Download full-text PDF Read

Search full-text
for publications, Download
researchers, citation
or questions Copy
or linkDiscover by subject area Recruit researchers Join for free Login

Article PDF Available

Impact of humic acids on the colonic

microbiome in healthy volunteers Download full-text PDF

February 2017 · World Journal of Gastroenterology 23(5):885

Read full-text
DOI:10.3748/wjg.v23.i5.885
License · CC BY-NC 4.0

Authors:

Alexander Swidsinski
Charité Universitätsmedizin Berlin

Yvonne Dörffel Vera Loening-Baucke

Christoph Gille

Show all 9 authors

Citations (17) References (23)

Abstract

Discover the world's

AIM To test the effects of humic acids on innate microbial communities of the
colon. METHODS We followed the effects of oral supplementation with humic
research
acids (Activomin®) on concentrations and composition of colonic microbiome
20+ million members
in 14 healthy volunteers for 45 d. 3 × 800 mg Activomin® were taken orally for
10 d followed by 3 × 400 mg for 35 d. Colonic microbiota were investigated 135+ million publication
using multicolor fluorescence in situ hybridization (FISH) of Carnoy fixated and pages
paraffin embedded stool cylinders. Two stool samples were collected a week
prior to therapy and one stool sample on days 10, 31 and 45. Forty-one FISH 2.3+ billion citations
probes representing different bacterial groups were used. RESULTS The sum
concentration of colonic microbiota increased from 20% at day 10 to 30% by Join for free
day 31 and remained stable until day 45 (32%) of humic acid supplementation
(P < 0.001). The increase in the concentrations in each person was due to
growth of preexisting groups. The individual microbial profile of the patients
remained unchanged. Similarly, the bacterial diversity remained stable.
Concentrations of 24 of the 35 substantial groups increased from 20% to 96%.
Two bacterial groups detected with Bac303 (Bacteroides) and Myc657 (mycolic
acid-containing Actinomycetes) FISH probes decreased (P > 0.05). The others
remained unaffected. Bacterial groups with initially marginal concentrations (<
0.1 × 10⁹/mL) demonstrated no response to humic acids. The concentrations of
pioneer groups of Bifidobacteriaceae, Enterobacteriaceae and Clostridium
difficile increased but the observed differences were statistically not significant.
CONCLUSION Humic acids have a profound effect on healthy colonic
microbiome and may be potentially interesting substances for the development
of drugs that control the innate colonic microbiome.

Public Full-text 1

Available via license: CC BY-NC 4.0

Content may be subject to copyright.

Submit a Manuscript: http://www.wjgnet.com/esps/ World J Gastroenterol 2017 February 7; 23(5): 885-890

Help Desk: http://www.wjgnet.com/esps/helpdesk.aspx ISSN 1007-9327 (print) ISSN 2219-2840 (online)
DOI: 10.3748/wjg.v23.i5.885 © 2017 Baishideng Publishing Group Inc. All rights reserved.

ORIGINAL ARTICLE

ta
Obser
vional Study
Impact of humic acids on the colonic microbiome in healthy
volunteers

Alexander Swidsinski, Yvonne Dörffel, Vera Loening-Baucke, Christoph Gille, Anne Reißhauer, Önder Göktas,
Monika Krüger, Jürgen Neuhaus, Wieland Schrödl

Alexander Swidsinski, Vera Loening-Baucke, Christoph Gille, the use is non-commercial. See: http://creativecommons.org/
Anne Reißhauer, Önder Göktas, Laboratory for Molecular licenses/by-nc/4.0/
Genetics, Polymicrobial Infections and Biofilms, Department
of Medicine, Section of Gastroenterology, Hepatology and Manuscript source: Invited manuscript
Endocrinology, Charité Universitätsmedizin Berlin, 10098 Berlin,
Germany Corresp ondence to: Dr. Alexander Swidsinski, Laboratory
for Molecular Genetics, Polymicrobial Infections and Biofilms,
Yvonne Dörffel , Outpatient Clinic, Luisenstr. 11-13, Charité Department of Medicine, Section of Gastroenterology, Hepatology
Universitätsmedizin Berlin, 10117 Berlin, Germany and Endocrinology, Charité Universitätsmedizin Berlin, 10098
Berlin, Germany. alexander.swidsinski@charite.de
Monika Krüger, Jürgen Neuhaus, Wieland Schrödl, entre for Telephone: +49-30-450514003
Infectious Diseases, Faculty of Veterinary Medicine, University Fax: +49-30-450514933
of Leipzig, 04103 Leipzig, Germany
Received: October 4, 2015
Author contributions: Swidsinski A, Krüger M and Schrödl W Peer-review started: October 7, 2015
designed the study; Dörffel Y, Neuhaus J conducted the study; First decision: October 28, 2015
Loening-Baucke V and Göktas Ö critically revised and wrote the Revised: November 13, 2016
manuscript; Reißhauer A performed FISH, and Gille C analyzed Accepted: January 3, 2017
the data staistically. Article in press: January 3, 2017
Published online: February 7, 2017
Institutional review board statement: The investigations were
approved by the ethical review board of the Charite Hospital and
Institut für Bakteriologie und Mykolo gie An den Tierklini ken
Leipzig.
Abstract
Informed consent statement: All study participants were AIM
healthy volunteers working in the laboratory provided informed To test the effects of humic acids on innate microbial
written consent prior to study enrollment. communities of the colon.

