PMID- 35305414
OWN - NLM
STAT- MEDLINE
DCOM- 20220408
LR  - 20220503
IS  - 1873-3336 (Electronic)
IS  - 0304-3894 (Linking)
VI  - 432
DP  - 2022 Jun 15
TI  - Stalking Mycobacterium bovis in the total environment: FLOW-FISH & FACS to 
      detect, quantify, and sort metabolically active and quiescent cells in complex 
      matrices.
PG  - 128687
LID - S0304-3894(22)00476-9 [pii]
LID - 10.1016/j.jhazmat.2022.128687 [doi]
AB  - Mycobacterium bovis causes tuberculosis (TB) at the human-wildlife-livestock 
      interface. Environmental persistence of M. bovis excreted by infected hosts may 
      cause indirect transmission to other animals. However, methodological constrains 
      hamper assessment of M. bovis viability and molecular signature in environmental 
      matrices. In this work, an innovative, modular, and highly efficient single-cell 
      workflow combining flow cytometry (FLOW), fluorescence in situ hybridization 
      (FISH), and fluorescence-activated cell sorting (FACS) was developed, allowing 
      detection, quantification, and sorting of viable and dormant M. bovis cells from 
      environmental matrices. Validation with spiked water and sediments showed high 
      efficiency (90%) of cell recovery, with high linearity between expected and 
      observed results, both in cell viability evaluation (r(2) =0.93) and 
      FISH-labelled M. bovis cells quantification (r(2) >/=0.96). The limit of detection 
      was established at 10(5) cells/g of soil in the cell viability step and 10(2) 
      cells/g of soil in the taxonomical labelling stage. Moreover, FACS efficiency 
      attained noteworthy recovery yield (50%) and purity (60% viable cells; 70% 
      taxonomically labelled M. bovis). This new methodology represents a huge step for 
      M. bovis assessment outside the mammal host, offering the rapid quantification of 
      M. bovis cell load and cell viability, including viable but non-culturable cells, 
      and further downstream cell analyses after FACS. Subsequent environmental data 
      integration with the clinical component will expand knowledge on transmission 
      routes, promising new paths in TB research and an intervention tool to mitigate 
      the underlying biohazard.
CI  - Copyright (c) 2022 Elsevier B.V. All rights reserved.
FAU - Pereira, Andre C
AU  - Pereira AC
AD  - Centre for Ecology, Evolution and Environmental Changes (cE3c), Faculdade de 
      Ciencias, Universidade de Lisboa, Lisboa, Portugal; Biosystems & Integrative 
      Sciences Institute (BioISI), Faculdade de Ciencias, Universidade de Lisboa, 
      Lisboa, Portugal.
FAU - Tenreiro, Ana
AU  - Tenreiro A
AD  - Biosystems & Integrative Sciences Institute (BioISI), Faculdade de Ciencias, 
      Universidade de Lisboa, Lisboa, Portugal.
FAU - Tenreiro, Rogerio
AU  - Tenreiro R
AD  - Biosystems & Integrative Sciences Institute (BioISI), Faculdade de Ciencias, 
      Universidade de Lisboa, Lisboa, Portugal.
FAU - Cunha, Monica V
AU  - Cunha MV
AD  - Centre for Ecology, Evolution and Environmental Changes (cE3c), Faculdade de 
      Ciencias, Universidade de Lisboa, Lisboa, Portugal; Biosystems & Integrative 
      Sciences Institute (BioISI), Faculdade de Ciencias, Universidade de Lisboa, 
      Lisboa, Portugal. Electronic address: mscunha@fc.ul.pt.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220312
PL  - Netherlands
TA  - J Hazard Mater
JT  - Journal of hazardous materials
JID - 9422688
RN  - 0 (Soil)
SB  - IM
MH  - Animals
MH  - Animals, Wild/microbiology
MH  - Flow Cytometry
MH  - In Situ Hybridization, Fluorescence
MH  - Mammals
MH  - *Mycobacterium bovis/genetics
MH  - Soil
OTO - NOTNLM
OT  - Contaminated environment
OT  - Flow cytometry
OT  - Fluorescence in situ hybridization
OT  - Fluorescence-activated cell sorting
OT  - Indirect transmission
OT  - Mycobacterium bovis
EDAT- 2022/03/20 06:00
MHDA- 2022/04/09 06:00
CRDT- 2022/03/19 20:08
PHST- 2022/01/29 00:00 [received]
PHST- 2022/02/27 00:00 [revised]
PHST- 2022/03/09 00:00 [accepted]
PHST- 2022/03/20 06:00 [pubmed]
PHST- 2022/04/09 06:00 [medline]
PHST- 2022/03/19 20:08 [entrez]
AID - S0304-3894(22)00476-9 [pii]
AID - 10.1016/j.jhazmat.2022.128687 [doi]
PST - ppublish
SO  - J Hazard Mater. 2022 Jun 15;432:128687. doi: 10.1016/j.jhazmat.2022.128687. Epub 
      2022 Mar 12.