| Autor: |
Goh CBS; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu.; Tropical Medicine and Biology Multidisciplinary Platform, Monash University Malaysia, Bandar Sunway, 47500 Malaysia., Goh CHP; School of Medical and Life Sciences, Sunway University, Bandar Sunway, 47500 Malaysia., Wong LW; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu., Cheng WT; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu., Yule CM; School of Science and Engineering, University of the Sunshine Coast, Queensland, 4556, Australia., Ong KS; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu., Lee SM; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu.; Tropical Medicine and Biology Multidisciplinary Platform, Monash University Malaysia, Bandar Sunway, 47500 Malaysia., Pasbakhsh P; School of Engineering, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. pooria.pasbakhsh@monash.edu., Tan JBL; School of Science, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. tan.ban.lee@monash.edu.; Tropical Medicine and Biology Multidisciplinary Platform, Monash University Malaysia, Bandar Sunway, 47500 Malaysia. |
| Abstrakt: |
The full plethora of environmental bacteria is often poorly represented in vitro as the majority remain difficult, if not impossible, to culture under standard laboratory settings. These bacteria often require native conditions for the formation of cell masses that collectively have higher chances of survival. With that, a 3D-printed version of the isolation chip (iChip) was used to cultivate bacteria from a tropical peat swamp in situ prior to growth and maintenance in vitro . Briefly, plates made from either acrylonitrile butadiene styrene (ABS), polylactic acid (PLA), or epoxy resin were tested in terms of their usability and durability under acidic conditions similar to those of peat matter. The epoxy resin plates were then found to be most optimal for the sampling conditions. Peat soil samples were collected from the base of a Koompassia malaccensis tree and reconstituted in molten 10% (wt/vol) tryptone soy agar (TSA) prior to inoculation. The iChips were subsequently assembled and buried in the site of origin. As a comparison, bacteria from the same soil sample were cultivated directly on TSA and incubated at 28 °C for two weeks. Thereafter, agar plugs from the iChip were transferred to TSA plates to allow microcolonies within each plug to grow. Each pure isolate from both cultivation approaches that grew was then pooled and extracted for total DNA prior to 16S rRNA gene amplification and sequencing via Illumina MiSeq. Taxonomic abundance comparison revealed that the bacterial taxa at the level of order were significantly different between the two approaches, particularly in the orders, Burkholderiales , Xanthomonodales , Enterobacteriales , and Actinomycetales (differences of 12.0, 7.1, 8.0, and 4.2%, respectively). This indicated that the 3D-printed iChips present a possible low-cost tool for the isolation of bacterial genera that may not be able to grow on media directly in vitro . |
