Extremophiles are microorganisms which have adapted so that they can survive and even thrive in conditions that are normally fatal to most life-forms. For example, some species have been found in the following extreme environments: Temperature: as high as 130 °C (266 °F),as low as −17 °C (1 °F) Acidity/alkalinity: less than pH 0, up to pH 11.5 Salinity: up to saturation Pressure: up to 1,000-2,000 atm, down to 0 atm (e.g. vacuum of space) Radiation: up to 5kGy Extremophiles are significant in different ways. They extend terrestrial life into much of the Earth's hydrosphere, crust and atmosphere, their specific evolutionary adaptation mechanisms to their extreme environment can be exploited in bio-technology, and their very existence under such extreme conditions increases the potential for extraterrestrial life. • Grow at o °c. • Optimum temperature 15 °c or lower. • Maximum 20 °c Habitats :Isolated Artic and Antartic habitats. (90% of the ocean is 5°C or colder) Examples :• Arthrobacter sp., • Psychrobacter sp. PSYCHROTROPHS: • Legionella. Psychrophiles Halophiles can be found anywhere with a concentration of salt five times greater than the salt concentration of the ocean, • Great Salt Lake (Utah) • Owens Lake (California) • Dead Sea, • Evaporation Ponds. Halobacterium sp. strain NRC-1, each cell about 5 μm in length. • Adapted extreme hypertonic environment. • Grow optimally,Presents of Nacl or other salts. • Sea water contain 35% mixes with fresh water- nearly 0%. • E.x: Halobacterium. Halococcus Halobacterium- Halococcus. Phylum: Extreme Halobacterium Halobacterium sp. strain NRC-1, each cell about 5 μm in length. Domain: Kingdom: Scientific classification Archaea Euryarchaeota Phylum: Euryarchaeota Class: Order: Halobacteria Halobacteriales Family: Halobacteriaceae Genus: Halobacterium • • • • H. jilantaiense H. noricense H. salinarum H. piscisalsi Binomial name Halobacterium Elazari-Volcani 1957 Species In taxonomy, Halobacterium is a genus of the Halobacteriaceae Domain: Kingdom: Phylum: Class: Order: Family: Genus: • • • • • • • Halococcus Scientific classification Archaea Euryarchaeota Euryarchaeota Halobacteria Halobacteriales Halobacteriaceae Halococcus Binomial name Halococcus Schoop 1935 Species H. dombrowskii H. hamelinii H. morrhuae H. qingdaogense H. saccharolyticus H. salifodinae H. thailandensis Halococcus is a genus of the Halobacteriaceae. • Adapted completely hypertonic saline condition • Require high level Nacl Salinity: It is remarkably constant throughout the deep sea. Some minor differences in salinity, but none that are ecologically significant, except in the Mediterranean & Red seas. • Below the thermocline, the water mass of the deep ocean is cold and far more homogeneous. • temperature of the epipelagic zone, is above 20°C. • based on the epipelagic, it drops over several hundred meters to 5 or 6°C at 1,000 meters. • Affects growth of microbs • High temperature damages microbes by denaturing enzymes ,transport carriers and other proteins • Microorganisms can placed in 5 clanes based on temperature ranges THERMOPHILES: Grow at 55 ° c or higher. Minimum -45 ° c . Optimum-55 c to 65 °c. • Present in a planet’s surface from which Geothermally “Heated Water Issues”. • Commonly found near volcanically active places, ocean basins & hotspots. A colony of thermophiles in the outflow of Mickey Hot Springs, • It forms some features in under the sea called Oregon, “Black Smokers”. the water temperature is approximately 60°C. • • • • HYPERTHERMOPHILES: Grow at 90°c. Maxima above 100°c. Don ‘t grow below 55 °c.Microorganisms in deep-sea hydrothermal plumes:• Hydrothermal vents vary considerably, from relatively low-temperature (<25 °C) fluid discharges to the spectacular high-temperature (~350 °C) • black smokers1–3. • The high-temperature vents give rise to buoyant plumes which can be detected hundreds of kilometres away from ridge crests4,5. BLACK SMOKERS: It found on sea bed, typically in the abyssal & hadal zones. In the immediate vicinity of hydrothermal vents, chemoautotrophic bacteria are present in vent fluids, attached to rock surfaces9,10, and as endosymbionts in certain macro fauna11 WHITE SMOKERS: • Contain barium, calcium & silicon. • Vent organism depend on chemosynthetic bacteria for food. • It contains huge number of bacteria. PRESSURE: Prokaryotes live in deep sea. Hydrostatic pressure -600 to 1100 atm and temperature -2 to 3 °c. Greatest environmental factors acting on deep sea organism. it increases 1 atmosphere (atm) for each 10 m in depth. In deep sea is under pressures between 200 and 600 atm, the range of pressure is from 20 to 1,000 atm. Microbes live on land or surface water – 1atm Can play a major role in nutrient recycling in deep sea. E.g : Photobacterium , Shewanella. • Existing below the “thermocline” & above seabed. • It is very icy and dark at the bottom. • Sunlight can’t