DETECTION OF BILIARY FUNCTIONAL PROBLEMS USING MICROWAVES

This article reports a new in vitro bile analysis based on the measurement of the dielectric properties at microwave frequencies. The measurements were made using rectangular cavity perturbation technique at the S-band of microwave frequency with the different samples of bile obtained from healthy persons as well as from patients. It is observed that an appreciable change in the dielectric properties of patient’s samples with the normal healthy samples and these measurements were in good agreement with clinical analysis. These results prove an alternative in-vitro method of detecting bile abnormalities based on the measurement of the dielectric properties of bile samples using microwaves without surgical procedure.

HEAVY METAL TOXICITY IN BIVALVE- HISTOLOGICAL AND HISTOCHEMICAL ENQUIRY

Division of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Cochin University of Science and Technology

Biochemical & Pharmacological evaluation of liver oils of selected deep sea sharks and chimaeras of the Indian EEZ

With a seacoast of 8,1 18 km, an exclusive economic zone (EEZ) of 2 million square km, and with an area of about 30,000 square km under aquaculture, lndia produces close to six million tonnes of fish, over 4 per cent of the world fish production. While the marine waters upto 50m depth have been fully exploited, those beyond, remain unexplored. There is an ever increasing demand for fishery resources as food. The coastal fishery resources of the country are dwindling at a rapid pace and it becomes highly imperative that we search for alternate fishery resources for food. The option we have is to hunt for marine fishery resources. Studies pertaining to proximate composition, amino acid and fatty acid composition are essential to understand the nutraceutical values of these deep sea fishery resources. The present study was aimed to carry out proximate composition of deep sea fishery resources obtained during cruises onboard the FORV Sarise Sampada, to identify fishery resources which have appreciable lipid content and thereby analyse the bioactive potentials of marine lipids, to study the amino acid profile of these fishery resources, to understand the contents of SPA, MUFA and PUFA and to calculate the n3/n6 fatty acid contents. Though the presence of nutraceuticals was identified in the marine fishery resources their use as potential food resources deserve further investigation. So the study were carried out to calculate the hepatosomatic indices of sharks & chimaeras and conduct biochemical characterisation of liver oils of Apristurus indicus, Cenlrophorus scalprams, Centroselachus crepidater, Neoharriotta raleighana, and Harriotta pinnata obtained during cruises onboard the FORV Sugar Sampada.Therapeutic use of shark liver oil is evident from its use for centuries as a remedy to heal wounds and fight flu (Neil er al. 2006). Japanese seamen called it 'samedava' or "cure all". Shark liver oil is being promoted worldwide as a dietary supplement to boost the immune system, fight infections, to treat cancer and to lessen the side effects of conventional cancer treatment. These days more emphasis is laid on the nutritive benefits of shark liver oils especially on the omega 3 polyunsaturated fatty acids ( PUFAs) (Anandan er al. 2007) and alkylglycerols (AKGs) (Pugliese er al. I998) contained in them due to the high rise of inflammatory disorders such as arthritis, asthma and neurodegenerative diseases like Alzheimer’s, Parkinson’s and Schizophrenia. So the present study also evaluate the pharmacological properties with respect to analgesic, anti-inflammatory, anti pyretic and anti-ulcer effects of four different liver oils of sharks belonging to the Indian EEZ and to identify the components of oil responsible for these activities.The analgesic and anti-inflammatory activities of liver oils from Neoharriotra raleighana (NR), Centrosymnus crepidater (CC), Apristurus indicus (AI), and Centrophorus sculpratus (CS) sharks caught from the Arabian Sea and the Indian Ocean were compared. The main objectives also include determination of the cholesterol lowering effects of liver oils of Neoharriotra raleighana (NR) and Centrophorus sculpratus (CS) on the high fat diet induced dyslipidemia and to compare the impact of four isolipidemic diets, on levels of serum diagnostic marker enzymes, on lipid profile of blood and liver and antioxidant status of heart in male Albino rats. And also to study the efficacy of Centrophorus sculpratus (CS) liver oil against Complete Freund’s Adjuvant-induced arthritis and to compare the anti-inflammatory activity of this oil with a traditionally used anti-inflammatory substance gingerol (oleoresin extracted from ginger.). The results of the present study indicated that both (Centrophorus sculpratus liver oils as well as gingerol extracts proved to be effective natural remedies against CFA-induced arthritis in Albino rats.

