BIOL/CSES 4684 - Pseudomonas

http://filebox.vt.edu/users/chagedor/biol_4684/Microbes/nitro.html

1. IDENTIFYING CHARACTERISTICS Cells stain gram negative. Aerobic. Obligate chemolithotroph. Short to long rods, appear in pairs or short chains, coccoid forms often observed. Motile (polar flagella) or non-motile. Has cytomembranes that occur in flattened vesicles in peripheral regions of cytoplasm. Growth range 5-30 degrees C. Optimum pH range 5.8-9.5. The percent G+C ranges from 47 to 50%. Oxidizes ammonia to nitrite (1st step in two step process of nitrification). Habitats: soil, sewage, freshwater and marine ecosystems, brackish environments. The above picture shows Nitrosomonas europaea under phase contrast microscopy (2,200X).

2. TAXONOMIC DESCRIPTION Nitrosomonas species belong to the family Nitrobacteraceae which contains two groups responsible for nitrification. The first group oxidizes ammonia to nitrite (Nitrosomonas belongs to this group), the second group oxidizes nitrite to nitrate. Nitrosomonas species grow chemolithotrophically using reduced inorganic nitrogen compounds as a source of energy. Some species of Nitrosomonas utilize urea by way of urease as an ammonia source. Metabolism is aerobic, NH4+ is the electron donor and O2 is the electron acceptor. Most species are motile with flagella located in their polar regions but some species are nonmotile. DNA homology experiments are used to determine the presence of further species. Distinguishing characteristics for the species that are used in identification are the G+C content of DNA, the shape and size of cells, salt requirement, urea utilization, the presence of carboxysomes, and cell protein patterns. The above picture showsNitrosomonas spp.under electron microscopy (39,000X).

3. ISOLATION AND ECOLOGY Isolation requires a selective media with ammonia as an electron donor and bicarbonate as the sole carbon source. Media requirements include an extensively washed high purity agar or a silica gel agar. The media has to be completely inorganic because growth is inhibited by the presence of organic materials. Also, growth is slow which enable overgrowth of contaminants. Isolation is extremely difficult and colonies formed are very small in diameter. Visible turbidity may not be seen even after extensive nitrification so the best means for growth detection is to assay for the production of nitrite. One to two weeks is needed for incubation, then the chemical assays will reveal successful enrichment and an attempt to isolate pure cultures can be made. Nitrosomonas species are found widely distributed in soil and water. They are present where considerable amounts of ammonia are available. Species grow well in lakes and streams that receive inputs of untreated and treated sewage. They also strive more in neutral and alkaline habitats because acidic environments inhibit nitrification. Ammonia tends to accumulate in anaerobic habitats since oxygen is required for ammonia oxidation, but nitrifying bacteria tend to develop at the thermocline in stratified lakes where ammonia and oxygen are both present. The above picture shows Nitrosomonas europea under electron microscopy (39,600X).

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http://classroom.sdmesa.edu/eschmid/Lecture15-Microbio.htm
Examples of Nitrifying Bacteria (a: Nitrobacter winogradskyi-2,500x; b: N. winogradskyi cytomembranes -213,000x; c: Nitrosomonas europaea-2,500x; d: cytoplasmic membranes in N. europaea)

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