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Assessmen o An ibac e ial P ope ies o Me hanolic Lea Ex ac s and Thei
F ac ions om Beninese Medicinal Plan s
Amandine NOUDAMADJO 1, Bien enu GLINMA 1, 2, *, Sèdami MEDEGAN FAGLA 1, 2, Bénédic a KPADONOU-
KPOVIESSI 1, Du and Dah-NOUVLESSOUNON 3 and Salomé Dossou Sika KPOVIESSI 1
1 Depa men o Chemis y, Facul y o Sciences and Technology (FAST), O ganic Chemis y, Na u al Subs ances and
Applica ions Labo a o y (LaCOSNA). Uni e si y o Abomey-Cala i (UAC), 02 BP 69 Bohicon, Benin.
2 Depa men o Chemis y, Facul y o Heal h Sciences (FSS), Pha malab D ug Resea ch and De elopmen Cen e, Uni e si y
o Abomey-Cala i (UAC), Abomey-Cala i, Benin.
3 Depa men o Biochemis y, Facul y o Sciences and Technology (FAST), Labo a o y o biology and molecula yping in
mic obiology (LBTMM), Uni e si y o Abomey-Cala i (UAC), Abomey-Cala i, Benin.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
Publica ion his o y: Recei ed on 11 June 2025; e ised on 16 Augus 2025; accep ed on 01 Sep embe 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.3.3002
Abs ac
The use o plan s in disease ea men is an ancien p ac ice ha laid he g oundwo k o mode n pha macology. This
s udy assessed he an ibac e ial ac i i ies o c ude ex ac s and sol en ac ions om Flueggea i osa and Newbouldia
lae is, wo species ex ensi ely used in A ican adi ional medicine. Me hanolic lea ex ac s we e ac iona ed wi h
hexane, dichlo ome hane, e hyl ace a e, and wa e . An ibac e ial sc eening was pe o med ia aga well di usion and
b o h mic odilu ion me hods o de e mine inhibi ion diame e s (ID), minimum inhibi o y concen a ions (MIC), and
minimum bac e icidal concen a ions (MBC). Toxici y was assessed using A emia salina la ae. Phy ochemical
sc eening e ealed he p esence o di e se seconda y me aboli es. Ex ac s and ac ions om F. i osa exhibi ed
s onge an ibac e ial e ec s, pa icula ly agains S aphylococcus au eus and En e ococcus aecalis, wi h IDs o 7–15 and
7–13.5 mm, espec i ely. The me hanolic ex ac (F M) showed MIC alues o 2.5 and 5 mg/mL, while ac ions F 1 and
F 2 eco ded MICs o 10 and 2.5 mg/mL, espec i ely, on he wo bac e ia. MBC alues anged om 7.5 o 10 mg/mL,
indica ing good bac e icidal po ency (P = 1–4). Con e sely, N. lae is displayed weak o no inhibi ion, excep o ac ion
Nl2, which signi ican ly inhibi ed S. au eus (ID = 13 mm, MIC = 2.5 mg/mL, MBC = 10 mg/mL, P = 4). Nei he plan
showed an ibac e ial ac i i y agains Esche ichia coli. All es ed ex ac s and ac ions we e non- oxic o A. salina (LC50
> 0.30 mg/mL). The indings p esen ed he signi ican an ibac e ial po en ial o plan , likely linked o syne gis ic
phy ocompounds, p o iding scien i ic suppo o i s e hnomedicinal applica ion. Fu he bio-guided ac iona ion and
compound isola ion a e ecommended o iden i y hei ac i e an ibac e ial cons i uen s.