Toxicity, accumulation and depuration of heavy metalsin the brown mussel perna indica

In this thesis certain important aspects of heavy metal toxicity have been worked out. Recent studies have clearly shown that when experimental media contained more than one heavy metals, such metals could conspicuously influence the toxic reaction of the animals both in terms of quantity and nature. The experimental results available on individual metal toxicity show that, in majority of such results, unrealistically high concentrations of dissolved metals are involved. A remarkable number of factors have been shown to influence metal toxicity such as various environmental factors particularly temperature and salinity, the condition of the organism and the ability of some of the marine organisms to adapt to metallic contamination. Further, some of the more sensitive functions like embryonic and larval development, growth and fecundity, oxygen utilization and the function of various enzymes are found to be demonstrably sensitive in the presence of heavy metals. However, some of the above functions could be compensated for by adaptive process. If it is assumed that the presence of a single metal in higher concentrations could affect the life function of marine animals, more than one metal in the experimental media should manifest such effects in a greater scale. Commonly known as synergism or more than additivity, majority of heavy metals bring about synergistic reaction .The work presented in this thesis comprises lethal and sublethal toxicities of different salt forms of copper and silver on the brown mussel Perna indica. during the present investigation sublethal concentrations of copper and silver in their dent effects on survival, oxygen consumption, filtration, accumulation and depuration on Perna indica. The results are presented under different sections to make the presentation meaningful .

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.

Provenance, Isolation and Characterisation of Organic Matter in the Cochin Estuarine Sediment-“a Diagenetic Amino Acid Marker Scenario

