Laser Induced Fluorescence Based Optical Fiber Probe for Analyzing Bacteria

Optical fiber based laser induced fluorescence (LIF) measurements were carried out using Rhodamine B to analyze two different species of bacteria , a Gram-positive bacteria namely Bacillus smithii , and fibrin alginolvticus, a Gram- negative bacteria . The fiber sensor was clearly able to distinguish between the two species of bacteria . Quenching effect of the dye Rhodamine B by Bacillus smithii was observed . The effect of dye on the samples was also studied in detail.

Heterotrophic Bacteria Associated with Healthy and Moribund Larvae of Penaeus monodon H.Milne Edwards

Heterotrophic bacterial flora of Pmonadon from an apparently healthy hatchery system as well as a pool with heavy mortality were isolated and studied. In the healthy systems comparatively higher generic diversity with Pseudomonas, Acinetobacter, Bacillus, Micrococcus, members of the family Enterobacteriaceae and coryneform group in the diminishing order of dominance was recorded. Meanwhile from the moribund larvae and rearing water Aeromonas and Pseudomonas could be isolated in almost equal proportions. Strikingly, Aeromonas could not be isolated from the apparently healthy larval rearing system and its exclusive occurrence in the sick culture system in comparatively higher percentage suggested its possible role in the mortality. They were found to be highly halophilic exhibiting growth at 10% NaCl. On testing their sensitivity to twenty antibiotics, four of them (Streptomycin, Gentamycin, Methamine mandelate and Cloramphenicol) were found to be effective on all the isolates of Aeromonas and Pseudomonas suggesting their possible application in the hatchery system in times of emergency. While doing so, Streptomycin would do comparatively better than the others as the minimum inhibitory dose required was comparatively lower (200ppm) within a period of 24 hours

Laser induced fluorescence based optical fiber probe for analyzing bacteria

Optical fiber based laser induced fluorescence (LIF) measurements were carried out using Rhodamine B to analyze two different species of bacteria , a Gram-positive bacteria namely Bacillus .cmithii , and fibrin alginolvticus, a Gram-' negative bacteria . The fiber sensor was clearly able to distinguish between the two species of bacteria . Quenching effect of the dye Rhodamine B by Bacillus smitltii was observed . The effect of dye on the samples was also studied in detail.

Studies on immobilization of Bacteria

Cell immnhilizatinn technology in a rapidly expanding arna in the endeavour of microbial fnrmentatiwn.During the lnmt 15 years anveral prnceafinn have been developed and more are in developmental atage of approaching commercial utilizatinn.In the present programme it was planned to develop an optimized process for the innobilization of alpha amylase producing Bacillus polymyxa (CBTB 25) an isolate obtained from Cochin University campus primarily for the production of alpha-amylase.Optimal concentration of support material that attributaa stability and maximal activity to the immobilized cell beads was determined using different concentrations of sodium aliginate as support and estimation of amylase production.An overeall assessment of the data obtained for the various studies conducted denotes that immobilized cells synthesize alpha-amylase at comparable rates with free cells and produce reducing sugara at a higher level than free cells.Results indicated that both phosphate and citrate buffers could be used for disrupting the immobilized beads since they enforced maximal release of cells through leaching from the beads within one hour.On comparative analysis it was observed that immobilized cells could synthesize alpha amylase at similar levels with free cells of B.polymyxa.On Co-immobilization of B.Polymyxa with S.cerevisiae,the co-immobilizate beads could effeciently convert starch directly to ethanol with a yield of 14.8% at 1 : 2 ratio.

L-Glutaminase production by Marine Bacteria

L-glutaminases (L—glutamine amidohydrolase EC.3.5.l.2) is proposed as a prospective candidate for enzyme therapy cnf cancer and also as zui important additive during enzymatic digestion of shoyu koji since it could enhance glutamate content of soysauce. Commercial production of glutaminase could make possible its wide application in these areas, which would demand availability of potential sources and suitable fermentation techniques. The ‘present investigation highlighted marine environment as a potential source of efficient glutaminase producing bacteria mainly species of pseudomonas, aeromonas ,vibrio,alcaligenes, acinetobacter bacillus and planococci.Among them pseudomonas fluorescens ACMR 267 and v.cholerae ACMR 347 were chosen as the ideal strains for glutaminase production.Extracellular glutaminase fraction from all strains were in higher titres than intracellular enzymes during growth in mineral media, nutrient broth and nutrient broth added with glutamine.Glutaminase from all strains were purified employing (NH4)2SO4 fractionation followed tnr dialysis and ion exchange chromatography. The purified glutaminase from all strains were observed to be active and stable over a wide range of gfii and temperature.Optimization studies cflf environmental variables that normally influence time yiehi of glutaminase indicated that the optimal requirements of these bacteria for maximal glutaminase production remained stable irrespective of the medium, they are provided with for enzyme production. However, solid state fermentation technique was observed to be the most suitable process for the production of Glutaminase.

