Biochemical and molecular characterization of pathogenic vibrios from hatcheries and aquaculture farms

Vibrio are important during hatchery rearing. aquaculture phase and post-harvest quality of shrimps. Vibrio spp are of concern to shrimp farmers and hatchery operators because certain species can cause Vibriosis. Vibrio species are of concern to humans because certain species cause serious diseases.With the progress in aquaculture, intensive systems used for shrimp aquaculture create an artificial environment that increases bacterial growth. To maintain the productivity of such an intensive aquaculture, high inputs of fish protein have to be employed for feeding together with high levels of water exchange and the massive use of antibiotics/ probiotics / chemicals. It seems that the combination of these conditions favours the proliferation of vibrios and enhances their virulence and disease prevalence. The risk of a microbial infection is high, mainly at larval stages. The effect and severity are related to Vibrio species and dose, water, feed, shrimp quality and aquaculture management.Consumption of seafood can occasionally result in food-bome illnesses due to the proliferation of indigenous pathogens like Vibrio.Of the l2 pathogenic Vibrio species, 8 species are known to be directly food associated. Strict quality guidelines have been laid by the importing nations, for the food products that enter their markets. The microbiological quality requirement for export of frozen shrimp products is that V.cholerae, V.parahaemolyticus and V. vulnificus should be absent in 25g of the processed shrimp (Export Inspection Council of India, 1995). The mere presence of these pathogenic Vibrios is sufficient for the rejection of the exported product.The export rejections cause serious economic loss to the shrimp industry and might harm the brand image of the shrimp products from the countiy.There is a need for an independent study on the incidence of different pathogenic vibrios in shrimp aquaculture and investigate their biochemical characteristics to have a better understanding about the growth and survival of these organisms in the shrimp aquaculture niche. PCR based methods (conventional PCR, duplex PCR, multiplex-PCR and Real Time PCR) for the detection of the pathogenic Vibrios is important for rapid post-harvest quality assessment. Studies on the genetic heterogeneity among the specific pathogenic vibrio species isolated from shrimp aquaculture system provide; valuable information on the extent of genetic diversity of the pathogenic vibrios, the shrimp aquaculture system.So the present study was undertaken to study the incidence of pathogenic Vibrio spp. in Penaeus monodon shrimp hatcheries and aquaculture farms, to carry out biochemical investigations of the pathogenic Vibrio spp isolated from P. monodon hatchery and. aquaculture environments, to assess the effect of salt (NaCl) on the growth and enzymatic activities of pathogenic Vibrio spp., to study the effect of preservatives, and chemicals on the growth of pathogenic Vibrio spp. and to employ polymerase chain reaction (PCR) methods for the detection of pathogenic V ibrio spp.Samples of water (n=7) and post-larvae (n=7) were obtained from seven Penaeus monodon hatcheries and samples of water (n=5), sediment (n=5) and shrimp (n=5) were obtained from five P. monodon aquaculture farms located on the East Coast of lndia. The microbiological examination of water, sediment, post-larvae and shrimp samples was carried out employing standard methods and by using standard media.The higher bacterial loads were obtained in pond sediments which can be attributed to the accumulation of organic matter at the pond bottom which stimulated bacterial growth.Shrimp head. (4.78 x 105 +/- 3.0 x 104 cfu/g) had relatively higher bacterial load when compared to shrimp muscle 2.7 x 105 +/- 1.95 x 104 cfu/g). ln shrimp hatchery samples, the post-larvae (2.2 x 106 +/- 1.9 x 106 cfu/g) had higher bacterial load than water (5.6 x 103 +/- 3890 cfu/ml).The mean E.coli counts were higher in aquaculture pond sediment (204+/-13 cfu/g) and pond water (124+/-88 cfu/ml). Relatively lower Escherichia coli counts were obtained from shrimp samples (12+/-11 to 16+/-16.7 cfu/g). The presence of E.coli in aquaculture environment might have been from the source water. E.coli was not detected in hatchery waters and post-larvae.

Studies on improved practices of prawn farming for higher production in central Kerala

The thesis entitled "Studies on improved practices of prawn farming for higher production in central Kerala" prepared by the author describes various practices prevailing in the study area in order to elucidate their relative merits. The study on semi-intensive farming at Mundapuram, Kannur was also carried out and included in the thesis for comparison.The author felt it important to make a critical study of the existing culture practices in the central Kerala, a region where it has been existing since time immemorial.Careful analysis of data accrued by the author has helped him to identify strength, weakness, opportunities and threats confronting the shrimp farming. As a result it was possible to evolve an appropriate management technology taking into consideration the various ecological (location specific), social and economical conditions prevalent in the vast study area.