Conflict-of-interest statement: There are no conflicts of METHODS

interest to report.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was
selected by an in-house editor and fully peer-reviewed by external
reviewers. It is distributed in accordance with the Creative
Commons Attribution Non Commercial (CC BY-NC 4.0) license,
which permits others to distribute, remix, adapt, build upon this
work non-commercially, and license their derivative works on
different terms, provided the original work is properly cited and
We followed the effects of oral supplementation
®
with humic acids (Activomin ) on concentrations
and composition of colonic microbiome in 14 healthy
®
volunteers for 45 d. 3 × 800 mg Activomin were
taken orally for 10 d followed by 3 × 400 mg for 35 d.
Colonic microbiota were investigated using multicolor
fluorescence in situ hybridization (FISH) of Carnoy
fixated and paraffin embedded stool cylinders. Two
stool samples were collected a week prior to therapy
and one stool sample on days 10, 31 and 45. Forty-

WJG|www.wjgnet.com 885 February 7, 2017|Volume 23|Issue 5|

Swidsinski A et al . Humic acids and colonic microbiome

one FISH probes representing different bacterial groups

were used.
RESULTS
The sum concentration of colonic microbiota in-
creased from 20% at day 10 to 30% by day 31 and
remained stable until day 45 (32%) of humic acid
supplementation (P < 0.001). The increase in the
concentrations in each person was due to growth of
preexisting groups. The individual microbial profile
of the patients remained unchanged. Similarly, the
bacterial diversity remained stable. Concentrations of
24 of the 35 substantial groups increased from 20%
to 96%. Two bacterial groups detected with Bac303
(Bacteroides ) and Myc657 (mycolic acid-containing
Actinomycetes ) FISH probes decreased (P > 0.05).
The others remained unaffected. Bacterial groups with
9 and mi crob ial ac tivi ty t hat tra nsfo rms the de ad
initially marginal concentrations (< 0.1 × 10 /mL)
demonstrated no response to humic acids. The con-
centrations of pioneer groups of Bifidobacteriaceae ,
Enterobacteriaceae and Clostridium difficile increased
but the observed differences were statistically not
significant.
CONCLUSION
Humic acids have a profound effect on healthy colonic
microbiome and may be potentially interesting sub-
stances for the development of drugs that control the
innate colonic microbiome.
Key words: Fluorescence in situ hybridization; Colonic
INTRODUCTION
All great ancient cultures were based on agriculture
for which soil quality and prevention of its exhaustion
were absolutely critical. Humus as an organic fecundity
substrate of the earth excited thinkers from the
ancient times and stimulated both solid research and
charlatanry. First descriptions of medical applications
can be found in Sanskrit and also ancient writings of
Rome and China. Despite nearly mystic reverence and
enormous interest, it is not before the early 1800s that
chemical characterization and description of humic
acids took place.
Humic substances are complex organic sub-
stances of soil, which are formed in the process of
humification. Humification involves natural chemical
remains of living things into humic substances. It is
the second greatest organic process on earth after
photosynthesis and is responsible for fossil coal, oil
deposits and others. Microorganisms utilize and break
down organic substances and lead to accumulation
of recalcitrant molecules. When microorganisms die,
they are themselves broken down and added to the
recalcitrant humic mass. The concurrent chemical-
physical polymerization modies humic substances in
an unpredictable matter. In all, the genesis of humic
substances can take hundreds or even thousands of
years and leads to high variety, unique composition
and extreme difculties in characterization of these .
[1,2]

microbiota; Colonic bioreactor; Humic acids; Healthy
volunteers; Oral supplementation
© The Author(s) 2017. Published by Baishideng Publishing
Group Inc. All rights reserved.
Core tip: Modern patients are increasingly interested
in natural medicinal products, which are often not
scientifically evaluated. Humins arise from organic
microbial degradation and are an important mediator
of microbial interactions in nature. Although used
for medical indications since ancient times, no
data exist on the impact of humins on the human
microbiome. Our investigations in healthy volunteers
show that orally applied humic acids increase the sum
concentrations of preexisting colonic microbiota from
20% to 30% without changes in the bacterial diversity
of the individual microbiome and may be a serious
amendment/alternative to fecal transplantation or
probiotics.
Swidsinski A, Dörffel Y, Loening-Baucke V, Gille C, Reißhauer
A, Göktas Ö, Krüger M, Neuhaus J, Schrödl W. Impact of humic
acids on the colonic microbiome in healthy volunteers. World J
Gastroenterol 2017; 23(5): 885-890 Available from: URL: http://
www.wjgnet.com/1007-9327/full/v23/i5/885.htm DOI: http://
dx.doi.org/10.3748/wjg.v23.i5.885
The growing interest of the modern society for
environmental and biological welfare refreshed the
attractiveness for implementation of humins. Gastro-
enterologists are often confronted with a wish of
patients to be treated with “natural” products and
asked for opinion on humic acids. The study of the
scientic literature reveals a large number of medical
tri al s wi th di et ary s upp le men ts of h umi c ac ids
conducted all over the world. The reported effects
include different, partially incoherent properties such
as anti-inammatory and immune-stimulatory as well
as analgesic, antimicrobial, antiviral/anti-HIV activity,
[1-3]
antioxidant and even stroke protective effe cts .
The striking eclecticism of the ndings and the lack of
systematic studies make it difcult to build an unbiased
opinion. Furthermore humic substances are distributed
under a wide variety of trade names and descriptions
in an unregulated market.
The colon is a central bio-fermenting organ degrading
digestive leftovers. Since microbial activity is central
in genesis and processing of humic acids, the innate
human microbial communities should be the main object
on which the effects of humic acids will be apparent.
Astonishingly, we found no data to this topic in the
literature. In order to close this gap, we investigated
®
the impact of orally applied Activomin (Pharmawerk
Weinboehla, Weinboehla, Germany) on concentrations

