Production of Alkaline Protease by Free and Immobilised Cells of VIBRIO sp. Under Different Fermentation Systems and Its Application on Deproteinisation of Prawn Shell Waste for Chitin Recovery

This thesis presents a detailed account of a cost - effective approach towards enhanced production of alkaline protease at profitable levels using different fermentation designs employing cheap agro-industrial residues. It involves the optimisation of process parameters for the production of a thermostable alkaline protease by Vibrio sp. V26 under solid state, submerged and biphasic fermentations, production of the enzyme using cell immobilisation technology and the application of the crude enzyme on the deproteinisation of crustacean waste.The present investigation suggests an economic move towards Improved production of alkaline protease at gainful altitudes employing different fermentation designs utilising inexpensive agro-industrial residues. Moreover, the use of agro-industrial and other solid waste substrates for fermentation helps to provide a substitute in conserving the already dwindling global energy resources. Another alternative for accomplishing economically feasible production is by the use of immobilisation technique. This method avoids the wasteful expense of continually growing microorganisms. The high protease producing potential of the organism under study ascertains their exploitation in the utilisation and management of wastes. However, strain improvement studies for the production of high yielding variants using mutagens or by gene transfer are required before recommending them to Industries.Industries, all over the world, have made several attempts to exploit the microbial diversity of this planet. For sustainable development, it is essential to discover, develop and defend this natural prosperity. The Industrial development of any country is critically dependent on the intellectual and financial investment in this area. The need of the hour is to harness the beneficial uses of microbes for maximum utilisation of natural resources and technological yields. Owing to the multitude of applications in a variety of industrial sectors, there has always been an increasing demand for novel producers and resources of alkaline proteases as well as for innovative methods of production at a commercial altitude. This investigation forms a humble endeavour towards this perspective and bequeaths hope and inspiration for inventions to follow.

Application of Fermentation Techniques in the Utilization of Prawn Shell Waste

The present study aimed at the utlisation of microbial organisms for the production of good quality chitin and chitosan. The three strains used for the study were Lactobacillus plantarum, Lactobacililus brevis and Bacillus subtilis. These strains were selected on the basis of their acid producing ability to reduce the pH of the fermenting substrates to prevent spoilage and thus caused demineralisation of the shell. Besides, the proteolytic enzymes in these strains acted on proteinaceous covering of shrimp and thus caused deprotenisation of shrimp shell waste. Thus the two processes involved in chitin production can be affected to certain extent using bacterial fermentation of shrimp shell.Optimization parameters like fermentation period, quantity of inoculum, type of sugar, concentration of sugar etc. for fermentation with three different strains were studied. For these, parameters like pH, Total titrable acidity (TTA), changes in sugar concentration, changes in microbial count, sensory changes etc. were studied.Fermentation study with Lactobacillus plantarum was continued with 20% w/v jaggery broth for 15 days. The inoculum prepared yislded a cell concentration of approximately 108 CFU/ml. In the present study, lactic acid and dilute hydrochloric acid were used for initial pH adjustment because; without adjusting the initial pH, it took more than 5 hours for the lactic acid bacteria to convert glucose to lactic acid and during this delay spoilage occurred due to putrefying enzymes active at neutral or higher pH. During the fermentation study, pH first decreased in correspondence with increase in TTA values. This showed a clear indication of acid production by the strain. This trend continued till their proteolytic activity showed an increasing trend. When the available sugar source started depleting, proteolytic activity also decreased and pH increased. This was clearly reflected in the sensory evaluation results. Lactic acid treated samples showed greater extent of demineralization and deprotenisation at the end of fermentation study than hydrochloric acid treated samples. It can be due to the effect of strong hydrochloric acid on the initial microbial count, which directly affects the fermentation process. At the end of fermentation, about 76.5% of ash was removed in lactic acid treated samples and 71.8% in hydrochloric acid treated samples; 72.8% of proteins in lactic acid treated samples and 70.6% in hydrochloric acid treated samples.The residual protein and ash in the fermented residue were reduced to permissible limit by treatment with 0.8N HCI and 1M NaOH. Characteristics of chitin like chitin content, ash content, protein content, % of N- acetylation etc. were studied. Quality characteristics like viscosity, degree of deacetylation and molecular weight of chitosan prepared were also compared. The chitosan samples prepared from lactic acid treated showed high viscosity than HCI treated samples. But degree of deacetylation is more in HCI treated samples than lactic acid treated ones. Characteristics of protein liquor obtained like its biogenic composition, amino acid composition, total volatile base nitrogen, alpha amino nitrogen etc. also were studied to find out its suitability as animal feed supplement.Optimization of fermentation parameters for Lactobacillus brevis fermentation study was also conducted and parameters were standardized. Then detailed fermentation study was done in 20%wlv jaggery broth for 17 days. Also the effect of two different acid treatments (mild HCI and lactic acid) used for initial pH adjustment on chitin production were also studied. In this study also trend of changes in pH. changes in sugar concentration ,microbial count changes were similar to Lactobacillus plantarum studies. At the end of fermentation, residual protein in the samples were only 32.48% in HCI treated samples and 31.85% in lactic acid treated samples. The residual ash content was about 33.68% in HCI treated ones and 32.52% in lactic acid treated ones. The fermented residue was converted to chitin with good characteristics by treatment with 1.2MNaOH and 1NHCI.Characteristics of chitin samples prepared were studied and extent of Nacetylation was about 84% in HCI treated chitin and 85%in lactic acid treated ones assessed from FTIR spectrum. Chitosan was prepared from these samples by usual chemical method and its extent of solubility, degree of deacetylation, viscosity and molecular weight etc were studied. The values of viscosity and molecular weight of the samples prepared were comparatively less than the chitosan prepared by Lactobacillus plantarum fermentation. Characteristics of protein liquor obtained were analyzed to determine its quality and is suitability as animal feed supplement.Another strain used for the study was Bacillus subtilis and fermentation was carried out in 20%w/v jaggery broth for 15 days. It was found that Bacillus subtilis was more efficient than other Lactobacillus species for deprotenisation and demineralization. This was mainly due to the difference in the proteolytic nature of the strains. About 84% of protein and 72% of ash were removed at the end of fermentation. Considering the statistical significance (P