Fermentation Of Rice And Related Products By Bacillus Sp.

Recently’ recognition cnf immobilization ‘technology for the rapid conversion of several substrates into metabolites and repeated reuse of the biocatalysts have drawn the attention of the fermentation scientists and technologists to try these new technologies for the rapid production of pnxkmt and enhancement of the efficiencies of the systems Hence in the present study rice was selected,as a substrate since it is a rich source of starch, available and cultivated throughout the year almost in all part of our country. Rice although known for its use as a staple food in many forms as rice, idli, dosai etc., has not been used in industry extensively. However, it ii; a potential resource for’ the production of alcohol, high protein food anui for sugar and sugar syrups as it is evidenced by the few reports mentioned in the review of literature. Of the several microorganisms available, Bacillus sp, is a known candidate for the production of amylases. Hence in the present study Bacillus sp, was desired for its known efficiencies in starch conversion

Heterotrophic bacteria from the continental slope sediments of arabian sea

The marine environment is indubitably the largest contiguous habitat on Earth. Because of its vast volume and area, the influence of the world ocean on global climate is profound and plays an important role in human welfare and destiny. The marine environment encompasses several habitats, from the sea surface layer down through the bulk water column, which extends >10,000 meters depth, and further down to the habitats on and under the sea floor. Compared to surface habitats, which have relatively high kinetic energy, deep-ocean circulation is very sluggish. By comparison, life in the deep sea is characterized by a relatively constant physical and chemical environment. Deep water occupying the world ocean basin is a potential natural resource based on its properties such as low temperature, high pressure and relatively unexplored properties. So, a judicious assessment of the marine resources and its management are essential to ensure sustainable development of the country’s ocean resources. Marine sediments are complex environments that are affected by both physiological and biological factors, water movements and burrowing animals. They encompass a large extent of aggregates falling from the surface waters. In aquatic ecosystems, the flux of organic matter to the bottom sediments depend on primary productivity at the ocean surface and water depth. Over 50% of the earth’s surface is covered by deep-sea sediments that are primarily formed through the continual deposition of particles from the productive pelagic waters (Vetriani et al., 1999). These aggregates are regarded as ‘hot spots’ of microbial activity in the ocean (Simon et al., 2002). This represents a good nutritional substrate for heterotrophic bacteria and favours bacterial growth

Probiotic effect of Bacillus NL110 and Vibrio NE17 on the survival, growth performance and immune response of Macrobrachium rosenbergii (de Man)

Eight hundred and eighty-¢ve strains of bacterial isolates fromvarious samples associatedwith the natural habitat ofMacrobrachiumrosenbergii were screened for their probiotic potential. Two putative probionts namely Bacillus NL110 and Vibrio NE17 isolated from the larvae and egg samples, respectively, were selected for experimental studies and were introduced to the juveniles of M. rosenbergii (0.080 0.001g) through di¡erent modes such as through feed, water and both. The probiotic potential of the above bacteria in terms of improvements inwater quality, growth, survival, speci¢c growth rate (SGR), feed conversion ratio and immune parameters was evaluated. The treatment groups showed a signi¢cant improvement in SGR and weight gain (Po0.001). Survival among di¡erent treatment groups was better than that in the control group. There were also signi¢cant improvements in the water quality parameters such as the concentration of nitrate and ammonia in the treatment groups (Po0.05). Improvements in immune parameters such as the total haemocyte count (Po0.05), phenoloxidase activity and respiratory burst were also signi¢cant (Po0.001). It is concluded that screening of the natural micro£ora of cultured ¢sh and shell¢sh for putative probionts might yield probiotic strains of bacteria that could be utilized for an environment-friendly and organic mode of aquaculture.

