Development of Cell Culture Systems from Selected Species of Fish and Prawns

The present work deals with the development of primary cell culture and diploid cell lines from two fishes, such as Poecilia reticulata and Clarias gariepinus. The greatest difficulty experienced was the avoidance of bacterial and fungi contamination. Three types of cell cultures are commonly developed, primary cell culture, diploid cell lines and heteroploid cell lines. Primary cell culture obtained from the animal tissues that have been cultivated in vitro for the first time. They are characterized by the same chromosome number as parent tissue, cultivated in vitro for the first time, have wide range of virus susceptibility, usually not malignant, six chromatin retarded and do not grow as suspension cultures. Diploid cell lines arise from a primary cell culture at the time of subculturing. Diploid cell lines commercially used in virology are W1-38 (human embryonic lung), W1-26 (human embryonic lung) and HEX (Human embryonic kidney). Heteroploid cell lines have been subcultivated with less than 75% of the cells in the population having a diploid chromosome constitution. Tissue cultures have been extensively used in biomedical research. The main applications are in three areas, Karyological studies, Identification and study of hereditary metabolic disorders and Somatic cell genetics. Other applications are in virology and host-parasite relationships. In this study an attempt was made to preserve the ovarian tissue at low temperature in the presence of cryoprotectants so that the tissue can be retrieved at any time and a cell culture could be developed.

Studies on Copepod Parasites on Elasmobranchs of Kerala Coast

The present study of the parasitic copepods gives the taxonomic description of thirty one species of parasites collected from various elasmobranch fishes of Kerala coast. Copepods parasitic on fishes include three sub orders, viz. poecilostomatoida, cyclopoida and siphonostomatoida. Parasitic copepods of elasmobranch fishes for the present study were collected from different fish landing centres of Kerala and by undertaking regular fishing trips. The collected parasites are identified to the species level and described. It is found that out of thirty one species, fifteen are new to science. They belong to the genera viz. Nothobomolochus Vervoot, 1962, Caligus Muller, 1785, Alebion, Kroyer, 1863, Gloipotes Steenstrup and Lutken, 1861, Pandarus Leach,1819, Perissopus Steenstrup and Lutken, 1861, Echthrogaleus Steenstrup and Lutken, 1861 and Kroyeria van Beneden, 1853. Fifteen new host records were reported. Two genera viz. Echthrogaleus and Entepherus were reported for first time from Indian waters. A new genus called Penicillus was erected. The general observations made during this study revealed certain interesting aspects of host-parasite relationship, host specificity, site specificity, adaptive modifications and geographical distribution.

Molecular Characterization and Gene Expression Profiling of Antimicrobial Peptides in Penaeid Shrimps

The constitutive production of AMPs in shrimps ensures that animals are able to protect themselves from low-level assaults by pathogens present in the environment. As these molecules play important roles in the shrimp immune defense system, the expression level of these AMPs are possible indicators of the immune state of shrimps. The present study also indicates the antiviral property of AMPs, especially ALF, stressing the importance of their up-regulation through the application of immunostimulants/probiotics as a prophylactic strategy in aquaculture. The present study shows that shrimp defense system is equipped enough to evade WSSV infection to a certain extent, when the animals were maintained on marine yeast and probiotic diet, whereas the control diet fed group succumbed to WSSV infection. This study reveals that marine yeast and probiotic supplemented diet can delay the process of WSSV infection and confer greater protection to the animals. Particularly, the protection conferred by marine yeast, C. haemulonii S27 and Bacillus MCCB101 were highly promising imparting greater hope to the aquaculture community to overcome the prevailing disease problems in aquaculture. It may be inferred from the present study that up-regulation of AMP genes could be effected by the application of immunostimulants and probiotics. Also, AMP expression profile could be used as an effective tool for screening immunostimulants and probiotics for application in shrimp culture. Ultimately, it is likely that no single compound or strategy will provide a solution to the problem of disease within aquaculture and that, in reality, a suite of techniques will be required including the manipulation of the rearing environment, addition of probionts as a matter of routine during culture, and the use of immunostimulants and other supplements during vulnerable growth phases. Finally, the development of good management practices, the control of environmental variables, genetic improvement in the penaeid species, understanding of host-virus interaction, modulation of the shrimp immune system, supported by functional genomics and proteomics of this crustacean, as a whole suggests that the control of WSSV is not far.

