Studies on vibrio spp. in juveniles of penaeus indicus in culture systems

The present study is carried out to understand (i) the incidence and occurrence of species of Vibrio in different culture systems in and around Cochin, (ii) characteristics of vibrios isolates, their ecology including growth response to various hydrological parameters, sensitivity to about 40 antibiotics, and (iii) role of physico-chemical parameters in pathogenicity of vibrios, etc. and the results emerged from the investigations are important and encouraging for better understanding of the 'vibriosis' in the culture systems to develop remedial measures to control diseases. The Thesis begins with an “Introduction” followed by “A review of literature” on diseases of penaeid shrimps with particular reference to 'vibriosis' and “Material and methods” which details with the methods and procedures followed in the experiments and analyses of data. This Thesis consists of three chapters. Chapter Ideals with the incidence and ecology of Vibrio spp. in water, sediment and in juveniles of the Indian white prawn Penaeus indicus in the culture systems. In Chapter 2, characteristics of vibrio isolates including growth response to various levels oftemperature, salinity and pl 1, sensitivity to 40 antibiotics and minimal inhibitory concentration tests are detailed.e out-breaks. The Chapter 3 discusses the role of physico-chemical parameters in the incidence, seasonal abundance of Vibrio spp. and in 'vibriosis'. A summary of the whole work and list of references are also included at the end. This study gives a detailed information regarding the incidence and ecology of vibrios in the culture systems. their characteristics and pathogenicity

Studies on lipid nutrition in larvae and juveniles of the indian white prawn penaeus indicus h. milne edwards

Recent studies with prawns indicate that their growth, metamorphosis, maturation and moulting are affected by the typo and level of lipids supplied in the diets. Despite the recognition of the importance of lipids in the diets of prawns there is no information on the essentiality and quantitative lipid requirements of Indian penaeid prawns. Therefore during the present study about 24 laboratory experiments were conducted to determine the essentiality and dietary requirements of total lipids, phoapholipida, fatty acids cholestrol, and to ascertain the nutritional value of natural lipid sources for the larvae, post-larvae tad juveniles of one of the most suitable cultivable species of panaeid prans. All the experiments were conducted in the laboratory following standard procedures, using isonitrogen and approaximately isocaloric purified diets. Changes were made in tha ingridients as required for specific requirements. For the larvae diets of particle size < 37) were fed, For the postlarvae and juveniles pellet feed was given. while data on survival and growth of larvae and pout-larvae 1-10 were recorded, data were collected on the survival, growth, food conversion ratio, protein efficiency ratio and biochemical composition at the body for post-larva 11-25 and juveniles. The influence ot fatty acid pattern of dietary lipid sources on the fatty acids profile of prawns were also studied in the case of juvenile prawns. Analysis of variance andleast significant differences test were employed to determine the significant differences between treatments in the observed parameters with the help of a newlett Packard master computer.

Culture and Nutritional Enrichment of the Rotifer Brachionus Rotundiformis ( Tschugunoff ) for the Rearing of Marine Fin Fish and Shrimp Larvae

The present study is an attempt to standardize the environmental condition like pH, salinity and photoperiod, and also the feed for the maximum production of rotifers. Considering the deficiency of essential fatty acids in rotifers, enrichment experiments were carried out and fatty acids profile were analysed. Attempts were made to improve the production of clown fish (Amphiprion sebae) juveniles using enriched rotifers. Attempts were also made to rear various larval stages of Penaeus monodon with enriched rotifers as a substitute for Artemia nauplii.

Studies on the Deep - Water Crab Charybdis (Goniohellenus ) Smithii Macleay From the Seas Around India

The present study deals with a general introduction which outlines the objective of the study providing an exhaustive review of works on crabs with particular reference to deep-sea forms. In the first section, Taxonomy and Geographical disribution of the crab are dealt with. The species is described in detail based on several male and female specimens obtained from the pelagic and bottom collections, and its identity in Indian waters is established. It is also distinguished from a closely allied species so far not reported from Indian waters. The second section comprises the biology of the species and it is dealt with under four subheading, namely Habit and Habitats, Reproduction, Food and feeding and Proximate composition. The different habitats occupied by juveniles, subadults and adults of the species have been described and discussed in the light of available information on differential distribution of other related species. The reproductive biology is described in various details touching on gross anatomy and histology of the reproductive systems, spermatogenesis, oogenesis, size at maturity, ovarian maturation process, fecundity, egg carriage and breeding. The food and feeding habits of the species have been studied with reference to the different life stages such as juveniles, subadults and adults during the different phases of life based on stomach content analysis. The percentage of meat recovery and protein, carbohydrate and lipid content of meat have been described in the section dealing with proximate composition. In section three the distribution and abundance of the crab for the entire Indian EEZ and some contiguous ares have been described and illustrated in detail separately for pelagic and benthic realms. The size frequency disrtibution, sex ratios, length weight relationship and relative abundance of breeding population in the experimental catches have been dealt with in detail and discussed.

