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COMPARATIVE POTENTIAL OF ENDOPHYTIC EXTRACTS
AS NATURAL ALTERNATIVES TO ANTIBACTERIAL DRUGS
B.B. Tojiye 1, U.K. Yusupo 2, D.M. Ruziye a3
PhD s uden , Ins i u e o Mic obiology, Academy o Sciences o he Republic o Uzbekis an1
Ins i u e o Mic obiology, Academy o Sciences o he Republic o Uzbekis an2
PhD in Biological Sciences, Senio Resea che , Ins i u e o Mic obiology, Academy o Sciences
o he Republic o Uzbekis an3
h ps://doi.o g/10.5281/zenodo.17674479
Abs ac . The aim o his s udy was o compa e he an ibac e ial ac i i y o he endophy ic
ex ac o he Al e na ia 8S sp. s ain isola ed om Ginkgo biloba L. and he AV4L isola e ob ained
om Aloe e a. The assays demons a ed ha he Al e na ia 8S sp. s ain exhibi s a mo e
p onounced bac e icidal e ec agains es cul u es such as S aphylococcus au eus, Pseudomonas
ae uginosa, Bacillus sub ilis, Esche ichia coli, and Candida albicans compa ed o AV4L, which showed
mode a e bac e icidal ac i i y. Di e ences in he deg ee o an ibac e ial e ec we e obse ed
based on p elimina y esul s, while he composi ion o ac i e compounds and hei mechanisms o
ac ion equi e u he in es iga ion. The ob ained indings suppo he po en ial o endophy ic
ungi as p omising sou ces o na u al an ibac e ial agen s.
Keywo ds: al e na ia 8S sp., Aloe e a, AV4L, an ibac e ial ac i i y, endophy es.
In oduc ion
I has been no iced ecen ly ha he numbe o pa hogenic mic oo ganisms esis an o
al eady exis ing an ibio ics has no iceably inc eased. This ac is conside ed o be a se ious h ea
o human heal h and equi es he u gen sea ch o new sou ces o an imic obial compounds. One
o he mos p omising ends in mode n mic obiology and bio echnology is he esea ch o na u al
compounds syn hesized by mic oo ganisms and plan s ha ha e demons a ed p onounced
an ibac e ial ac i i y. O pa icula in e es is a g oup o endophy ic mic oo ganisms, which
includes bac e ia and ungi ha li e in plan issues wi hou causing pa hological changes.
Plan me aboli es s ill ep esen he mos signi ican sou ce o disco e y o new and
po en ially aluable pha maceu icals. Howe e , ex ac ion om plan sou ces is limi ed due o
season abili y, he in luence o clima ic condi ions, and he need o cul i a ion a eas. An
al e na i e sou ce ha is highly p omising, inexhaus ible, and enewable o high- alue bioac i e
p oduc s is endophy ic ungi, ha asymp oma ically inhabi a ious pa s o all exis ing plan
species [1].
These mic oo ganisms a e capable o syn hesizing a wide a ie y o seconda y me aboli es
simila in hei ac i i y o plan -de i ed compounds, o en mo e s able and e ec i e. Special
men ion should be gi en o ep esen a i es o he genus Al e na ia, which ha e been epo ed o
exhibi an agonis ic ac i i y agains a ious pa hogenic bac e ia and ungi. On he o he hand, Aloe
e a AV4L con inues o be one o he mos s udied medicinal plan , he ex ac s o which se e as
na u al an isep ics and sou ces o biologically ac i e subs ances. Howe e , ew s udies ha e been
conduc ed on he compa a i e an ibac e ial ac ion o endophy ic ungi and plan ex ac s especially
unde condi ions o local isola ion and es ing [2].
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In his espec , he goal o he cu en s udy was o pe o m a compa a i e analysis o he
an ibac e ial ac ion o he Al e na ia 8S sp. s ain and he endophy ic ex ac AV4L om Aloe
e a agains mul iple pa hogenic mic oo ganisms.
Ma e ials and me hods
Ex ac ion o Seconda y Me aboli es om Endophy ic Fungi. To de e mine biological
ac i i y, he ex ac ion o seconda y me aboli es om he biomass o endophy ic ungi was ca ied
ou acco ding o me hodologies desc ibed in se e al in e na ional scien i ic s udies, wi h
labo a o y modi ica ions in oduced as needed [3]. App oxima ely 5 g o ozen ungal biomass
we e ho oughly g ound in a mo a wi h s e ile glass sand, a e which 50 mL o e hyl ace a e
(E OAc) was added as he ex ac ion sol en . The esul ing suspension was ans e ed in o a
conical lask and incuba ed on a shake a oom empe a u e o 24 hou s. The mix u e was hen
il e ed h ough Wha man №1 il e pape , and anhyd ous sodium sul a e (Na₂SO₄) was added o
emo e esidual mois u e. Subsequen ly, he il a e was e apo a ed o d yness using a o a y
e apo a o unde educed p essu e [4]. The emaining esidue was dissol ed in 1 mL o dime hyl
sul oxide (DMSO) o ob ain a s ock ex ac , which was s o ed a +4 °C un il u he use. The
ob ained ex ac s we e used o assess an ibac e ial and an i ungal ac i i ies [5].
