial activity of tissue and associated bacteria from benthic sea anemone Stichodactyla haddoni against microbial pathogens.

Associated bacteria from Stichodactyla haddoni are found maximum in tentacle tissues than the body tissue. There are eight associated bacterial species viz., Alcaligenes sp, Corynebacterium sp, Aeromonas sp, Sporosarcina sp, Renibacterium sp, Camobacterium sp1, Camobacterium sp2 and Salinococcus sp were recorded. The culture extracts from the associated bacterial species showed sensitivity against human bacterial and fungalpathogens. However, the hexane tissue extract of sea anemone showed maximum sensitivity (24 mm dia.) against the fish bacterial pathogen Aeromonas hydrophila than the other chosen pathogens. Comparatively the tissue extracts showed promising antimicrobial sensitivity than the cell free extracts of associated bacteria, and hence, the tissue samples from the sea anemone Stichodactyla haddoni is recommended for further exploration of novel antimicrobial drugs than the associated bacteria.

Growth of Avicennia marina and Ceriops decandra seedlings inoculated with halophilic azotobacters.

Inoculation of azotobacter has significant positive effects on the growth characteristics and pigments in mangrove seedlings of Avicennia marina and Ceriops decandra. The bacterial inoculation significantly increased the root dry biomass at the maximum of 75.8% at 30 gl(-1) salinity in Ceriops decandra. But in Avicennia marina, the shoot dry biomass was increased significantly at the maximum of 56.12% at 30 gl(-1) salinity in general, the Azotobacter beijerinkii improved the growth characteristics better in both species of mangroves preferably at higher salinity levels in A. marina and at a range of salinity in C. decandra. The results recommend this forraising vigorous seedlings under nursery conditions.

Making artificial honey using yeast cells from salivary glands of honey bees.

The salivary glands of a honey bee, Apis cerana and the yeast cells isolated from these glands were studied for their effects on sucrose solution. This solution exhibited lowered pH and increased levels of fructose and total amino acids as the time of incubation proceeded. The solution thus made was similar to the natural honey.

How to alleviate degradation of mangroves?

This work has experimentally proved that hyper salinity, a major cause for degradation of coastal mangrove habitats, can be alleviated by flushing of hyper saline soil with tidal water and/or with rainwater. Over a period of three years after digging the creeks to flush hyper saline soil with tidal water, an appreciable reduction in soil salinity and a moderate increase in colonization of mangroves are observed. Soil analysis showed a significant reduction in salinity after 2 months of storage of rainwater with a significant and concomitant increase of heterotrophic bacterial counts and nutrients. This study raises the possibility of converting degrading mangrove habitats to luxuriant ones through man-made efforts.

Polythene and plastic-degrading microbes in an Indian mangrove soil

Biodegradation of polythene bags and plastic cups was analyzed after 2, 4, 6, and 9 months of incubation in the mangrove soil. The biodegradation of polythene bags was significantly higher (up to 4.21 in 9 months) than that of plastic cups (up to 0.25 in 9 months). Microbial counts in the degrading materials were recorded up to 79.67 x 10(4) per gram for total heterotrophic bacteria, and up to 55.33 x 10(2) per gram for fungi. The microbial species found associated with the degrading materials were identified as five Gram positive and two Gram negative bacteria, and eight fungal species of Aspergillus. The species that were predominant were Streptococcus, Staphylococcus, Micrococcus (Gram +ve), Moraxella, and Pseudomonas (Gram -ve) and two species of fungi (Aspergillus glaucus and A. niger). Efficacy of the microbial species in degradation of plastics and polythene was analyzed in shaker cultures. Among the bacteria, Pseudomonas species degraded 20.54 of polythene and 8.16 of plastics in one-month period. Among the fungal species, Aspergillus glaucus degraded 28.80 of polythene and 7.26 of plastics in one-month period. This work reveals that the mangrove soil is a good source of microbes capable of degrading polythene and plastics.

How do mangrove forests induce sedimentation?

The mangrove forests play a significant role as sediment traps. They reduce tidal flows and induce sedimentation of soil particles at low tide. However, there are no such processes taking place in the non-mangrove areas. Site of Avicennia-Rhizophora interphase is more efficient than Avicennia and or Rhizophora zones, in trapping the sediment by 30, 25 and 20% respectively at low tide as compared to high tide.

Growth of a mangrove (Rhizophora apiculata) seedlings as influenced by GA3, light and salinity

The growth performance of Rhizophora apiculata Blume (mangrove) seedlings in the presence and absence of exogenous gibberellic acid (GA3) under different combinations of salinity and light was analyzed. Root and shoot growth responses of 75-day old seedlings in liquid-culture, were measured. It was concluded that light exhibited a significant inhibitory effect on all the growth parameters-number of primary roots, primary root length, shoot elongation, number of leaves, total leaf area; and, the GA3 treatment singly or in combinations with light, showed a significant influence on the total leaf area and primary root length.