Study on Solid Waste Management : An Economic Analysis with Respect to Ernakulam District

This thesis analyzed waste generation and waster disposal problems in municipalities and Cochin Corporation in Ernakulam district.Then the potential of resource recovery and recycling from biodegradable and non bio-degradable waste is established.The study further focused on the need for segregation of waste at the source as biodegradable and non biodegradable solid waste.The potential of resource recovery is explained in detail through the case study.The thesis also highlights the economically viable and environmental friendly methods o f treatment of waste.But the problem is that concerted and earnest attempts are lacking in making use of such methods.In spite of the health problems faced,people living near the dump sites are forced to stay there either because of their weak economic background or family ties.The study did not calculate the economic cost of health problems arising out of unscientific and irresponsible methods of waste disposal.

Resource abundance and Survival of indigenous ornamental fishes of central Kerala with emphasis on handling and packing stress in Puntius filamentosus(VALENCIENNES)

This thesis Entitled Resource abundance and survival of indigenous ornamental fishes of central kerala with emphasis on handling and packing stress in puntius filamentosus (valenciennes).Kerala state is endowed with 41 west flowing and three east flowing rivers originating in the Western Ghats. These rivers and their vast network of tributaries and distributaries harbour rich and diversified fish fauna. Most of the freshwater fishes available in Kerala are highly appreciated as ornamental fishes in the national and international markets.Today the ornamental fish industry is one of the largest industries all over the world. The demand for ornamental fishes has been increasing steadily with the enlargement of the industry, such that the current demand for indigenous ornamental fishes have exceeded the supply. This has led to serious concern about the resources available in the country that can be utilised judiciously for the economic benefit of the state. With an aim to fill up the lacuna, a database of freshwater ornamental fishes of Kerala was created as part of the present study. Ornamental fishes destined for export marketing should thrive well in the aquarium conditions.The study reiterates fishes caught from different environmental conditions and feeding habits have a greater ability to adapt and acclimatise to an entirely new environment and food habits. Marketing studies based on the statistics available with Marine Products Export Development Authority show that these species are not being exported at the required level over the past 6 years, when compared to the availability in the water bodies of Kerala. Sustainable utilisation of these resources from the wild using modern management principles and code of conduct for responsible fishing are advisable until captive breeding technology is popularised.

Waste Water Treatment Using Low Cost Natural Adsorbents

Heavy metals are major toxic pollutants with severe health effects on humans. They are released into the environment from a variety of industrial activities. Cadmium, lead, zinc, chromium and copper are the most toxic metals of widespread use in industries such as tanning, electroplating, electronic equipment manufacturing and chemical processing plants. Heavy metals contribute to a variety of adverse health environmental effects due to their acute and chronic exposure through air, water and food chain. Conventional treatment methods of metal removal are often limited by their cost and ineffectiveness at low concentrations. Adsorption, the use of inactivated biomass as adsorbents offers an attractive potential alternative to their conventional methods. Mango peel and Alisma plantago aquatica are naturally occurring and abundant biomass can offer an economical solution for metal removal.The Cd(II), Pb(II), Zn(II), Cr(III) and Cu(II) adsorption by milled adsorbents of mango peel and Alisma plantago aquatica were evaluated in batches.