Keywo ds: Medicinal plan s; Ex ac s and ac ions; An ibac e ial ac i i y; Mic o-o ganisms
1. In oduc ion
Medicines a e essen ial o ea ing a ious diseases. Bu hey a e o en used inco ec ly [1-3]. I a ional use o
medicines can cause many p oblems. These include inc eased mo ali y and mo bidi y. They can also esul in poo
ea men ou comes. In addi ion, i can lead o was age o heal hca e esou ces [1, 4]. An ibio ics a e pha maceu icals
employed in he ea men o bac e ial in ec ions, including, bu no limi ed o, pneumonia, b onchi is, ea in ec ions,
meningi is, u ina y ac in ec ions, sep icemia and sexually ansmi ed diseases. This is one o medicine's mos
signi ican disco e ies, sa ing millions o li es [5-8]. An ibio ics kill bac e ia o s op hem om ep oducing, allowing
he body's na u al de enses o elimina e hem. Howe e , hey a e ine ec i e agains i uses and mos o he ypes o
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
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in ec ion [9]. The o e use o an ibio ics has led o an inc ease in bac e ial esis ance o an imic obial agen s [10]. This
esis ance can a ise ei he endogenously, h ough mu a ion and selec ion wi hin he pa hogen i sel , o exogenously,
h ough ho izon al gene ansmission (HGT) om en i onmen al o ganisms (e.g. an ibio ic p oduce s, commensals and
non-human pa hogens) o human pa hogens [11-14]. In ec ions caused by esis an bac e ia can lead o se ious
consequences, including longe illnesses and hospi alisa ions, inc eased mo ali y, and educed pa ien p o ec ion
du ing su ge y and o he medical p ocedu es [10, 15]. An imic obial esis ance (AMR) is one o he op global public
heal h and de elopmen h ea s. I is es ima ed ha bac e ial AMR was di ec ly esponsible o 1.27 million global
dea hs in 2019 and con ibu ed o 4.95 million dea hs [16]. The wo ld aces an an ibio ics pipeline and access c isis.
The e is an inadequa e esea ch and de elopmen pipeline in he ace o ising le els o esis ance, and u gen need o
addi ional measu es o ensu e equi able access o new and exis ing accines, diagnos ics and medicines [17]. In addi ion
o dea h and disabili y, an imic obial esis ance (AMR) has signi ican economic cos s. The Wo ld Bank es ima es ha
AMR could esul in US$ 1 illion addi ional heal hca e cos s by 2050, and US$ 1 illion o US$ 3.4 illion g oss
domes ic p oduc (GDP) losses pe yea by 2030 [18]. Faced wi h inc easing an ibio ic esis ance and a sho age o new
an ibio ics, i is essen ial o use an ibio ics a ionally [10, 19, 20]. A wide ange o human in ec ions a e caused by he
ollowing pa hogens : S aphylococcus au eus, En e ococcus aecalis and Esche ichia coli. They a e esponsible o a
numbe o p oblems ha a ec people's li es and mobili y. Fu he mo e, he de elopmen o esis ance o exis ing
ea men s, he high cos o an ibio ics, and hei inaccessibili y o mos o he popula ion make i essen ial o ind mo e
accessible solu ions. Thus, u gen ac ions need o be aken o e ec i ely con ol human, animal and plan AMR
pa hogens. Plan s a e cu en ly o pa amoun impo ance in all ci ilisa ions ha use hem, whe he wild o cul i a ed,
o ood, de ense and clo hing [21]. The he apeu ic use o plan s o ea human diseases is e y old [22] and has
e ol ed alongside humani y h oughou his o y [23]. S aphylococcus au eus, En e ococcus aecalis and Esche ichia coli
a e impo an pa hogens ha cause a wide ange o human in ec ions. T adi ional heale s use medicinal plan s o ea
a ious diseases, including mala ia, ulce s, diabe es and cance …. Unlike mode n medicine, his o m o medicine is
accessible o all and less expensi e [24-26]. In p e ious wo k, we explo ed he phy ochemical sc eening and biological
ac i i ies o ex ac s om plan s Flueggea i osa and Newbouldia lae is. This pape s udies he an ibac e ial ac i i ies o
hese wo plan s on h ee s ains S aphylococcus au eus, En e ococcus aecalis and Esche ichia coli, as well as hei la al
oxici y.
2. Ma e ial and me hods
2.1. Ma e ial
2.1.1. Plan ma e ial
The plan ma e ial consis s solely o lea es (Figu e 1). A e ha es ing, hey we e iden i ied, ce i ied and au hen ica ed
by he Na ional He ba ium o he Uni e si y o Abomey-Cala i unde he numbe s YH1060/HNB and YH1061/HNB.
Figu e 1 Le «Flueggea i osa» (lea es and ui s) ; Righ «Newbouldia lae is» (lea es and s ems)
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
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2.1.2. Chemical ma e ial
The a ious sol en s we e pu chased di ec ly om he manu ac u e s and a e used wi hou u he pu i ica ion. These
a e n-hexane (Me ck), dichlo ome hane (ReAgen ), and e hyl ace a e (Sigma-Ald ich), iodo-ni o e azolium chlo ide
(Sigma-Ald ich).