WJG|www.wjgnet.com 886 February 7, 2017|Volume 23|Issue 5|

Swidsinski A et al . Humic acids and colonic microbiome

and diversity of the human colonic microbiome.
®
Activomin is the only registered and standardized
humic acids preparation in Germany.
MATERIALS AND METHODS
Patients, subjects and samples
Fourteen healthy volunteers from the Laboratories of
Centre for Infectious Diseases, Faculty of Veterinary
Medicine, University of Leipzig and Laboratory
for Polymicrobial Infections and Biofilms, Charité
Universitäts Medizin Berlin (24-64 years of age, mean
39 years, 5 males and 9 females) have taken 3 ×
®
2 capsules (3 × 800 mg) Activomin orally for 10 d
followed by 3 × 1 capsule (3 × 400 mg) for 35 d. Two
stool samples were collected a week prior to therapy
and one stool sample on days 10, 31 and 45.
The study was approved by the ethics commission
of University of Leipzig. The collection of fecal samples
for uorescence in situ hybridization (FISH) diagnosis
of dysbiosis was approved by the ethics commission of
the Charité University Hospital.
FISH
Coloni c mic robiot a were invest igated using FISH
analysis of Carnoy fixated and paraffin embedded
[4]
stool cylinders . Multicolor FISH simultaneously using
3 differently stained FISH probes (C3 - orange, FITC-
dobe - g reen, C5 - dark red) and count erstained
with DAPI for DNA structures was performed on 4
µm longitudinally cut sections of punched-out stool
cylinders. Sections were placed on SuperFrost plus
slides.
A Nikon e600 fluorescence microscope was used.
The images were photo-documented with a Nikon
DXM 1200F color camera and software (Nikon, Tokyo,
Japan).
Bacteria were quantified using group specific C3
probes. The FITC marked universal probe was used
in each hybridization to evaluate the number of all
bacteria, C5 marked probes with a different to C3
probes specificity were used to exclude unspecific
binding. Only signals that hybridized with a specific
FISH probe and the universal FISH probe, but did not
hybridize with specific FISH probes from unrelated
bacterial groups, were evaluated.
Bacterial concentrations of homogeneous po-
pulations were enumerated visually in one of the 10
× 10 fields of the ocular raster corresponding to 10
µm × 10 µm of the section surface at magnication of
1000. This number was assigned to concentration of 1
9
× 10 bacteria/mL, which was most equivalent to the
[4]
calculation formula, which we had used previously .
In case of uneven distribution of bacteria over the
microscopic eld, the positive signals were enumerated
in ten elds of the ocular raster along the gradient of
distribution and divided by ten.
Investigated bacterial groups/FISH probes
Forty-one bacterial FISH probes were applied,
Table 1. The exact specification of the FISH probes
and hybridization conditions are available in public
[5]
resources . The names of the FISH probes are listed
according to abbreviations of the probeBase online
resource (http://www.microbial-ecology.net/probebase/
[6]
credits.asp). The Fprau probe is described in 6 .
The FISH probes were arranged in Table 1 to four
functional groups described previously: essential bacteria,
individual pioneer bacteria, individual substantial and
[7]
individual marginal or accidental bacteria .
Bacteria detected with EREC (mainly Roseburia),
Bac303 (Ba cte roi des ), Fp rau (F ae cal iba cte ri um
prausnitzii) probes are always present in healthy
human subjects and together contribute about half of
the colonic microbiome. They are obviously essential
for colonic bio-fermentation.
All other bacterial groups are individual, present only
in some of the subjects in substantial concentrations
9
(mean ≥ 0.1 × 10 /mL) or marginal concentrations
9
(mean < 0.1 × 10 /mL).
Four FISH probes including Bif153 (Bifidobac-
teriaceae), Cdif198 (Clostridium difficile), Ebac1790
(Enterobacteriaceae) and Clit135 (Clostridium litu-
seburense) represent individual bacterial groups
with pioneer function, which are found prevalent in
newborns, after antibiotic treatment and convalescence
patients, but are seldom found in low concentration in
healthy persons.
Statistical analysis
Differences between groups were evaluated using
the twosided t-Student U test. Data are presented
as mean ± SD, P <0.05 was considered statistically
signicant.
RESULTS
All participants completed the stool collection, even the
one man, who developed loose stools and bloating. No
other side effects were reported.
Humic acids induced changes of the microbiome
Table 1 summarizes changes in the mean concen-
trations of single bacterial groups prior to and during
®
supplementation with Activomin . Bacteria in the Table
1 are arranged to sets of essential, individual pioneer,
individual substantial and individual marginal bio-
fermenting groups.
The mean microbial concentration after 45 d of
supplementation of humic acids increased 14% in the
essential groups (P < 0.01), 28% (NS) in the individual
pioneer groups and 41% (P < 0.002) in the individual
substantial groups. The accidental bacterial groups
with initially marginal concentrations demonstrated no
response to humic acids.

WJG|www.wjgnet.com 887 February 7, 2017|Volume 23|Issue 5|

Swidsinski A et al . Humic acids and colonic microbiome

Table 1 Mean microbial concentrations (± SD) as detected with applied uorescence in situ hybridization probes (109 bacteria/mL)

