First development and characterisation of polyclonal and monoclonal antibodies to the emerging fresh water toxin cylindrospermopsin

As increasing incidences in the occurrence of cylindrospermopsin (CYN) appear, in addition to further research on its toxicological nature, improved rapid methods to detect this toxin are required. Antibody based assays are renowned for their ability to provide rapid, portable, simple to use tests. As yet however there are no publications outlining how an antibody to CYN can be produced. A range of chemical approaches was investigated to synthesise CYN immunogens for antibody production but failed to generate a response. Finally, a modified Mannich reaction for immunogen synthesis was employed to couple the toxin to two carrier proteins. Both protein conjugates were successfully used to raise both polyclonal and monoclonal antibodies of high sensitivity to CYN. These antibodies were characterised employing competitive indirect ELISA and an optical biosensor assay. By ELISA the sensitivity achieved ranged from 27 to 131. pg/mL and by SPR 4.4 to 11.1. ng/mL thus demonstrating that the selection of immunoassay platform is important for the detection level required by the end user for their application. Low cross-reactivity to the much less toxic metabolite deoxyCYN was observed. This is the first reported production of antibodies to this toxin. © 2013 Elsevier B.V.

Syntrophic relationships between algae and bacteria in a fresh-water stream and in culture /

Interactions between freshwater algae and bacteria were examined in a natural stream habitat and a laboratory model. Field observations provided circumstantial evidence, in statistical correlation for syntrophy between the microbial populations. This relation is probably subject to control by the temperature and pH of the aquatic environment. Several species of a pond community were isolated in axenic culture and tests were performed to determine the nature of mixed species interactions. Isolation procedures and field studies indicated that selected strains of Chlorella and Azotobacter were closely associated in their natural habitat. With the suspected controlling parameters, pH and temperature, held constant, mixed cultures of algae and bacteria were compared to axenic cultures of the same organisms, and a mutual stimulation of growth was observed. A mixed pure culture apparatus was designed in this laboratory to study the algal-bacterial interaction and to test the hypothesis that such an interaction may take place through a diffusable substance or through certain medium-borne conditions, Azotobacter was found to take up a Chlorella-produced exudate, to stimulate protein synthesis, to enhance chlorophyll production and to cause a numerical increase in the interacting Chlorella population. It is not clear whether control is at the environmental, cellular or genetic level in these mixed population interactions. Experimental observations in the model system, taken with field correlations allow one to state that there may be a direct relationship governing the population fluctuations of these two organisms in their natural stream surroundings.

Development of Innovative Low Cost Larviculture Technologies of the Giant Fresh Water Prawn,Macrobrachium Rosenbergii(De Man)

School of Industrial Fisheries, Cochin University of Science and Technology

Distribution, Extracellular Virulence Factors and Drug Resistance of Motile Aeromonads in Fresh Water Ornamental Fishes and Associated Carriage Water

During last decades there has been a continuous growth of aquaculture industries all over the world and taking into consideration the spurt in freshwater ornamental fish aquaculture and trade in Kerala, the present study was aimed to assess the prevalence of various motile Aeromonas spp. in fresh water ornamental fishes and associated carriage water. The extracellular virulence factors and the antibiogram of the isolates were also elucidated. Various species of motile aeromonads such as Aeromonas caviae, A. hydrophila, A. jandaei, A. schubertii, A. sobria, A. trota and A. veronii were detected. Aeromonas sobria predominated both fish and water samples. Extracellular enzymes and toxins produced by motile aeromonds are important elements of bacterial virulence. The production of extracellular virulence factors - proteases, lipase, DNase and haemolysin by the isolates were studied. All the isolates from both fish and water samples produced gelatinase and nuclease but the ability to produce lipase, caseinase and haemolysins was found to vary among isolates from different sources. Among the 15 antibiotics to which the isolates were tested, all the isolates were found to be sensitive to chloramphenicol, ciprofloxacin and gentamicin and resistant to amoxycillin. Local aquarists maintain the fish in crowded stressful conditions, which could trigger infections by the obligate/ opportunistic pathogenic members among motile aeromonads

Antibiotic resistance pattern of motile aeromonads from farm raised fresh water fish

Motile aeromonads isolated from the intestines of farm-raised freshwater fish such as Catla catla, Labeo rohita and Ctenopharyngodon idella have been characterized to species level. Morphological and physiological grouping revealed 61% Aeromonas hydrophila, 30% Aeromonas caviae, 7% Aeromonas sobria and 2% which remained unidentified. Hemolytic activity was detected mostly in A. hydrophila, while only half of the A. sobria and A. caviae showed this activity. Antibiotic resistance patterns of the strains revealed that they had acquired a relatively higher resistance to oxytetracycline, amoxycillin, ampicillin, novobiocin and polymixin-B, implicating possible use of these antibiotics in the aquaculture systems.