reach there but most of the deep sea produced light that can be seen easily in dark • deep sea or deep layer in the ocean existing below the thermoline and allow the seabed depth of 1000 or more • Most organisms falling organic matter produced in the photic zone Definition: Nitrification is the biological oxidation of ammonia with oxygen into nitrite followed by the oxidation of these nitrites into nitrates. Nitrification in the marine environment: • In the marine environment, nitrogen is often the limiting nutrient • The nitrification step of the cycle is of particular interest in the ocean because it creates nitrate, the primary form of nitrogen responsible for “new” production. • Furthermore, as the ocean becomes enriched in anthropogenic CO2, the resulting decrease in pH could lead to decreasing rates of nitrification. Nitrification as stated above is formally a two-step process. First Step : ammonia is oxidized to nitrite, Nitrification is a process of nitrogen compound oxidation : NH3 + 11/2 O2 + Nitrosomonas -------→ NO2- + H2O + H+ NO2- + 1/2O2 + Nitrobacter ------------→ NO3- NH3 + O2 -----------------------------------→ NO2− + 3H+ + 2e− NO2− + H2O ------------------------------→ NO3− + 2H+ + 2e− Second Step : nitrite is oxidized to nitrate. Different microbes are responsible for each step in the marine environment. Several groups of ammonia oxidizing bacteria (AOB) are known in the marine environment Example::Bacteria :Nitrosomonas, Nitrospira, and Nitrosococcus. Nitrospina and Nitrobacter are known to carry out this step in the ocean. All contain the functional gene ammonia monooxygenase (AMO) which, as its name implies, is responsible for the oxidation of ammonia. Definition:Denitrification is a microbially facilitated process of nitrate reduction that may ultimately produce molecular nitrogen (N2) through a series of intermediate gaseous nitrogen oxide products. Measurement:denitrification rates were measured in sediment cores from a MARINE of the Chesapeake Bay using high precision membrane inlet mass spectrometry. Denitrification was independent of salinity over the range of 1-13 ppt and directly dependent on nitrate concentration over the range of 0-200 µM in the overlying water. Denitrification was observed when the water colunm nitrate concentration was <1 µM, indicating that nitrification in the sediments was occurring. 4KNO3+502---------- 2K2 O + 2N2 molecules of oxygen are consumed for each molecule of nitrogen evolved, but the amount of oxygen liberated at each stage is different: 1. Nitrate to nitrite. 2KNO3+ 02 - --- + 2KNO2 (ii) Nitrite to hyponitrite. 2KNO2+ 02 -- -- +2 KN202. (iii) Hyponitrite to nitrogen. 2K2N202 + 02 - -- + 2K2O + 2N2 • Methanogenesis or biomethanation is the formation of methane by microbes known as methanogens. • Organisms capable of producing methane have been identified only from the kingdom Archaea, a group phylogenetically distinct from both eukaryotes and bacteria, although many live in close association with anaerobic bacteria. • The production of methane is an important and widespread form of microbial metabolism. • In most environments, it is the final step in the decomposition of biomass. Strains of methanogens • • • • • • • • • • • • • • • • • • • • Methanobacterium bryantii Methanobacterium formicum Methanobrevibacter arboriphilicus Methanobrevibacter gottschalkii Methanobrevibacter ruminantium Methanobrevibacter smithii Methanocalculus chunghsingensis Methanococcoides burtonii Methanococcus aeolicus Methanococcus deltae Methanococcus jannaschii Methanococcus maripaludis Methanococcus vannielii Methanocorpusculum labreanum Methanoculleus bourgensis (Methanogenium olentangyi & Methanogenium bourgense) Methanoculleus marisnigri Methanofollis liminatans Methanogenium cariaci Methanogenium frigidum Methanogenium organophilum • • • • • • • • • • • • • • • • Methanogenium wolfei Methanomicrobium mobile Methanopyrus kandleri Methanoregula boonei Methanosaeta concilii Methanosaeta thermophila Methanosarcina acetivorans Methanosarcina barkeri Methanosarcina mazei Methanosphaera stadtmanae Methanospirillium hungatei Methanothermobacter defluvii (Methanobacterium defluvii) Methanothermobacter thermautotrophicus (Methanobacterium thermoautotrophicum) Methanothermobacter thermoflexus (Methanobacterium thermoflexum) Methanothermobacter wolfei (Methanobacterium wolfei) Methanothrix sochngenii Ammonification: When a marine plant or animal dies, or an animal expels waste, the initial form of nitrogen is organic. Bacteria, or fungi in some cases, convert the organic nitrogen within the remains back into ammonium (NH4+), a process called ammonification or mineralization. Enzymes Involved: GS: Gln Synthetase (Cytosolic & PLastid) GOGAT: Glu 2-oxoglutarate aminotransferase (Ferredoxin & NADH dependent) GDH: Glu Dehydrogenase: Minor Role in ammonium assimilation. Important in amino acid