The an ibac e ial p ope ies o he ex ac s we e de e mined using he well-di usion
me hod agains he ollowing es cul u es: g am-posi i e bac e ia (S. au eus and B. sub ilis), g am-
nega i e bac e ia (E. coli and P. ae uginosa), as well as yeas (C. albicans). Mea -pep one aga
(HiMedia, India) was used o es ing he ex ac s.
Pe i dishes con aining he nu ien medium we e inocula ed wi h a 24-hou suspension o
he es cul u es p epa ed in physiological saline a a concen a ion o 1.5 × 10⁸ CFU/mL,
co esponding o he McFa land s anda d. A e sligh d ying, wells wi h a diame e o 5 mm we e
cu in o he aga , and 100 μL o each ex ac was added in o he wells in iplica e. The pla es we e
incuba ed a 37 °C o 24–48 hou s.
Ce iaxone (a hi d-gene a ion cephalospo in an ibio ic, 30 μg/disc) se ed as he posi i e
con ol, while dime hyl sul oxide (DMSO), he sol en o he seconda y me aboli e ex ac s o
endophy ic ungi, was used as he nega i e con ol. The esul s we e e alua ed by measu ing he
diame e s (in millime e s) o he inhibi ion zones o med a ound he wells [1].
Resul s and discussion
The an ibac e ial ac i i y o he biomass ex ac s o Al e na ia 8S sp. and AV4L was
e alua ed using he aga di usion me hod agains i e es cul u es: S aphylococcus au eus,
Pseudomonas ae uginosa, Bacillus sub ilis, Esche ichia coli, and Candida albicans (Table 1).
Sample
S. au eus
P. ae uginosa
B. sub ilis
E. coli
C. albicans
Al e na ia 8S sp.
29 mm
24 mm
28 mm
35 mm
21 mm
AV4L
16 mm
20 mm
13 mm
16 mm
16 mm
Table 1. An ibac e ial ac i i y o Al e na ia 8S sp. and AV4L ex ac s agains selec ed es
mic oo ganisms.
The ob ained da a on he an ibac e ial ac i i y o he Al e na ia 8S sp. and AV4L ex ac s
clea ly demons a e a dis inc ad an age o he ungal isola e o e he plan -de i ed ex ac . The
Al e na ia 8S sp. ex ac exhibi ed p onounced inhibi o y ac i i y agains all i e es cul u es,
including bo h g am-posi i e (S. au eus, B. sub ilis) and g am-nega i e (E. coli, P. ae uginosa)
bac e ia, as well as C. albicans. The maximum inhibi ion zones (35 mm o E. coli and 29 mm o
S. au eus) a e compa able o, o e en exceed, he alues epo ed in se e al published s udies.
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Wi h ega d o plan -de i ed ex ac s o Ginkgo biloba, da a om se e al s udies indica e
ha hey also demons a e an imic obial ac i i y agains bo h g am-posi i e and g am-nega i e
mic oo ganisms. Howe e , he inhibi ion zones ypically ange om 12 o 25 mm, making he
an imic obial ac i i y o Ginkgo biloba compa able o ha o AV4L. In con as , he Al e na ia
8S sp. isola e exhibi s a signi ican ly s onge an agonis ic e ec , highligh ing i s po en ial alue
as a sou ce o biologically ac i e compounds [6-7].
Conclusion
O e all, hese indings indica e ha Al e na ia 8S sp. demons a es an ibac e ial ac i i y
ha is compa able o o su passes he ac i i y o Ginkgo biloba and Aloe e a ex ac s epo ed in
he li e a u e. While plan ex ac s such as Aloe and Ginkgo adi ionally con ain a wide spec um
o phenolic compounds wi h mode a e an imic obial e ec s, endophy ic ungi a e capable o
syn hesizing mo e speci ic and po en seconda y me aboli es—such as al e na iol, al e na iol
monome hyl e he , enuazonic acid, and o he de i a i es—which possess s ong bac e icidal
p ope ies. This likely explains he supe io ac i i y o Al e na ia 8S sp. compa ed o AV4L and
he plan -de i ed ex ac s desc ibed in p e ious s udies.
Fu he mo e, he compa a i e analysis shows ha he ungal ex ac no only p o ides a
b oad spec um o ac ion bu also achie es a high deg ee o inhibi ion, while Aloe e a and Ginkgo
biloba ex ac s, despi e hei p o en biological ac i i y, exhibi subs an ially milde and
p edominan ly bac e ios a ic e ec s. This unde sco es he high po en ial o endophy ic ungi as
al e na i e sou ces o po en na u al an imic obial agen s.
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