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Activities

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Marine Algal Biotechnology

Marine microalgae are primary producers capable of converting solar energy into energy rich organic bio-molecules which indeed forms the basis of energy cycle of ocean. They produce an array of bio-chemicals which has potential applications in food, medical and industry. Harnessing these resources through biotechnology and engineering approach will be of great use to meet the food and industrial requirement of the mankind. In order to harness this marine microalgal bioresources a large number of marine microalgal strains were isolated, from the near shore and offshore water of Indian sea and potential candidate species for production of lipid and nutracuticals has been identified under the marine microalgal biotechnology programme. Different types of mass culture systems, namely Tubular photobioreactor, Bubble column photobioreactor and Solar power operated raceway culture system were developed and tested for mass culture of marine microalgae. Pilot scale outdoor mass scale experiments for four species of high lipid yielding marine microalgae namely, Chlorella vulgaris,Chlorella sorokiniana, Chlorella pyrenoidosa and Neochlorisaquatica were carried out and a maximum biomass productivity of 0.76 g/L/d, with 25.53% lipid was achieved in C. sorokiniana., A cost effective DC powered electro-flocculation method with 98 % efficiency with a power consumption rate of 0.32 kWh/Kg of microalgal was developed for harvesting the microalgae.

A process for production of lutein from indigenous, high lutein producing marine Chlorella vulgaris cultured under mixotrophic culture condition in raceway pond with a maximum biomass productivity of 0.6 g/L/d and 12 mg/g lutein content was developed. Lutein is a commercially important nutraceutical used for preventing age related macular degeneration.. A simple method of production of pharmacoactive nutrient with significant nutritional value and potent biological activity against central nervous system and depression was also standardized from Chlorella vulgaris. A fatty acids-carotenoid complex (Such as oleic, linoleic acids and carotenoids like canthaxanthin and neoxanthin and cryptoxanthin and echinenone) with significant anti-tuberculosis effect (IC50- 5 µg/mL) against the multidrug strain (MDR) of Mycobacteriumtuberculosis was isolated from Chlorella vulgaris. Biodiesel (B100 and B10) produced by transesterification process from the algal lipid (oil) extracted for Chlorella vulgaris were tested for efficiency, engine performance as per ASTM standards. Test run of biodiesel (B-10) powered vehicle from Nellore to Chennai was successfully completed. Pilot scale paddle wheel operated 25 tonnes capacity raceway ponds was developed in the NIOT seafront facility at Pamanji AP for standardization of mass scale algal culture and downstream processes for production of nutraceuticals.

Marine micro algae biomass		Lutein

Distribution of marine minerals		Pharmacoactive nutrient		Oil extracted from algae

Marine Microbial  Biotechnology

Deep sea microbes are primitive form of life which lives under high hydrostatic pressure, low temperature, and sustain life without sunlight. Due to their unique adaptation, they are capable of producing a wide range of novel molecules which will be of medical, industrial and environmental importance. In the ever-expanding search for sources of novel biologically active molecules to combat the menace of emerging diseases and increasing drug resistance in human pathogens, the deep sea microbes have emerged as a new frontier of the resource. To explore the deep sea microbes the scientist has to negotiate with the extreme pressure and temperature that exist in the deep sea. The deep sea bacteria are primarily barophilic in nature and they cannot tolerate a drastic change in pressure. To circumvent this, the group has successfully designed and developed a novel 350 bar pressure rated Deep-sea Microbial Culture Facility consisting of high pressure retainable water sampler, and a pressure, temperature controlled serial dilution and fermenter system by which the deep sea barophilic microbes can be collected, cultured and studied for their native biochemical production in their in-situ living conditions. The group has isolated 348 barotolerantdeep sea microbes which includes bacteria, fungi, and actinomyces from 1000 – 4500 m depth and has successfully cultured species of the genus such as Streptomyces, Bacillus, Nigrospora, and Ascotricha at 100 - 250 bar pressure in the high pressure culture system.