Bioremediation of Zinc Using Bacillus sp. Isolated from Metal- Contaminated Industrial Zone

The production of heavy metals has increased quickly since the industrial revolution. Heavy metals frequently form compounds that can be toxic, carcinogenic, or mutagenic, even in very small concentrations. The usual techniques of removing metals from wastewaters are in general expensive and have many restrictions. Alternative methods of metal removal and recovery based on biological materials have been measured. Among various agents, the use of microbes for the removal of metals from industrial and municipal wastewater has been proposed as a promising alternative to conventional heavy metal management strategies in past decades. Thus, the present study aims to isolate and characterize bacteria from soil, sediment, and waters of metal-contaminated industrial area to study the zinc resistance patterns and the zinc bioaccumulation potential of the selected microorganism. Zinc analysis of the samples revealed that concentrations varying from 39.832 m g/L to 310.24 m g/L in water, 12.81 m g/g to 407.53 m g/g in soil, and 81.06 m g/g to 829.54 m g/g in sediment are present. Bacterial zinc resistance study showed that tolerance to Zn was relatively low (<500 m g/ml). Ten bacterial genera were represented in soil and 11 from water, while only 5 bacterial genera were recorded from sediment samples. Bacillus, Pseudomonas , and Enterobacter were found in soil, sediment, and water samples. Highly zincresistant Bacillus sp. was selected for zinc removal experiment. Zinc removal studies revealed that at pH 5 about 40% reduction occurs; at pH 7, 25% occurs; and at pH 9, 50% occurs. Relatively an increased removal of Zinc was observed in the fi rst day of the experiment by Bacillus sp. The metal bioaccumulative potential of the selected isolates may have possible applications in the removal and recovery of zinc from industrial ef fluents.

Probiotic effect of Bacillus NL110 and Vibrio NE17 on the survival, growth performance and immune response of Macrobrachium rosenbergii (de Man)

Eight hundred and eighty-¢ve strains of bacterial isolates fromvarious samples associatedwith the natural habitat ofMacrobrachiumrosenbergii were screened for their probiotic potential. Two putative probionts namely Bacillus NL110 and Vibrio NE17 isolated from the larvae and egg samples, respectively, were selected for experimental studies and were introduced to the juveniles of M. rosenbergii (0.080 0.001g) through di¡erent modes such as through feed, water and both. The probiotic potential of the above bacteria in terms of improvements inwater quality, growth, survival, speci¢c growth rate (SGR), feed conversion ratio and immune parameters was evaluated. The treatment groups showed a signi¢cant improvement in SGR and weight gain (Po0.001). Survival among di¡erent treatment groups was better than that in the control group. There were also signi¢cant improvements in the water quality parameters such as the concentration of nitrate and ammonia in the treatment groups (Po0.05). Improvements in immune parameters such as the total haemocyte count (Po0.05), phenoloxidase activity and respiratory burst were also signi¢cant (Po0.001). It is concluded that screening of the natural micro£ora of cultured ¢sh and shell¢sh for putative probionts might yield probiotic strains of bacteria that could be utilized for an environment-friendly and organic mode of aquaculture

"Alkaline Lipase Production by Marine Bacillus smithii BTMS 11"

Considering the potential of marine environment present study was designed for the screening and isolation of a potential salt tolerant. alkaline and thennotolerant lipase producing bacteria from the costal belts of South India and consequent development of ideal bioprocess for industrial production, purification characterisation and evaluation of the potential of the lipase enzyme for various industrial applications 1. Screening and isolation of a potential lipase producing bacteria. 2. Optimization of various physicochemical factors in Submerged fennentation for the production of alkaline lipase 3. Purification ofthe lipase enzyme 4. Characterisation of the enzyme 5. Evaluation of the enzyme for various industrial applications

Heterotrophic Bacteria Associated with Healthy and Moribund Larvae of Penaeus monodon H.Milne Edwards