Day 0 Day 10 Day 31 Day 45 Change in % P value

from day 0 to
day 45
Mean sum concentrations of all detected bacteria 85.4 ± 25.6 107.4 ± 15.6 123.7 ± 34.1 126.1 ± 50.1 ↑ 32% < 0.001
Essential all (n = 3) 36.2 ± 14.7 44.0 ± 5.1 42.7 ± 7.7 42.8 ± 9.0 ↑ 14% < 0.01
Erec (Eubacterium rectale, Clostridium coccoides group) 11.7 ± 6.9 17.1 ± 2.5 19 ± 4 17.7 ± 4.8 ↑ 30% < 0.001
Bac303 (Bacteroides) 12.9 ± 5.3 12.2 ± 5.7 9.5 ± 4.4 9.9 ± 5.0 ↓ 30% ns
Fprau (Faecalibacterium prausnitzii) 11.6 ± 5.9 14.7 ± 3.8 14 ± 3.6 14.7 ± 6.9 ↑ 21% ns
Individual pioneer
All (n = 4) 7.8 ± 5.0 11.5 ± 9.3 9.9 ± 6.8 10.9 ± 8.5 ↑ 28% ns
Ebac1790 Enterobacteriaceae 0.25 ± 0.8 0.6 ± 2.1 1.2 ± 2.3 1.1 ± 2.0 ↑ 72% ns
Cdif198 Clostridium difcile 0.04 ± 0.09 0.01 ± 0.03 0.3 ± 0.93 0.10 ± 0.03 ↑ 96% ns
Bif153 Genus Bidobacterium 7.1 ± 5.5 9.1 ± 8.6 7.7 ± 5.3 9.7 ± 7.2 ↑ 27% ns
Clit135 Clostridium lituseburense group including C. difcile 0.5 ± 0.86 0.7 ± 1.05 0.4 ± 1.08 0.4 ± 0.8 ↔ ns
Individual substantial mean > 0.1 × 109/mL
All (n = 28) 41.7 ± 17.3 51.4 ± 14.0 70.4 ± 28.8 71.6 ± 36.8 ↑ 41% < 0.002
ACI623 Acidaminococcaceae sp. (not the Selenomonas species) 1.4 ± 1.9 0.7 ± 0.8 1.6 ± 2.2 1.2 ± 1.6 ↔ ns
AKK406 Akkermansia 2.3 ± 3.7 2.8 ± 4.6 1.9 ± 2.9 2.6 ± 4.1 ↔ ns
Ato291 Atopobium cluster 3.8 ± 2.9 4.9 ± 3.5 6.1 ± 4.5 6.4 ± 3.6 ↑ 41% 0.01
Bbif186 B. bidum 0.3 ± 0.9 0.3 ± 0.6 0.2 ± 0.5 0.3 ± 0.5 ↔ ns
Blon1004 B. longum 0.7 ± 1.2 0.9 ± 1.5 1.0 ± 1.5 0.6 ± 0.9 ↔ ns
Bputre698 Bacteroides putredinis 0.8 ± 1.6 0.8 ± 1.3 1.8 ± 2.1 1.6 ± 1.8 ↑ 50% ns
Burkho Burkholderia spp. 0.7 ± 0.7 1.4 ± 1.0 1.3 ± 1.4 1.3 ± 1.2 ↑ 46% 0.01
Ceut705 C. eutactus, Coprococcus sp. 3.0 ± 4.4 4.5 ± 5.1 5.6 ± 6.7 4.1 ± 5.2 ↑ 32% ns
Chis150 Clostridium histolyticum 0.6 ± 1.2 1.4 ± 3.6 2.5 ± 4.5 1.5 ± 2.1 ↑ 60% ns
Cor653 Coriobacterium group 0.5 ± 0.8 0.8 ± 1.0 1.2 ± 2.2 1.2 ± 1.5 ↑ 42% ns
Cvir1414 Clostridium viride group 1.9 ± 2.1 3.4 ± 2.2 4.2 ± 2.5 4.0 ± 2.1 ↑ 53% < 0.001
Ecyl387 Eubacterium cylindroides 0.7 ± 0.5 0.7 ± 0.4 1.4 ± 1.1 1.2 ± 0.7 ↑ 42% 0.01
Ehal1469 Eubacterium hallii 0.6 ± 0.9 0.6 ± 1.1 0.7 ± 0.8 0.7 ± 0.8 ↔ ns
Eram997 Eubacterium ramulus 0.3 ± 1.3 0.03 ± 0.04 0.7 ± 1.4 1.0 ± 1.5 ↑ 70% ns
Lab158 Lactobacillus sp., Enterococcus sp. 0.1 ± 0.2 0.8 ± 1.1 1.7 ± 3.0 0.5 ± 0.9 ↑ 80% 0.02
Muc1437 Akkermansia muciniphila 2.8 ± 3.9 1.8 ± 3.2 6.5 ± 7.4 7.8 ± 8.9 ↑ 64% 0.015
Myc657 Mycobacterium subdivision (mycolic 3.1 ± 1.5 2.5 ± 1.3 1.6 ± 1.1 1.9 ± 1.9 ↓ 39% ns
acid-containing Actinomycetes)
Phasco741 Phascolarctobacterium faecium 0.6 ± 0.9 0.9 ± 0.8 0.8 ± 0.8 1.1 ± 1.1 ↑ 45% ns
Pnig657 Prevotella nigrescens 2.2 ± 3.7 0.7 ± 1.3 2.6 ± 3.1 1.7 ± 2.3 ↔ ns
ProCo1264 Ruminococcus productus 0.7 ± 2.0 1.5 ± 2.6 1.4 ± 2.3 1.9 ± 3.7 ↑ 63% ns
Ra729 Ruminococcus albus 2.2 ± 3.2 5.5 ± 5.0 4.7 ± 5.0 3.9 ± 4.6 ↑ 44% 0.02
SFB1 Segmented lame(14%ntous bacteria 2.3 ± 3.3 1.6 ± 2.7 2.3 ± 1.6 2.9 ± 1.9 ↔ ns
SNA Sphaerotilus natans 4.3 ± 3.7 6.1 ± 5.4 6.8 ± 5.9 5.9 ± 5.8 ↑ 27% ns
Strc493 most Streptococcus spp. 1.3 ± 3.3 0.5 ± 1.1 1.9 ± 3.9 3.7 ± 4.9 ↑ 65% ns
SUBU1237 Burkholderia spp., Sutterella spp. 1.7 ± 2.6 3.4 ± 2.6 5.6 ± 4.4 5.6 ± 4.2 ↑ 69% 0.001
Urobe63a Ruminococcus obeum-like 1.6 ± 2.3 2.2 ± 2.3 2.5 ± 2.9 3.2 ± 2.4 ↑ 50% 0.05
Veil223 Veilonella 0.1 ± 0.3 0.1 ± 0.4 0.6 ± 1.4 0.9 ± 2.1 ↑ 88% ns
Ver620 Verrucomicrobium 1.7 ± 3.9 0.5 ± 1.6 1.1 ± 3.2 1.9 ± 5.3 ↔ ns
Individual marginal or accidental mean 150 (n = 6)
Cperf191 Clostridium perfringens 0.001 0 0 0.001 ↔ ns
Efaec Enterococcus faecalis 0.01 ± 0.02 0.01 ± 0.03 0.01 ± 0.02 0.01 ± 0.03 ↔ ns
MIB724 mouse intestinal bacteria 0.01 ± 0.06 0.001 ± 0.002 0.01 ± 0.03 0.07 ± 0.1 ↔ ns
Pce Burkholderia spp. 0.09 ± 0.30 0.03 ± 0.10 0.3 ± 0.7 0.07 ± 0.20 ↔ ns
Rbro730 Clostridium sporosphaeroides, Ruminococcus 0.04 ± 0.20 0.08 ± 0.30 0.8 ± 3.0 0.1 ± 0.5 ↑ ns
bromii, Clostridium leptum
Urobe63b Ruminococcus obeum-like 0.01 ± 0.04 0.0001 0.001 0.6 ± 1.1 ↔ ns