catabolism. cycle • Sulfur is one of theSulfur constituents of many proteins, vitamins and hormones. It recycles as in other biogeochemical cycles. • The essential steps of the sulfur cycle are: • Mineralization of organic sulfur to the inorganic form, hydrogen sulfide: (H2S). • Oxidation of sulfide and elemental sulfur (S) and related compounds to sulfate. • Reduction of sulfate to sulfide. • Microbial immobilization of the sulfur compounds and subsequent incorporation into the organic form of sulfur. Dimethylsulfoniopropionate (DMSP):- Formula:(CH3)2S+CH2CH2COO−. This zwitterionic metabolitefound :Marine Phytoplankton, Seaweeds, Species Of Terrestrial And Aquatic Vascular Plants. Functions:Osmolyte Physiological and environmental roles. Degradation:DMSP is broken down by marine microbes to form two major volatile sulfur products. Ismethanethiol (CH3SH), Dimethyl Sulfide (CH3SCH3; DMS). Its major breakdown product Ismethanethiol (CH3SH), assimilated by bacteria into protein sulfur. Its second volatile breakdown product :Dimethyl Sulfide (CH3SCH3; DMS) DMSP DMSP lyase DMS Most DMS in seawater is cleaved from DMSP by the enzyme DMSP lyase, although many non-marine species of bacteria convert methanethiol to DMS DMS is also taken up by marine bacteria, but not as rapidly as methanethiol. Although DMS usually consists of less than 25% of the volatile breakdown products of DMSP, the high reactivity of methanethiol makes the steady-state DMS concentrations in seawater approximately 10 times those of methanethiol (~3 nM vs. ~0.3 nM). Curiously, there have never been any published correlations between the concentrations of DMS and methanethiol. This is probably due to the non-linear abiotic and microbial uptake of methanethiol in seawater, and the comparatively low reactivity of DMS. However, a significant portion of DMS in seawater is oxidized to dimethyl sulfoxide (DMSO). Relevant to global climate, DMS is thought to play a role in the Earth's heat budget by decreasing the amount of solar radiation that reaches the Earth's surface. DMSP has also been implicated in influencing the taste and odour characteristics of various products. For example:DMSP is odourless and tasteless, it is accumulated at high levels in some marineherbivores or filter feeders. Increased growth rates, vigour and stress resistance among animals cultivated on such diets have been reported. DMS, is responsible for repellent, 'off' tastes and odours that develop in some seafood products because of the action of bacterial DMSP-lyase, which cogenerates acrylate. Formula:- (CH3)2SO • This colorless liquid is an important polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible in a wide range of organic solvents • organosulfur compound • It penetrates the skin very readily, giving it the unusual property for many individuals of being secreted onto the surface of the tongue after contact with the skin and causing a garlic-like taste in the mouth. Synthesis and production:• It was first synthesized in 1866 by the Russian scientist Alexander Zaytsev • Oxidation of dimethyl sulfide with oxygen or nitrogen dioxide gives DMSO Biological Use:• DMSO is used in PCR to inhibit secondary structures in the DNA template or the DNA primers. • It is added to the PCR mix before reacting, where it interferes with the selfcomplementarity of the DNA, minimizing interfering reactions. • DMSO may also be used as a cryoprotectant, added to cell media to prevent cell death during the freezing process. Medicine:Use of DMSO in medicine dates from around 1963, when an Oregon Health & Science University Medical School team, headed by Stanley Jacob, discovered it could penetrate the skin and other membranes without damaging them and could carry other compounds into a biological system. In medicine, DMSO is predominantly used as a topical analgesic, a vehicle for topical application of pharmaceuticals, as an anti-inflammatory, and an antioxidant. Because DMSO increases the rate of absorption of some compounds through organic tissues, including skin, it can be used as a drug delivery system. It is frequently compounded with antifungal medications, enabling them to penetrate not just skin but also toe and fingernails. Veterinary medicine:DMSO is commonly used in veterinary medicine as a liniment for horses, alone or in combination with other ingredients. In the latter case, often, the intended function of the DMSO is as a solvent, to carry the other ingredients across the skin. Also in horses, DMSO is used intravenously, again alone or in combination with other drugs. It is used alone for the treatment of increased intracranial pressure and/or cerebral edema in horses. In cryobiology DMSO has been used as a cryoprotectant and is still an important constituent of cryoprotectant vitrification mixtures used to preserve organs, tissues, and cell suspensions. Without it, up to 90% of frozen