Polyphosphate Accumulation by Marine Bacteria

Phosphate (Pi) is one among the most important essential residues in maintenance and inheritance of life, with far diverse physiological role as structural, functional and energy transduction. Phosphate accumulation in wastewaters containing run off of fertilizers and industrial discharges is a global problem that results in algal blooms in bays, lakes and waterways. Currently available methods for removing phosphates from wastewater are based primarily on polyP accumulation by the activated sludge bacteria. PolyP plays a critical role in several environmental and biotechnological problems. Possible relation of interaction between polyP accumulation phenomenon, the low biomass, low Pi uptake, and varying results obtained in response to the impact of sodium chloride, pH, temperature, various inorganic salts and additional carbon sources studied, are all intriguing observations in the present investigation. The results of the present study have evidenced very clearly the scope for potential strains of bacteria from both sea water and marine sediments which could be exploited both for Pi removal in wastewater released by industries and intensive aquaculture practices in to the aquatic environment as well as to harness the potential strains for industrial production of polyP which was wide range of applications.

Reviving the Macrobrachium rosenbergii (de Man) Fishery in Vembanad Lake, India

In Vembanad Lake and its confluent rivers (Kerala, India), the catches of Macrobrachium rosenbergii (de Man) were reported to have dwindled to a mere 39 t in the 1980s from average landings of 300 t during the 1960s. This decline is due to the impact of a number of human interventions affecting the ecosystem and, hence, the stocks of M. rosenbergii. Monitoring of landings in 1994-1995 and 1995-1996 indicates an improvement in catches. This paper discusses the reasons for the decline and revival in stocks and suggestions for their replenishment.

Studies on influance of environmental parameters affecting the biology of oyster crassostrca madrasensis(Preston)

The Indian edible oyster Crassostrea madrasensis (Preston) is known to be a highly suitable candidate species for culture. Though Q, madrasensis has been subjected to intensive research, there has been no significant attempt to culture this oyster commercially. One major reason for the lack of interest in oyster culture could be the disparity in growth, survival and production reported by earlier workersf from different regions along the Indian coast. Greater predictability of production can create confidence and encourage entrepreneurs interested in oyster culture. The present study, which is a detailed investigation on the influence of various environmental variables on growth and reproduction of Q, madrasensis, is not confined to the impact of only hydrological parameters but is also extended to study the effect of different degrees of aerial exposure on growth and survival. The main objective of the study is to develop a background for subsequent development of a site suitability index for culture of Q, madrasensis along the Indian coast. Two sets of experiments were conducted during the present study. Details of the experiments are presented in the thesis under two major chapters comprising four sections each. Each chapter has a separate introduction, materials and methods, results and discussion. .

“Population Characteristics, Fishery and Post larval distribution of Macrobrachium rosenbergii (de Man) and M. idella (Hilgendorf) of Vembanad lake”

Macrobrachium rosenbergii (de Man) and M.ide77a (Hilgendorf) form two commercially important Palaemonid prawns of the Vembanad lake (9 28' and 10 10’ N and 76 13' and 76 31' E). Both of them were known to have contributed to a very lucrative fishery during the sixties, however, in recent years these natural resources have badly depleted owing to the impact of many man made alterations brought about in the ecosystem such as habitat reduction, physical obstruction imposed in the migratory pathway of these species, pollution hazards, etc,. Changed environmental conditions and increased fishing pressures caused persistent alterations in the stock size of these prawns during the past so many years, however, no serious attempt was made to monitor the stock size from time to time and also to bring out the resource characteristics. Though, the morphotypic differentiation in grow out male population of M.rosenbergii has been documented, no similar studies were conducted with regard to natural male and female population. Based on the data collected during fishery cruise surveys conducted in Vembanad lake from March '94 to February '96, population characteristics. postlarval distribution, fishery and population dynamics of M.rosenbergii and M.ide77a of the lake were studied in detail.

Dayglow Emissions on Mars and Venus

An attempt has been made in this thesis to model some of the emissions observed by SPICAM and SPICAV on Mars and Venus, respectively, viz., CO Cameron band, CO+ 2 ultraviolet doublet, N2 triplet bands, atomic oxygen green (5577 A), red doublet (6300, 6364 A), and ultraviolet (2972 A) emissions. One of major sources of these emissions is photoelectron impact ionization/excitation. In this thesis, an electron degradation model based on Monte Carlo technique has been developed to calculate the production/excitation rates of above mentioned emissions due to electron impact. The limb brightness pro les of emissions are calculated and compared with the observations wherever available. The e ect of various model input parameters on dayglow emissions intensities is also evaluated

Characterization and Pathogenicity of Vibrio cholerae and Vibrio vulnificus from Marine environments