Metagenomic analysis of uncultivable bacterial communities in 2000 m deep-sea sediment off Barren Island in the Andaman Sea using High-throughput 16S rRNA gene sequencing revealed the occurrence of more than 44,000 species-level OTUs representing high bacterial diversity belonging to Actinobacteria, Firmicutes, and Proteobacteria. The group has optimized the production of novel extracellular glutaminase-free L-asparaginase from marine actinobacteria Nocardiopsisalba and the yield was increased by two-fold using newly designed NIOT ASP media. Lipopeptide surfactant biosynthesis gene cluster from Bacillus licheniformis was heterologously expressed in E.coli and the production was increased to three-fold over that of the original strain. Novel bioactive secondary metabolites such as multiple prodiginines and tripyrrole producing Streptomyces species; Grisiofulvin and spirobenzofuran producing Nigrospora species: Pyrazine and piperazinedione producing Streptomyces olivaceus; Sequesterpenes derivatives producing Ascotricha species; Cyclic peptide producing Streptomyces fenghuangensis; Antimicrobial peptide producing Bacillus subtilis and Ectoine enzyme producing Bacillus clausii were isolated from the deep sea. The group further developed a Multiplex PCR kit for the real-time detection of three major virulent genes in Enterococcus faecalis from the environmental and food samples and developed a novel deep-sea bacterial consortium for the bioremediation of hydrocarbons.

High pressure retainable water sampler rossette		High pressure retainable water sampler

Deep sea yeast Hortaea werneckii N129A8 (LM)		Deep sea yeast Hortaea werneckii N129A8 (SEM)

Acinetobacter sp. isolated from deep sea		Actinobacteria isolated from deep sea

Fungus sp. isolated from deep sea		Actinobacteria isolated from deep sea

Open Sea Cage Culture

The marine capture fishery in India is nearing its maximum potential with 3.9 million productions against the estimated Maximum Sustainable Yield (MSY) of 4.45 mmt. There is a growing concern about the ability to produce enough sea foods to meet the widening demand. Ocean farming and cage culture is the one of the viable option to meet the increasing demand for fish protein. In order to develop and test reliable open sea cage suitable for India sea conditions, HDPE floating collar cages of 9 m diameter with multipoint mooring system were developed and tested in different environmental conditions namely protected bay North Bay at Port Blair, semi enclosed bay Olaikuda near Rameshwaram and open sea near Kothachathram and Tuplipalem, Nellore, A.P. In absence of large sale nursery rearing facility and availability of large number of stockable size fish seeds, nursery rearing of seabass and cobia fish fingerling in the indigenously developed 2 m diameter nursery cages deployed within the open sea cages was standardized. The grow-out protocols of 6 species (seabass, cobia, milk fish, parrot fish, carangids and siganids) of marine finfishes till harvest were standardized. Open sea cage culture demonstrations were conducted by involving traditional and marginal fishers at Olaikuda (Tamil Nadu), Kothachathram (Andhra Pradesh), North Bay (Port Blair) and Tuplipalam (Andhra Pradesh). The open sea cage systems at deployed at Tuplipalem successfully withstood the fury of cyclone ‘Vardha’ which made the landfall near the culture site with a wind velocity of 100-120 km/hr. NIOT is also in the process of developing and testing of advanced semi rigid and rigid submersible cages suitable of rough Indian sea condition. The group has identified potential suitable area for open sea cage culture through Geospatial analysis of Indian seas using 10 vital environmental parameters.

Open sea cage at Nort Bay – Port Blair		Cobia culture in open sea cages

Nursery cage in growout cages		Nursery rearing of sea bass in cages

Cobia harvested at Olaikuda, TN		Sea bass harvest- Olaikuda, TN

Parrot fish harvest- Olaikuda, TN		Rabbit fish harvest – Olaikuda, TN

Sea bass harvest at Kothachathram, A.P.		Cobia harvest at Tupilipalem, A.P.