Haematological responses of Penaeus monodon to environmental alterations and pathogenic invasion

This thesis Entitled Haematological responses of penaeus monodon to environmental alterations and pathogenic invasion. Thesis concluded from the present study that stress is accompanied by alterations in haemolymph metabolic variables and immune responses that influences the susceptibility of P. monodon to infection. Acute salinity variations were proved to be a stress condition that enhances the susceptibility of P. monodon to V. harveyi and WSSV infection. Ambient Cu at 0.1 mg 1" and ambient Zn at 1.0 mg 1" proved immunostimulatory in increasing the immunocompetence of P. monodon to WSSV infection and higher concentrations of Cu and Zn proved immunosuppressive. Haemolymph total protein, total carbohydrates and total lipids showed the highest relation with immune responses. THC, PO, ACP and ALP that greatly correlated with the survival rate proposed as reliable biomarkers of health in P. monodon. The study highlights the need for proper management practices and regular health monitoring to be adopted to avoid mass mortality in shrimp culture ponds.

Population Characteristics of Prawns in Natural and Selective Stocking System

present work deals with the various aspects of population characteristics of penaeus indicus ,Metapenaeus dobsoni and metapenaeus monoceros during their nursery phase in tidal ponds and adjacent backwaters.Importance of the present study is to suggest scientific basis for the management of penaeid resources in tidal ponds and backwaters based on their biological characteristics to ensure better yield.Seasonal closure of fishing will be effective in improving the size of the shrimp at harvest.Hydrology of tidal ponds varied with location, but showed a common seasonal pattem.Seasonal variation in temperature was very small. It fluctuated between 27.5 to 32.3°C in tidalponds and 26.9 to 29.9°C in open backwaters.Improvement of nursery habitats with due consideration for biological requirements of the resource will ensure better growth, survival and abundance of the stock.The recruitment, growth and emigration data of prawns from their nurseries can be used successfully for fishery forecasting. projecting juvenile growth forward through time, it is possible to establish, which cohort contributes to offshore fishery each year. So, by interpreting the recruitment and growth data of species in their nurseries with offshore catch data, fishery can be forecasted successfully.

A marine candida sake as source of immunostimulants to fenneropenaeus indicus

Aquaculture has developed rapidly over the last three decades to become an important activity worldwide.The Food and Agricultural Organization (FAO) of the UN acknowledge that global fishery output must be increased by at least 50% to offset projected shortfalls in dietary protein by 2030.LAquaculture has developed rapidly over the last three decades and has become an importat industry as today’s demand for fish exceeds the natural supply.lmmunostimulants are chemical compounds that activate the immune system of animals and render them more resistant to infections by viruses, bacteria, fungi, and parasites. lmmunostimulants have been obtained from diverse natural sources where, microbial cell wall acts as the main source.The salient findings of the study are summariseSeven marine yeasts were screened for growth promoting and immunostimulant property in F. indicus. Candida sake S165 was found to be best in terms of its support for growth and protection against white spot virus infection.The study revealed that marine yeast Candida sake can be effectively used as potential source of immunostimulants for application in penaeid prawns culture systems. The study emphasise the fact that the dose and frequency of application of immunostimulants are to be standardised and validated before commercialisation to achieve optimum stimulation of the immune system and to avoid immune fatigue die to verdose.Marine yeast (whole cell) was found to support better immunostimulation compared to its cell wall component B-1,3-glucan. This study shows that administration of marine yeast (whole cell) or B-1,3-glucan as immunostimulants in aquaculture would definitely help in protection of the stock to a few more days even though total protection is not being imparted. This partial protection itself would be highly helpful to the farming industry so that they can get sufficient time to plan for a safe harvest and save the crop from cent percent mortality.

Some studies on the reproductive endocrinology of tiger prawn Penaeus monodon Fabricius

The growing demand for quality prawn seed from the farmers‘ and entrepreneurs, coupled with uncertainity of their availability from nature at the appropriate time in required quantities has prompted‘ research on problems connected with prawn seed production. Endocrine control of reproduction in the penaeid shrimp _P_. monodon has been investigated in detail by adopting a comprehensive approach to the problem. The major aspects of the study included in depth investigations of the cytological details of the reproductive and neuroendocrine organs in correlation with the process of gonadal maturation. Based on the conclusions drawn from such ultrastructural studies various endocrine manipulations were carried out to see their effects on gonadal maturation. Besides that studies on the reproductive quality of male P_. monodon and the cryopreservation of spermatophores form a part of the present investigation. The shrimp 3; Inonodon used in the present study were collected from the offshore waters of Cochin, Madras and Mandapam and from the culture ponds of Vypeen Island near Cochin (Kerala) . The entire investigation on the cytological aspects were carried out using standard histological and electron-microscopic methods. Endocrine manipulations and cryopreservation studies is also carried out using the standard methods.