Routing protocol enhancement for handling node mobility in wireless sensor networks

In wireless sensor networks, the routing algorithms currently available assume that the sensor nodes are stationary. Therefore when mobility modulation is applied to the wireless sensor networks, most of the current routing algorithms suffer from performance degradation. The path breaks in mobile wireless networks are due to the movement of mobile nodes, node failure, channel fading and shadowing. It is desirable to deal with dynamic topology changes with optimal effort in terms of resource and channel utilization. As the nodes in wireless sensor medium make use of wireless broadcast to communicate, it is possible to make use of neighboring node information to recover from path failure. Cooperation among the neighboring nodes plays an important role in the context of routing among the mobile nodes. This paper proposes an enhancement to an existing protocol for accommodating node mobility through neighboring node information while keeping the utilization of resources to a minimum.

Efficacy of fermented prawn shell waste as a feed ingredient for Indian white prawn, Fenneropenaeus indicus

Prawn shell waste collected from shrimp-processing plants in Cochin, India, was subjected to fermentation using 20 chitinoclastic and proteolytic/non-proteolytic bacterial strains. The products generated were analysed for protein, lipid, total sugars, N-acetyl glucosamine, free amino acids and ash. Shrimp diets were prepared using these 20 fermented products and a control diet using raw prawn shell waste. Feeding experiment was conducted with postlarvae (PL21) of Indian white prawn, Fenneropenaeus indicus for a period of 21 days. Biogrowth parameters such as mean weight gain, feed conversion ratio, specific growth rate and protein efficiency ratio were estimated and the animals were challenged with white spot virus orally via diet. Enhanced growth could be observed in prawns fed F134 and F124, incorporated with the fermentation products generated using Bacillus spp., C134 and C124 respectively. The percentage survival of prawns after 7 days of challenge was found to be highest for groups fed diet F111 incorporated with fermentation product generated using Bacillus sp. These products of bacterial fermentation hold promise as growth enhancers and immunostimulants in aquaculture. KEY WORDS: biogrowth parameters, feed

Routing Protocol Enhancement for handling Node Mobility in Wireless Sensor Networks

In wireless sensor networks, the routing algorithms currently available assume that the sensor nodes are stationary. Therefore when mobility modulation is applied to the wireless sensor networks, most of the current routing algorithms suffer from performance degradation. The path breaks in mobile wireless networks are due to the movement of mobile nodes, node failure, channel fading and shadowing. It is desirable to deal with dynamic topology changes with optimal effort in terms of resource and channel utilization. As the nodes in wireless sensor medium make use of wireless broadcast to communicate, it is possible to make use of neighboring node information to recover from path failure. Cooperation among the neighboring nodes plays an important role in the context of routing among the mobile nodes. This paper proposes an enhancement to an existing protocol for accommodating node mobility through neighboring node information while keeping the utilization of resources to a minimum.

Handling OOV Words in Phrase-Based Statistical Machine Translation for Malayalam

Statistical Machine Translation (SMT) is one of the potential applications in the field of Natural Language Processing. The translation process in SMT is carried out by acquiring translation rules automatically from the parallel corpora. However, for many language pairs (e.g. Malayalam- English), they are available only in very limited quantities. Therefore, for these language pairs a huge portion of phrases encountered at run-time will be unknown. This paper focuses on methods for handling such out-of-vocabulary (OOV) words in Malayalam that cannot be translated to English using conventional phrase-based statistical machine translation systems. The OOV words in the source sentence are pre-processed to obtain the root word and its suffix. Different inflected forms of the OOV root are generated and a match is looked up for the word variants in the phrase translation table of the translation model. A Vocabulary filter is used to choose the best among the translations of these word variants by finding the unigram count. A match for the OOV suffix is also looked up in the phrase entries and the target translations are filtered out. Structuring of the filtered phrases is done and SMT translation model is extended by adding OOV with its new phrase translations. By the results of the manual evaluation done it is observed that amount of OOV words in the input has been reduced considerably

Routing Protocol Enhancement for handling Node Mobility in Wireless Sensor Networks

In wireless sensor networks, the routing algorithms currently available assume that the sensor nodes are stationary. Therefore when mobility modulation is applied to the wireless sensor networks, most of the current routing algorithms suffer from performance degradation. The path breaks in mobile wireless networks are due to the movement of mobile nodes, node failure, channel fading and shadowing. It is desirable to deal with dynamic topology changes with optimal effort in terms of resource and channel utilization. As the nodes in wireless sensor medium make use of wireless broadcast to communicate, it is possible to make use of neighboring node information to recover from path failure. Cooperation among the neighboring nodes plays an important role in the context of routing among the mobile nodes. This paper proposes an enhancement to an existing protocol for accommodating node mobility through neighboring node information while keeping the utilization of resources to a minimum.

