Supplemented culture medium to promote the growth and isolation of Spiroplasma melliferum

The study of microorganisms requires controlled conditions that allow their growth outside their natural host, making culture media fundamental in microbiology. These media provide essential nutrients for microbial survival, proliferation, and isolation. Spiroplasmas, are helical and motile bacteria belonging to the class Mollicutes, exhibit a marked metabolic dependence due to the absence of a cell wall and the loss of multiple biosynthetic pathways. For this reason, there in vitro culture it is possible to use media supplemented with animal serum and complex extracts, such as LD8 medium and its variants, which supply compounds that these bacteria cannot synthesize. Supplementation of culture media with yeast extract, serum, and cholesterol is necessary to ensure cell membrane integrity, viability, and growth of fastidious microorganisms such as Spiroplasmas. The use of enriched media has enabled the isolation and physiological, genomic, and molecular characterization of Mollicutes, facilitating the understanding of their symbiotic interactions with hosts. However, culturing Spiroplasmas remains a challenge due to their high nutritional requirements. The present study aimed to evaluate different precursors to promote the growth and isolation of Spiroplasma melliferum.