PMID- 33592223
OWN - NLM
STAT- MEDLINE
DCOM- 20211001
LR  - 20211001
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 182
DP  - 2021 Mar
TI  - Visualization of probiotics via epifluorescence microscopy and fluorescence in 
      situ hybridization (FISH).
PG  - 106151
LID - S0167-7012(21)00019-1 [pii]
LID - 10.1016/j.mimet.2021.106151 [doi]
AB  - Aerobic plate counts, the standard for bacterial enumeration in the probiotic 
      industry, can be biased towards fast-growing organisms that replicate on 
      synthetic media and can significantly underestimate total bacterial abundance. 
      Culture-independent approaches such as fluorescence in situ hybridization (FISH) 
      hold promise as a means to rapidly and accurately enumerate bacteria in probiotic 
      products. In addition, FISH has the potential to more accurately represent 
      bacterial growth dynamics in the environment in which products are applied 
      without imposing additional growth constraints that are required for enumeration 
      via plate counts. In this study, we designed and optimized three new FISH probes 
      to visualize and quantify Bacillus amyloliquefaciens, Bacillus pumilus, and 
      Bacillus licheniformis within probiotic products. Microscopy-based estimates were 
      consistent or higher than label claims for Pediococcus acidilactici, Pediococcus 
      pentosaceus, Lactobacillus plantarum, Bacillus subtilis, Bacillus 
      amyloliquefaciens, Bacillus licheniformis and Bacillus pumilus in both a direct 
      fed microbial (DFM) product as well as a crop microbial biostimulant (CMB) 
      product. Quantification with FISH after a germination experiment revealed the 
      potential for this approach to be used after application of the product.
CI  - Copyright (c) 2021 Elsevier B.V. All rights reserved.
FAU - Pasulka, Alexis L
AU  - Pasulka AL
AD  - Center for Applications in Biotechnology, California Polytechnic State 
      University, 1 Grand Ave., San Luis Obispo, CA 93407, USA; Biological Sciences 
      Department, California Polytechnic State University, 1 Grand Ave., San Luis 
      Obispo, CA 93407, USA. Electronic address: apasulka@calpoly.edu.
FAU - Howes, Amy L
AU  - Howes AL
AD  - Biological Sciences Department, California Polytechnic State University, 1 Grand 
      Ave., San Luis Obispo, CA 93407, USA.
FAU - Kallet, Julia G
AU  - Kallet JG
AD  - Biological Sciences Department, California Polytechnic State University, 1 Grand 
      Ave., San Luis Obispo, CA 93407, USA.
FAU - VanderKelen, Jennifer
AU  - VanderKelen J
AD  - Center for Applications in Biotechnology, California Polytechnic State 
      University, 1 Grand Ave., San Luis Obispo, CA 93407, USA.
FAU - Villars, Clayton
AU  - Villars C
AD  - Biological Sciences Department, California Polytechnic State University, 1 Grand 
      Ave., San Luis Obispo, CA 93407, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210214
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
SB  - IM
MH  - Bacillus/*ultrastructure
MH  - Fluorescence
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Microscopy/*methods
MH  - *Probiotics
OTO - NOTNLM
OT  - Bacterial enumeration
OT  - Epifluorescence microscopy
OT  - Fluorescence in situ hybridization
OT  - Probiotics
EDAT- 2021/02/17 06:00
MHDA- 2021/10/02 06:00
CRDT- 2021/02/16 20:09
PHST- 2021/01/07 00:00 [received]
PHST- 2021/01/28 00:00 [revised]
PHST- 2021/01/28 00:00 [accepted]
PHST- 2021/02/17 06:00 [pubmed]
PHST- 2021/10/02 06:00 [medline]
PHST- 2021/02/16 20:09 [entrez]
AID - S0167-7012(21)00019-1 [pii]
AID - 10.1016/j.mimet.2021.106151 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2021 Mar;182:106151. doi: 10.1016/j.mimet.2021.106151. Epub 
      2021 Feb 14.