STANDARDIZATION OF PROCESS PARAMETERS FOR READY TO EAT FISH PRODUCTS IN INDIGENOUS POLYMER COATED TIN FREE STEEL CANS

In the present work, Indigenous polymer coated Tin Free Steel cans were analyzed fortheir suitability for thermal processing and storage of fish and fish products following standard methods. The raw materials used for the development of ready to eat thermally processed fish products were found to be of fresh condition. The values for various biochemical and microbiological parameters of the raw materials were well within the limits. Based on the analysis of commercial sterility, instrumental colour, texture, WB-shear force and sensory parameters, squid masala processed to F0 value of 8 min with a total process time of 38.5 min and cook value of 92 min was chosen as the optimum for squid masala in tin free steel cans while shrimp curry processed to F0 7 min with total process time of 44.0 min and cook value of 91.1 min was found to be ideal and was selected for storage study. Squid masala and shrimp curry thermally processed in indigenous polymer coated TFS cans were found to be acceptable even after one year of storage at room temperaturebased on the analysis of various sensory and biochemical parameters. Analysis of the Commission Internationale d’ Eclirage L*, a* and b* color values showed that the duration of exposure to heat treatment influenced the color parameters: the lightness (L*) and yellowness (b*)decreased, and the redness (a*) significantly increased with the increase in processing time or reduction in processing temperature.Instrumental analysis of texture showed that hardness-1 & 2 decreased with reduction in retort temperature while cohesiveness value did not show any appreciable change with decrease in temperature of processing. Other texture profile parameters like gumminess, springiness and chewiness decreased significantly with increase of processing time. W-B shear force values of mackerel meat processed at 130 °C were significantly higher than those processed at 121.1 and 115 °C. HTST processing of mackerel in brine helped in reducing the process time and improving the quality.The study also indicated that indigenous polymer coated TFS cans with easy openends can be a viable alternative to the conventional tin and aluminium cans. The industry can utilize these cans for processing ready to eat fish and shell fish products for both domestic and export markets. This will help in reviving the canning industry in India.

Biochemical effects of the pesticide Chlorpyrifos on the fish Oreochromis mossambicus (Peters)

Man uses a variety of synthetic material for his comfortable materialistic life. Thus human interactions may become harmful for various terrestrial and aquatic lives. This is by contaminating their habitat and by becoming a threat to organisms itself. Thus the application and dispersal of several organic pollutants can lead to the development of several mutated forms of the species when exposed to sublethal concentrations of the pollutants. Otherwise, a decrease in number or extinction of these exposed species from earth's face may happen. Pesticides, we use for the benefit of crop yield, but its persistence may become havoc to non-target organism. Pesticides reaching a reservoir can subsequently enter the higher trophic levels. Organophosphorus compounds have replaced all other pesticides, due to its acute toxicity and non-persistent nature.Hence the present study has concentrated on the toxicity of the largest market-selling and multipurpose pesticide, chlorpyrifos on the commonly edible aquatic organism, fish. The euryhaline cichlid Oreochromis mossambicus was selected as animal model. The study has concentrated on investigating biochemical parameters like tissue-specific enzymes, antioxidant and lipid-peroxidation parameters, haematological and histological observations and pesticide residue analysis.Major findings of this work have indicated the possibility of aquatic toxicity to the fish on exposure to the insecticide chlorpyrifos. The insecticide was found as effective to induce structural alteration, depletion in protein content, decrease in different metabolic enzyme levels and to progress lipid peroxidation on a prolonged exposure of 21 days. The ion-transport mechanism was found to be adversely affected. Electrophoretic analysis revealed the disappearance of several protein bands after 21days of exposure to chlorpyrifos. Residue, analysis by gas chromatography explored the levels of chlorpyrifos retaining on the edible tissue portions during exposure period of 21days and also on a recovery period of 10 days.