2.1.3. Mic obiological ma e ial
All biological ma e ial is acqui ed and used unde he condi ions o he Labo a o y o Biology and Molecula Typing in
Mic obiology.
2.2. Me hods
Ou p e ious wo k [27, 28] desc ibed he ha es ing o plan lea es ; he p epa a ion o me hanol ex ac s and hei
phy ochemical analysis. This pape desc ibed he p ocedu e o ob aining ac ions om hese ex ac s and some
bac e ial es s. Two me hods we e used o ac iona e he ex ac s, depending on he quan i y o c ude ex ac s a ailable.
2.2.1. F ac iona ion me hod o F. i osa c ude ex ac
A o al o 211 g o c ude ex ac ob ained by mace a ing F. i osa powde in me hanol, was ea ed wi h a me hanol-
wa e mix u e (20:80, / ) o sol en ac iona ion (Figu e 2). The sol en s used we e o inc easing pola i y: hexane,
dichlo ome hane (DCM), e hyl ace a e. Each sol en was added o he ex ac h ee imes in succession: 500 mL o
hexane, e hyl ace a e and dichlo ome hane. Each o hese ac ions was concen a ed using a o a y e apo a o and hen
d ied. Fou ac ions we e ob ained: hexane (F 1 = 26 g), dichlo ome hane (F 2 = 14 g), e hyl ace a e (F 3 = 49 g), and
he aqueous esidue (F 4 = 47 g).
Figu e 2 P ocedu e o ac iona ing o F. i osa c ude ex ac
2.2.2. F ac iona ion me hod o N. lae is c ude ex ac
70 g o he c ude ex ac o N. lae is by mace a ion we e subjec ed o liquid-liquid pa i ioning using sol en s o
inc easing pola i y (Figu e 3). Using he same p ocedu e as desc ibed abo e, ou ac ions we e ob ained such as
ac ion o hexane (Nl1 = 30 g), dichlo ome hane (Nl2 = 9 g), e hyl ace a e (Nl3 = 6 g) and he aqueous esidue (Nl4 =
20 g).
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
34
Figu e 3 P ocedu e o ac iona ing o N. lae is c ude ex ac
2.3. An ibac e ial es
The ollowing mic oo ganisms we e used in he s udy: wo G am-posi i e (S aphylococcus au eus ATCC 29213 and
En e ococcus aecalis ATCC 29212) and one G am-nega i e (Esche ichia coli ATCC 25922). An imic obial ac i i y was
assessed by sensi i i y es ing o ex ac s and ac ions on bac e ial s ains. Indica o s o ac i i y will be minimum
inhibi o y concen a ions (MICs) and minimum bac e icidal concen a ions (MBCs).
The es s we e pe o med using a s anda d me hodology ha has been adap ed o use in ou labo a o y.
2.3.1. Sensi i i y es
The Mülle -Hin on (MH) solid medium di usion me hod, as desc ibed by Anani e al. [29], will be used o es he
sensi i i y o mic obial s ains o ex ac s. The p e ious day's bac e ial p e-cul u e (one colony in one millili e o liquid
Muelle -Hin on) will be dilu ed o achie e a u bidi y o 0.5 on he McFa land scale (i.e. 10⁸ CFU/mL), hen educed o
10⁶ CFU/ml in s e ile dis illed wa e . This bac e ial suspension (1000 µL) will be used o lood a Pe i dish con aining
Muelle -Hin on aga medium (Bio-Rad, F ance) [30-32]. Using a pe o a o , 6 mm diame e pape disks will be made.
The s e ile disks will be deposi ed unde asep ic condi ions on o pla es ha ha e p e iously been looded wi h bac e ial
cul u e. 30 µL o he ex ac o be es ed will be inocula ed on o he disks unde asep ic condi ions. Fo each ex ac , he
expe imen will be duplica ed and a nega i e con ol will be pe o med using he sol en ins ead o he ex ac . The
pla es a e hen le o 15–30 minu es a oom empe a u e be o e being incuba ed a 37 °C in an o en o 24 and 48
hou s [33]. A e he incuba ion imes o 24 and 48 hou s, inhibi ion diame e s a e measu ed using a g adua ed ule
[34].