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.

Systematics, Life history traits, abundance andstock assessment of cobia, Rachycentron canadum ( Linnaeus, 1766) occurring in Indian waters with special reference to the north west coast in India

This thesis entitled Systematics,life history traits ,abundance and stock assessment of cobia rachycentron canadum (linnaeus ,1766) occurring in indian waters with special reference to the northwest coast of india.Cobia, Rachycentron canadum is a fast growing pelagic fish belonging to the monotypic family Rachycentridae. They show worldwide distribution in tropical and sub tropical waters. Cobia is exploited commercially in various countries like Taiwan, Pakistan, India, United State of America, Australia, Gulf of Mexico and the Caribbean. Recreational fishery of Cobia exists in different parts of the world. In India Cobia is caught as bycatch of trawlers, gillnet and hook and line fishery.This study also focuses on to bring out the distribution pattern and also to assess the biomass and estimate sustainable yield of Cobia inhabiting in Indian EEZ. In addition to above, present study standardised live collection methods and also reviewed culture prospects.Results of osteological study and morphological studies indicate its close resemblance to Remora and support the view that Cobia was evolved from Dolphin fishes and remoras followed it. Study also confirmed that Cobia does not have any relative or similar species and is a monotypic species belonging to the family Rachycentridae. Re description of the species was done based on the characters identified. In this study, feeding intensity was also assessed following methods like Gastrosomatic index, Mean index of feeding intensity and Index of fullness. Sex wise, month wise and length group wise fluctuations in the feeding intensity also were studied. Cobia actively fed during post monsoon period. In general, adult fed actively than the juveniles. Trophic level value estimated (4.36) indicates that Cobia occupy top level position in the food chain.

Studies on the bacteria associated with Penaeus indicus in a culture system

Ocean persists as a rich and renewable source of cheap protein for the whole world. Among the prawns/shrimps landed from the Indian Ocean and her backwaters, more than 90% are exported to affluent countries. The Indian white prawn Penaeus indicus, constitutes the major portion of the frozen shrimps exported from India every year. The present study is aimed at gathering information on the total heterotrophic bacteria (THB) associated with B. indicus, with special reference to eggs, nauplii, zoeae, mysis, and post larvae in hatchery, and juveniles and adults in culture pond. Simultaneously, IHB associated with E. indicus in its natural habitat also is studied for comparison. It is envisaged that this information will be highly useful for modifying the existing hatchery and pond management-practices.

Nutritional studies in juvenile penaeus indicus with fererence to protein and vitamin requirements

Man in his quest to explore food has turned towards the oceans and during the last five decades there has been phenomenal increase in the exploitation of the marine fisheries resources. The present study is carried out to determine the requirement of protein and water-soluble vitamins in the diet of juveniles of the Indian white prawn p.indicus using purified diets and to evaluate the nutritive value of a few plant and animal protein sources for the same species. A total of ten statistically designed experiments are conducted in the laboratory undermost identical conditions and following similar methodologies. The study also tries to suggest the essential and optimal requirements for protein as well as water-soluble vitamins in the diet of juvenile p.indicus

"|mpact of trawling on sea bottom and its living communities along the inshore waters of southwest coast of India'’

Mechanized fishing started in Indian waters in mid —fifties and large-scale operation of trawl fishing began in the mid sixties by the surfeit of individual entrepreneurs. The southwest coast of India especially the coastal waters of Kerala are the most productive area in the subcontinent and the state has been in the forefront in marine fish production (Kurup, 2001a). Though the coastline of Kerala is one tenth of the coastline of India, the state occupies the foremost position in the marine fish production of the country, accounting for more than 30% of the marine fish landings (Thomas, 2000). The coastal waters of Kerala have rich and diversified fishery resources, which are prone to heavy exploitation by a unprecedently high number of fishing gears, among them, mechanized bottom trawlers with a numerical strength of 4550 (Kurup, 2001a) against the permissible number of 1145 (Kalawar, et al., 1985) are the most destructive. Trawling operations during monsoon periods in Kerala has been a subject of controversy between traditional fishermen and trawl fishers on a subject that trawl fishing destroys large amount of juveniles and young ones of fishes since this period is the major breeding season of most of the fish and prawns (John, 1996). Therefore Government of Kerala imposed a ban on bottom trawling activities from 1988 onwards for a period varying from 21-70 days, which usually commences from June 15th. Though many studies revealed that large amount of non-target groups were destroyed in the commercial trawl fishing in the Indian waters, no concerted study has been conducted so far to evaluate the real impact of bottom trawling on the sea bottom and its living communities. The present study was conducted to assess the impact of excessive bottom trawling exerted on the sea bottom habitat and its living communities, which would be useful in impressing up on the seriousness of habitat degradation and biotic devastation, enabling the concerned to adopt relevant conservation and management steps to conserve the resources for sustainable exploitation

