Re iew
The an ibio ic c isis: How bac e ial p eda o s can help
Juana Pé ez
a
, F ancisco Ja ie Con e as-Mo eno
a
, F ancisco Ja ie Ma cos-To es
b
,
Au elio Mo aleda-Muñoz
a
, José Muñoz-Do ado
a,
⇑
a
Depa amen o de Mic obiología, Facul ad de Ciencias, A da. Fuen enue a s/n, Uni e sidad de G anada, 18071 G anada, Spain
b
Depa men o Cell and Molecula Biology, Uppsala Uni e si y, Uppsala 751 24, Sweden
a icle in o
A icle his o y:
Recei ed 29 July 2020
Recei ed in e ised o m 7 Sep embe 2020
Accep ed 8 Sep embe 2020
A ailable online 15 Sep embe 2020
Keywo ds:
An ibio ic c isis
Bac e ial p eda o s
BALOs
Myxobac e ia
summa y
Disco e y o an imic obials in he pas cen u y ep esen ed one o he mos impo an ad ances in public
heal h. Un o una ely, he massi e use o hese compounds in medicine and o he human ac i i ies has
p omo ed he selec ion o pa hogens ha a e esis an o one o se e al an ibio ics. The cu en an ibio ic
c isis is c ea ing an u gen need o esea ch in o new biological weapons wi h he abili y o kill hese
supe bugs. Al hough a p ope solu ion equi es his p oblem o be add essed in a a ie y o ways, he
use o bac e ial p eda o s is eme ging as an excellen s a egy, especially when used as whole cell he -
apeu ic agen s, as a sou ce o new an imic obial agen s by awakening silen me abolic pa hways in axe-
nic cul u es, o as biocon ol agen s. Mo eo e , s udies on hei p ey a e unco e ing mechanisms o
esis ance ha can be sha ed by pa hogens, ep esen ing new a ge s o no el an imic obial agen s. In
his e iew we discuss po en ial o he s udies on p eda o -p ey in e ac ion o p o ide al e na i e solu-
ions o he p oblem o an ibio ic esis ance.
Ó2020 The Au ho s. Published by Else ie B.V. on behal o Resea ch Ne wo k o Compu a ional and
S uc u al Bio echnology. This is an open access a icle unde he CC BY-NC-ND license (h p://c ea i e-
commons.o g/licenses/by-nc-nd/4.0/).
Con en s
1. In oduc ion . . . ..................................................................................................... 2547
2. P eda o y bac e ia: wha a e hey? How do hey kill? . . . . . . . ............................................................... 2548
3. BALOS as biocon ol agen s . . . . . . . . . . .................................................................................. 2549
4. Myxobac e ia as sou ces o new an ibio ics, bioac i e p oduc s and ly ic enzymes . . . ............................................ 2551
5. Summa y and ou look . . . . . . . . . . . . . . .................................................................................. 2553
Decla a ion o Compe ing In e es . . . . . . .................................................................................. 2554
CRediT au ho ship con ibu ion s a emen . . . . . . . . . . . . . . . . . . ............................................................... 2554
Acknowledgmen s . . . . . . . . . . . . . . . . . .................................................................................. 2554
Appendix A. Supplemen a y da a . . . . . . . .................................................................................. 2554
Re e ences . . . . ..................................................................................................... 2554
1. In oduc ion
An ibio ics e olu ionized medicine a e he se endipi ous dis-
co e y o penicillin by Fleming in he 1920s [1], he subsequen
de elopmen o syn he ic an imic obials [2], he es ablishmen o
use ul pla o ms o sc eening and isola ion o an ibio ic-
h ps://doi.o g/10.1016/j.csbj.2020.09.010
2001-0370/Ó2020 The Au ho s. Published by Else ie B.V. on behal o Resea ch Ne wo k o Compu a ional and S uc u al Bio echnology.
This is an open access a icle unde he CC BY-NC-ND license (h p://c ea i ecommons.o g/licenses/by-nc-nd/4.0/).
Abb e ia ions: AR, an ibio ic esis ance; ARB, an ibio ic- esis an bac e ia; ARG,
an ibio ic- esis an gene; MDRB, mul i-d ug esis an bac e ia; WHO, Wo ld Heal h
O ganiza ion; HGT, ho izon al gene ans e ; SM, seconda y me aboli e; BALOs,
Bdello ib io and like o ganisms; OSMAC, one s ain many compounds; BGC,
biosyn he ic gene clus e ; NRPS, non ibosomal pep ide syn he ase; PKS, polyke ide
syn hase; OMV, ou e memb ane esicle.
⇑
Co esponding au ho .
E-mail add ess: [email p o ec ed] (J. Muñoz-Do ado).
Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
jou nal homepage: www.else ie .com/loca e/csbj
p oducing mic oo ganisms [3], and he de elopmen o he pha -
maceu ical indus y. These an ibac e ial d ugs ha e sa ed millions
o li es, no only by comba ing a al in ec ious diseases, bu also by
enabling physicians o make ad ances in su ge y, in o gan ans-
plan a ion, in cance chemo he apy, and in he use o a i icial
de ices [4,5]. Fo hese easons, he disco e y and use o an ibio ics
in public heal h is one o he main pilla s o mode n medicine, con-
side ed one o he g ea achie emen s o he 20 h cen u y.
An ibio ics a e na u al p oduc s p oduced by mic oo ganisms
o hei semisyn he ic de i a i es. These compounds ha e been a
ea u e o he en i onmen o a long ime, so ha bac e ia ha e
needed o e ol e some o ms o an ibio ic esis ance (AR) o su -
i e. Fo ins ance, bac e ia inac i a e an ibio ics by p oducing
enzymes ha modi y hem, bu hey can also al e hei a ge o
ac ion, o p e en he an ibio ics om accumula ing, ei he
because hey a e expelled using e lux pumps o by al e ing he
pe meabili y o he memb ane [6,7]. The an ibio ic- esis an genes
(ARGs) a e a o ed in na u e due o he selec i e p essu e o na u-
al p oduc ion o an ibio ics by bac e ia. Howe e , he use, o e use,
o misapplica ion o hese d ugs, no only o he apeu ic pu poses,
bu also in animal eed and ag icul u e, ha e added an addi ional
selec i e p essu e and accele a ed he abili y o bac e ia o e ol e
and, consequen ly, led o he ine i able eme gence and sp ead o
AR [8–11]. Fu he mo e, bac e ia can collec mul iple esis ance
mechanisms, and his ac has p omo ed he appea ance o he
so-called supe bugs o MDRB (mul i-d ug esis an bac e ia),
which a e s ains ha ha e acqui ed esis ance o a wide a ie y
o an ibio ics. The Wo ld Heal h O ganiza ion (WHO) has issued
he p io i y pa hogens lis wi h hi een human pa hogens ha
ha e de eloped high le els o esis ance ac oss he wo ld [12,13].
Membe s o he so-called ESKAPE pa hogens cons i u e an espe-
cially u gen h ea o human heal h [14].
