Systems, devices, and methods for separating and washing microorganisms
The present disclosure provides materials and methods for separating organisms from various other components of a composition. In particular, the present disclosure provides devices, systems, and methods for separating and washing helminths from contaminants such as bacteria.
1. A method of separating microorganisms, the method comprising:
a) applying a composition comprising a plurality of microorganisms and one or more contaminants to a device comprising:
a column having a first end for receiving the composition and a second end distal to the first end,
a fluid inlet port proximal to the second end of the column for receiving washing fluid, and
a waste outlet port proximal to the first end of the column;
b) applying a washing fluid to the device via the fluid inlet port; and
c) actively transporting the washing fluid and contaminants contained therein to the waste outlet port using a pump as the plurality of microorganisms passes from the first end to the second end of the column counter to the flow of the washing fluid, thereby separating the plurality of microorganisms from contaminants in the composition.
2. The method of claim 1 , wherein the first end comprises a first opening for receiving the composition and wherein the second end comprises a second opening for collecting the plurality of organisms, wherein the first opening has a diameter greater than the second opening.
3. The method of claim 1 , wherein the column comprises:
a slow flow section proximal to the first end of the column, wherein the slow flow section is defined by a first diameter,
a rapid flow section proximal to the second end of the column, wherein the rapid flow section is defined by a second diameter that is less than the first diameter; and
a tapered wash hold section positioned between the slow flow section and the rapid flow section.
4. The method of claim 3 , wherein the tapered wash hold section comprises a single tapered segment.
5. The method of claim 3 , wherein the tapered wash hold section comprises two or more tapered segments that taper towards the second end of the column.
6. The method of claim 1 , wherein the device further comprises a valve positioned between the fluid inlet port and the second end.
7. The method of claim 1 , wherein the device further comprises a removable cap that covers the first end.
8. The method of claim 1 , wherein the device further comprises a pump in fluid connection with the fluid inlet port for actively transporting the washing fluid from the fluid inlet port towards the waste outlet port.
9. The method of claim 1 , wherein the device further comprises a removable collection vial positioned at the second end of the column.
10. The method claim 1 , wherein the device further comprises a solid support for holding the column in a vertical position.
11. The method of claim 1 , wherein the organism separation device is comprised in a system including a plurality of organism separation devices, each device comprising components a), b), and c) as described in claim 1 .
12. The method of claim 11 , wherein the devices in the system are arranged in parallel.
13. The method of claim 11 , wherein the system further comprises a pump in fluid connection with the fluid inlet port for two or more of the devices in the system, wherein the pump actively transports the washing fluid from the fluid inlet ports to the waste outlet ports of the two or more devices.
14. The method of claim 1 , comprising:
a) applying the composition to the first end of the column while the washing fluid is actively transported from the fluid inlet port to the waste outlet port at a first flow rate;
b) reducing the flow rate from the fluid inlet port to a second flow rate; and
c) collecting the plurality of organisms from the second end of the column;
wherein the plurality of organisms have been separated from at least one other component in the washing fluid.
15. The method of claim 14 , wherein the washing fluid comprises at least 0.5% protein (w/w).
16. The method of claim 14 , wherein the plurality of organisms are helminths.
17. The method of claim 16 , wherein the helminths are Hymenolepis diminuta cysticercoids (HDCs).
18. The method of claim 16 , wherein the plurality of helminths collected from the second end of the column are substantially free of bacteria.
19. A composition comprising separated helminths obtained by the method of claim 14 .