Bioactivity Profile of Polyunsaturated Fattyacid extracts from Sardinella longiceps and Sardinella fimbriata – A comparative study

The oceans have proved to be an interminable source of new and effective drugs. Innumerable studies have proved that specific compounds isolated from marine organisms have great nutritional and pharmaceutical value. Polyunsaturated fattyacids (PUFA) in general are known for their dietary benefits in preventing and curing several critical ailments including Coronary heart disease (CHD) and cancers of various kinds. Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) are two PUFA which are entirely marine in origin – and small Clupeoid fishes like sardines are known to be excellent sources of these two compounds. In this study, we selected two widely available Sardine species in the west coast, Sardinella longiceps and Sardinella fimbriata, for a comparative analysis of their bioactive properties. Both these sardines are known to be rich in EPA and DHA, however considerable seasonal variation in its PUFA content was expected and these variations studied. An extraction procedure to isolate PUFA at high purity levels was identified and the extracts obtained thus were studied for anti-bacterial, anti-diabetic and anti-cancerous properties.Samples of both the sardines were collected from landing centre, measured and their gut content analysed in four different months of the year – viz. June, September, December and March. The fish samples were analyzed for fattyacid using FAME method using gas chromatography to identify the full range of fattyacids and their respective concentration in each of the samples. The fattyacids were expressed in mg/g meat and later converted to percentage values against total fatty acids and total PUFA content. Fattyacids during winter season (Dec-Mar) were found to be generally higher than spawning season (June-Sept). PUFA dominated the profiles of both species and average PUFA content was also higher during winter. However, it was found that S. longiceps had proportionately higher EPA as compared to S. fimbriata which was DHA rich. Percentage of EPA and DHA also varied across months for both species – the spawning season seemed to show higher EPA content in S. longiceps and higher DHA content in S. fimbriata. Gut content analysis indicate that adult S. fimbriata is partial to zooplanktons which are DHA rich while adult S. longiceps feed mainly on EPA rich phytoplankton. Juveniles of both species, found mainly in winter, had a gut content showing more mixed diet. This difference in the feeding pattern reflect clearly in their PUFA profile – adult S. longiceps, which dominate the catch during the spawn season, feeding mostly on phytoplankton is concentrated with EPA while the juveniles which are found mostly in the winter season has slightly less EPA proportion as compared to adults. The same is true for S. fimbriata adults that are caught mostly in the spawning season; being rich in DHA as they feed mainly on zooplankton while the juveniles caught during winter season has a relatively lower concentration of DHA in their total PUFA.Various extraction procedures are known to obtain PUFA from fish oil. However, most of them do not give high purity and do not use materials indicated as safe. PUFA extracts have to be edible and should not have harmful substances for applying on mice and human subjects. Some PUFA extraction procedures, though pure and non-toxic, might induce cis-trans conversions during the extraction process. This conversion destroys the benefits of PUFA and at times is harmful to human body. A method free from these limitations has been standardized for this study. Gas Chromatography was performed on the extracts thus made to ensure that it is substantially pure. EPA: DHA ratios for both samples were derived - for S. longiceps this ratio was 3:2, while it was 3:8 for S. fimbriata.Eight common strains of gram positive and gram negative bacterial strains were subjected to the PUFA extracts from both species dissolved in acetone solution using Agar Well Diffusion method. The activity was studied against an acetone control. At the end of incubation period, zones of inhibition were measured to estimate the activity. Minimum inhibitory concentration for each of the active combinations was calculated by keeping p < 0.01 as significant. Four of the bacteria including multi-resistant Staphylococcus aureus were shown to be inhibited by the fish extracts. It was also found that the extracts from S. fimbriata were better than the one from S. longiceps in annihilating harmful bacteria.Four groups of mice subjects were studied to evaluate the antidiabetic properties of the PUFA extracts. Three groups were induced diabetes by administration of alloxan tetra hydrate. One group without diabetes was kept as control and another with diabetes was kept as diabetic control. For two diabetic groups, a prescribed amount of fish extracts were fed from each of the extracts. The biochemical parameters like serum glucose, total cholesterol, LDL & HDL cholesterol, triglycerides, urea and creatinine were sampled from all four groups at regular intervals of 7 days for a period of 28 days. It was found that groups fed with fish extracts had marked improvement in the levels of total LDL & HDL cholesterol, triglycerides and creatinine. Groups fed with extracts from S. fimbriata seem to have fared better as compared to S. longiceps. However, both groups did not show any marked improvement in blood glucose levels or levels of urea.Cell lines of MCF-7 (Breast Cancer) and DU-145 (Prostate Cancer) were used to analyse the cytotoxicity of the PUFA extracts. Both cell lines were subjected to MTT Assay and later the plates were read using an ELISA reader at a wavelength of 570nm. It was found that both extracts had significant cytotoxic effects against both cell lines and a peak cytotoxicity of 85-90% was apparent. IC50 values were calculated from the graphs and it was found that S. longiceps extracts had a slightly lower IC50 value indicating that it is toxic even at a lower concentration as compared to extracts from S. fimbriata.This study summarizes the bioactivity profile of PUFA extracts and provides recommendation for dietary intake; fish based nutritional industry and indigenous pharmaceutical industry. Possible future directions of this study are also elaborated.