The genus Vibrioof the family Vibrionaceae are Gram negative, oxidasepositive, rod- or curved- rodshaped facultative anaerobes, widespread in marine and estuarine environments. Vibrio species are opportunistic human pathogens responsible for diarrhoeal disease, gastroenteritis, septicaemia and wound infections and are also pathogens of aquatic organisms, causing infections to crustaceans, bivalves and fishes. In the present study, marine environmental samples like seafood and water and sediment samples from aquafarms and mangroves were screened for the presence of Vibrio species. Of the134 isolates obtained from the various samples, 45 were segregated to the genus Vibrio on the basis of phenotypic characterization.like Gram staining, oxidase test, MoF test and salinity tolerance. Partial 16S rDNA sequence analysis was utilized for species level identification of the isolates and the strains were identified as V. cholerae(N=21), V. vulnificus(N=18), V. parahaemolyticus(N=3), V. alginolyticus (N=2) and V. azureus (N=1). The genetic relatedness and variations among the 45 Vibrio isolates were elucidated based on 16S rDNA sequences. Phenotypic characterization of the isolates was based on their response to 12 biochemical tests namely Voges-Proskauers’s (VP test), arginine dihydrolase , tolerance to 3% NaCl test, ONPG test that detects β-galactosidase activity, and tests for utilization of citrate, ornithine, mannitol, arabinose, sucrose, glucose, salicin and cellobiose. The isolates exhibited diverse biochemical patterns, some specific for the species and others indicative of their environmental source.Antibiogram for the isolates was determined subsequent to testing their susceptibility to 12 antibiotics by the disc diffusion method. Varying degrees of resistance to gentamycin (2.22%), ampicillin(62.22%), nalidixic acid (4.44%), vancomycin (86.66), cefixime (17.77%), rifampicin (20%), tetracycline (42.22%) and chloramphenicol (2.22%) was exhibited. All the isolates were susceptible to streptomycin, co-trimoxazole, trimethoprim and azithromycin. Isolates from all the three marine environments exhibited multiple antibiotic resistance, with high MAR index value. The molecular typing methods such as ERIC PCR and BOX PCR revealed intraspecies relatedness and genetic heterogeneity within the environmental isolatesof V. cholerae and V. vulnificus. The 21 strains of V. choleraewere serogroupedas non O1/ non O139 by screening for the presence O1rfb and O139 rfb marker genes by PCR. The virulence/virulence associated genes namely ctxA, ctxB, ace, VPI, hlyA, ompU, rtxA, toxR, zot, nagst, tcpA, nin and nanwere screened in V. cholerae and V. vulnificusstrains.The V. vulnificusstrains were also screened for three species specific genes viz., cps, vvhand viu. In V. cholerae strains, the virulence associated genes like VPI, hlyA, rtxA, ompU and toxR were confirmed by PCR. All the isolates, except for strain BTOS6, harbored at least one or a combination of the tested genes and V. choleraestrain BTPR5 isolated from prawn hosted the highest number of virulence associated genes. Among the V. vulnificusstrains, only 3 virulence genes, VPI, toxR and cps, were confirmed out of the 16 tested and only 7 of the isolates had these genes in one or more combinations. Strain BTPS6 from aquafarm and strain BTVE4 from mangrove samples yielded positive amplification for the three genes. The toxRgene from 9 strains of V. choleraeand 3 strains of V. vulnificus were cloned and sequenced for phylogenetic analysis based on nucleotide and the amino acid sequences. Multiple sequence alignment of the nucleotide sequences and amino acid sequences of the environmental strains of V. choleraerevealed that the toxRgene in the environmental strains are 100% homologous to themselves and to the V. choleraetoxR gene sequence available in the Genbank database. The 3 strains of V. vulnificus displayed high nucleotide and amino acid sequence similarity among themselves and to the sequences of V. cholerae and V. harveyi obtained from the GenBank database, but exhibited only 72% homology to the sequences of its close relative V. vulnificus. Structure prediction of the ToxR protein of Vibrio cholerae strain BTMA5 was by PHYRE2 software. The deduced amino acid sequence showed maximum resemblance with the structure of DNA-binding domain of response regulator2 from Escherichia coli k-12 Template based homology modelling in PHYRE2 successfully modelled the predicted protein and its secondary structure based on protein data bank (PDB) template c3zq7A. The pathogenicity studies were performed using the nematode Caenorhabditiselegansas a model system. The assessment of pathogenicity of environmental strain of V. choleraewas conducted with E. coli strain OP50 as the food source in control plates, environmental V. cholerae strain BTOS6, negative for all tested virulence genes, to check for the suitability of Vibrio sp. as a food source for the nematode;V. cholerae Co 366 ElTor, a clinical pathogenic strain and V. cholerae strain BTPR5 from seafood (Prawn) and positive for the tested virulence genes like VPI, hlyA, ompU,rtxA and toxR. It was found that V. cholerae strain BTOS6 could serve as a food source in place of E. coli strain OP50 but behavioral aberrations like sluggish movement and lawn avoidance and morphological abnormalities like pharyngeal and intestinal distensions and bagging were exhibited by the worms fed on V. cholerae Co 366 ElTor strain and environmental BTPR5 indicating their pathogenicity to the nematode. Assessment of pathogenicity of the environmental strains of V. vulnificus was performed with V. vulnificus strain BTPS6 which tested positive for 3 virulence genes, namely, cps, toxRand VPI, and V. vulnificus strain BTMM7 that did not possess any of the tested virulence genes. A reduction was observed in the life span of worms fed on environmental strain of V. vulnificusBTMM7 rather than on the ordinary laboratory food source, E. coli OP50. Behavioral abnormalities like sluggish movement, lawn avoidance and bagging were also observed in the worms fed with strain BTPS6, but the pharynx and the intestine were intact. The presence of multi drug resistant environmental Vibrio strainsthat constitute a major reservoir of diverse virulence genes are to be dealt with caution as they play a decisive role in pathogenicity and horizontal gene transfer in the marine environments.