The i s s ep in AR ansmission is he ans e o ARGs o en i-
onmen al bac e ia h ough mobile gene ic elemen s by ho izon al
gene ans e (HGT) o human o animal commensal bac e ia o
pa hogens, gene a ing esis an clones ca ying ARGs ha can hen
be sp ead in he en i onmen [15,16]. The in e connec ion
be ween humans, animals and hei ecosys ems leads o he ans-
mission o esis ance. Al hough AR in hospi als o heal h cen e s
ep esen s a majo conce n, he dissemina ion is no es ic ed o
clinical se ings. An ibio ics elimina ed by humans o animals a e
discha ged in o u ban, ag icul u al o a m was e wa e , and can
be inco po a ed by ood c ops h ough i iga ion wi h hose con-
amina ed wa e s, inc easing he selec i e p essu e [17]. Mo eo e ,
was e wa e ea men plan s, which ecei e a g ea a ie y o bac-
e ia, including an ibio ic- esis an bac e ia (ARB) om a ious
sou ces, ac as ese oi s and o e con enien condi ions o he
in e ac ion and exchange o ARGs by HGT, con ibu ing o a
b oade dissemina ion [18]. O he ypes o indus ial con ami-
nan s, such as me als, can con ibu e o AR, since many esis ance
mechanisms ha p o ec agains hem, such as mul id ug e lux
pumps, also con e esis ance o an ibio ics [19–21]. Finally, phys-
ical o ces con ibu e o hei dissemina ion. The hype -connec ion
o he cu en wo ld p omo es he wo ldwide sp ead o AR. This
has se ious consequences, no only in he ea men o clinical
and/o e e ina y in ec ions, bu also in main aining he balance
o mic obial communi ies ac oss he biosphe e.
Cu en ly he e a e 700,000 dea hs pe yea due o ARB. I is
es ima ed ha by 2025 many o he cu en an ibio ics will be ine -
ec i e [22], and ha by 2050 supe bugs may cause 10 million
dea hs annually wo ldwide [23]. The p oblems de i ed om AR
in heal hca e could cos he wo ld a illion USD pe yea in heal h-
ca e, which would lead o a educ ion o 2% o 3.5% in g oss domes-
ic p oduc [23,24].
Many public heal h agencies and economic and poli ical ins i u-
ions ac oss he wo ld ecognize ha AR is one o he g ea chal-
lenges o he 21s cen u y. Many expe s conside ha we a e
ini ia ing he pos -an ibio ic e a, in which we will see he e u n
o epidemics p e alen du ing he p e-an ibio ic e a [5]. Bu he
an ibio ic c isis ha we ha e expe ienced in he las decade is
caused no only by he appea ance o MDRB, bu also by he sca -
ci y o esou ces dedica ed o he sea ch o new p oduc s, and, in
his scena io, he pipeline o he disco e y o new an ibio ics is
unning dange ously low [4,5,12,24–27]. Since he de elopmen
o esis ance is ine i able, e en i we make good and spa ing use
o an ibio ics, he sea ch o new an ibio ics and al e na i e he a-
pies is c i ical. The WHO, in a ecen epo , p oposes se e al
guidelines o e ec i ely deal wi h AR o a oid d i ing back o a
p e-an ibio ic e a. These leading s a egies, along wi h p ocedu es
o educe an ibio ic p essu e and ansmission o AR and ARGs, a e:
i) de eloping new ideas ha ques ion p e-exis ing o ms o knowl-
edge; ii) p o ec ing esea ch om pu ely comme cial de e mi-
nan s; iii) c ea ing new in e p e a ions and s a egies o add ess
esis ance; and i ) eo ien a ing esea ch o be e unde s and
he ole o bac e ia and human-mic obe ela ionships [22]. In his
con ex , labo a o ies a ound he wo ld a e de eloping new he a-
peu ic app oaches o educing he AR bu den, which include
mic obio a ansplan p ocedu es o displace ARB, gene ic engi-
nee ing o in e up esis ance genes (such as edi ing based on
CRISPR/Cas), me agenomic enginee ing, phage cock ails agains
ARB, accines agains pa hogens, an imic obial pep ides, he use
o me als (Cu, Ag, Ga), de elopmen o molecules ha in e e e
wi h he abili y o bac e ia o communica e and g oup oge he
in an ibio ic- esis an bio ilms, he sea ch o new an imic obial
p oduc s, and he use o bac e ia as he apeu ic agen s [16,29].
P eda o y bac e ia play an essen ial ole in hese new lines o
esea ch [16,28–31]. The ac ha hese mic op eda o s a e
designed o lyse and kill o he bac e ia, including MDRB and hose
pa hogens ha o m bio ilms, has led o hem being p oposed as a
easonable an ibio ic al e na i e o be explo ed. The aim o his
e iew is o collec and discuss he ad ances in he use o p eda-
o y bac e ia in his igh agains AR, ei he as biocon ol agen s
(‘‘li ing an ibio ics”) o as biological esou ces o inno a i e
an imic obial p oduc s (‘‘mic o ac o ies”).
2. P eda o y bac e ia: wha a e hey? How do hey kill?
P eda o y bac e ia use o he bac e ia o yeas s as a ood sou ce;
hence, hey a e able o ac i ely hun and kill hei neighbo s o
la e consume hei mac omolecules as nu ien s. Va ious s udies
ha e shown ha hese p eda o s a e widely dis ibu ed in many
na u al and a i icial en i onmen s (such as a i icial mic ocosms,
expe imen s in labo a o y, was e wa e s ea men plan s, o aqua-
cul u e plan s) whe e hey play an impo an ole no only in
main aining mic obial di e si y, bu also in shaping ecosys ems
[32–39].
In he con ex o igh ing AR, p eda o y bac e ia in na u e con-
ibu e o limi ing he sp ead o ARGs in di e en en i onmen s,
because hey can p ey upon ARB and deg ade hei DNA [40,41],
educing he ARG pools and hei sp ead o o he bac e ia by
HGT ia conjuga ion o ans o ma ion [42]. Mo eo e , hey can
also limi he possibili y o he ans e o ARGs ia ansduc ion
by p eying on bac e ia a ge ed by phage- esis an bac e ia [43].
Bac e ial hun ing s a egies can be ca ego ized in wo majo
g oups: epibio ic and endobio ic p eda ion [44]. In he epibio ic
s a egy, p eda o s a ack and consume he p ey om he ou side.
Two di e en subg oups can be di e en ia ed wi hin his g oup:
epibio ic ‘‘soli a y hun e s”, which, s ongly a ached o he p ey
cell en elope, suck hei con en s ou be o e di iding in o wo
daugh e cells; and he epibio ic ‘‘coope a i e hun e s”, whe e
indi idual cells coope a e wi hin he communi y by sha ing a com-
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2548
bina ion o di usible hyd oly ic enzymes and seconda y me abo-
li es (SMs), including an ibio ics, ha kill and decompose he p ey
in a non-species-speci ic manne be o e consuming he mac o-
molecules [44]. Whe he hun ing is uly coope a i e o me ely
communal is no clea [45]. In he epibio ic lone-hun e subg oup
he mos s udied gene a a e Mica ib io, Vampi ococcus and Vam-
pi o ib io, and he species Bdello ib io exo o us, [32]. The coope a-
i e (o communal) hun ing s a egy subg oup includes bac e ia
whose mode o a ack is based on lysis a a dis ance ha does
no equi e con ac wi h he p ey, such as he genus S ep omyces
[46,47], and hose ha equi e close p oximi y o he p ey cells
such as myxobac e ia [44,48–52].