The response to humic acids of single bacterial
groups was principally the same as in all functional
sets of substantial bacteria. The concentrations of
most bacterial groups within essential (2 of 3) pioneer
(3 of 4) and individual substantial groups (19 of 28)
increased in rates of 20% to 60%. In most cases,
the increase was observed already at day 10 and
continued to day 45. In groups with comparatively
low initial mean concentrations (Ebac1790, Cdif198,
Chis150, Eram997, Lab158, Veil223) an increase
could be higher than 70% and up to 96%, but the
contribution of these groups to the overall bacterial
numbers was relatively low. Only the concentrations
of bacteria detected with Bac303 (Bacteroides) and
Myc657 (mycolic acid-containing Actinomycetes) FISH
probes decreased under humic acids supplementation,
but was statistically not signicant, because of the high
variance and low number of probands.
The increase in concentrations of microbiota was
caused by preexisting groups, and not due to emerging
new microorganisms. The individual microbial profile
remained constant. In none of the test persons did

WJG|www.wjgnet.com 888 February 7, 2017|Volume 23|Issue 5|

Swidsinski A et al . Humic acids and colonic microbiome

the ratio of positive/negative individual groups change
more than 5%.
Humic acid supplementation did not affect microbial
diver sity. Me an p ercen t of sub stant ial ind ividu al
bacterial groups positive for bacteria for each person
was nearly the same over time with 72%; 74%; 76%;
72% at the control days accordingly.
The patterns in distribution of single bacterial groups
over the stool cylinder differed depending on the
species but remained the same in the mucus close
transient zone and in the center of the fecal cylinder
regardless of humic acids supplementation.
of the essential bacterial groups observed in our study
was due to suppression of Bacteroides, and probably
further resulted from the fact, that essential bacterial
groups are already normally maximal promoted by the
host and their growth cannot be endlessly boosted.
Aside of the numeric impact on the microbiome,
we do not know which clinical effects humic acids
promoted, since all test persons were healthy and all,
®
except one, tolerated Activomin without negative or
apparent positive health effects.
However, reduced diversity and concentrations of
colonic microbiota were demonstrated in IBD , IBS
[9]