Heterotrophic bacterial flora of Pmonadon from an apparently healthy hatchery system as well as a pool with heavy mortality were isolated and studied. In the healthy systems comparatively higher generic diversity with Pseudomonas, Acinetobacter, Bacillus, Micrococcus, members of the family Enterobacteriaceae and coryneform group in the diminishing order of dominance was recorded. Meanwhile from the moribund larvae and rearing water Aeromonas and Pseudomonas could be isolated in almost equal proportions. Strikingly, Aeromonas could not be isolated from the apparently healthy larval rearing system and its exclusive occurrence in the sick culture system in comparatively higher percentage suggested its possible role in the mortality. They were found to be highly halophilic exhibiting growth at 10% NaCl. On testing their sensitivity to twenty antibiotics, four of them (Streptomycin, Gentamycin, Methamine mandelate and Cloramphenicol) were found to be effective on all the isolates of Aeromonas and Pseudomonas suggesting their possible application in the hatchery system in times of emergency. While doing so, Streptomycin would do comparatively better than the others as the minimum inhibitory dose required was comparatively lower (200ppm) within a period of 24 hours

Characterization of Bacteriocins from Bacillus licheniformis strain BTHT8 and Bacillus subtilis strain BTFK101 isolated from marine sediment

Pathogenic microorganisms such as Bacillus cereus, Listeria Monocytogenes and Staphylococcus sp have caused serious diseases, and consequently contributed to considerable economic loss in the food and agricultural industries. Antibiotics have been practically used to treat these pathogens since penicillin G was discovered more than half a century ago. Many different types of antibiotics have been discovered or synthesized to control pathogenic microorganisms. Repetitive use and misuse of antibiotics by the agricultural and pharmaceutical industries have caused the emergence of multidrug-resistant microorganisms, even to the strongest antibiotics currently available; therefore, the rapid development of more effective antimicrobial compounds is required to keep pace with demand. Bacteria were isolated from marine water and sediment samples collected from various locations off the coast of Cochin and salt pans of Tuticorin using pour plate technique. One hundred and twelve isolates were obtained. Seventeen isolates exhibiting antimicrobial activity were segregated after primary screening. The secondary screening which was aimed at selection of bacteria that produce proteinaceous inhibitory compounds, helped to select five strains viz. BTFK101, BTHT8, BTKM4, BTEK16 and BTSB22. The five isolates inhibited the growth of six Gram positive test organisms viz. B. cereus, B. circulans, B. coagulans, B. pumilus, Staphylococcus aureus and Clostridium perfringens. After quantitative estimation of the bacteriocin production, the two strains BTFK101 and BTHT8 were selected for further study.

Antibody Based Diagnostic for Detection Of Vibrios and their Biological Control using Antagonistic Bacteria in Macrobrachium Rosenbergii Larval Rearing Systems

The main objective of the work undertaken here was to develop an appropriate microbial technology to protect the larvae of M.rosenbergii in hatchery from vibriosis. This technology precisely is consisted of a rapid detection system of vibrios and effective antagonistic probiotics for the management of vibrios. The present work was undertaken with the realizations that to stabilize the production process of commercial hatcheries an appropriate, comprehensive and fool proof technology is required primarily for the rapid detection of Vibrio and subsequently for its management. Nine species of Vibrio have been found to be associated with larvae of M. rosenbergii in hatchery. Haemolytic assay of the Vibrio and Aeromonas on prawn blood agar showed that all isolates of V. alginolyticus and Aeromonas sp., from moribund, necrotized larve were haemolytic and the isolates of V.cholerae, V.splendidus II, V.proteolyticus and V.fluvialis from the larvae obtained from apparently healthy larval rearing systems were non-haemolytic. Hydrolytic enzymes such as lipase, chitinase and gelatinase were widespread amongst the Vibrio and Aeromonas isolates. Dominance of V.alginolyticus among the isolates from necrotic larvae and the failure in isolating them from rearing water strongly suggest that they infect larvae and multiply in the larval body and cause mortality in the hatchery. The observation suggested that the isolate V. alginolyticus was a pathogen to the larvae of M.rosenbergii. To sum up, through this work, nine species of Vibrio and genus Aeromonas associated with M.rosenbergii larval rearing systems could be isolated and segregated based on the haemolytic activity and the antibodies (PA bs) for use in diagnosis or epidemiological studies could be produced, based on a virulent culture of V.alginolyticus. This could possibly replace the conventional biochemical tests for identification. As prophylaxis to vibriosis, four isolates of Micrococcus spp. and an isolate of Pseudomonas sp. could be obtained which could possibly be used as antagonistic probiotics in the larval rearing system of M.rosenbergii.

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.