The endobio ic s a egy is used by lone p eda o y bac e ia ha
ac i ely mo e o encoun e hei p ey, s ick o hei ou side, pe o-
a e and modi y he p ey cell wall by sec e ing hyd oly ic enzymes,
and pene a e he pe iplasmic o he cy oplasmic space, de ou ing
i om wi hin. Once he cycle is inished, p eda o s mo e on o
a ack neighbo ing cells. A ep esen a i e species o his g oup is
Bdello ib io bac e io o us [32,53–55].
Biochemical and mic obiological s udies ca ied ou in se e al
labo a o ies and analyses o he genomes and ansc ip omes du -
ing p eda ion (p eda osomes) o obliga e epibio ic p eda o s such
as Mica ib io ae uginosa o us,Vampi o ib io chlo ella o us, and B.
exo o us [56–58], and endobio ic B. bac e io o us and o he BALOs
(Bdello ib io and like o ganisms), ha e e ealed ha hey a e able
o syn hesize and sec e e an unusual numbe o hyd oly ic p o-
eins, such as lipases, glycanases, pep idases and p o eases, which
a e p obably in ol ed in damaging and diges ing p ey cell s uc-
u es [53,58–61]. They a e, howe e , non-compe en an ibio ic
p oduce s [62], since hei killing s a egies a e based on he ac i e
sea ch and ecogni ion o he p ey, he speci ic binding o he
ex e nal s uc u es, and i s deg ada ion. This pa icula way o li e
has led esea che s o mainly ocus on i s possible applica ion in
al e na i e li e he apies in which he p eda o s’ whole cells can
be used o kill o con ol he g ow h o o he ARB; ha is, he
use o p eda o s in bac e ial he apy as biocon ol agen s [53,61].
Howe e , he coope a i e acul a i e epibio ic p eda o s, which
include gene a o he wo mos impo an g oups o bac e ial SM
and hyd oly ic enzyme p oduce s, Ac inobac e ia and myxobac e-
ia,a e by a he mos p omising esou ces o inno a i e bioac-
i e na u al p oduc s. Wi hin Ac inobac e ia, he genus
S ep omyces has been ho oughly in es iga ed o many yea s o
he isola ion o an ibio ics [63–65], bu i has only ecen ly been
desc ibed as being a bac e ial p eda o ha kills o he soil bac e ia
by sec e ing hyd oly ic enzymes and SMs a away om he p ey
[46,47]. Myxobac e ia include b oad p ey ange p eda o s such as
Myxococcus xan hus, Myxococcus la escens,Myxococcus i escens,
Myxococcus mac ospo us,Co allococcus co alloides,S igma ella
au an iaca,Chond omyces apicula us and Chond omyces c oca us
[44]. Compa ed wi h Bdello ib io and BALOs, which exhibi a
es ic ed p ey spec um, myxobac e ia a e able o kill a b oad
ange o bac e ia, including clinically ele an species [66–78]. This
is p obably due o hei capaci y o sec e e a ple ho a o ly ic
me aboli es which, ac ing ei he in isola ion o syne gis ically,
a e able o a ack a wide a ie y o p ey. They a e able o p oduce
cell wall-deg ading enzymes, lipases, nucleases, polysaccha idases,
p o eases and, unlike many o he an ibio ic p oduce s, a ious
classes o an ibio ics, some o which a e ha dly e e ound as
mic obial SMs [79-82]. Al hough hey ha e been unde es ima ed
in he pas , in he las decade hey ha e eme ged as excellen ‘‘mi-
c o ac o ies”, wi h a high capaci y o p oduce an unlimi ed numbe
o p e iously unknown p omising an imic obials [82].
The use o epibio ic p eda o s in al e na i e he apies in medi-
cine o as biocon ol in di e en ields should be done in such a
way ha hey con ibu e o he igh agains MDBR, wi hou
inc easing he accumula ion o an ibio ics in he en i onmen ha
may gene a e AR.
3. BALOS as biocon ol agen s
Endobio ic p eda o s, such as B. bac e io o us, o he BALOs, and
some lone hun e s ha ollow an epibio ic s a egy, such as M.
ae uginosa o us, a e gene a ing in e es o he ea men o
in ac able, an ibio ic- esis an in ec ions. Mul iple s udies, using
human cells o se um, o animal models such as chicks, zeb a ish,
abbi s, guinea pigs, mice, and a s, and many in i o s udies ha e
shown ha p eda o whole cells o hei enzymes can be used as
he apeu ic agen s and as al e na i e o complemen a y applica-
ions in biological con ol [83–89] (Fig. 1 and Supplemen a y
Table 1).
The po en ial o hese p eda o y bac e ia as a p e e ed o sup-
plemen a y ea men o mul id ug- esis an bac e ial in ec ions is
suppo ed by a ange o e idence.
Fi s , p eda o s such as Bdello ib io and Mica ib io ha e he abil-
i y o kill human G am-nega i e pa hogens such as Acine obac e ,
Ae omonas,Bo de ella,Bu kholde ia,Ci obac e ,En e obac e ,
Esche ichia,Klebsiella,Lis onella,Mo ganella,P o eus,Salmonella,
Se a ia,Shigella,Vib io,Ye sinia,Helicobac e pylo i, and Legionella
ha ha e acqui ed, o a e a isk o acqui ing, esis ance o an ibi-
o ics [86,90–95]. These p eda o s also educe he bac e ial bu den
o d ug- esis an membe s o he ESKAPE g oup such as K. pneumo-
niae,A. baumannii [86,96–98] o P. ae uginosa [99],S eno ophomo-
nas mal ophilia [100] and o he MDRB G am-nega i e clinical
pa hogens such as Shigella [101]. The abili y o kill all hese
G am-nega i e bac e ia makes BALOS o pos ula e as an al e na i e
o igh agains mos o he MDBR iden i ied as a c i ical p io i y by
WHO. Howe e , he e a e also se e al G am-posi i e bac e ia in
his lis , wha makes in e es ing o explo e he capaci y o BALOs
o kill hem. In ac , i has ecen ly been epo ed ha B. bac e i-
o o us seems o a ack S. au eus cys ic ib osis isola es by using
an unusual epibio ic s a egy [99].
These p eda o s may also al e he commensal mic obio a, and
i is known ha hey inc ease he numbe o G am-posi i e species
in he gu mic obiome o chickens and a s [94,102]. The conse-
quences o his imbalance on animal heal h emain o be in es i-
ga ed. Iebba e al.[103] epo ed ha Bdello ib io is p esen and
abundan in he human gu mic obio a popula ions o heal hy indi-
iduals and sugges ed ha he use o p eda o y bac e ia as p obi-
o ics could es o e he balance in he in es inal ecosys em o
con ol dysbio ic e en s. Howe e , s udies by Sha zkes e al.
[102] on he e ec on a gu mic obio a showed mixed e ec s,
since hey ound ha in a ec al inocula ion o Bdello ib io con-
ibu es o heal h, while M. ae uginosa o us has po en ial ad e se
consequences. The e o e, al hough u he s udies a e equi ed o
deciphe he consequences o changes in he mic obio a due o
he adminis a ion o p eda o y bac e ia, i is wo h bea ing in
mind ha an ibio ics also may ha e ad e se e ec s on commensal
bac e ia.