Establishment of Ballast Water Treatment Technologies – Test Facility

The spread of invasive species is one of the greatest threats to many ecosystems in the world. In the marine realm, most of the bio-invasions have been attributed to ballast water used to stabilize ocean-going vessels. Marine discharge of ballast water causes spread of viruses, bacteria, microbes, small invertebrates, eggs, cysts and larvae of various species to the new habitats. Considering the gravity of the impact of ballast water in spreading bio-invasion, the International Maritime Organization (IMO) has initiated preventive measures to treat ballast water to remove potentially harmful biological material before discharging into a new habitat. As part of this, the Ballast Water Management Convention is effective since September 08, 2017. In the present scenario, there exists no land-based Ballast Water Treatment Technologies - Test Facility in this very important region of the Indian Ocean. National Institute of Ocean Technology is in the process of establishing the Ballast Water Treatment Technologies - Test facility on the east coast of India in NIOT Sea Front facility at Pamanji village, Vakadu Mandal, Nellore District, Andhra Pradesh. NIOT has already completed a design for the intake system and the test facility based on the guidelines suggested by IMO for the establishment of land-based ballast water test facility.

Fish aggregating Devices (FADs):

FADs are popular form of fisheries resource enhancement and management in many parts of the world. FADs help in increasing the fish catch and at the same time reducing the operational time and fuel cost. In view of this, NIOT developed FADs using high density polyethylene (HDPE) pipes of 710 mm OD and deployed 28 oceanic FADs at a depth of 1000 m to 2000 m around Lakshadweep group of Islands (the largest deployment of Oceanic FADs reported so far)and 18 number in Andaman and Nicobar Island. Subsequently, an increase in fish landing and fish variety was reported by the Lakshadweep administration.

FADs (28 Nos.) deployed at Lakshdweep		Assembling of FAD at NIOT Chennai

Emplacement of Artificial Reef along Odisha Coast

Artificial reef (AR) are man-made, underwater structure, typically built for the purpose of promoting marine life in areas of generally featureless bottom. Artificial reefs may also serve to improve hydrodynamics for surfing or to control beach erosion. AR could play an important role in conservation of biodiversity and fishery resources in near shore areas severely affected by destructive fishing activities like trawling and use of other destructive gears. Furthermore AR provides new fishing grounds for small scale artisanal fisheries and sport fishing. NIOT has designed and deployed 750 numbers of artificial reef (AR) structures (1 m x 1 m size) and deployed successfully in three places namely Penthakata, Chandrabhaga and Balinolcha along the Odisha coast for the Department of Fisheries, Government of Orissa through UNDP funding. Underwater survey indicated dense colonization of invertebrates on AR structures and congregation of large sized groupers, sweet lips and other reef fishes. The interaction with the traditional fishermen revealed the consistent increase in fish catch around the AR deployment sites near to the shore and thereby reduction in fuel usage and time consumption in search of fishes. The reef supports the traditional fishers and discourages trawling, thereby protecting the turtles and their nesting grounds.

Deployment of artificial reef in Odisha		Congregation of Grouper in artificial reef

Island resource information and marine environmental impact assessment

The developmental programmes to be undertaken in the islands necessitate striking a balance between ecology and resource use. The island resource information programme is primarily focused on the development of Geographical Information System (GIS) based island-wise resource inventory to provide a powerful platform to integrate multiple resource information in the form of multi-layered Geo-database. This will help the resource managers and policy makers to evaluate the natural resources in a holistic manner through a user-friendly and interactive interface to take appropriate decisions for various developmental activities in the islands. It can also be used as a powerful tool to synthesize all the data, for comprehensive interpretation and forecasting of marine environmental ecosystems and biodiversity of the islands.

Barren Island – DEM		Coral coverage-North Bay, Port Blair

Training:

The group has also conducted number of training programmes on cage culture of lobster, crabs, and marine fin fish, fabrication and deployment and maintenance of cages and SCUBA diving training programmes to coastal fisher and scientific community.