Cochin estuarine system is among the most productive aquatic environment along the Southwest coast of India, exhibits unique ecological features and possess greater socioeconomic relevance. Serious investigations carried out during the past decades on the hydro biogeochemical variables pointed out variations in the health and ecological functioning of this ecosystem. Characterisation of organic matter in the estuary has been attempted in many investigations. But detailed studies covering the degradation state of organic matter using molecular level approach is not attempted. The thesis entitled Provenance, Isolation and Characterisation of Organic Matter in the Cochin Estuarine Sediment-“ A Diagenetic Amino Acid Marker Scenario” is an integrated approach to evaluate the source, quantity, quality, and degradation state of the organic matter in the surface sediments of Cochin estuarine system with the combined application of bulk and molecular level tools. Sediment and water samples from nine stations situated at Cochin estuary were collected in five seasonal sampling campaigns, for the biogeochemical assessment and their distribution pattern of sedimentary organic matter. The sampling seasons were described and abbreviated as follows: April- 2009 (pre monsoon: PRM09), August-2009 (monsoon: MON09), January-2010 (post monsoon: POM09), April-2010 (pre monsoon: PRM10) and September- 2012 (monsoon: MON12). In order to evaluate the general environmental conditions of the estuary, water samples were analysed for water quality parameters, chlorophyll pigments and nutrients by standard methods. Investigations suggested the fact that hydrographical variables and nutrients in Cochin estuary supports diverse species of flora and fauna. Moreover the sedimentary variables such as pH, Eh, texture, TOC, fractions of nitrogen and phosphorous were determined to assess the general geochemical setting as well as redox status. The periodically fluctuating oxic/ anoxic conditions and texture serve as the most significant variables controlling other variables of the aquatic environment. The organic matter in estuary comprise of a complex mixture of autochthonous as well as allochthonous materials. Autochthonous input is limited or enhanced by the nutrient elements like N and P (in their various fractions), used as a tool to evaluate their bioavailability. Bulk parameter approach like biochemical composition, stoichiometric elemental ratios and stable carbon isotope ratio was also employed to assess the quality and quantity of sedimentary organic matter in the study area. Molecular level charactersation of free sugars and amino acids were carried out by liquid chromatographic techniques. Carbohydrates are the products of primary production and their occurrence in sediments as free sugars can provide information on the estuarine productivity. Amino acid biogeochemistry provided implications on the system productivity, nature of organic matter as well as degradation status of the sedimentary organic matter in the study area. The predominance of carbohydrates over protein indicated faster mineralisation of proteinaceous organic matter in sediments and the estuary behaves as a detrital trap for the accumulation of aged organic matter. The higher lipid content and LPD/CHO ratio pointed towards the better food quality that supports benthic fauna and better accumulation of lipid compounds in the sedimentary environment. Allochthonous addition of carbohydrates via terrestrial run off was responsible for the lower PRT/CHO ratio estimated in thesediments and the lower ratios also denoted a detrital heterotrophic environment. Biopolymeric carbon and the algal contribution to BPC provided important information on the better understanding the trophic state of the estuarine system and the higher values of chlorophyll-a to phaeophytin ratio indicated deposition of phytoplankton to sediment at a rapid rate. The estimated TOC/TN ratios implied the combined input of both terrestrial and autochthonous organic matter to sedimentsAmong the free sugars, depleted levels of glucose in sediments in most of the stations and abundance of mannose at station S5 was observed during the present investigation. Among aldohexoses, concentration of galactose was found to be higher in most of the stationsRelative abundance of AAs in the estuarine sediments based on seasons followed the trend: PRM09-Leucine > Phenylalanine > Argine > Lysine, MON09-Lysine > Aspartic acid > Histidine > Tyrosine > Phenylalanine, POM09-Lysine > Histadine > Phenyalanine > Leucine > Methionine > Serine > Proline > Aspartic acid, PRM10-Valine > Aspartic acid > Histidine > Phenylalanine > Serine > Proline, MON12-Lysine > Phenylalanine > Aspartic acid > Histidine > Valine > Tyrsine > MethionineThe classification of study area into three zones based on salinity was employed in the present study for the sake of simplicity and generalized interpretations. The distribution of AAs in the three zones followed the trend: Fresh water zone (S1, S2):- Phenylalanine > Lysine > Aspartic acid > Methionine > Valine ῀ Leucine > Proline > Histidine > Glycine > Serine > Glutamic acid > Tyrosine > Arginine > Alanine > Threonine > Cysteine > Isoleucine. Estuarine zone (S3, S4, S5, S6):- Lysine > Aspartic acid > Phenylalanine > Leucine > Valine > Histidine > Methionine > Tyrosine > Serine > Glutamic acid > Proline > Glycine > Arginine > Alanine > Isoleucine > Cysteine > Threonine. Riverine /Industrial zone (S7, S8, S9):- Phenylalanine > Lysine > Aspartic acid > Histidine > Serine > Arginine > Tyrosine > Leucine > Methionine > Glutamic acid > Alanine > Glycine > Cysteine > Proline > Isoleucine > Threonine > Valine. The abundance of AAs like glutamic acid, aspartic acid, isoleucine, valine, tyrosine, and phenylalanine in sediments of the study area indicated freshly derived organic matter.

Is Microtox R toxicity related to potentially harmful algae proliferation in Mediterranean salt marshes?