Banana waste as substrate for a-amylase production by Bacillus subtilis (CBTK 106) under solid-state fermentation

Bacillus subtilis CBTK 106, isolated from banana wastes, produced high titres of a-amylase when banana fruit stalk was used as substrate in a solid-state fermentation system. The e¤ects of initial moisture content, particle size, cooking time and temperature, pH, incubation temperature, additional nutrients, inoculum size and incubation period on the production of a- amylase were characterised. A maximum yield of 5 345 000 U mg~1 min~1 was recorded when pretreated banana fruit stalk (autoclaved at 121 ¡C for 60 min) was used as substrate with 70% initial moisture content, 400 lm particle size, an initial pH of 7.0, a temperature of 35 ¡C, and additional nutrients (ammonium sulphate/sodium nitrate at 1.0%, beef extract/peptone at 0.5%, glucose/sucrose/starch/maltose at 0.1% and potassium chloride/sodium chloride at 1.0%) in the medium, with an inoculum-to-substrate ratio of 10% (v/w) for 24 h. The enzyme yield was 2.65-fold higher with banana fruit stalk medium compared to wheat bran

Utilization of prawn waste for chitinase production by the marine fungus Beauveria bassiana by solid state fermentation

Prawn waste, a chitinous solid waste of the shell®sh processing industry, was used as a substrate for chitinase production by the marine fungus Beauveria bassiana BTMF S10, in a solid state fermentation (SSF) culture. The process parameters in¯uencing SSF were optimized. A maximum chitinase yield of 248.0 units/g initial dry substrate (U/gIDS) was obtained in a medium containing a 5:1 ratio (w/v) of prawn waste/sea water, 1% (w/w) NaCl, 2.5% (w/w) KH2PO4, 425±600 lm substrate particle size at 27 °C, initial pH 9.5, and after 5 days of incubation. The presence of yeast extract reduced chitinase yield. The results indicate scope for the utilization of shell®sh processing (prawn) waste for the industrial production of chitinase by using solid state fermentation.

Solid Waste Management: A Case Study of Arppukara Grama Panchayat Of Kottayam District, Kerala (India)

The rising pressure of Population, together with the constantly changing technologies, and development perspectives, contribute to the ever increasing volumes of wastes in different forms. The solid and liquid wastes generated in the urban areas were considered a burden to the society, and hazardous to the environment. The fact is that the growth of consumerist culture and aimless throwing of refuse by the people created the outbreak of environmental pollution. Unhygienic environment and solid waste accumulation coincided with mosquito breeding which causes, the spread of most epidemics. The rationale behind most of the diseases is the unhygienic pattern followed by the people both in rural and urban areas. As an environmental package, the disposal of solid waste from different sources, such as house holds markets, commercial areas, slaughter houses, hospitals and industries, therefore assumed crucial importance. So as a part of the theory and practice, a study on the area, solid waste management of Arppukara Grama Panchayat of Kottayam district is taken into consideration. The study conducted here proposes, to examine the quality and quantity of the solid waste generated in the panchayat and also it's impact on the existing social, economical, environmental and ecological systems

Waste plastic as a stabilizing additive in Stone Mastic Asphalt

The present study investigates the benefits of stabilizing the stone mastic asphalt (SMA) mixture in flexible pavement with shredded waste plastic. Conventional (without plastic) and the stabilized SMA mixtures were subjected to performance tests including Marshall Stability, tensile strength and compressive strength tests. Triaxial tests were also conducted with varying percentage bitumen by weight of mineral aggregate (6% to 8%) and by varying percentage plastic by weight of mix (6% to 12% with an increment of 1%). Plastic content of 10% by weight of bitumen is recommended for the improvement of the performance of Stone Mastic Asphalt mixtures. 10% plastic content gives an increase in the stability, split tensile strength and compressive strength of about 64%, 18% and 75% respectively compared to the conventional SMA mix. Triaxial test results show a 44% increase in cohesion and 3% decrease in angle of shearing resistance showing an increase in the shear strength. The drain down value decreases with an increase in plastic content and the value is only 0.09 % at 10% plastic content and proves to be an effective stabilizing additive in SMA mixtures

Selection of Marine Yeasts for the Generation of Single Cell Protein from Prawn-shell Waste

Marine yeasts (33 strains) were isolated from the coastal and offshore waters off Cochin. The isolates were identified and then characterized for the utilization of starch, gelatin, lipid, cellulose, urea, pectin, lignin, chitin and prawn-shell waste. Most of the isolates were Candida species. Based on the biochemical characterization, four potential strains were selected and their optimum pH and NaCI concentration for growth were determined. These strains were then inoculated into prawn-shell waste and SCP (single cell protein) generation was noted in terms of the increase in protein content of the final product.