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.

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

Probiotic Effects of Lactic Acid Bacteria Against Vibrio Alginolyticus in Penaeus (Fenneropenaeus) Indicus (H. Milne Edwards)

Cell free extracts of four strains of Lactic acid bacteria (LAB) viz. Lactobacillus. acidophilus, Streptococcus.cremoris, Lactobacillus bulgaricus –56 and Lactobacillus bulgaricus –57 inhibited growth of Vibrio alginolyticus in nutrient broth. The antagonism of LAB to Vibrio alginolyticus was further confirmed by streak plating wherein suppression of growth of Vibrio was obtained. Juveniles of Penaeus indicus (average weight 0.985 ± 0.1 g) on administering orally a moist feed base containing 5 × 106 cells·g of the four LAB probionts for a period of four weeks showed better survival (56 to 72%) when challenged with V. alginolyticus by intra-muscular injection of 0.1 ml containing 3 × 109 cells·ml. Animals maintained on a diet devoid of bacterial biomass exhibited 80% mortality. No external or internal pathological changes were observed in shrimp fed with the LAB incorporated diets. Results showed inhibition of V. alginolyticus by LAB and stimulation of the non-specific immune response resulting in resistance to disease in the shrimp fed on LAB incorporated diets.

Application of bacterins and yeast Acremonium dyosporii to protect the larvae of Macrobrachium rosenbergii from vibriosis

The fresh water prawn, Macrobrachium rosenbergii, has proven potential for use as an aquaculture species (Hanson & Goodwin, 1997; Kurup, 1984). In India alone, culture of this species of prawn in low saline areas requires about 200 million seed per year (Kurup, 1984). In hatcheries poor survival rate has been associated with vibriosis at di#erent stages of the larval cycle. Members of the family Vibrionaceae associated with the larvae of M. rosenbergii were shown to be pathogenic under laboratory conditions (Bhat et al., 2000, in press). Vibrios have been associated with mortality of penaeid prawns by several workers (Aquacop, 1977; Hameed, 1993; Karunasagar et al., 1994). Two methods have been suggested to protect both the larvae and juveniles from vibriosis; one is the administration of bacterins prepared from pathogenic strains (Itami et al., 1989, 1991; Adams, 1991; Song & Sung, 1990; Sung et al., 1991) and the other is the utilization of yeast 1-3 and 1-6 glucans as immunostimulants for enhancing the non-specific defense system (Sung et al., 1994; Song et al., 1997). In the light of these observations it was hypothesised that bacterins and yeast glucans may also be e#ective in protecting the larvae of M. rosenbergii from vibriosis as has been achieved in the case of penaeids. To examine this hypothesis, the ability of bacterins and an extracellular glucan-producing yeast to increase the overall survival and metamorphosis of larvae in a hatchery, as well as to protect against an experimental challenge under laboratory conditions, was evaluated

Habitat ecology and food and feeding of the herring bow crab Varuna litterata (Fabricius, 1798) of Cochin backwaters, Kerala, India

Habitat ecology and food and feeding of the herring bow crab, Varuna litterata of Cochin Backwaters, Kerala, India were investigated for a period of one year (April 2011-March 2012). Among the 15 stations surveyed, the crabs were found to occur only in 4 stations, which had a close proximity to the sea. Sediment analysis of the stations revealed that the substratum of these stations is sandy in nature and is rich in organic carbon content (0.79% to 1.07%). These estuarine crabs is euryhaline and are found to be distributed in areas with a sandy substratum, higher organic carbon content and more tidal influx. The stomach contents analysis of crabs examined showed that their diet included crustacean remains, plants, sand and debris, fishes, miscellaneous group and unidentified matter. In adults and sub-adults, crustaceans formed the dominant food group, while in juveniles, sand and debris formed the dominant group. From the present study, V. litterata was found to be a predatory omnivore capable of ingesting both animal and plant tissues