cells will become inactive. It is particularly important in the freezing and long-term storage of embryonic stem cellsand hematopoietic stem cells, which are often frozen in a mixture of 10% DMSO, Media and 30% fetal bovine serum. In the cryogenic freezing of heteroploid cell lines (MDCK, VERO, etc.) a mixture of 10% DMSO with 90% EMEM (70% EMEM + 30% fetal bovine serum + antibiotic mixture) is used. As part of an autologous bone marrow transplant the DMSO is re-infused along with the patient's own hematopoietic stem cells. In a 1978 study at the Cleveland Clinic Foundation in Cleveland, Ohio, researchers concluded that DMSO brought significant relief to the majority of the 213 patients with inflammatory genitourinary disorders that were studied. They recommended DMSO for all inflammatory conditions not caused by infection or tumor in which symptoms were severe or patients failed to respond to conventional therapy. DMSO has been examined for the treatment of numerous conditions and ailments, but the U.S. Food and Drug Administration (FDA) has approved its use only for the symptomatic relief of patients with interstitial cystitis. In interventional radiology, DMSO is used as a solvent for Ethylene Vinyl Alcohol in the Onyx liquid embolic agent, which is used in embolisation, the therapeutic occlusion of blood vessels. Safety:DMSO by itself has low toxicity. DMSO's use in reducing brain tissue swelling following traumatic brain injury. DMSO exposure to developing mouse brains can produce brain degeneration. This neurotoxicity could be detected at doses as low as 0.3 mL/kg, a level exceeded in children exposed to DMSO during certain medical treatments. Glove selection is important when working with DMSO. Thick rubber gloves are ecommended. Nitrile gloves, which are very commonly used in chemical laboratories, have been found to dissolve rapidly with exposure to DMSO. Because DMSO easily penetrates the skin, substances dissolved in DMSO may be quickly absorbed. For instance, a solution of sodium cyanide in DMSO can cause cyanide poisoning through skin contact. it contains Dimethyl sulfoxide can produce an explosive reaction when exposed to acyl chlorides; at a low temperature, this reaction produces the oxidant for Swern oxidation. DMSO disposed into sewers can also cause odor problems in municipal effluents: waste water bacteria transform DMSO under hypoxic (anoxic) conditions into dimethyl sulfide (DMS) that has a strong disagreeable odor, similar to rotten cabbage.[20] However, chemically pure DMSO is odorless because of the lack of C-S-C (sulfide) and C-S-H (mercaptan) linkages. Deodorization of DMSO is achieved by removing the odorous impurities Marine Microbiology- 2Mark Questions Explain ammonification process Is the process by which the organically bound nitrogen of microbial, plant, and animal biomass is recycled after their death. Ammonification is carried out by a diverse array of microorganisms that perform ecological decay services, and its product is ammonia or ammonium ion. Dimethyl Sulfonio Propionate (DMSP) Is a zwitterionic metabolite found in marine phytoplankton, seaweeds and some species of terrestrial and aquatic vascular plants. DMSP is broken down by marine microbes to form two major volatile sulfur products, each with distinct effects on the environment. Its major breakdown product is methanethiol (CH3SH) which is assimilated by bacteria into protein sulfur. Dimethyl Sulfoxide (DMSO) • Is an abundant but poorly understood methylated sulfur compound in the marine environment. One potentially significant loss pathway for DMSO is through its biological reduction to dimethylsulfide (DMS), which has been documented in a number of organisms, most notably bacteria. Nitrogen cycle • The nitrogen cycle is the process by which nitrogen is converted between its various chemical forms. This transformation can be carried out via both biological and non-biological processes. Important processes in the nitrogen cycle include fixation, mineralization, nitrification, and denitrification. cycle • Sulfur is one of theSulfur constituents of many proteins, vitamins and hormones. It recycles as in other biogeochemical cycles. The essential steps of the sulfur cycle are: • Mineralization of organic sulfur to the inorganic form, hydrogen sulfide: (H2S). • Oxidation of sulfide and elemental sulfur (S) and related compounds to sulfate. • Reduction of sulfate to sulfide. • Microbial immobilization of the sulfur compounds and subsequent incorporation into the organic form of sulfur. Organisms that can fix atmospheric nitrogen • Free living bacteria: Aerobic (Azotobacter, Klebsiella); Anaerobic (Desulfovibrio, some • Clostridium species) • Cyanobacteria: (Nostoc common in lichen) • Symbionts = live in symbiosis with certain plants :with legumes (Rhizobium); with other plants (Frankia, Azospirillium) REFERENCES: • Net reference.