Studies on some aspects of the reproductive physiology of the female grey mullet mugil cephalus l.

Aquaculture is the dynamic pursuit of production of organisms from water a process analogous to agriculture on land. The field of aquaculture is an emerging bioindustry, based upon the culture and husbandry of economically utilizable aquatic organisms. Of late, there has been a global upsurge for aquaculture, the main reasons for which include the requirement of protein source for the increasing world population, the decision by various world nations to increase the fish yield by developing unutilized or partially utilized water bodies and depletion of natural stock which is evident in recent years due to excessive exploitation .The present study has been taken up on the reproductive physiology of the female grey mullet, M. cephalus. The thesis is presented in seven chapters. In the present study, variations in the major biochemical parameters namely, moisture, proteins, lipids, carbohydrates cholesterol, carotenoid, ash, calcium and iron in four tissues E. muscle, liver, ovary and bloodserum of cephalus have been analysed at different maturity stages.

Investigation on key molecular targets responsible for antidiabetic properties of Symplocos cochinchinensis (Lour.) S. Moore

Diabetes mellitus is a heterogeneous metabolic disorder characterized by hyperglycemia with disturbances in carbohydrate, protein and lipid metabolism resulting from defects in insulin secretion, insulin action or both. Currently there are 387 million people with diabetes worldwide and is expected to affect 592 million people by 2035. Insulin resistance in peripheral tissues and pancreatic beta cell dysfunction are the major challenges in the pathophysiology of diabetes. Diabetic secondary complications (like liver cirrhosis, retinopathy, microvascular and macrovascular complications) arise from persistent hyperglycemia and dyslipidemia can be disabling or even life threatening. Current medications are effective for control and management of hyperglycemia but undesirable effects, inefficiency against secondary complications and high cost are still serious issues in the present prognosis of this disorder. Hence the search for more effective and safer therapeutic agents of natural origin has been found to be highly demanding and attract attention in the present drug discovery research. The data available from Ayurveda on various medicinal plants for treatment of diabetes can efficiently yield potential new lead as antidiabetic agents. For wider acceptability and popularity of herbal remedies available in Ayurveda scientific validation by the elucidation of mechanism of action is very much essential. Modern biological techniques are available now to elucidate the biochemical basis of the effectiveness of these medicinal plants. Keeping this idea the research programme under this thesis has been planned to evaluate the molecular mechanism responsible for the antidiabetic property of Symplocos cochinchinensis, the main ingredient of Nishakathakadi Kashayam, a wellknown Ayurvedic antidiabetic preparation. A general introduction of diabetes, its pathophysiology, secondary complications and current treatment options, innovative solutions based on phytomedicine etc has been described in Chapter 1. The effect of Symplocos cochinchinensis (SC), on various in vitro biochemical targets relevant to diabetes is depicted in Chapter 2 including the preparation of plant extract. Since diabetes is a multifactorial disease, ethanolic extract of the bark of SC (SCE) and its fractions (hexane, dichloromethane, ethyl acetate and 90 % ethanol) were evaluated by in vitro methods against multiple targets such as control of postprandial hyperglycemia, insulin resistance, oxidative stress, pancreatic beta cell proliferation, inhibition of protein glycation, protein tyrosine phosphatase-1B (PTP-1B) and dipeptidyl peptidase-IV (DPPxxi IV). Among the extracts, SCE exhibited comparatively better activity like alpha glucosidase inhibition, insulin dependent glucose uptake (3 fold increase) in L6 myotubes, pancreatic beta cell regeneration in RIN-m5F and reduced triglyceride accumulation in 3T3-L1 cells, protection from hyperglycemia induced generation of reactive oxygen species in HepG2 cells with moderate antiglycation and PTP-1B inhibition. Chemical characterization by HPLC revealed the superiority of SCE over other extracts due to presence of bioactives (beta-sitosterol, phloretin 2’glucoside, oleanolic acid) in addition to minerals like magnesium, calcium, potassium, sodium, zinc and manganese. So SCE has been subjected to oral sucrose tolerance test (OGTT) to evaluate its antihyperglycemic property in mild diabetic and diabetic