2.3.2. De e mina ion o he minimum inhibi o y concen a ion (MIC)
In addi ion o es ing bac e ial sensi i i y o a ious mix u es, he MICs we e de e mined. The MIC o an ex ac o a
s ain is he lowes concen a ion a which no isible g ow h occu s wi hin 24 hou s. In his s udy, hey we e de e mined
using he mic o-dilu ion me hod and iodo-ni o e azolium (INT) as a bac e ial iabili y indica o [35]. The me hod
in ol ed p epa ing he bac e ial inoculum wi h 24-hou -old young colonies, dis ibu ing 50 µL o s e ile MH b o h in o
96 wells o a mic opla e (excep he i s ow), adding 50 µL o he s ock ex ac solu ion o he i s wo columns o he
mic opla e, c ea ing a wo old dilu ion om he i s well o he second ow o he las well, dispensing 50 µL o inoculum
in o each well, incuba ing he mic opla es o 24 hou s a 37 °C, adding 20 µL o 0.01% INT solu ion o each well; Re-
incuba e o 30 minu es a 37°C ; Take he eading. Wells u ning pink indica e bac e ial g ow h.
The MIC co esponds o he i s well in which no pink colo a ion is obse ed due o he p esence o INT.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
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2.3.3. De e mina ion o he minimum bac e icidal concen a ion (MBC)
The MBC will be de e mined on he basis o he MIC esul s. A e iden i ying he MIC, all he o he ubes om he MIC
o he high concen a ions will be inocula ed on o Pe i dishes con aining MH aga medium using a loop. The pla es will
be examined a e an incuba ion pe iod o 24 hou s a 37°C. The MBC is he ex ac concen a ion a which no bac e ial
g ow h is obse ed [36].
2.3.4. De e mina ion o he an imic obial po ency o ex ac s and ac ions
An ibac e ial po ency (AP) is de ined as he a io o he minimum bac e icidal concen a ion (MBC) o he minimum
inhibi o y concen a ion (MIC). This p o ides in o ma ion on he bac e icidal and bac e ios a ic p ope ies o a p oduc
[37].
AP =MBC
MIC
I he an ibac e ial powe (AP) is less han o equal o ou (AP ≤ 4), he p oduc es ed is bac e icidal. I AP is g ea e
han ou , he p oduc es ed is bac e ios a ic [38].
A bac e ios a ic e ec means ha bac e ia s op mul iplying wi hou necessa ily being des oyed. I in ol es he
e e sible inhibi ion o ce ain biological unc ions necessa y o mic obial me abolism, g ow h and mul iplica ion
wi hou a ec ing all i al unc ions [39]. The bac e icidal e ec esul s in he de ini i e des uc ion o he mic obe o e
a pe iod o ime. This des uc ion is linked o an i e e sible lesion ha ende s u he p oli e a ion impossible [40].
2.4. Toxici y sc eening
La al oxici y was assessed using he me hod ou lined by Dougnon e al. [41]. A emia salina Leach eggs we e incuba ed
in seawa e o 48 hou s un il he young la ae ha ched. A se ies o second-o de dilu ions o he ex ac we e made o
ob ain a ange o concen a ions. Six een la ae we e placed in 1 mL o seawa e and hen ans e ed o each dilu ion
o he ex ac . All dilu ions, as well as he con ol solu ion con aining no ex ac , we e le unde agi a ion o 24 hou s.
The numbe o dead la ae in each solu ion was coun ed unde a mic oscope o assess he la al cy o oxici y o he
ex ac . Dose- esponse da a we e exp essed as loga i hms o base 10 and he mean le hal concen a ion (LC₅₀) was
de e mined by linea eg ession. Finally, la al oxici y was assessed using he Mousseux scale [42]. An LC50 alue
g ea e han o equal o 0.1 mg/mL indica es ha he ex ac is non- oxic, while a alue be ween 0.1 mg/mL and 0.05
mg/mL indica es low oxici y. An LC50 alue be ween 0.05 mg/mL and 0.01 mg/mL indica es mode a e oxici y, while
an LC50 alue below 0.01 mg/mL indica es high oxici y.
3. Resul s and discussion
3.1. Phy ochemical sc eening
In ou p e ious wo k, i was demons a ed ha ex ac s om plan lea powde we e ich in a ious seconda y
me aboli es. Thei ex ac s we e sc eened o he p esence o alkaloids, ca bohyd a es, phenols, gums and mucilage,
la onoids, s e oids, p o eins, annins, and saponins using s anda d quali a i e me hods ( able 1). Se e al
phy ochemicals esponsible o pha macological ac i i ies, such as an imic obial, an ipa asi ic, analgesic, and an i-
in lamma o y p ope ies, ha e been iden i ied.