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

Pyocyanin (5-methyl-1-hydroxyphenazine) produced by Pseudomonas aeruginosa as antagonist to vibrios in aquaculture: overexpression, downstream process and toxicity

Pyocyanin is a versatile and multifunctional phenazine, widely used as a bio-control agent. Besides its toxicity in higher concentration, it has been applied as bio-control agents against many pathogens including the Vibrio spp. in aquaculture systems. The exact mechanism of the production of pyocyanin in Pseudomonas aeruginosa is well known, but the genetic modification of pyocyanin biosynthetic pathways in P. aeruginosa is not yet experimented to improve the yield of pyocyanin production. In this context, one of the aims of this work was to improve the yield of pyocyanin production in P. aeruginosa by way of increasing the copy number of pyocyanin pathway genes and their over expression. The specific aims of this work encompasses firstly, the identification of probiotic effect of P. aeruginosa isolated from various ecological niches, the overexpression of pyocyanin biosynthetic genes, development of an appropriate downstream process for large scale production of pyocyanin and its application in aquaculture industries. In addition, this work intends to examine the toxicity of pyocyanin on various developmental stages of tiger shrimp (Penaeus monodon), Artemia nauplii, microbial consortia of nitrifying bioreactors (Packed Bed Bioreactor, PBBR and Stringed Bed Suspended Bioreactor, SBSBR) and in vitro cell culture systems from invertebrates and vertebrates. The present study was undertaken with a vision to manage the pathogenic vibrios in aquaculture through eco-friendly and sustainable management strategies with the following objectives: Identification of Pseudomonas isolated from various ecological niches and its antagonism to pathogenic vibrios in aquaculture.,Saline dependent production of pyocyanin in Pseudomonas aeruginosa originated from different ecological niches and their selective application in aquaculture,Cloning and overexpression of Phz genes encoding phenazine biosynthetic pathway for the enhanced production of pyocyanin in Pseudomonas aeruginosa MCCB117,Development of an appropriate downstream process for large scale production of pyocyanin from PA-pUCP-Phz++; Structural elucidation and functional analysis of the purified compoundToxicity of pyocyanin on various biological systems.

Larval biology of the spiny lobsters of the genus panulirus

Lobsters are renowned the world over as tasty delicacies and are in great demand for Epicurian gourests. As a result these animals found in different parts of the world from countries like India they are mostly frozen and exported thus earning considerable foreign exchange for the country. Importance of this crustacean is thus well known and whatever harvested from nature are utilized to the maximum extent. The demand being much more than what is being produced the need for artificial culture becomes evident. In this context a complete knowledge of the lifecycle ,larval biology and their culture is very essential. Thus with the object of studying the life history of the commercially important lobsters fished from Indian waters the larval biology of panulirus homarus has been taken up as the project for the present study

Echno economic studies on industrial fishing in the upper east coast of India

The present scenario of industrial fishing in India is that most of large trawlers are based at Visakhapatnam and congregate in the potential shrimp ground in the upper East coast of India commonly known as the Sandheads. These are outriggcr vessels operating two or four trawl nets along with a testing trawl called try net. In the early Seventies these vessels were operating on a very high economic return which was evident from the steady increase in number of outriggers over a period of twenty years. Since the total allowable catch has to be shared by all vessels including the increasing fleet, reduction per vessel output is bound to happen. Therefore some of them could not survive the competition and withdrew from the scene. The number of outriggers did not increase subsequently. However, there arose a doubt whether the existing fleet of about 180 vessels are fishing economically or whether there is any scope for further introduction of industrial vessels in the region. This study is focussing to the techno economic aspects of industrial fishing in the upper East coast of India

Studies on osmoregulation in the penaeid prawn Metapenaeus dobsoni (Miers)

Metapenaeus dobsoni (Miers) is the most abundant species along the coast of Kerala. It is cultured extensively by adoption of traditional farming practices. The geographical location and water source determines the seasonal and annual environmental fluctuations the prawn farming systems experiences. The life cycle of the shrimp includes its migration to the coastal deeper waters for spawning and the immigration of larvae to the estuaries for growth. The survival of the species in such complex ecosystems is thus critical to its life cycle. The animal adapts itself to different environments through a physiological process known as osmoregulation. The present study on osmoregulation in the penaeid prawn Metapenaeus dobsoni was thus undertaken to understand the mechanism adopted by this species to survive in different environments. A number of experimental work have been conducted to understand the effect of salinity on the internal variations. However the effect of the complex environmental conditions as existent in nature on the osmotic variations in this species has not been dealt with in any of the earlier studies.