animal models. SCE showed significant antihyperglycemic activity in in vivo diabetic models. Chapter 3 highlights the beneficial effects of hydroethanol extract of Symplocos cochinchinensis (SCE) against hyperglycemia associated secondary complications in streptozotocin (60 mg/kg body weight) induced diabetic rat model. Proper sanction had been obtained for all the animal experiments from CSIR-CDRI institutional animal ethics committee. The experimental groups consist of normal control (NC), N + SCE 500 mg/kg bwd, diabetic control (DC), D + metformin 100 mg/kg bwd, D + SCE 250 and D + SCE 500. SCEs and metformin were administered daily for 21 days and sacrificed on day 22. Oral glucose tolerance test, plasma insulin, % HbA1c, urea, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total protein etc. were analysed. Aldose reductase (AR) activity in the eye lens was also checked. On day 21, DC rats showed significantly abnormal glucose response, HOMA-IR, % HbA1c, decreased activity of antioxidant enzymes and GSH, elevated AR activity, hepatic and renal oxidative stress markers compared to NC. DC rats also exhibited increased level of plasma urea and creatinine. Treatment with SCE protected from the deleterious alterations of biochemical parameters in a dose dependent manner including histopathological alterations in pancreas. SCE 500 exhibited significant glucose lowering effect and decreased HOMA-IR, % HbA1c, lens AR activity, and hepatic, renal oxidative stress and function markers compared to DC group. Considerable amount of liver and muscle glycogen was replenished by SCE treatment in diabetic animals. Although metformin showed better effect, the activity of SCE was very much comparable with this drug. xxii The possible molecular mechanism behind the protective property of S. cochinchinensis against the insulin resistance in peripheral tissue as well as dyslipidemia in in vivo high fructose saturated fat diet model is described in Chapter 4. Initially animal were fed a high fructose saturated fat (HFS) diet for a period of 8 weeks to develop insulin resistance and dyslipidemia. The normal diet control (ND), ND + SCE 500 mg/kg bwd, high fructose saturated fat diet control (HFS), HFS + metformin 100 mg/kg bwd, HFS + SCE 250 and HFS + SCE 500 were the experimental groups. SCEs and metformin were administered daily for the next 3 weeks and sacrificed at the end of 11th week. At the end of week 11, HFS rats showed significantly abnormal glucose and insulin tolerance, HOMA-IR, % HbA1c, adiponectin, lipid profile, liver glycolytic and gluconeogenic enzyme activities, liver and muscle triglyceride accumulation compared to ND. HFS rats also exhibited increased level of plasma inflammatory cytokines, upregulated mRNA level of gluconeogenic and lipogenic genes in liver. HFS exhibited the increased expression of GLUT-2 in liver and decreased expression of GLUT-4 in muscle and adipose. SCE treatment also preserved the architecture of pancreas, liver, and kidney tissues. Treatment with SCE reversed the alterations of biochemical parameters, improved insulin sensitivity by modifying gene expression in liver, muscle and adipose tissues. Overall results suggest that SC mediates the antidiabetic activity mainly via alpha glucosidase inhibition, improved insulin sensitivity, with antiglycation and antioxidant activities.

Studies on Modified Atmosphere Packaging of Pearl Spot (Etroplussuratensis) With Special Emphasis to Biochemical and Microbiological Parameters

A modified atmosphere may be defined as a packaging or storage of a perishable product in an atmosphere other than that of air. A modified atmosphere (MA) applies to food packaged products changes continuously throughout the storage period. The pearl spot (Etroplus suratensis) is an important brackish water fish belonging to the family Cichlidae. The present work was carried out to see the effect of modified atmosphere packaging on the shelf life fresh pearl spot stored in ice to extent the shelf life. The objectives of the present study are to study the suitability of Thermoformed Trays for modified atmosphere packaging, to standardize the most suitable gas mixture for modified atmosphere packaging pearl spot based on sensory evaluation, to find out the effect of modified atmosphere packaging in comparison to air packaging, to study the biochemical, microbiological, sensory and textural characteristics during storage, to study the safety concern regarding the Clostridium botulinum during modified atmosphere packaging, to find out the most suitable chemical quality indices for modified atmosphere stored pearl spot

Ecological and Biochemical Studies On Cyanobacteria of Cochin Estuary and Their Application as Source of Antioxidants