Effect of probiotics on bacterial population and health status of shrimp in culture pond ecosystem.

The artificially manufactured probiotics having beneficial bacteria, Bacillus spp. was applied regularly in a modified extensive shrimp (Penaeus monodon) culture pond, located on the bank of Vellar estuary, Parangipettai. The populations of total heterotrophic bacteria (THB), beneficial bacteria (Bacillus spp.) and pathogenic bacteria (vibrios) were monitored in water and sediment of the pond. The results were compared with a control pond, situated in the same location having same water spread area, stocking density, species managed with same technologies and optimum environmental parameters in which no probiotic was applied. The populations of THB and Bacillus spp. in the experimental pond increased and the vibrios decreased after each application of probiotics. But the result of the control pond showed an increasing trend of the populations of THB, Bacillus spp. and vibrios towards days of culture. The control pond had lower levels of THB and Bacillus spp. and higher levels of vibrios than the probiotic applied (experimental) pond. Also the probiotics maintained optimum transparency and low organic load in the experimental pond as compared to control. In general, water and sediment had almost equal number of Bacillus spp. and vibrios, but sediment had higher THB load than water. The applications of probiotics lesser pathogenic vibrios and enhance beneficial bacilli in the culture leading to improved water quality, promoted growth and survival rates and increased the health status of the shrimp without stress and disease outbreaks. Thus the application of probiotics could lead to disease-free and profitable shrimp culture operations which will be helpful for shrimp farmers as most of them are now-a-days severely affected by microbial diseases.

The effects of electric impulse on growth of Rhizophora mucronata seedlings (Rhizophorales: Rhizophoraceae)

Exposure of mangrove seedlings to electric current was studied at an intensity of 10, 20 and 30 mA to 0.5, 1, 3, 6, 12 and 24 h in different combinations, either to plumule or radicle or both parts of the propagules of Rhizophora mucronata Poir (sample size = 30). The electric current-treated propagules and untreated control were cultured hydroponically in sea water for 75 days, and root and shoot growth parameters measured. The root elongation was significantly influenced by duration of exposure, site of propagule treated and combined effects of current intensity x site of propagule treated. Leaf number and area were significantly influenced by intensity of electric current and duration of exposure; whereas shoot elongation was affected by the duration of exposure. The root number was significantly affected by duration of exposure. In general, the electric impulse improved the growth characteristics of shoot and root.

A survey of some Indian medicinal plants for anti-human immunodeficiency virus (HIV) activity.

BACKGROUND & OBJECTIVES: India has an extensive area of forest enriched with plant diversity. Several of these plants have been used as folklore medicines. However, the medicinal plants have rarely been investigated for anti-human immunodeficiency virus activity. Hence, some Indian medicinal plants were screened in vitro against human immunodeficiency virus (HIV). METHODS: The inhibitory effect of plant extracts on HIV replication was monitored in terms of inhibition of virus induced cytopathogenicity in MT-4 cells. The MT-4 cells were infected with HIV. The HIV infected or mock infected MT-4 cells were incubated at 37 degrees C in a CO2 incubator in the presence of the plant extracts. After five days, cell viability was measured by tetrazolium based colorimetric assay. RESULTS & INTERPRETATION: Of the 69 plant species screened, 16 were effective against HIV-1 and 4 were against both HIV-1 and HIV-2. The most effective extracts against HIV-1 and HIV-2 are respectively Cinnamomum cassia (bark) and Cardiospermum helicacabum (shoot + fruit). The findings provide a rationale for further studies on isolation of active principles and pharmacological evaluation.

In vitro anti human immunodeficiency virus activity of mangrove plants.

Mangrove plant extracts were screened in vitro against human immunodeficiency virus (HIV) on MT-4 cells. HIV infected MT-4 cells were cultured for five days in the presence of the extract, at various concentrations. Anti-HIV activity was evaluated by tetrazolium-based colorimetric assay. Seven extracts were found effective. Five of the active extracts completely inhibited the virus adsorption to the cells. Most of the active plants are from the family Rhizophoraceae.

In vitro anti-vaccinia virus activity of some marine plants.

Seventy three marine plant extracts were tested in vitro in chick embryo fibroblast cell culture and their anti-vaccinia virus activity was evaluated in terms of reduction in number of plaques by the extracts. Only seven extracts were found to show the activity. Sargassum wightii, a seaweed had the highest activity, reducing 65 per cent of plaques formed by vaccinia virus.