Second, hese p eda o s a e able o inhibi he o ma ion o , o
educe, p e o med G am-nega i e and e en G am-posi i e bio-
ilms h ough a ple ho a o sec e ed enzymes, pa icula ly p o-
eases and nucleases [86,92,104–108]. Bio ilms can be ound in
many na u al en i onmen s and on indus ial equipmen , was e
wa e ea men plan s, hospi al su aces, heal h ca e se ings
and ins umen s. Bac e ia a e able o pe sis as ese oi s wi hin
hese mul icellula s uc u es, and bio ilms inside he hos allow
pa hogens o o e come inna e immune de enses. This capaci y o
dis up bio ilms o ele an pa hogens and o emo e en i onmen-
al and indus ial bio ilms opens he way o using hese p eda o s
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2549
o hei enzymes no only in medicine, bu also in many biocon ol
p ocesses ha will be discussed below [100,109,110].
Thi d, opical applica ion, inges ion, injec ion, o in anasal
inocula ion o whole cells ha e no appa en cy o oxici y, nei he
in e ms o pa hological e ec s, no in e ms o diminu ion in cell
iabili y in in i o cell cul u e models [111–113] o in i o animal
models [94,96,101,102,113–116].
Fou h, hey do no inci e a sys emic o sus ained immune
esponse, mos likely due o he special s uc u e o hei
lipopolysaccha ide, which lacks he ypical nega i ely cha ged
phospha e g oups, esul ing in only low binding a ini y o he
lipopolysaccha ide ecep o s in human immune cell pa hogens
[101,117]. They a e passi ely engul ed by mac ophages and a e
able o pe sis inside hese cells o e 24–48 h as non- eplica i e
o ms, al hough hey e ain hei p eda o y compe ence. Al hough
hey do no a ec hos cell iabili y, hey s imula e mode a e cy o-
kine esponses [111–113,115]. P eda o pe sis ence inside mac o-
phages o su icien ime o p ey on pa hogens opens he way o
using p eda o y bac e ia o elimina e in acellula pa hogens, such
as Salmonella, Klebsiella o F ancisella species. Mo eo e , he ela-
i ely benign occupancy o mac ophages by Bdello ib io could p e-
en o he in acellula pa hogens om en e ing. Howe e , his
passi e up ake sugges s ha mac ophages migh p esen p eda-
o y bac e ia o an ibody- o ming cells and, consequen ly, humans
could de elop immune eac ions agains hem a e epea ed expo-
su es [30]. In ac , Raghuna han e al.[118] de ec ed, al hough a
low le els, IgG and IgM an ibodies agains wo B. bac e io o us
s ains in mo e han 90% o human se um samples om a biobank,
sugges ing ha he p eda o y ea men could be used only once.
On he o he hand, expe imen s in immune-comp omised zeb a-
ish inocula ed wi h B. bac e io o us HD100 demons a ed ha
coope a ion be ween he hos immune sys em and bac e ial p eda-
ion is impo an , and i mus be aken in o conside a ion in o de
o maximize an ibac e ial he apy bene i s [101]. Al hough a lo o
p og ess has been made in ecen yea s, mo e esea ch is needed o
unde s and no only he in e ac ions and a e o hese p eda o y
bac e ia wi hin human immune cells, bu also o elucida e he p o-
cesses in ol ed in hei up ake by phagocy ic cells, how long bac-
e ia su i e in he hos cells, and he mechanisms induced by he
p eda o y bac e ia.
Fi h, Bdello ib io is an ae obic p eda o , condi ioning i o be an
e ec i e ea men in ae obic en i onmen s such as supe icial
bu ns o wounds, eyes and lungs [85,96,119,120]. Ne e heless,
his p eda o is able o ole a e mic oae ophilic condi ions, which
widens i s a eas o use o ea ing gas oin es inal and pe iodon al
in ec ions [83,85,121–124].
Six h, one o he main ad an ages o using hese p eda o s as
he apeu ic agen s is he ailu e o p ey bac e ia o de elop
induced esis ance agains p eda ion. The main eason o his is
ha hese p eda o s a e no an ibio ic p oduce s, and hei killing
mechanisms do no a ge speci ic ecep o p o eins ha can
e ol e esis ance. Howe e , p eda ion can weaken due o di e en
adap a ions a ising in he p ey bac e ia. Fo ins ance, ansien
esis ance has been obse ed in some p ey simila o bac e ial pe -
sis ence [83,125]. As a consequence, Bdello ib io p eda ion ne e
comple ely kills o he a acked popula ion. To esol e his di i-
cul y, he combined use o B. bac e io o us and an ibio ics in a
co- he apy has been p oposed [85]. This combined he apy has
he addi ional ad an age o expanding he indi idual killing spec-
a o bo h he an ibio ics and he p eda o . Im e al. [126] ha e
con i med ha a combina ion o Bdello ib io and iolacein is e ec-
i e agains G am-posi i e bac e ia, mul id ug- esis an pa ho-
Fig. 1. Applica ions o Bdello ib io and BALOs in medicine and as biocon ol agen s.
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2550
gens, and G am-posi i e and nega i e mixed species popula ions.
Howe e , o an e icien de elopmen o hese co- he apies i is
necessa y o de e mine he co ec p eda o -an ibio ic combina-
ion o inc ease he an imic obial e iciency, while a oiding de el-
opmen o d ug esis ance. Wi h his in mind, esea che s a e
making e o s o de e mine he sensi i i y o Bdello ib io agains
an ibio ics [127], and mo e esea ch mus be ca ied ou o de e -
mine i s abili y o g ow on an ibio ic-inac i a ed cells and he AR
mechanisms de eloped by he p eda o . Recen ly, expe imen s
using Bdello ib io and p ey-speci ic bac e iophages e adica ed
E. coli when p eyed upon by bo h p eda o s, demons a ing he
po en ial o his inno a i e co- he apy as a u u e al e na i e
an ibac e ial ea men ha would educe he selec ion o single
p eda o -speci ic esis ance [128].
And se en h, he e is a lack o up ake o p ey gene ic ma e ial
du ing p eda ion, and i has been shown ha , e en a e egula
epea ed high exposu e o he p ey genome, he e is no inco po a-
ion o any majo pa hogenici y island(s) o o any po en ially i -
ulen genes om he p ey [129]. The exchange o gene ic ma e ial
would ha e an addi ional nega i e impac because p ey could also
inco po a e ARGs, which could con ibu e o he sp ead o esis-
ance. The absence o DNA in e change u he s eng hens he
possibili y o using hese bac e ia in he u u e as a sa e al e na i e
in he igh agains AR.
Besides clinical uses (Fig. 1,Supplemen a y Table 1), BALOs a e
also being explo ed as a easible al e na i e o an ibio ics as bio-
con ol agen s in ho icul u e, aquacul u e, li es ock a ming and
ood p ocessing. In addi ion, hey a e being assayed o hei appli-
ca ion in o he bio echnological p ocesses such as was e wa e
ea men , ex ac ion o bioplas ics, educ ion o ho izon al ans-
e o ARGs, and o he en i onmen ally- iendly applica ions in
which he educ ion o unwan ed bac e ia would be posi i e
[43,85,130–132] (Fig. 1,Supplemen a y Table 1).
The use o epibio ic p eda o s as biocon ol agen s emains la -
gely unexplo ed. Howe e , he abili y o myxobac e ia (besides
hei p eda o y beha io s) o glide on solid su aces, p oduce a
wide a ie y o an ibio ics, and o m myxospo es, make hese bac-
e ia excellen , comme cially iable candida es o he biocon ol
o phy opa hogenic bac e ia and ungi [69,73,133–135] (Supple-
men a y Table 1).
