Journal
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume 237, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.molbiopara.2020.111279
Keywords
Plasmodium; Malaria; Liver stage; Sporozoites; Elizabethkingia; In vitro culture
Categories
Funding
- National Institute of Allergy and Infectious Diseases, USA [R21 AI137748]
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Studies of Plasmodium sporozoites and liver stages require dissection of Anopheles mosquitoes to obtain sporozoites for experiments. Sporozoites from the rodent parasite P. yoelii are routinely used to infect hepatocytes for liver stage culture, but sometimes these cultures become contaminated. Using standard microbiological techniques, a single colony type of Gram-negative rod-shaped bacteria was isolated from contaminated cultures. Mass spectrometry and sequencing of the bacterial 16S ribosomal RNA gene identified the contaminant as Elizabethkingia spp. Based on sequence comparison and published studies of the Anopheles microbiome, the best match was E. anophelis. Culture contamination was not ameliorated by density gradient purification of sporozoites. However, the addition of vancomycin to the culture media consistently reduced contamination and improved culture outcomes as measured by liver stage parasite size. Thus, mosquito salivary gland-derived E. anophelis is identified a potential contaminant of Plasmodium liver stage cultures that can be mitigated by the addition of antibiotics.
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