Lobster fattening Tharuvaikulam, T.N		Lobster fattening trainingTharuvaikulam

Mud crab fattening pens at Pulicate lake		Mud crab fattening training- Odisha

Mud crab fattening at Port Blair		Mud crab fattening training Havelock

Cage cutlture training at Tuplipalem		Cage cutlture training at Kollam

SCUBA diving training at Andaman		SCUBA diving training at Lakshdweep

FACILITIES:

Marine Biotechnology division at NIOT Chennai has various laboratories like, microalgal, micro biology, molecular biology, biochemistry, cell culture, ballast water treatment and testing, seawater chemistry, and isotope handling facility. These laboratories are equipped with state-of-the-art sophisticated equipments, such as HPLC, GC, GC-MS, LC-MS, FTIR, TOC Analyzer, Fast Column Chromatography, Beta and Gama counters, Fluorescent, UV/Vis Spectrophotometer, ELISA Reader, Gel documentation system, Gradient and Real Time PCR, Cryo microtome, ultra centrifuge, Industrial sonicator, Lyophylizer, High pressure fermentor, Photobiorector, Water purification systems, Phase contrast, Fluorescent up-right and inverted microscopes, Atomic Force Microscope, Scanning Electron Microscope, etc to carry out cutting edge research in the field of marine biology, molecular biology and biotechnology.

Atal Center for Ocean Science and Technology for Island is the field cum marine biology laboratory situated in sprawling 20 hectares campus encompassing 16 hectares aquaculture demonstration farm, and 2500 square meters of administrative cum laboratory complex at Minnie Bay, Port Blair. The center has seawater intake facility, jetty, OBM boats, various marine biological sampling gears, SCUBA diving equipment’s, and advance analytical equipment’s and laboratory for carrying out cutting edge research in marine biotechnology

Andaman and Nicobar Centre for Ocean Science and Technology (ANCOST) were renamed as “Atal Centre for Ocean Science and Technology for Islands” (ACOSTI), OSTI - NIOT was dedicated to the nation, by Hon’ble Minister Dr. Harsh Vardhan on 15^th September 2018.

Major Achievements:

Marine Algal Biotechnology

• Isolated more than 200 marine microalgal strains from the coastal and oceanic waters of Indian sea including Andaman and Nicobar and Lakshadweep Islands and screened for biomass, lipid and nutracueticals production.
• Designed, developed and tested Continuous Flow Bubble Column Reactor, Tubular Photobioreactor and solar powered paddle wheel operated Raceway system for mass culture of microalgae and standardised outdoor mass culture of three high lipid yielding marine microalgae, Chlorella vulgaris, C. sorokiniana and C. pyrenoidosa.
• Developed and optimized electro flocculation technique for harvesting biomass from large scale marine microalgae culture.
• Establishment of large scale algal culture facility at Pamanji village in Andhra Pradesh.
• Developed and optimized a process for production of lutein from marine microalgae Chlorella sorokiniana.
• Developed and standardized a process for production of pharmacoactive nutrient from the wet biomass of marine microalgae Chlorella vulgaris.