4. Myxobac e ia as sou ces o new an ibio ics, bioac i e
p oduc s and ly ic enzymes
Al hough Bdello ib io and BALOs a e mainly being explo ed
wi hin he concep o whole cell he apy, o he in es iga ions a e
being ca ied ou ha aim o make use o he la ge amoun o bac-
e ially des uc i e ly ic enzymes ha p eda o s employ o kill
hei p ey [59,61,130,131,136,137]. Those applica ions include
some in e es ing uses such as ools o an ibody analysis o o
con olling he sp ead o ARGs in mixed mic obial communi ies
h ough he deg ada ion o cell- ee DNA o inac i a ion o phage
pa icles, hus limi ing wo o he gene ic DNA ans e p ocesses
ha dic a e he ou come o sp ead: ans o ma ion and ansduc-
ion [43].
Fo he disco e y o new an imic obial agen s, coope a i e
p eda o s a e he mos p omising bac e ia. In he pas , he p oduc-
e s o an ibio ics and o he bioac i e p oduc s we e mainly iso-
la ed om soil and es ed by adi ional di usion me hods using
cul u e il a es o ex ac s [4,138]. The ad en o he pos -
genomic e a, wi h he inc ease in genome sequencing and anno a-
ion, nex -gene a ion sequencing echnologies, mode n p o eomic,
ansc ip omic and me abolomic ools, and scien i ic and echno-
logical ad ances in gene ics, biology, obo ics, chemis y, me a-
bolic enginee ing, and bioin o ma ics, has ampli ied he ange o
me hodologies used o he isola ion and iden i ica ion o p oduc-
e s and has opened new doo s o he iden i ica ion o hei
‘‘an ibio ic’omes”, de ined as he speci ic subse o mic obial na u-
al p oduc s wi h an ibio ic ac i i y [65,139–141].
Al hough isola ion o new bac e ial p oduce s is one o he p i-
o i y lines in an ibio ic disco e y, ano he app oach is he use o
new echnologies o exploi he knowledge accumula ed o e he
yea s abou well ecognized ‘‘mic o ac o ies”. The e m ‘‘OSMAC”
(one s ain many compounds) was in oduced a he beginning
o his cen u y [142], and i e e s o he ac ha a single bac e ial
s ain is capable o p oducing a di e se collec ion o s uc u ally
di e en SMs. Bac e ia ne e p oduce he ull collec ion o com-
pounds encoded by hei genomes a he same ime unde a speci-
ic en i onmen al condi ion, since his would be ene ge ically
cos ly. To a oid unnecessa y cos s, hey modi y hei ansc ip-
omes, p o eomes, and me abolomes o su i e unde changing
condi ions. Fo ha eason, biosyn hesis pa hways a e i mly con-
olled by complex egula o y ne wo ks ha espond o in e nal
and/o ex e nal signals, many o which a e s ill unknown. The
OSMAC app oach ocuses on he sys ema ic al e a ion o cul u ing
pa ame e s, such as medium componen s (sal s, amino acids, ca -
bon sou ce), empe a u e, pH, cul u e ae a ion, ype o cul u e es-
sel, e c., o imi a e in he labo a o y hose na u al en i onmen s
ha op imize he p oduc ion o use ul compounds [143]. The
de ailed s udy o e e y single s ep o he complex me abolic pa h-
ways o he biosyn hesis o SMs, hei egula i e c oss alk, and he
signals ha ac i a e he egula o y bac e ial senso s is acili a ing
he disco e y o no el p oduc s om one bac e ium o ungus
[144]. Myxobac e ia a e pe ec candida es o be conside ed om
he OSMAC poin o iew [82,142]. The Helmhol z-Cen e o In ec-
ion Resea ch in Ge many has a collec ion o myxobac e ia wi h
mo e han 9,000 s ains, and mo e han 1,000 biosyn he ic gene
clus e s (BGCs) associa ed wi h myxobac e ia a e deposi ed in
he an isMASH da abase (an ismash.seconda yme aboli es.o g).
M. xan hus DK1622, besides being a p eda o , is also a model
o ganism o s udying p oka yo ic de elopmen . O e many yea s,
nume ous g oups ha e accumula ed a deep knowledge abou he
p oduc ion o SMs and ly ic enzymes, mul icellula beha io ,
in a- and ex acellula signaling, mo emen mechanisms, adap a-
ions o changing en i onmen s, e c. [145]. This p eda o holds a
la ge genome wi h an unp eceden edly high numbe o egula o y
mechanisms [146–148] and an ou s anding biosyn he ic capaci y
o deg ada i e enzymes and SM p oduc ion, including 18 non ibo-
somal pep ide syn he ases (NRPS), 22 polyke ide syn hases (PKS),
and six mixed PKS/NRPS, which conco ds well wi h i s p eda o y
li es yle [146,149]. Analyses using he an isMASH se e calcula e
ha hose clus e s co espond o a o al o 14.5% o i s 9.1 Mb gen-
ome [150]. Se e al o hese na u al p oduc s ha e been isola ed
[80,82,151–158] (Fig. 2) and e en exp essed in he e ologous hos s
[159]. Howe e , many BGCs a e no ye assigned o hei co e-
sponding hypo he ical SMs, p obably because some o hese genes
emain silenced (c yp ic genes) o a e exp essed a low le els,
yielding low quan i ies o he espec i e compounds in he g ow h
condi ions assayed [82,149,160]. The same si ua ion is obse ed in
S ep omyces, a genus no o ious o i s abili y o p oduce a la ge
quan i y and a ie y o an ibio ics and SMs [65,161].S ep omyces
genomes a e also la ge and ypically con ain mo e han 20 BGCs
dedica ed o specialized me abolism such as NRPSs and PKSs. In
he case o S ep omyces coelicolo A3, hese enzymes a e ound
in up o 27 clus e s, which ep esen 10.6% o i s 8.7 Mb genome
[162,163]. Only a small ac ion o hem a e ansc ip ionally ac i e
unde labo a o y condi ions and mos o hei p oduc s ha e ne e
been cha ac e ized [161,164].
This huge un apped biosyn he ic po en ial makes he unlocking
o he c yp ic pa hways o hose coope a i e p eda o s ano he
good s a egy o an ibio ic disco e y. I is easonable o hink ha
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2551
Fig. 2. The coope a i e p eda o M. xan hus as a sou ce o an ibio ics and new p oduc s. A. Du ing p eda ion, M. xan hus sec e es ly ic enzymes, an ibio ics and o he
seconda y me aboli es, some o hem included in OMVs. B. An ibio ics and bioac i e p oduc s iden i ied in M. xan hus species. 1: myxoch omide; 2: myxalamid; 3:
myxo i escin TA; 4: ci ilin A; 5: myxop incomide; 6: al hiomycin; 7: sa amycin Mx1.
Fig. 3. De ense mechanisms in he p ey wi h bio echnological applica ions.A. Silenced an ibio ics a e induced in he p ey du ing he p eda o y p ocess. In he pic u es, M.
xan hus (Mx, p eda o ) induces in S ep omyces coelicolo (Sc, p ey) he blue an ibio ic ac ino hodin, in solid and liquid media. B. No el an ibio ic esis ance mechanism ha e
been disco e ed in B. hu ingensis: myxo i escin TA glucosyla ion. C. O he physical/chemical de enses mechanisms induced by M. xan hus p eda ion in di e en bac e ia.