Mediterranean salt marshes are ecosystems that are highly influenced by sea changes and freshwater inputs from runoff. In these ecosystems, toxic and non-toxic algae blooms often produce large and unpredictable biomasses of phytoplankton. The Microtox R test has been described as a successful, quick method for detecting toxicity in various phytoplankton taxa. Ourstudy sought to test the efficiency of Microtox R in detecting toxic HAB in Mediterranean salt marshes. The results showed that the Microtox R test was able to detect toxic substances in the particulate matter of several lagoons in the Empordà salt marshes. This Microtox R toxicity coincided with periods when potentially harmful cyanobacteria, dinoflagellates and haptophytes had a high biomass. The results suggest that potentially harmful phytoplankton cannot be ruled out as a source of Microtox R

Fluvial biofilms: a pertinent tool to assess β-blockers toxicity

Among increasingly used pharmaceutical products, β-blockers have been commonly reported at low concentrations in rivers and littoral waters of Europe and North America. Little is known about the toxicity of these chemicals in freshwater ecosystems while their presence may lead to chronic pollution. Hence, in this study the acute toxicity of 3 β-blockers: metoprolol, propranolol and atenolol on fluvial biofilms was assessed by using several biomarkers. Some were indicative of potential alterations in biofilm algae (photosynthetic efficiency), and others in biofilm bacteria (peptidase activity, bacterial mortality). Propranolol was the most toxic β-blocker, mostly affecting the algal photosynthetic process. The exposure to 531 μg/L of propranolol caused 85% of inhibition of photosynthesis after 24 h. Metoprolol was particularly toxic for bacteria. Though estimated No-Effect Concentrations (NEC) were similar to environmental concentrations, higher concentrations of the toxic (503 μg/L metoprolol) caused an increase of 50% in bacterial mortality. Atenolol was the least toxic of the three tested β-blockers. Effects superior to 50% were only observed at very high concentration (707 mg/L). Higher toxicity of metoprolol and propranolol might be due to better absorption within biofilms of these two chemicals. Since β-blockers are mainly found in mixtures in rivers, their differential toxicity could have potential relevant consequences on the interactions between algae and bacteria within river biofilms

The use of pulse amplitude modulated fluorescence techniques for metal toxicity assessment in fluvial biofilms

Metal pollution in rivers is in great concern with human activities in the fluvial watershed. This thesis aims to investigate the potential use of chl-a fluorescence parameters as biomarkers of metal toxicity, and to find cause-effect relationships between metal exposures, other environmental factor (i.e. light), and functional and structural biofilm responses. This thesis demonstrates that the use of chl-a fluorescence parameters allows detect early effects on biofilms caused by zinc toxicity, both in the laboratory as in polluted rivers. In microcosm experiments, the use of chl-a fluorescence parameters allows evaluates structural changes on photosynthetic apparatus and in algal groups’ composition of biofilms long-term exposed to zinc. In order to evaluate the effects of chronic metal pollution in rivers, it is recommended the use of biofilm translocation experiments and the use of a multi-biomarker approach.

Contribution of antioxidant enzymes to toxicity assessment in fluvial biofilms

Per tal d’avaluar l’impacte de la contaminació en els ecosistemes aquàtics, aquesta tesi es centra en una aproximació multi-biomarcador en els biofilms. En complement dels biomarcadors clàssics, es va demostrar que les activitats dels enzims antioxidants (AEA): catalasa, ascorbat peroxidasa i glutatió reductasa eren biomarcardors d'estrès oxidatiu en els biofilms. Tot i que les AEA poden veure's influenciades amb la mateixa mesura per factors naturals (edat del biofilm, llum de colonització o d'exposició) i contaminants (herbicides i farmacèutics), aquestes AEA permeten entendre millor l'efecte dels contaminants. Cal remarcar que assajos de toxicitat aguda es poden utilitzar per comparar la capacitat antioxidant entre comunitats i conèixer la seva pre-exposició a l'estrès oxidatiu. Aquesta aproximació multi-biomarcador a nivell de comunitat és especialment interessant per avaluar la toxicitat dels contaminants emergents (β-blockers) sobre espècies no-diana. Per tal de millorar-la, també es va verificar la possibilitat de mesurar l'expressió gènica en biofilms.

Evaluaton of a genetic marker within the organ of Corti of transgenic mice

This paper reviews a study to evaluate a genetic marker within the organ of Corti in transgenic mice for potential use in cell lineage studies.