Mesozooplankton of the Arabian Sea and the Bay of Bengal with special reference to planktonic ostracods

The Arabian Sea and the Bay of Bengal are both highly dynamic ecosystems, due to the seasonally reversing monsoon winds, but the processes affecting the mesozooplankton community remain poorly understood. These are important basins exhibiting enhanced biological production as a result of upwelling, winter cooling and other episodic events such as eddies and gyres. Zooplankters are primarily the prey for almost all fish larvae. Seasonal changes in the biogeochemical processes can strongly affect zooplankton density and distribution, which in turn, strongly affect the larval growth, and consequently, the pelagic fish recruitment. It is clear that plankton biomass and biogeochemical fluxes are not in steady state. Acoustic data on mesozooplankton abundance suggests that they also exist in the mesopelagic zone. Earlier studies were confined only to the upper 200 m and hence the structure of mesozooplankton community in the deeper layers was not well known. Copepods are the dominant mesoplankton group, and therefore the majority of the studies were focused on them. The planktonic ostracods are the second major crustacean group and at times, their swarms can outnumber all other planktonic groups. The understanding of the community structure of the ostracods is essential to establish their role in the marine food web. Mesozooplankton is responsible for the vertical flux of organic matter produced by phytoplankton and is assumed to be equivalent to new production (Eppley & Peterson, 1979). Since the fate of newly produced organic matter depends upon their consumers, the zooplankton biomass must be estimated in size fractions or taxonomic components to understand the vertical flux of organic carbon. It is thus important to update our knowledge on different groups of zooplankton on the basis of seasonal and temporal distribution. The distribution in space and time is essential for modeling the carbon cycling that structure the marine ecosystems

studies on Prawn Gill Nets of The Kerala Coast

In the present study. extensive investigations were carried out on various factors affecting the selectivity of prawn gill nets with reference to material, mesh size, coefficient of hanging secolouration. Effect of tidal current on fishing height of prawn gill net and seasonal variation of catch during the course of these investigations were also studied.

Studies on sphaeroma terebrans bate of the port of Cochi

Sphaeroma terebrans Bate, one of the most destructive wood-boring isopod crustacean occurring in the Cochin harbour region was taken up for investigations on its habitat, breeding, attack on timber, salinity tolerance and respiration. The respiration of the animal under different stresses was noted using an instrument fabricated in the laboratory. Of the various hydrographic conditions of the Cochin harbour region studied, salinity is found to be the most fluctuating factor and the variations are largely influenced by the seasonal rain fall from one side and the tidal currents on the other.In spite of the great fluctuations in salinity, Sphaeroma terebrans occurs throughout the year in the Cochin harbour region, though their number varies in different months. Exposure tests show that Sphaeroma terebrans can remain for about 20 to 24 hours outside water in a damp atmosphere.

Studies On Pobtunid Crabs [Crustacea Decapoda Brachyura)

True crabs are the most fascinating group of organisms among the decapod crustaceans. Great importance is attached at present to the increased exploitation of these animals and therefore there is great scope for further development of their fishery. They have a broad and hard carapace, massive chelate legs, bent abdomen and exhibit high degree of adaptation to the environment. They show pelagic, benthic, intertidal, burrowing and terrestrial modes of life. Their commensal association with other invertebrates, their breeding behaviour and life history are of great interest to biologists. More than six hundred species of crabs are known to occur in Indian waters and among them about eight species form a regular fishery along the entire stretch of peninsular India (Rao §§_al., 1973) round the year. Crab fishery in India is fast developing and there is vast scope for them as there are many more potential species. Among the various crustacean diets, crabs are celebrated for deliciousness and for nutritional richness. In recent days, crab food items have become more popular and gained global reception. These resources can also be augmented further by culturing them in ponds in the future. Information on biology and ecology of constituent species go a long way not only in effective exploitation and regulation of the respective fishery resources but also helps in evolving a suitable gear for their capture. Information collected on the national level in various aspects as reproduction, growth rate, larval development, parasites, diseases, nutritive values etc. will be of help in evolving a national policy for the effective utilisation and conservation of this resource. They also provide the baseline information for undertaking any purposeful and meaningful culture activities. Information on the various aspects mentioned above is very much restricted in true crabs and hence the present study