Improving Mycoplasma ovipneumoniae culture medium by a comparative transcriptome method

Abstract

Mycoplasma ovipneumoniae (Mo) is difficult to culture, resulting in many difficulties in related research and application. Since nucleotide metabolism is a basic metabolism affects growth, this study conducted a "point-to-point" comparison of the corresponding growth phases between the Mo NM151 strain and the Mycoplasma mycoides subsp. capri (Mmc) PG3 strain. The results showed that the largest difference in nucleotide metabolism was found in the stationary phase. Nucleotide synthesis in PG3 was mostly de novo, while nucleotide synthesis in NM151 was primarily based on salvage synthesis. Compared with PG3, the missing reactions of NM151 referred to the synthesis of deoxythymine monophosphate. We proposed and validated a culture medium with added serine to fill this gap and prolong the stationary phase of NM151. This solved the problem of the fast death of Mo, which is significant for related research and application.

Isolation and Characterization of Mycoplasma ovipneumoniae Infecting Goats with Pneumonia in Anhui Province, China

“…Mycoplasmas in the samples were isolated from nasal swabs through cultivation on pleuro-pneumonia-like organism (PPLO) agar and broth using previously described methods [29,30]. Nasal swabs and pleural fluid samples were cultivated in 15 mL conical tubes containing PPLO broth (21 g/L; Qingdao Hopebio Biotechnology Co., Ltd., Qingdao, China) supplemented with heat-inactivated horse serum (200 mL/L; Gibco, Grand Island, NY, USA), 25% fresh yeast extract (100 mL/L; Oxoid, Hampshire, UK), polymyxin B sulfate (3 units/mL; Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China), penicillin G (200,000 IU/L; Shanghai Macklin Biotechnology Co., Ltd., Shanghai, China), D-glucose (2 g/L), pyruvic acid sodium salt (2 g/L), and 0.4% phenol red (0.18 mL; Beijing Solarbio Technology Co., Ltd., Beijing, China).…”

DNA hybridization-induced fluorescence variation in ThT: a new strategy of developing aqueous sensors for MO genes

“…7 However, traditional microbial culture techniques suffer from limitations such as complicated operation and high consumption of time and labor. 8,9 Therefore, serologic testing via the enzyme-linked immunosorbent assay has gradually replaced these techniques because of its rapidity, sensitivity, simplicity, and ease of standardization. Unfortunately, its specificity is controversial.…”