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Open Access Highly Accessed Research article

A multi-laboratory profile of Mycoplasma contamination in Lawsonia intracellularis cultures

Jeong-Min Hwang1, Ji-Hye Lee2 and Jung-Yong Yeh2*

Author Affiliations

1 Veterinary Research Center, Green Cross Veterinary Products Co., Ltd., 227-5, Kugal-dong, Giheung-gu, Yongin-si, Gyeonggi-do 446-903, Republic of Korea

2 Foreign Animal Diseases Division, Animal, Plant, and Fisheries Quarantine and Inspection Agency, Anyang-ro 175, Manan-gu, Anyang-si, Gyeonggi-do 430-824, Republic of Korea

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BMC Research Notes 2012, 5:78  doi:10.1186/1756-0500-5-78

Published: 27 January 2012

Abstract

Background

During the routine laboratory cultivation of Lawsonia intracellularis, Mycoplasma contamination has been a frequent problem. When Mycoplasma contamination occurs in laboratories that study L. intracellularis, the cultures must be discarded for 4 reasons: 1) Mycoplasma is inevitably concentrated along with L. intracellularis during the passage of L. intracellularis; 2) Mycoplasma inhibits the growth of L. intracellularis; and 3) it is impossible to selectively eliminate Mycoplasma in L. intracellularis cultures. In this study, we observed the contamination of Mycoplasma species during L. intracellularis cultivation among multiple laboratories.

Results

The presence of a Mycoplasma infection in the L. intracellularis cultures was verified using polymerase chain reaction (PCR), and a sequence analysis of the partial 16S rRNA and 23S rRNA genes was performed. A PCR-based assay using genus-specific universal primers revealed that 29 (85.3%) of the 34 cultures were contaminated with Mycoplasma, including 26 with M. hyorhinis (89.2%), 2 with M. orale (6.9%), and 1 with M. fermentans (3.4%). The Mycoplasma contamination was not the result of infection with material of pig origin. McCoy cells, which are required for the cultivation of L. intracellularis, were also ruled out as the source of the Mycoplasma contamination.

Conclusions

In this study, M. hyorhinis was identified as the most common mollicute that contaminated L. intracellularis cultures. Whether L. intracellularis enhances the biological properties of Mycoplasma to promote infection in McCoy cells is not known. Because the McCoy cell line stocks that were used simultaneously were all negative for Mycoplasma, and the same worker handled both the McCoy cells to maintain the bacteria and the L. intracellularis cultures, it is possible that the L. intracellularis cultures are more vulnerable to Mycoplasma contamination. Taken together, these results suggest that continuous cultures of L. intracellularis must be tested for Mycoplasma contamination at regular intervals.

The GenBank accession numbers for the sequences reported in this paper are JN689375 to JN689377.