Spirillum Cultivation

Spirillum cells are tough, helical, 0.25-1.7 microns in diameter, with less than one to many turns of helix. Most species have polyhydroxybutyrate particles in their bodies. The genus moves in tufted polar hairs. All but one species (S. delicatum) have bipolar flagella.

Spirillum is Gram-negative. It belongs to the organic chemotrophic bacteria, and carries out strict respiratory metabolism, and oxygen is the final electron acceptor. A few species can live in the presence of nitrates.

Figure 1. Spirillum. (From wikipedia)

The genus generally does not produce acid from sugars, and even in rare cases positive reactions are evident only in medium with low peptone concentrations. Sugar is usually not the only carbon source. The genus does not hydrolyze lorin and hippurate. It is negative for indole, sulfatase, amylase and phenylalanine deaminase. In addition, this genus does not normally liquefy gelatin, but can utilize cysteine to produce hydrogen sulfide. The genus was positive for oxidase and contactase. A few species are urease positive. Sometimes contact enzymes are weakly positive. Phosphatase is usually positive.

Nearly one-half of the species produce fluorescent water-soluble pigments with a yellowish-green color. Nutritional requirements are simple, and most species do not require amino acids, vitamins, purines or pyrimidines. Various organic acids, alcohols or amino acids can be used as the sole carbon source. For marine bacteria, a certain amount of seawater should be added to the medium. Spirillum has an optimum temperature of 30°C and does not grow at 10°C or 45°C. Freshwater bacteria do not grow when the NaCl concentration is above 3%. And some do not grow even at a low concentration of 0.3%.

Spirillum is commonly found in fresh and salt water containing organic matter.

Spirillum Culture Service

Creative Biogene offers customizable Spirillum strain culture services. This service can help you obtain Spirillum cultures for subsequent scientific research. This service allows you to skip the complicated and tedious groping of culture conditions, which helps to speed up research on this strain.

Spirillum volutans

Spirillum volutans is the largest spirochete. Bacteria cultured in peptone succinate (PSS) nutrient medium for 18 hours were observed by phase contrast microscopy, and the cell diameter was 1.4-1.7μm; wavelength 16-28μm; helix diameter 5-8 μm; helix length 14 -60μm, the helix number of cells can be less than one turn to a maximum of five turns. Dark granules of polyhydroxybutyrate were particularly evident in the cytoplasm.

Spirillum volutans can move. Viewing live cells with microscopy, it is easy to see that each end has about 75 bundles of flagella. The flagellar bundles at the front and rear of the thallus exhibited high-speed rotation, forming directional conical turns. Reversing this orientation is accompanied by cell movement in the opposite direction. Oriented anterior and posterior flagellar bundles cooperate with each other, and the coordination is deregulated by various compounds.

It is difficult to isolate tumble spirochetes, and so far it can only be isolated by Rittenberg's capillary method. The bacteria grow poorly on nutrient gravy. Flocculent abundant, cloudy growth in peptone succinate (PSS) nutrient solution.

Until now, it has not been possible to cultivate with a fully defined medium. When the peptone component in the PSS medium was replaced with 2.5 g of acid hydrolyzed casein without vitamins and salts, a rich culture could be grown. Synthetic casein hydrolyzate cannot replace acid hydrolyzate, even if added with vitamins, synthetic peptides or trace inorganic salts. Growth was inhibited when the concentration of phosphate added to PSS liquid medium was greater than 0.003 mol.

Spirillum volutans are obligate micro-oxygen, requiring an atmosphere containing 1-9% oxygen. If there is no such condition, 0.15% agar can be used to solidify the liquid layer and the growth can also be obtained.

Optimum temperature for Spirillum volutans 30°C; poor growth at 25°C and 37°C; no growth at 10°C or 42°C. It was isolated from a mixed culture collected from stagnant ponds in Virginia, USA and from the cooling tower water of a beet sugar factory in England.

Spirillum giesbergeri

Using phase contrast microscopy to observe the culture in PSS nutrient solution for 18 hours, the diameter of Spirillum giesbergeri cells is 1.1-1.4μm, the diameter of the helix is 0.2-5.0μm; the length of the helix is 4-40μm, and the spiral pattern of the cells is less than one circle. Dark granules of tight hydroxybutyrate were particularly evident in the cytoplasm. When observed in vivo with dark field, the flagellar tufts at both ends of the cells can be seen. Growth was moderate in PSS nutrient solution, with sediment at the bottom, clear supernatant, and slow growth. Colonies on PSS agar (1.5% agar) were round, white, raised and 1.0 mm in size. No growth on most of the various individual carbon sources in defined inorganic salts medium; however, in defined medium containing succinate with the addition of 0.15% agar, the subsurface of the medium A long band is formed at 3-6mm. Growth bands did not migrate to the surface even after continued incubation.

With heat-labile antigens, this species was sero-reactive with S. graniferum and S. sinuosum. The bacteria can grow aerobic, but the most suitable conditions are microaerobic. Its optimum temperature is 30°C; moderate growth at 25°C; poor growth at 37°C; no growth at 10°C or 42°C. The bacteria were isolated from pond water.

Microbial GMP Production

Creative Biogene's fermentation platform has Good Manufacturing Practices (GMP) and can provide customers with a wide range of high-quality microbial fermentation products such as active pharmaceutical ingredients, enzymes and various fine chemicals. In addition, our microbiology experts have completed the transformation and innovation of traditional processes through continuous breakthroughs in key technologies of microbial fermentation processes, and fully contributed to the smooth delivery of the project.

Production Capacity

Creative Biogene builds a world-class microbial fermentation technology platform, providing a variety of services from strain screening and optimization to fermentation production and product purification. We have many years of rich experience and provide good technical support for microbial GMP production.

Facility Display

As a leader in microbial production, Creative Biogene has comprehensive production process technology and high-volume manufacturing capabilities. Our goal is to help our customers develop streamlined and controlled manufacturing processes and to support customers throughout the entire product development process, from the R&D stage to market launch.

Device Example:

• Fermentation, centrifugation and filtration upstream process equipment;
• Fully automatic fermenters ranging in volume from 4,000L to 12,000L with a total capacity of over 100,000 liters;
• From industrial-scale chromatography systems, membrane systems to larger-scale continuous centrifuges;
• Recycling and Downstream Equipment;
• Waste treatment equipment.

Why Choose Us?

The culture of Spirillum requires specific formulations of growth media for use in cloning, plasmid DNA preparation, and protein expression. Creative Biogene offers a selection of bacterial growth media and custom services for your specific application. If you are interested in our microbial anaerobic and aerobic culture platform, please contact us for more details.