Mangrove plant Ceriops tagal as a potential source of anti White Spot Syndrome Virus preparations for Penaeus monodon

White spot syndrome virus (WSSV) is the deadliest virus among crustaceans ever discovered having several unique and novel features. Recent developments in genomics and proteomics could elucidate the molecular process involved in the WSSV infection and the host pathogen interaction to some extent. Until now no fool proof treatment or prophylactic measure has been made available to control WSSV out breaks in culture system. Even though there are technologies like application of immunostimulants, vaccines, RNAi and several antiviral natural products none of them has been taken to the level of clinical trials. However, there are several management options such as application of bioremediation technologies to maintain the required environmental quality, maintenance of zero water exchange systems coupled with application of probiotics and vaccines which on adoption shall pave way for successful crops amidst the rapid spread of the virus. In this context the present work was undertaken to develop a drug from mangrove plants for protecting shrimp from WSSV.Mangroves belong to those ecosystems that are presently under the threat of destruction, diversion and blatant attack in the name of so called ‘developmental activities’. Mangrove plants have unique ecological features as it serves as an ecotone between marine and terrestrial ecosystem and hence possess diversity of metabolites with diverse activities. This prompted them being used as remedial measures for several ailments for ages. Among the mangrove plants Ceriops tagal, belonging to the family Rhizophororaceae was in attention for many years for isolating new metabolites such as triterpenes, phenolic compounds, etc. Even though there were attempts to study various plant extracts to develop anti-viral preparations their activity against WSSV was not investigated as yet.

Studies on The Copepod Fish Parasites of Kerala Coast

The present study of the parasitic copepods gives an account of the taxonomic description of seventy seven species of parasites collected from the food fishes of the Kerala coast. Out of the seventy seven species described, fourteen are new to science, two new records for the Indian waters and ten new host records. The males of Parapetalus longipinnatus Rangnekar and Lerna~thropus indicus Pillai were collected and described for the first time. The parasites described belong to the suborders Cyclopoida, Caligoida and Lernaeopodoida. The available description of many species of this locality is reviewed and supplemented with the help of the present detailed study. The general observations made during this study reveale certain interesting aspects of the host parasite relationship, host specificity, adaptive modification and geographical distribution. A brief discussion of these observations made is also presented.

Studies on the freshwater copepod fish parasites of Kerala

Studies on parasitic copepods from freshwater fishes are still in its infancy. In recent years, there- is a renewed enthusiasm in the study of freshwater fish parasites due to rapidly increasing aquaculture practices. The importance of diseases and their control assumes great significance because of the adverse impact of diseases on fish production and its economy. Copepods are one of the most harmful parasites of freshwater fishes. Reports on the damages caused by copepod parasites from different parts on the world are increasing alarmingly. But the information on parasitic copepods of freshwater fishes in India is quite meagre. Knowledge regarding this group of parasites, their Biology and pathology from Kerala. is lacking. The thesis consists of five chapters. The first chapter is a general introduction which deals with the review of literature on various aspects of parasitic copepods viz; systematics, life history, host-parasite relationship, ecology, pathogenicity, prophylaxis and control measures. Systematics of parasitic copepods of freshwater fishes collected during the present study forms the second chapter. The third chapter deals with the life cycle study of the new Lernaeid copepod, Lernaea osphronemi. The fourth chapter contains host-parasite relationship. ecology and treatment of ‘the’ new species of Lernaea On Osphronemus goramy. General observations and a summary of the entire work constitute the fifth chapter

Parasite distribution and histopathological studies on certain commercially important fishes of cochin area

In order to check the damage caused by the parasites, even though it is difficult in open waters, a proper understanding of the seasonal variation in the distribution of the parasite and other factors like age of the host, sex of the host, which affect distribution of parasite is a must. Although several workers have carried out investigations on the taxonomy of metazoan parasites of marine and brackish water Fishes of India, very little attempt is made to correlate such investigations with the host and the environment. In this thesis such an attempt is made by the researcher. In chapter one the literature related to the prevalence, mean intensity of infection, and histopathological changes caused by the metazoan parasites, in particular by helminths, copepods and isopods, was reviewed. Chapter two contains observations on the distribution pattern of parasites in relation to the season, sex, and size of the host. It was found that the prevalence rate of radiorynchus_indicus infecting the alimentary canal of Tachysurugs mgacuglatus, Ergasilus sp. infecting the gills of T maculatus, and Lernaeeniicus ramosus found on the body surface of Qeimgipteirfugs jagonicus was higher during monsoon season. But agarna malayi found in the opercular chamber and Ehilometra cephalus infecting the gonads of valamugil speiglari showed a higher prevalence rate during the postmonsoon season. This was discussed on the basis of the hydrographical characteristics prevailing in the study area during the three different seasons. It was also observed that the sex of the host did not influence significantly the distribution pattern of the parasites. The reasons for this were also discussed. Invariably, the size of the host was found to influence the parasite distribution pattern. It was observed that the prevalence rate showed an increase with increase in size of the fish. This was discussed on the basis of food habits of the host, along with other aspects An attempt was made in chapter three to study the histopathological effects of‘ the various parasites on their respective sites of attachments on host Fishes. It was found that except Rhadinorhyhchugs indicus, all other parasites produced damages of varying intensity, in the form of hypertrophy, rhyperplasia, haemorrhage, tissue disruption and ulcers. Interestingly, E. indicus, an acantho— cephalid with a powerful proboscis for attachment was found not to cause any serious damage to the intestine of the host Fish. All these aspects are included in the third and final chapter of the thesis.