In the last episode of Vinyl Worm (Part I), we mainly discussed the identification of some Vinyloids samples as bacteria, and some of the Vinyloids samples identified as Mycoplasma. Here are two typical vinyl bug identification results to share with you:
Fig. 1 Colony of Mycoplasma hynohis on solid medium
Identification results: Samples 7, 12, 18 identified as Mycoplasma hyorhinis
Size: 0.15-0.3um
Infecting the host: It infects the epithelial cells of the upper respiratory system of pigs and is transmitted through the oral-nasal route. Piggies can be infected by sows or their companions (asymptomatic carriers).
Contaminated cell pathway: It is speculated that trypsin is mainly extracted from the pancreas of pigs, purified crystals, and then made into lyophilized preparations, improper filtration and sterilization of pancreatic enzyme solution may carry Mycoplasma hypis suis, it is recommended that you buy well-known brand pancreatic enzymes to reduce the risk of contamination.
Fig. 2 Mycoplasma bovine colonies on solid medium
Identification results: Sample 23 identified as Mycoplasma bovis (Mycoplasma bovine);
Size: 0.15-0.3um
Infection host: mainly parasitic in cattle, the infection site is extensive, infecting the respiratory tract of cattle, causing pneumonia, otitis media, arthritis, conjunctivitis.
Contamination of cellular pathways: One of the main nutrients for cell proliferation is fetal bovine serum. Fetal bovine serum is derived from animals, and it is mainly filtered and sterilized by 0.1um membrane. If the blood source and filtration and sterilization are improper, it will bring potential mycoplasma contamination to the cells, so it is recommended that you buy a well-known brand of fetal bovine serum.
Next, we analyze, in the previous episode we introduced the common pollution characteristics of vinyl worm are active swimming, spinning shuttle, shaking in place, these three most common pollution phenomena. Unlike bacteria, mycoplasma do not have flagella that can be used to swim and are obviously unlikely to be either active swimming or spinning shuttle. The sample identified as mycoplasma contamination is most likely the case with a vinyl worm sample—the vinyl worm we see trembling in situ in the liquid. And mycoplasma is the smallest known prokaryotes, to be observed under an electron microscope, we can see that the black spots trembling in situ are most likely mycoplasma colonies.
First of all, we can first watch a few sets of shaking in place videos that may be mistaken for "vinyl worms":
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Video 1 Small black dots tremble in situ (L02 hepatocytes line contaminated with vinyl worm video)
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Video 2 The small black dot trembles in place
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Video 3 The small black dot vibrates in place
From these "vinyloid" contamination videos, we can summarize the morphological characteristics of vinyloids:
Almost all have black elliptical dots, some large and some small; Free outside the cell, trembling, if you look closely, you can find that it trembles in place.
In addition, in the analysis of cell contamination for customers, we also found that the so-called "black glue worm" pollution is really "unique". Here's a screenshot of our chat analytics:
Figure 3 Picture of a cell in a screenshot of a customer chat
Enlarge this image as follows:
Figure 4 The black dots in the red circle in the figure are most likely mycoplasma colonies
Next, for further confirmation, the customer sent another video:
Figure 5 Cell video in customer chat screenshot
Let's zoom in on the video and observe:
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Video 6 The small black dot trembles in place
Through the video, we can see that many oval-shaped small black dots are shaking in place. Previously, in our cell communication group, we asked the customer if he had done mycoplasma testing, and the customer said that he had done it and determined that it was mycoplasma contamination. In a separate dialog with the customer, I further confirmed with the customer whether it was mycoplasma contamination, and the screenshot is as follows:
Figure 7 Screenshot of chat with customer
Through this case study, we speculate that the so-called "black gum worm" contamination commonly known as trembling in situ is most likely mycoplasma contamination, and these trembling dots are most likely mycoplasma colonies. In the future, we will collect a large number of samples of small black dots that tremble in situ, do sequencing identification, further confirm our speculations with big data, and share these interesting results with everyone.
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This explanation is here, if you have cell culture problems, welcome to scan the code to join our WeChat group communication.