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Treponema Cultivation

Treponema Cultivation

Treponema is a single-celled Helicobacter. Measured with an electron microscope, it is 5-20 microns in length and 0.09-0.5 microns in width; it has a tight regular or irregular helix. Cells have one or more axofilaments inserted into the ends of the protoplasmic cylinder. Macrovesicles and spheroids of spirochetes can be seen in old cultures.

Treponema is a motile bacterium and is Gram-negative. When dyed with the silver attack method and Ryu's stain, the dyeing effect is good. Most species are not easily stained by Giemsa's method. Best viewed under a darkfield or phase contrast microscope. Most of the descriptions of the following genera relate only to some of the species that can be cultured in the test tube.

Treponema is a chemoheterotroph. Through fermentative metabolism, it can utilize amino acids and/or carbohydrates.

Figure 1. Plaque samples of Treponema cells.Figure 1. Plaque samples of Treponema cells. (Xuedong Zhou, et al.; 2015)

Treponema is a strictly anaerobic bacteria. Contact enzyme, oxidase and urine enzyme were all negative, and V.P. reaction was negative. The bacteria do not reduce nitrate and are not inhibited by iodoacetic acid.

Treponema is commonly found in the oral, intestinal and genital areas of humans and some animals. Some species are pathogenic bacteria.

Treponema Culture Service

Creative Biogene offers customizable Treponema strain culture services. This service can help you obtain Treponema 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.

Treponema pallidum

Treponema pallidum has elongated helical cells, tightly coiled, 6-20μm long and 0.09-0.18μm wide. The average length is 10-13 microns, and the average width is 0.13-0.15 microns. The ends of the cells are pointed. Three axofilaments are inserted at both ends of the cell. The bacterium moves in a slow drifting and flexing manner, rarely turning.

Pathogenic to humans and apes. Toxic strains (Nichols pathogenic bacteria and Gand strains, etc.) are propagated in rabbit testes. Subcutaneous inoculation of voles, mice and guinea pigs had no apparent infection or visible lesions. The virulent strains were able to move for 4-7 days under anaerobic conditions at 25°C although in medium containing ultrafiltrate of protein, sodium bicarbonate, pyruvate, cysteine and serum.

This strain can cause congenital and sexual syphilis in humans. Common in syphilis lesions.

Treponema pertenue

Treponema pertenue is an elongated helical bacterium that is morphologically indistinguishable from Treponema pallidum. The bacteria can move.

Poisonous strains can reproduce by scratching the skin of a large vole or rabbit. Skin lesions were produced on inoculation sites in voles and rabbits but not guinea pigs or mice. Toxic strains could not be cultured in vitro.

Treponema pertenue can cause yaws in humans. It was found in the lesions of the Yaws case, which is an infectious disease transmitted by contact. The disease is common in tropical countries such as Africa, Southeast Asia, some islands in the Western Pacific, and some tropical countries in the Americas.

Treponema carateum

Treponema carateum has elongated helical cells and is morphologically similar to Treponema pallidum. The bacteria can move.

The virulent strains were not successfully cultivated in vitro. Experimental infection in humans and chimpanzees has been accomplished using subcutaneous inoculation and direct exposure of human ruptured foci to the lacerated skin area. The development of lesions takes 35-70 days.

Treponema carateum causes pinta or carate disease in humans. Commonly found in the lymph of other skin lesions, it is a contagious disease mainly confined to Mexico, Central America, subtropical parts of South America, West India and Cuba.

Treponema paraluis-cuniculi

Treponema paraluis-cuniculi has elongated helical cells and is morphologically similar to Treponema pallidum. The bacteria can move.

Treponema paraluis-cuniculi has not been cultured in vitro. The bacteria can be propagated by inoculation in the testis of rabbits. In mice, guinea pigs, and large voles can cause latent infection, and Treponema has been found in the lymph nodes. Skin lesions were found only in guinea pigs and rabbits. Produces mild sexual spiral disease in rabbits. The bacteria were isolated from genital and perineal lesions in rabbits. Although it usually occurs around the face, eyes, ears, and nose, it mainly involves the genitals.

Treponema phagedunis

The spiral-shaped cells of Treponema phagedunis are 5-15 μm long and 0.24-0.40μm wide. The widest cells reach 0.40 μm and show a double outline by darkfield observation. The ends of the cells are sharper. Three axofilaments are inserted at both ends of the cell. Axofilaments protruding from the ends of cells are seen in aged cultures.

Treponema macrodentium

Treponema macrodentium is Helicobacter elongated, 5-16 microns long and 0.1-0.25 microns wide. The ends of the cells are pointed, and an axofilament is inserted into the respective ends of the cells. Exercise in fairly rapid movements. Young cells rotate rapidly along their long axis.

Grow in peptone-yeast extract medium or PPLO medium (BBL) containing 10% serum or ascites fluid containing cocarboxylase 5 μg/ml, glucose 1 mg/ml and cysteine 1 mg/ml. Animal serum is required for this bacterial growth. Serum can also be used in place of this requirement with isobutyric acid 20 µg/ml, spermine 150 µg/ml and niacinamide 400 µg/ml. Supplementation of rumen fluid and cocarboxylase in the medium also allowed growth. During the cultivation process, a fermentable carbohydrate is required as an energy source.

Treponema macrodentium ferments carbohydrates, producing acid but not gas. The final pH in the dextrose broth was 5.0-5.4. The bacteria can ferment fructose, glucose, maltose, ribose, cellobiose, galactose and xylose and sucrose. The final product of glucose fermentation is large amounts of lactic acid, moderate amounts of formic and acetic acids, and small amounts of succinic acid.

The optimum temperature of the bacteria is 37°C, and the optimum growth pH is 7.0. The bacteria can be isolated from the crevices of person's gums.

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 Treponema 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.

References

  1. Bergey's Manual of Systematic Bacteriology Book Review Int. J. of Syst. Bact.1985, p. 216.
  2. Xuedong Zhou, et al.; Atlas of Oral Microbiology, Subgingival Microbes. Academic Press. 2015, Pages 67-93.
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