Galac oglucan (le pic u e) and melanin (middle pic u e) p o ec Sino hizobium melilo i om p eda ion. M. xan hus induces de elopmen in S ep omyces ( igh pic u e).
Pic u es om panel A and igh pic u e om panel C a e ep oduced om Pé ez e al. (2011) Mic ob Bio echnol 4: 175–183. Le pic u e in panel C is adap ed om Muñoz-
Do ado e al., (2016) F on Mic obiol 7: 781. Middle pic u e in panel C is om Con e as-Mo eno e al. (2020) F on Mic obiol 11: 94. (Fo in e p e a ion o he e e ences o
colou in his igu e legend, he eade is e e ed o he web e sion o his a icle.)
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2552
one o he ecological oles o an ibio ics in na u e is o be used as
weapons agains suscep ible bac e ia du ing p eda o y p ocesses
and, in ac , some o hem ha e been shown o be in ol ed in p e-
da ion [46,165]. I has been ecen ly ound ha al hough he genes
esponsible o SM biosyn hesis in M. xan hus a e exp essed du ing
g ow h, hei exp ession inc eases du ing de elopmen [148].
These exp ession p o iles sugges ha SMs a e used o p o ec
de eloping cells om o he mic obes in he soil, o de end spo es
inside he ui ing bodies o o elease nu ien s om p ey o p o-
mo e ge mina ion [148]. I is also ealis ic o in e ha p oduc ion
o such me aboli es by p eda o s migh be egula ed by con ac
wi h and/o p oximi y o he p ey. The e o e, i would be use ul
o design e icien sc eening me hods in he p esence o a a ie y
o pa hogens o he induc ion o no el an ibio ics agains hem.
Bu in e species c oss alk can also igge he p oduc ion by
p ey o speci ic bioac i e compounds o de ensi e pu poses and
o he esis ance mechanisms o coun e a ack such as bio ilm o -
ma ion [166], spo e di e en ia ion, o induc ion o new ARGs [167]
(Fig. 3). Co-cul i a ion expe imen s, ollowed by deep analysis o
he in luence on he seconda y me abolome, a e gaining inc easing
a en ion and a e al eady yielding exci ing esul s. Some s udies
ha e demons a ed ha he p esence in cul u es o o he bac e ia
s imula es S ep omyces o p oduce some SMs [144,164]. Mos o
hese pai wise in e ac ions ha e been assayed wi h o he ela ed
Ac inobac e ia o Bacillus. Mo eo e , M. xan hus is able o induce
he o e p oduc ion o he blue an ibio ic ac ino hodin while p ey-
ing on S. coelicolo (Fig. 3A) [168]. O he esul s in ol ing he p o-
duc ion o SMs du ing p eda o y ela ionships a e he induc ion o
bacillaene by B. sub ilis [76,169], o he biosyn hesis o melanin by
S. melilo i o p o ec agains p eda ion by M. xan hus (Fig. 3)
[77,170].
The i s ansc ip omic s udy du ing a p eda o y p ocess o M.
xan hus agains E. coli [171] indica ed ha M. xan hus does no pe -
cei e li e p ey as ood, and ha he p ey shows ex ensi e an-
sc ip omic changes when co-cul u ed wi h he p eda o .
Mo eo e , M. xan hus supe na an s and sec e ed ou e memb ane
esicles (OMVs, see below) also induced changes in he exp ession
o la ge numbe s o p ey genes. Ano he ansc ip omic analysis
con i med he ac ino hodin o e p oduc ion obse ed in S ep o-
myces by Pé ez e al [168], and con i med ha i is i on compe i ion
du ing his in e ac ion which leads o he ac i a ion o ac i-
no hodin BGC plus 21 o he SM BGCs [172] (Fig. 3A). This is a good
example o how basic esea ch on bac e ial p eda o -p ey in e ac-
ion is helping o disco e new ways o ac i a e c yp ic biosyn-
he ic pa hways and is imp o ing ou comp ehension o
awakening mechanisms.
Ano he aspec o be conside ed in he sea ch o inno a i e
applica ions o p eda o s a e he G am-nega i e OMVs. OMVs
a e sphe ical po ions o he bac e ial ou e memb ane wi h a
diame e o ~20–250 nm, which ep esen an al e na i e o gen-
e al sec e ion sys ems. The myxobac e ial OMVs con ain a highly
concen a ed dose o hyd oly ic enzymes and molecules associa ed
wi h an ibio ic ac i i y, and a e able o deli e hese compounds o
dis an and inaccessible loca ions, enabling mo e e icien epibio ic
coope a i e killing o p ey [50,51,173,174]. The OMVs can be used
as shu les o deli e hyd ophobic compounds wi h any desi ed
p ope ies (Fig. 2). Fo ins ance, hey ha e been con i med o be
good adju an s in accine deli e y [175]. The myxobac e ial OMVs
ha e ecen ly been explo ed as na u al an imic obial ca ie s able
o deli e hese p oduc s o a ge cells. They a e able o kill some
pa hogenic bac e ia such as E. coli o P. ae uginosa [176,177].
Recen ly, hey ha e also been con i med o be e icien ools o
igh ing in acellula bac e ial pa hogens such as S. au eus. Those
in ec ions a e di icul o ea because se e al classes o an ibio ics
a e unable o each he pa hogen, and highe concen a ions and a
longe he apy ime a e needed. In hese assays, OMVs om Cys o-
bac e ela us and So angineae species exhibi low oxici y in a -
ious cell lines and p ima y immune cells, and a e able o ake hem
up in o in ec ed cells [177,178]. F om hese esul s i is possible o
en ision he exploi a ion o OMVs om myxobac e ia as no el
he apeu ic deli e y sys ems o comba bac e ial in ec ions. The
epo s on M. xan hus OMV p o eomics show ha hey a e en iched
(along wi h oxic p o eins and an ibio ics) in hypo he ical p o eins
o unknown unc ion which emain o be s udied in dep h
[173,176,179,180]. Mo eo e , a ecen p o eomic s udy o he
OMV ca go om se e al M. xan hus s ains seems o indica e ha
gene ically simila s ains o myxobac e ia ha e di e se OMV p o-
eomes [51,181], gi ing ise o new esea ch challenges.
Finally, i has been shown ha myxobac e ial p eda o y ac i i y
a ies depending on he p ey [77,78,182]. Mo eo e , Su on e al.
[183], using genome-wide associa ion s udies wi h 29 myxobac e-
ial s ains, iden i ied di e en numbe s o p eda ion genes
depending on he p ey o ganisms. They sugges ed ha he b oad
p ey ange o myxobac e ia seems o be a consequence o he accu-
mula ion o a angemen s o p ey-speci ic p eda ion genes, a he
han possession o a se o gene al an ibio ic genes. These esul s
clea ly indica e ha he awakening o c yp ic genes should be
assayed in he p esence o pa hogens agains which we a e looking
o a d ug, since hey may ein o ce he posi i e esul s o he s a -
egy o es ing o he bac e ia as p ey and assaying new p oduc s
agains esis an bac e ia.