Distribution, extracellular virulence factors and antibiogram of motile aeromonads in fresh water ornamental fishes and immune response of Cyprinus carpio against Aeromonas hydrophila infection

The thesis deals with the prevalence and distribution of motile aeromonads in selected ornamental fishes. The presence of motile aeromonads in ornamental fishes and associated carriage water is well documented. Though aeromonads are a part of autochthonous flora of natural waters, disease outbreak occurs as a result of environmental stress on the cultured species and virulence of the pathogens. While ornamental aquaculture in many parts of the world is highly organized and practiced scientifically, it is highly unorganized in India. The culture ponds/tanks are often maintained in very poor manner and the fishes are subjected to high degree of stress during transportation from the production facility to retail vendors. The situation is no better at retail outlets, where fishes are maintained in crowded condition without proper aeration or food. All these could result in high prevalence of diseases caused by motile aeromonads. No systematic study has been carried out to understand the prevalence of motile aeromonads in ornamental fishes and carriage water . It also gives an account of the production of extracellular virulence factors and the antibiogram of the different species of motile aeromonads isolated. The growth characteristics and virulence potential of a representative strain of Aeromonas hydrophila is also studied. The nucleotide sequencing of the strain was carried out and sequences deposited in Genbank. Survival and immune response of Cyprinus carpio under different stress conditions and on probiotic treatment with Bacillus NL110, when challenged with A. hydrophila is also dealt within this thesis.

Systematic study of the impact of fresh water fluxes on the glacial carbon cycle

During glacial periods, atmospheric CO2 concentration increases and decreases by around 15 ppm. At the same time, the climate changes gradually in Antarctica. Such climate changes can be simulated in models when the AMOC (Atlantic Meridional Oceanic Circulation) is weakened by adding fresh water to the North Atlantic. The impact on the carbon cycle is less straightforward, and previous studies give opposite results. Because the models and the fresh water fluxes were different in these studies, it prevents any direct comparison and hinders finding whether the discrepancies arise from using different models or different fresh water fluxes. In this study we use the CLIMBER-2 coupled climate carbon model to explore the impact of different fresh water fluxes. In both preindustrial and glacial states, the addition of fresh water and the resulting slow-down of the AMOC lead to an uptake of carbon by the ocean and a release by the terrestrial biosphere. The duration, shape and amplitude of the fresh water flux all have an impact on the change of atmospheric CO2 because they modulate the change of the AMOC. The maximum CO2 change linearly depends on the time integral of the AMOC change. The different duration, amplitude, and shape of the fresh water flux cannot explain the opposite evolution of ocean and vegetation carbon inventory in different models. The different CO2 evolution thus depends on the AMOC response to the addition of fresh water and the resulting climatic change, which are both model dependent. In CLIMBER-2, the rise of CO2 recorded in ice cores during abrupt events can be simulated under glacial conditions, especially when the sinking of brines in the Southern Ocean is taken into account. The addition of fresh water in the Southern Hemisphere leads to a decline of CO2, contrary to the addition of fresh water in the Northern Hemisphere.

Climatic impacts of fresh water hosing under Last Glacial Maximum conditions: a multi-model study

Fresh water hosing simulations, in which a fresh water flux is imposed in the North Atlantic to force fluctuations of the Atlantic Meridional Overturning Circulation, have been routinely performed, first to study the climatic signature of different states of this circulation, then, under present or future conditions, to investigate the potential impact of a partial melting of the Greenland ice sheet. The most compelling examples of climatic changes potentially related to AMOC abrupt variations, however, are found in high resolution palaeo-records from around the globe for the last glacial period. To study those more specifically, more and more fresh water hosing experiments have been performed under glacial conditions in the recent years. Here we compare an ensemble constituted by 11 such simulations run with 6 different climate models. All simulations follow a slightly different design, but are sufficiently close in their design to be compared. They all study the impact of a fresh water hosing imposed in the extra-tropical North Atlantic. Common features in the model responses to hosing are the cooling over the North Atlantic, extending along the sub-tropical gyre in the tropical North Atlantic, the southward shift of the Atlantic ITCZ and the weakening of the African and Indian monsoons. On the other hand, the expression of the bipolar see-saw, i.e., warming in the Southern Hemisphere, differs from model to model, with some restricting it to the South Atlantic and specific regions of the southern ocean while others simulate a widespread southern ocean warming. The relationships between the features common to most models, i.e., climate changes over the north and tropical Atlantic, African and Asian monsoon regions, are further quantified. These suggest a tight correlation between the temperature and precipitation changes over the extra-tropical North Atlantic, but different pathways for the teleconnections between the AMOC/North Atlantic region and the African and Indian monsoon regions.