Marine Microbial Biotechnology

• Isolated and identified more than 348 piezo tolerant deep sea microbes which includes bacteria, fungi, and actinomyces from water and sediment samples collected from 1000-4500 m depth of the Arabian Sea, Bay of Bengal, Andaman Sea and Indian Ocean.
• Design and development of 350 bar pressure rated high pressure low temperature microbial culture facility with serial dilution system.
• Developed 600 bar rated high pressure retainable water sampler in collaboration with Royal Netherland Institute of Ocean Research (NIOZ).
• Antifungal and anticancer compounds (griseofulvin and spirobenzofuran derivatives) producing deep sea fungus Nigrospora sp. and low molecular weight (<6KD) broad spectrum anti-microbial peptide producing deep sea bacteria, Bacillus subtilis were successfully cultivated in high pressure and low temperature fermenter system up to 100 bar pressure
• The marine fungus Purpureocillium lilacinum which produce an anti-oxidant extracellular polysaccharide, an antifungal(4-methyl-5-thiazoleethanol) and an anticancer (5-pyrazine derivatives) compound, was isolated from 1000 m depth.
• Isolated a novel extracellular glutaminase-free L-asparaginase producing marine actinobacteria, Nocardiopsis alba and a lipopeptide surfactant producing sponge associated Bacillus licheniformis from the Andaman Sea. The surfactant production was increased three fold over that of the original strain by recombinant DNA technology.
• Developed novel consortia of deep sea microbes (Ruegeria sp., Oceanobacillus sp., Neisotobacter sp., Photobacterium sp., Pseudoalteromonas sp., Ruegeria sp., Exiguobacteriumsp.,Enterobacter sp., Haererehalobacter sp., and Acinetobacter sp.,) for biodegradation of complex hydrocarbons.
• Metabarcoding of deep sea sediment samples collected from 1000 - 3300 m depth to understand the microbial diversity using NGS approaches.

Open Sea Cage Culture

• Developed and deployed 9m diameter HDPE vertical collar type floating gravity cages with multipoint mooring in Olaikuda (Tamil Nadu), Kothachathram, Tuplipalem (Andhra Pradesh) and North Bay (Andaman Islands).
• Successfully demonstrated culture of marine finfishes such as the Asian Seabass (Latescalcarifer), cobia (Rachycentroncanadum), Pompano (Trachinotusblochii), Milkfish (Chanoschanos), Parrot fish (Scarusghobban)and Giant Travelly (Caranxigonobilis) in open sea cages.
• The concept of nursery rearing of marine finfish fingerlings in sea cages was introduced and demonstrated with sea bass seeds of 6g size with up to 90% survival and 24g weight gain over a period of 45 days.
• Geospatial mapping of the Indian coast has been carried out using satellite and ground verified data for identification of potential sites suitable for cage culture.
• Conducted hands on training on cage fabrication, deployment and culture of marine fin fish to fisherman self help group at Olaikuda Rameshwaram and Tuplipalem, Andhra Pradesh and Kollam Kerala.

Ballast water treatment technology-test facility

• Generation of baseline data of physico- chemical and biological parameters at sea front facility at the proposed Ballast water treatment test facility site at Pamanji village in Andhra Pradesh
• Obtained NABL accreditation for testing for sea water parameters for testing ballast water samples in accordance to the D2 standards.

Screening of Polyketides from Deep - Sea Microbes

• Polyketides belong to a diverse group of natural products with potent biological characteristics that includes antibacterial and antitumor activities. The groups of enzymes involved in synthesis of polyketides are polyketide synthases (PKSs), which are usually multi-enzyme complexes, vary according to their sequences, structures and catalytic mechanisms. Within other class of PKSs (Type I and Type II), Type III PKSs are simple homodimeric enzymes, which are structurally, and catalytically less complex yet produce diverse array of compounds, including pyrones, acridones, chalcones, stilbenes, phloroglucinols and alkyl resorcinols.
• To identity and annotate type III PKSs from sediment metagenome a local PKSDB was created. The Identical Protein Groups (IPGs) resource of NCBI contains a single entry for each protein in several sources at NCBI, including annotated coding regions in GenBank and RefSeq, also from SwissProt and PDB (Protein Data Bank). This allows us to get more targeted search results and quickly identify a protein of interest. A total of 7879 sequences of type III polyketide synthases (PKSs) were retrieved from IPG and a local database (PKSdb) was created to screen type III PKS from metagenome data. BLASTX of NIOT_S9 metagenomic sequences with locally created PKSdb reported significant hits for type III PKSs.
• Please CLICK here to download the PKSDB