5. Summa y and ou look
De elopmen o AR by many pa hogens is one o he main chal-
lenges o esea che s in o de o a oid a new e a in which he
numbe o dea hs om in ec ious diseases ha we e hough o
be unde con ol inc eases o a le el ha ep esen s an unaccep -
able heal h and economic cos . Finding new al e na i es o kill
hese pa hogens equi es a ocus on a ious di e en s a egies,
and one o hese p omising s a egies is lea ning abou he beha -
io s o bac e ial p eda o s and p ey in na u e. Unde s anding he
s a egies de eloped by p eda o y bac e ia du ing e olu ion is
helping esea che s o emula e and use hem in di e en an imi-
c obial app oaches. Resea ch on he use o p eda o s as li ing
an ibio ics is yielding p omising esul s, and al hough mo e
esea ch is equi ed o demons a e hei e icacy in i o, hey ep-
esen a se ious al e na i e o be conside ed. Cu en ly, inding
s uc u al genes and egula o y elemen s in ol ed in p eda ion
and he induc ion o silenced and c yp ic genes o BGCs du ing
hese dynamic p eda o y p ocesses is no only s a ing o help in
he disco e y o new an ibio ics wi h no el mechanisms o ac ion
and new classes o bioac i e na u al p oduc s, bu also acili a ing
he es ablishmen o he bes labo a o y condi ions o op imal
scaled-up indus ial p oduc ion o me aboli es. Finally, om he
s udies o he an agonis ic p eda o –p ey ela ionships, esea ch-
e s a e lea ning abou he a ack s a egies used by he hun e s,
which a e opening inno a i e esea ch lines such as he use o
OMVs as an imic obial ca ie s. The mechanisms de eloped by
he p ey o de end i sel a e also p o ing use ul in inding ou abou
new de e minan s o AR. All o he s udies e iewed he e abou
bac e ial p eda o s a e no only opening new doo s o he disco -
e y o new an ibio ics, bu also ha nessing he eno mous po en ial
o bac e ial p eda o s as whole cell he apeu ic agen s. Taken
oge he , hese s udies will help o p epa e us o a nea u u e
when many an ibio ics ail o ea MDRB. Howe e , hey equi e
mo e suppo om go e nmen ini ia i es, along wi h changes in
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2553
egula ion o pa e he way o aluable, e icacious, highly a -
ge ed, pa hogen-speci ic an imic obial he apies.
Decla a ion o Compe ing In e es
None.
CRediT au ho ship con ibu ion s a emen
Juana Pé ez: W i ing - o iginal d a , Concep ualiza ion. F an-
cisco Ja ie Con e as-Mo eno: W i ing - e iew & edi ing. F an-
cisco Ja ie Ma cos-To es: W i ing - e iew & edi ing. Au elio
Mo aleda-Muñoz: W i ing - e iew & edi ing, Funding acquisi ion.
José Muñoz-Do ado: W i ing - o iginal d a , Funding acquisi ion,
Concep ualiza ion.
Acknowledgmen s
This wo k has been suppo ed by he Spanish Go e nmen ,
g an BFU2016-75425-P o Au elio Mo aleda-Muñoz (70% unded
by FEDER). Funde has no in ol emen in s udy design; in he col-
lec ion, analysis and in e p e a ion o da a; in he w i ing o he
epo ; and in he decision o submi he a icle o publica ion.
We hank Lola Con e as-Mo eno o echnical assis ance wi h
Fig. 1.
Appendix A. Supplemen a y da a
Supplemen a y da a o his a icle can be ound online a
h ps://doi.o g/10.1016/j.csbj.2020.09.010.
Re e ences
[1] Fleming AG. Responsibili ies and oppo uni ies o he p i a e p ac i ione in
p e en i e medicine. Can Med Assoc J 1929;20:1–13.
[2] Domagk G. Ein bei ag zu chemo he apie de bak e iellen in ek ionen. Deu
Med Wochensch 1935;61:250–3.
[3] Scha z A, Bugie E, Waksman SA. S ep omycin, a subs ance exhibi ing
an ibio ic ac i i y agains G am-posi i e and G am-nega i e bac e ia. P oc
Soc Exp Biol Med 1944;55:66–9.
[4] Lewis K. Pla o ms o an ibio ic disco e y. Na Re D ug Disco
2013;12:371–87.
[5] Lewis K. The science o an ibio ic disco e y. Cell 2020;181:29–45.
[6] Blai JA, Webbe MA, Baylay AJ, Ogbolu DO, Piddock LJV. Molecula
mechanisms o an ibio ic esis ance. Na Re Mic obiol 2015;13:42–51.
[7] C o s TS, Gaspa ini AJ, Dan as G. Nex -gene a ion app oaches o unde s and
and comba he an ibio ic esis ome. Na u e 2017;15:422–34.
[8] Allen HK, Dona o J, Wang HH, Ka en A, Cloud-Hansen KA, e al. Call o he wild:
an ibio ic esis ance genes in na u al en i onmen s. Na u e Re Mic obiol
2010;8:251–9.
[9] Bush K, Cou alin P, Dan as G, Da ies J, Eisens ein B, e al. Tackling an ibio ic
esis ance. Na Re Mic obiol 2011;9:894–6.
[10] D’Cos a VM, King CE, Kalan L, Mo a M, Sung WWL, e al. An ibio ic esis ance
is ancien . Na u e 2011;477:457–61.
[11] Van Boeckel TP, B owe C, Gilbe M, G en ell BT, Le in SA, e al. Global ends
in an imic obial use in ood animals. P oc Na l Acad Sci USA
2015;112:5649–54.
[12] Wo ld Heal h O ganiza ion. An ibac e ial agen s in clinical de elopmen : an
analysis o he an ibac e ial clinical de elopmen pipeline, including
ube culosis. WHO/EMP/ IAU/2017.12. 2017; WHO: Gene a.
[13] Tacconelli E, Ca a a E, Sa oldi A, Ha ba h S, Mendelson M, e al. Disco e y,
esea ch, and de elopmen o new an ibio ics: he WHO p io i y lis o
an ibio ic- esis an bac e ia and ube culosis. Lance In ec Dis
2018;18:318–27.
[14] Bouche HW, Talbo GH, B adley JS, Edwa ds JE, Gilbe D, e al. Bad bugs, no
d ugs: no ESKAPE! an upda e om he In ec ious Diseases Socie y o Ame ica.
Clin In ec Dis 2009;48:1–12.
[15] Be endonk TU, Manaia CM, Me lin C, Fa a-Kassinos D, Cy yn E, e al. Tackling
an ibio ic esis ance: he en i onmen al amewo k. Na Re Mic obiol
2015;13:310–7.
[16] He nando-Amado S, Coque TM, Baque o F, Ma ínez JL. De ining and
comba ing an ibio ic esis ance om One Heal h and Global Heal h
pe spec i es. Na Mic obiol 2019;4:1432–42.
[17] Gudda FO, Waigi MW, Odinga ES, Yang B, Ca e L, Gao Y. An ibio ic-
con amina ed was ewa e i iga ed ege ables pose esis ance selec ion isks
o he gu mic obiome. En i on Pollu 2020;114752.
[18] Pazda M, Kumi ska J, S epnowski P, Mulkiewicz E. An ibio ic esis ance genes
iden i ied in was ewa e ea men plan sys ems – a e iew. Sci To al En i on
2019;697:134023.
[19] Nies DH. E lux-media ed hea y me al esis ance in p oka yo es. FEMS
Mic obiol Re 2003;27:313–39.
[20] Blanco P, He nando-Amado S, Reales-Calde ón J, Co ona F, Li a F, e al.
Bac e ial mul id ug e lux pumps: Much mo e han an ibio ic esis ance
de e minan s. Mic oo ganisms 2016;4:14.