The main objectives of the present investigation were to evaluate the qualitative and quantitative distribution of natural cyanobacterial population and their ecobiological properties along the Cochin estuary and their application in aquaculture systems as a nutritional supplement due to their nutrient-rich biochemical composition and antioxidant potential. This thesis presents a detailed account of the distribution of cyanobacteria in Cochin estuary, an assessment of physico-chemical parameters and the nutrients of the study site, an evaluation of the effect of physico-chemical parameters on cyanobacterial distribution and abundance, isolation, identification and culturing of cyanobacteria, the biochemical composition an productivity of cyanobacteria, and an evaluation of the potential of the selected cyanobacteria as antioxidants against ethanol induced lipid peroxidation. The pH, salinity and nutritional requirements were optimized for low-cost production of the selected cyanobacterial strains. The present study provides an insight into the distribution, abundance, diversity and ecology of cyanobacteria of Cochin estuary. From the results, it is evident that the ecological conditions of Cochin estuary support a rich cyanobacterial growth.

Biochemical effects of ethanol on oreochromis mossambicus (Peters)

The present study dealt with the haematological, biochemical and istopathological impacts of different sub lethal concentrations of ethanol on a euryhaline teleost Oreochromis In05.s‘ambicu.5' (Peters).Studies carried out using GC indicated an increase in blood ethanol oncentration of the fish which mainly arose due to fishes entering into a state of hypoxia which explains ethanol production as an ubiquitous “anaerobic” end product, which gets accumulated whenever metabolic demand exceeds the mitochondrial oxidative potential. The very low amount of ethanol detected in the control group ofO mossambicns was mainly due to the activity of microorganisms in the gut ofO. Nzossambicus.Oedcma observed in the present study, is a defense mechanism that reduces the branchial superficial area of the fish which comes in contact with the external milieu. These mechanisms also increase the diffusion barrier to the pollutant. Dilation of the blood vessels is due to increased permeability helping in the free passage of ethanol into the blood stream. Telangiectasis observed explains the state of asphyxia of the fish when subjected to ethanol toxicity indicating acute respiratory distress. Gill aneurysm observed indicates impaired respiratory function. This is related to the rupture of the pillar cells which results in an increased blood flow inside the lamellae, causing dilation of the blood vessel or even aneurysm of gill.The present findings warrant future studies to explore A'T'Pases as possible biomarkers of pollutant exposure in ecotoxicology. This study indicated that O. mossambicus when exposed for 7 and 21 days to ethanol was under tremendous stress and parameters employed in this study can be adapted for future investigations as biomarkers of damage caused by ethanol to aquatic organisms. The present study revealed that O. mossambicus is sensitive to sub lethal concentrations of ethanol.

Biochemical Effects of Petroleum Hydrocarbons on the Tropical Teleost Oreochromis mossambicus (Peters)

The present work is a base—|ine attempt to investigate and assess the toxicity of water-accommodated fractions (WAF) of Bombay High crude oil. The experimental animal selected for the present study is a euryhaline teleost, Oreochromis mossambicus (Peters), adapted to fresh water. The fish has been selected on account of its economic valve, abundant availability, experimental feasibility, ease of rearing and maintenance and also because it is one of the commonly cultured species in the South-East Asian countries.

“Effects of monospecific and mixed algal diets on survival, development and biochemical composition of Penaeus monodon larvae”

The main objective of the present work is to acquire information regarding the growth responses of P. monodon larvae (from PZ1 upto PL1) to various mono specific and mixed diets. Evaluate the nutritional quality of selected species of micro algae viz. Chaetoceros calcitrans, Dunaliella salina, Isochrysis galbana and Nannochloropsis salina, larvae at three cell concentrations 10x104 cells/ml, 25x104 cells/ml and 50x104 cells/ml. The P. monodon larvae were transported, at the Nauplius stage, to the laboratory. The larvae were stocked at density of 150 larvae per litre in 5 litre FRP tanks with 3 litres of sea water. The algal cell density given to the larvae varied. The larval stages were fed with increasing densities of algae to evaluate the relationship between the food densities, ingestion rates, development and growth of the larvae. The water quality parameters, the percentage of survival rate, the growth estimation and the algal cell count were done. Each experiment was carried out in triplicate with a control group of larvae fed with Chaetoceros calcitrans. For the estimation standard procedures were used.to P. monodon