and non-gastroenterological diseases such as obesity,

DISCUSSION
The dietary supplementation of humic acids for medical
purposes and for promotion of health is deeply rooted
in cultural traditions. The humus and its components
are regarded as something purely biological, nature
promoting and positive. However, the mechanisms
how humic acids may work are purely understood. The
sheer indenite number of chemically active functional
groups within the extreme complex chemical structure
of humic substances makes biochemical investigations
[1]
elaborate, costly and difficult to reproduce . Even
apparent effects of humic acids on the quality of soil
and its microbiome remain vague and general as
to ancient times and are up to now not disclosed in
[8]
specic veriable details .
Our data first demonstrate that the humic acids
are indeed global fertilizers of microbial growth as
proposed by traditional view and lead to an increase
of more than 30% in the mean concentrations of
the colonic microbiome (P < 0.001). The promotion
of microbial growth involved 24 of 35 investigated
substantial bacterial groups. The only investigated
microbial groups that were negatively affected
by hum ic aci ds we re Bacteroides ( Bac3 03) an d
Mycobacterium subdivision mycolic acid-containing
Actinomycetes (Myc657). All other investigated groups
were either increased or not affected.
In newborns, during stress, convalescence or
disease, pioneer bacteria increase exponentially up
to ranges otherwis e typical for es sential bacterial
[7]
groups . We did not observe such a reaction in our
study. The most profound increase in concentrations
to 41% (P < 0.002) was that of the individual sub-
stantial bacteria. The increase in concentrations of
the pioneer groups was lower (28%) and statistically
not significant, indicating that host stress and
convalescence of the colonic microbiome are not
present. Lack of functional stress is also supported
by the fact that the individual microbial proles in all
subjects remained stable over the observation period
and that the patterns in distribution of bacteria over
the fecal cylinder did not change under humic acids
application.
The comparatively low increase (14%, P = 0.02)
[10-13]
diabetes, rheumatism and multiple sclerosis . These
changes in the microbiome are claimed responsible
for pathogenies of multiple other diseases. To repair
the disordered microbiome, fecal transplantation and
probiotics have been recommended and clin ically
tested. However, such transfections are difficult to
control and do not guarantee that the transferred
microorganisms prevail, settle and proliferate in the
[14]
colon .
Humic acids exert profound effects on the colonic
microbiota and may be an interesting group of sub-
stances for the development of specic drugs, which
deliberately influence colonic fermentation in an
inflamed colon, obesity, rheumatic and neurologic
disorders.
COMMENTS
COMMENTS
Background
Patients demonstrate increasing interest in medical treatments that are not part
of mainstream medicine. Critical argumentation is important, but difcult to do
when not evaluated with scientic methods. Humins are a product of microbial
metabolism and an important mediator of microbial interactions and activity. As
natural fertilizers humins are used for medical indications since ancient times.
It is believed that the human microbiome is the main target of humic activity.
However no data exist on the impact of humins on the human microbiome.
Research frontiers
An evergrowing number of studies demonstrate the involvement of the colonic
microbi ome in obesity, digesti ve, endocri ne, inflamma tory and auto-immune
and neurologic disorders. Different approaches are proposed to consolidate
and improve the colonic microbiome. The research hotspot is to move beyond
description and to introduce substances and therapies with proven controlled
graduate effects on the microbiome.
Innovations and breakthroughs
The presented results show, that orally applied humic acids have a profound
effect on the healthy colonic microbiome. Although the effects on single
microbial groups were multidirectional, the sum concentrations of all colonic
microbiota increased 20% to 30%. The increase occurred in the preexisting
microbial groups without changes in the bacterial diversity of the microbiome.
Applications
The main message of our study is, that humic acids may be an interesting
substrate for the development of defined drugs, which deliberately control
colonic fermentation in conditions where it is suppressed (post-antibiotics,
convalescence) or altered (metabolic disorders, inammation, obesity etc.), and
are a serious amendment/alternative to fecal transplantation or probiotics.

WJG|www.wjgnet.com 889 February 7, 2017|Volume 23|Issue 5|

Swidsinski A et al . Humic acids and colonic microbiome

Terminology
FISH - fluorescence in situ hybridization Cy3, FITC, Cy5, DAPI - different
uorescent dyes corresponding to orange, green, dark red and blue colours.
Fluorescence in situ hybridization (FISH) combines the specic identication
of microorganisms and the morphological aspect and is as a consequence
especially helpful for these purposes. Each single bacterium possesses 103-105
ribosomes of which each ribosome owns the same copy of ribosomal RNA.
Some of the regions of the rRNA are strain-specific, others are universal
for species, families or even kingdoms. Oligonucleotides synthesized
complimentary to rRNA sequences and labelled with fluorescent dye are
called FISH probes. When added to samples containing bacteria, FISH probes
hybridize with the rRNA of the bacterial ribosomes. No additional enhancement
of uorescence is necessary and bacteria can be visualized directly due to the
large number of ribosomes in each bacterium.
Peer-review
Although the scientic literature reveals a large number of medi cal tria ls with
dietary supplements of humic acids conducted all over the world. None of the
previous studies investigated effects of humic acids on the colonic microbiome.
REFERENCES
1 de Melo BA, Motta FL, Santana MH. Humic acids: Structural
properties and multiple functionalities for novel technological
developments. Mater Sci Eng C Mater Biol Appl 201 6; 62:
967-974 [PMID: 26952503 DOI: 10.1016/j.msec.2015.12.001]
2 van Rensburg CE. The Antiinflammatory Properties of Humic
Subst ances: A Mini Review. Phytother Res 2015; 29: 791-795
[PMID: 25732236 DOI: 10.1002/ptr.5319]
3 Ozkan A, Sen HM, Sehitoglu I, Alacam H, Guven M, Aras AB,
Akman T, Silan C, Cosar M, Karaman HI. Neuroprotective effect
of humic Acid on focal cerebral ischemia injury: an experimental
study in rats. Inflammation 2015; 38 : 32-39 [PMID: 25173888
DOI: 10.1007/s10753-014-0005-0]
4 Swidsinski A, Loening-Baucke V, Verstraelen H, Osowska S,
Doerffel Y. Biostructure of fecal microbiota in healthy subjects
and patients with chronic idiopathic diarrhea. Gastroenterology
2008; 135 : 568-579 [PMID: 18570896 DOI: 10.1053/
j.gastro.2008.04.017]
5 Loy A, Maixner F, Wagner M, Horn M. probeBase--an online
resource for rRNA-targeted oligonucleotide probes: new features
2007. Nucleic Acids Res 2007; 35: D800-D804 [PMID: 17099228
DOI: 10.1093/nar/gkl856]
6 Suau A, Rochet V, Sghir A, Gramet G, Brewaeys S, Sutren M,
Rigottier-Gois L, Doré J. Fusobacterium prausnitzii and related
species represent a dominant group within the human fecal flora.
Syst Appl Microbiol 2001; 24: 139-145 [PMID: 11403393 DOI:
10.1078/0723-2020-00015]
7 Swid si ns ki A, Loening-Baucke V, Schulz S, Manowsky J,
Verstraelen H, Swidsi nski S. Functional anatomy o f th e co lonic
bioreactor: Impact of antibiotics and Saccharomyces boulardii
on bacterial composition in human fecal cylinders. Syst Appl
Microbiol 2016; 39: 67-75 [PMID: 26723852 DOI: 10.1016/
j.syapm.2015.11.002]
8 Martinez CM, Alvarez LH, Celis LB, Cervantes FJ. Humus-
re du ci n g mic ro o rga nis ms an d th ei r va lu abl e con tr i bu ti on
in environmental processes. Appl Microbiol Biotechnol
2013; 97 : 10293-10308 [PMID: 24220793 DOI: 10.1007/
s00253-013-5350-7]
9 Kostic AD, Xavier RJ, Gevers D. The microbiome in
inflammatory bowel disease: current status and the future ahead.
Gastroenterology 2014; 146: 1489-1499 [PMID: 24560869 DOI:
10.1053/j.gastro.2014.02.009]
10 Wu G D, Lewis JD. Analysis of the human gut microbiome and
association with disease. Clin Gastroenterol Hepatol 2013; 11:
774-777 [PMID: 23643636 DOI: 10.1016/j.cgh.2013.03.038]
11 Baothman OA, Zamzami MA, Taher I, Abubaker J, Abu-Farha
M. The role of Gut Microbiota in the development of obesity and
Diabetes. Lipids Health Dis 2016; 15: 108 [PMID: 27317359 DOI:
10.1186/s12944-016-0278-4]
12 Van de Wiel e T, Van Praet JT, Marzorati M, Drennan MB,
Elewaut D. How the microbiota shapes rheumatic diseases.
Nat Rev Rheumatol 2016; 12: 398-411 [PMID: 27305853 DOI:
10.1038/nrrheum.2016.85]
13 DeGruttola AK, Low D, Mizoguchi A, Mizoguchi E. Current
Understanding of Dysbiosis in Disease in Human and Animal
Models. Inflamm Bowel Dis 2016; 2 2: 1137-1150 [PMID:
27070911 DOI: 10.1097/MIB.0000000000000750]
14 Li SS, Zhu A, Benes V, Costea PI, Hercog R, Hildebrand F, Huerta-
Cepas J, Nieuwdorp M, Salojärvi J, Voigt AY, Zeller G, Sunagawa
S, de Vos WM, Bork P. Durable coexistence of donor and recipient
strains after fecal microbiota transplantation. Science 2016; 352:
586-589 [PMID: 27126044 DOI: 10.1126/science.aad8852]