[21] Li LG, Xia Y, Zhang T. Co-occu ence o an ibio ic and me al esis ance genes
e ealed in comple e genome collec ion. ISME J 2017;11:651–62.
[22] Ledingham K, Hinchli e S, Jackson M, Thomas F, Tomson G. An ibio ic
esis ance: using a cul u al con ex s o heal h app oach o add ess a global
heal h challenge. Copenhagen, Denma k: WHO Regional O ice o Eu ope;
2019.
[23] O’Neill J. An imic obial esis ance: ackling a c isis o he heal h and weal h o
na ions. The Re iew on An imic obial Resis ance. Wellcome T us and he UK
Depa men o Heal h. London, UK; 2014.
[24] O’Neill J. Tackling d ug- esis an in ec ions globally: inal epo and
ecommenda ions. The Re iew on an imic obial esis ance. Wellcome T us
and he UK Depa men o Heal h. London, UK; 2016.
[25] Coope M, Shlaes D. !!
[160] Findlay BL. The chemical ecology o p eda o y soil bac e ia. ACS Chem Biol
2016;11:1502–10.
[161] Robe sen HL, Musiol-K oll EM. Ac inomyce e-de i ed polyke ides as a
sou ce o an ibio ics and lead s uc u es o he de elopmen o new
an imic obial d ugs. An ibio ics 2019;8:157.
[162] Ben ley SD, Cha e KF, Ce deño-Tá aga AM, Challis GL, Thomson NR, e al.
Comple e genome sequence o he model ac inomyce e S ep omyces
coelicolo A3(2). Na u e 2002;9:141–7.
[163] Ikeda H, Ishikawa J, Hanamo o A, Shinose M, Kikuchi H, e al. Comple e
genome sequence and compa a i e analysis o he indus ial mic oo ganism
S ep omyces a e mi ilis. Na Bio echnol 2003;21:526–31.
[164] T axle MF, Kol e R. Na u al p oduc s in soil mic obe in e ac ions and
e olu ion. Na P od Rep 2015;32:956.
[165] Xiao Y, Wei X, Eb igh R, Wall D. An ibio ic p oduc ion by myxobac e ia plays
a ole in p eda ion. J Bac e iol 2011;193:4626–33.
[166] DePas WH, Syed AK, Si uen es M, Lee JS, Wa shaw D, e al. Bio ilm o ma ion
p o ec s Esche ichia coli agains killing by Caeno habdi is elegans and
Myxococcus xan hus. Appl En i on Mic obiol 2014;80:7079–87.
[167] Wang C, Liu X, Zhang P, Wang Y, Li Z, e al. Bacillus licheni o mis escapes om
Myxococcus xan hus p eda ion by deac i a ing myxo i escin a h ough
enzyma ic glucosyla ion. En i on Mic obiol 2019;21:4755–72.
[168] Pé ez J, Muñoz-Do ado J, B aña AF, Shimke s LJ, Se illano L, e al. Myxococcus
xan hus inducesac ino hodin o e p oduc ion and ae ial mycelium o ma ion
by S ep omycescoelicolo . Mic ob Bio echnol 2011;4:175–83.
[169] Mülle S, S ack SN, Ryan SE, Kea ns DB, Ki by JR. P eda ion by Myxococcus
xan hus induces Bacillus sub ilis o o m spo e- illed megas uc u es. Appl
En i on Mic obiol 2015;81:203–10.
[170] Con e as-Mo eno FJ, Muñoz-Do ado J, Ga cía-Tomsig NI, Ma ínez-Na ajas
G, Pé ez J, e al. Coppe and melanin play a ole in Myxococcus xan hus
p eda ion on Sino hizobium melilo i. F on Mic obiol 2020;11:94.
[171] Li ings one PG, Milla d AD, Swain MT, Whi wo h DE. T ansc ip ional
changes when Myxococcus xan hus p eys on Esche ichia coli sugges
myxobac e ial p eda o s a e cons i u i ely oxic bu egula e hei eeding.
Mic ob Genom 2018;4:e000152.
[172] Lee N, Kim W, Chung J, Lee Y, Cho S, e al. I on compe i ion igge s an ibio ic
biosyn hesis in S ep omyces coelicolo du ing cocul u e wi h Myxococcus
xan hus. ISME J 2020;14:1111–24.
[173] Be leman JE, Allen S, Danielewicz MA, Remis JP, Go u A, e al. The le hal
ca go o Myxococcus xan hus ou e memb ane esicles. F on Mic obiol
2014;5:474.
[174] Remis JP, Wei D, Go u A, Zemla M, Ha aga J, e al. Bac e ial social ne wo ks:
s uc u e and composi ion o Myxococcus xan hus ou e memb ane esicle
chains. En i on Mic obiol 2014;16:598–610.
[175] Gnopo YMD, Wa kins HC, S e enson TC, DeLisa MP, Pu nam D. Designe ou e
memb ane esicles as immunomodula o y sys ems- ep og amming bac e ia
o accine deli e y. Ad D ug Deli Re 2017;114:132–42.
[176] E ans AGL, Da ey HM, Cookson A, Cu inn H, Cooke-Fox G, e al. P eda o y
ac i i y o Myxococcus xan hus ou e -memb ane esicles and p ope ies o
hei hyd olase ca go. Mic obiology 2012;158:2742–52.
[177] Schulz E, Goes A, Ga cia R, Pan e F, Koch M, e al. Biocompa ible bac e ia-
de i ed esicles show inhe en an imic obial ac i i y. J Con ol Release
2018;290:46–55.
[178] Goes A, Lapuhs P, Kuhn T, Schulz E, Rich e R, e al. Myxobac e ia-de i ed
ou e memb ane esicles: po en ial applicabili y agains in acellula
in ec ions. Cells 2020;9:194.
[179] Kahn J, Aguiluz K, Koch J, T eune -Lange A, Kono alo a A, e al. P o iling he
ou e memb ane p o eome du ing g ow h and de elopmen o he social
bac e ium Myxococcus xan hus by selec i e bio inyla ion and analyses o
ou e memb ane esicles. J P o eome Res 2010;9:5197–208.
[180] Whi wo h DE, Slade SE, Mi onas A. Composi ion o dis inc sub- p o eomes
in Myxococcus xan hus: me abolic cos and amino acid a ailabili y. Amino
Acids 2015;47:2521–31.
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2554
[181] Zwa ycz AS, Li ings one PG, Whi wo h DE. Wi hin-species a ia ion in OMV
ca go p o eins: he Myxococcus xan hus OMV pan-p o eome. Mol Omics 2020.
h ps://doi.o g/10.1039/d0mo00027.
[182] Li ings one PG, Mo phew RM, Whi wo h DE. Genome sequencing and pan-
genome analysis o 23 Co allococcus spp. s ains e eal unexpec ed di e si y,
wi h pa icula plas ici y o p eda o y gene se s. F on Mic obiol
2018;9:3187.
[183] Su on D, Li ings one PG, Fu ness E, Swain MT, Whi wo h DE. Genome-wide
iden i ica ion o myxobac e ial p eda ion genes and demons a ion o
o maldehyde sec e ion as a po en ially p eda ion- esis an ai o
Pseudomonas ae uginosa. F on Mic obiol 2019;10:2650.
J. Pé ez e al. Compu a ional and S uc u al Bio echnology Jou nal 18 (2020) 2547–2555
2555