P- Reviewer: Lakatos PL, Teramoto-Matsubara OT

S- Editor: Gong ZM L- Editor: A E- Editor: Wang CH

WJG|www.wjgnet.com 890 February 7, 2017|Volume 23|Issue 5|

Published by Baishideng Publishing Group Inc

8226 Regency Drive, Pleasanton, CA 94588, USA
Telephone: +1-925-223-8242
Fax: +1-925-223-8243
E-mail: bpgofce@wjgnet.com
Help Desk: http://www.wjgnet.com/esps/helpdesk.aspx
http://www.wjgnet.com

I S S N 10 0 7 - 9 3 2 7
05

9 7 71 0 07 9 32 045

© 2017 Baishideng Publishing Group Inc. All rights reserved.

 Citations (17) References (23)

... Many patients request natural remedies from their gastroenterologist [20] and many IBS patients use
complementary and alternative medicine [21,22]. Humic acids are such natural remedies that have been used since
ancient times [20]. ...

... Many patients request natural remedies from their gastroenterologist [20] and many IBS patients use
complementary and alternative medicine [21,22]. Humic acids are such natural remedies that have been used since
ancient times [20] . At present, they are mostly used in veterinary medicine to treat intestinal diseases [23,24]. ...

... At present, they are mostly used in veterinary medicine to treat intestinal diseases [23,24]. Humic acids are
complex organic substances mainly present in soil, but also in ground water [20, 25,26]. They are generated through
humification, a process of decomposition of organic material by microbial transformation [20,27]. ...

Humic Acids in Patients with Diarrhoea-Predominant Irritable Bowel Syndrome: Results from A
Randomised Controlled Trial

Article Jan 2021

Ingolf Schiefke
View

... HSs have been utilized in humans for ages as nutritional supplements and therapeutic purposes. The first
mentions of the therapeutic applications of HS may be found in Sanskrit and ancient Chinese and Roman texts,
where they were assigned magical properties [1, 2]. Some of the therapeutic characteristics of HS are mentioned in
the Chinese Materia Medica Pharmacology Compendium, which On the other hand, the plethoric benefits that have
been widely documented in the immune response, antioxidant status, digestive microbiology, and nutrient
metabolism in humans and animals added with HS suggest that the mechanisms of action go far beyond its mucosal
protective effect. ...

Mechanisms of Action of Humic Substances as Growth Promoters in Animals

Chapter Full-text available Jul 2022

María de Lourdes Angeles · Sergio Gomez-Rosales · Guillermo Tellez

View Show abstract

... Humic substances are known to be generally safe for use in humans, as these cause no toxicity issues.
Preparations of humic acid have been used for treatment of stomach (gastroenteritis) (Swidsinski et al., 2017)
diseases and skin ailments. Activomin R , a commercial preparation, is already available in the market in Germany.
...

Value-Added Products From Soil, Brown Coal, and Composted City Solid Waste

Article Full-text available Dec 2021

S. V. Eswaran

View Show abstract

... Moreover, humic acid's molecules, which assume a negative charge in neutral to basic media, can inhibit virus
replication by binding cationic domains of the virus, which are necessary for virus attachment to the cell surface [13].
The humic acids are indeed global fertilizers of microbial growth, as proposed by the traditional view, and lead to an
increase of more than 30% in the mean concentrations of the colonic microbiome [14] . The potential of humic
substances to form chelate complexes with heavy metals (such as cadmium) enables them to be used for the
elimination of heavy metals from living organisms [1], with continuous exploration in medicine studies [15]. ...

Determination of Organic Compounds, Fulvic Acid, Humic Acid, and Humin in Peat and Sapropel
Alkaline Extracts

Article Full-text available May 2021 · MOLECULES

Laurynas Jarukas · Liudas Ivanauskas · Giedre Kasparaviciene · Jurga Bernatoniene

View Show abstract

... OMC may be acting directly within the GI tract, as well as exerting indirect effects, such as through the liver. In the
liver, fulvic acid can act as an antioxidant by uncoupling electron transport in liver mitochondria leading to a decrease
in reactive oxygen species (ROS) production [72] , which may explain the previous observation that OMC prevents
HFD-induced increases in ALT activity [21]. ...

Novel Organic Mineral Complex Prevents High-Fat Diet-Induced Changes in the Gut and Liver of
Male Sprague-Dawley Rats

Article Full-text available Dec 2020 · J Nutr Metab

Melisa Shaunte Crawford · Alex E Mohr · Karen Sweazea

View Show abstract

... The uncontrolled transfer of water, minerals, and organic matter in the soil and vadose zone is an adverse
consequence of standard silviculture management (Thurman, 1985;Hestir et al., 2015;Dunn et al., 2017;Han et al.,
2017;Van Zalinge et al., 2017;Altieri et al., 2018), creating a need to control the occurrence, structure, and
composition of polymicrobial biofilms in soils to enrich organic biodegradation and increase soil fertility (Swidsinski et
al., 2017; Swidsinski, 2019). ...

Thermodynamic mathematical model of the Kastanozem complex and new principles of sustainable
semiarid protective silviculture management

Article Mar 2021

Valery P. Kalinichenko · A P Glinushkin · Alexander V. Swidsinski · Sergey V Gudkov

View Show abstract

... The obtained results are consistent with the reported positive effect of HSs on the intestinal microbiota of humans
[34] and animals [35,36], which probably occurs due to prevention of bacterial stress and more effective formation
of resting cells. ...

Role of the Structure of Humic Substances in Increasing Bacterial Survival

Article Full-text available Jan 2020

Yury Nikolaev · Timur Kanapatskiy

View Show abstract

Sequentially Modified Carbon Felt for Enhanced p-Nitrophenol Biodegradation through Direct
Interspecific Electron Transfer

Article Feb 2023 · J HAZARD MATER

Yiwen Feng · Jianping Lu · Zhongjun Shen · Baogang Zhang

View Show abstract

Addition of Different Levels of Humic Substances Extracted from Worm Compost in Broiler Feeds

Article Full-text available Nov 2021

Alejandra Domínguez-Negrete · Sergio Gomez-Rosales · María de Lourdes Angeles · Guillermo Tellez

View Show abstract

Identification of Organic Matter Dispersions Based on Light Scattering Matrices Focusing on Soil
Organic Matter Management

Article Full-text available Dec 2020

Nikolay Bunkin · A P Glinushkin · A. V. Shkirin · Valery P. Kalinichenko

View Show abstract

Show more

Recommended publications Discover more

Conference Paper Full-text available Article Full-text available

Fertilization-induced changes of the soil Oral cavity contains distinct niches with dynamic
microbiome can improve or impair microbial communities
agroecosystem functioning... May 2014 · Environmental Microbiology
March 2018
Xin Xu · Jinzhi He · Jing Xue · [...] · Xuedong
Davide Francioli · Andrea Simpson · Elke Zhou
Schulz · [...] · Thomas Reitz
Microbes colonize human oral surfaces within hours after delivery.
During postnatal development, physiological changes, such as
Agroecosystems have been excessively exploited to meet the
the eruption of primary teeth and replacement of the primary
rising demand for food and other agricultural products. To
dentition with permanent dentition greatly alter the microbial
maintain fertility of agricultural soils and to prevent losses of
habitats, which, in return, may lead to community composition
agroecosystem services, the application of organic and mineral
shifts at different phases in people's lives. By profiling saliva,
fertilizers has become a common approach. However, nutrient
supragingival and ... [Show full abstract]
inputs also influence the soil microbiome, which plays a pivotal
role for the functioning of ... [Show full abstract]
View full-text
View full-text

Article Full-text available Article Full-text available

Prevotella and Porphyromonas spp. constituent
le core microbiome spécifique des lésions à
germes ana...
December 2015 · Annales de Dermatologie et de Vénéréologie
Hélène Guet-Revillet · Jean-Philippe Jais · Hélène
Coignard-Biehler · [...] · Olivier F Join-Lambert
Introduction La maladie de Verneuil ou hidrosadénite suppurée
(HS) est une maladie cutanée inflammatoire caractérisée par des
lésions récidivantes ou chroniques des grands plis du corps.
Nous avons montré qu’une vaste majorité de ces lésions est
associée à une flore anaérobie polymorphe, à des streptocoques
du groupe milleri et à des actinomycetes. Pour analyser de
manière exhaustive ces flores ... [Show full abstract]
View full-text
Species specificity of symbiosis and secondary
metabolism in ascidians
August 2014 · The ISME Journal
Ma. Diarey Tianero · Jason Christopher Kwan ·
Tom Wyche · [...] · Eric Schmidt
Ascidians contain abundant, diverse secondary metabolites,
which are thought to serve a defensive role and which have been
applied to drug discovery. It is known that bacteria in symbiosis
with ascidians produce several of these metabolites, but very little
is known about factors governing these 'chemical symbioses'. To
examine this phenomenon across a wide geographical and
species scale, we ... [Show full abstract]
View full-text

Last Updated: 31 Dec 2022

Company Support Business solutions

About us Help Center Advertising

News Recruiting

Careers

© 2008-2023 ResearchGate GmbH. All rights reserved. Terms · Privacy · Copyright · Imprint