These include phy ochemicals wi h an imic obial and an i-in lamma o y p ope ies, and se e al ecognised ac i i ies
o hese plan s a e epo ed in he li e a u e [27, 28, 43-46]. In las wo ks, MIC o me hanolic ex ac = 31.2 and 250
μg/mL agains S. au eus s ain ATCC 12600 we e ob ained om F. i oa and N. lae is espec i ely ; hyodex anolic
ex ac mode a ely impac s cell g ow h. This explains why me hanolic ex ac was chosen o ob ain he di e en
ac ions o his wo k.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
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Table 1 Seconda y me aboli es om phy ochemical sc eening
Seconda y me aboli es
F. i osa
lea es
N. lae is
lea es
Seconda y me aboli es
F. i osa
lea es
N. lae is
lea es
Alkaloids
+
+
T i e penes
+
-
Polyphenols
+
+
Ca denolides
-
-
Tannins
+
+
Cyanogenic de i a i es
-
-
Ca echin annins
+
+
Mucilages
+
+
Gallic annins
+
+
Couma ins
+
+
Fla onoids ( la ones)
+
+
Reducing compounds
+
+
An hocyanins
+
+
F ee an h acene
de i a i es
-
-
Leuco an hocyanins
-
-
An h acenic O-
he e osides
+
+
Quinonic de i a i es
-
+
An h acenic C-
he e osides
+
+
Saponins
+
-
3.2. An ibac e ial Ac i i y
The esul s o he sensi i i y es we e shown in he ollowing able. The in luence o he ex ac s and ac ions on he
bac e ial s ains es ed made i possible o measu e he diame e s o he inhibi ion zones o he samples eco ded in he
able abo e.
Table 2 Inhibi ion diame e s o ex ac s and ac ions used in he s udy
Inhibi ion diame e s (mm)
Ex ac s and ac ions
S. au eus ATCC 29213
E. oecalis ATCC 29212
E. coli ATCC 25922
F M
8±0
7±0
-
F 1
15±0
13.5±1,5
10±3
F 2
7±0
7±0
-
F 3
-
-
-
F 4
-
-
-
NlM
-
-
-
Nl1
-
10±0
-
Nl2
13±0
-
-
Nl3
-
-
-
Nl4
-
-
-
F M : me hanolic ex ac o F. i osa ; F 1 : hexane ac ion ; F 2 : DCM ac ion ; F 3 : AcOE ac ion and F 4 : aqueous esidue.
NlM : me hanolic ex ac o N. lae is ; Nl1 : hexane ac ion ; Nl2 : DCM ac ion ; Nl3 : AcOE ac ion and Nl4 : aqueous esidue
Resul s showed ha he s ains a e mo e sensi i e o he me hanolic ex ac and ac ions om F. i osa han o hose
om N. lae is, which showed no educ ion in sensi i i y. The me hanol ex ac F M and he dichlo ome hane ac ion
F 2 showed a ini y o he G am-posi i e bac e ia S. au eus and E. aecalis wi h inhibi ion diame e s o 07 mm and 08
mm, espec i ely. Meanwhile, he hexane ex ac F 1 showed pa icula sensi i i y o he h ee s ains, wi h inhibi ion
diame e s o 15, 13.5, and 10 mm o S. au eus, E. aecalis, and E. coli, espec i ely. Fu he mo e, he ac ion F 3 and
he esidue F 4 showed no e ec on he s ains s udied. Rega ding Newbouldia lae is, only he hexane (Nl1) and
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
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dichlo ome hane (Nl2) ac ions had an e ec on he E. aecalis and S. au eus s ains, espec i ely. The e o e, he
me hanolic ex ac and he o he ac ions showed no esul s. No e ec was obse ed on he s ains, ei he o he e hyl
ace a e ac ions o o he aqueous esidues om he plan s. Ou esul s a e consis en wi h hose o Amenu e al. [47],
which a y depending on he mic oo ganisms s udied. In hei esea ch, i was demons a ed ha signi ican ac i i y
agains S. au eus and E. coli was exhibi ed by he e hanol ex ac om S. i osa oo s, wi h a iable inhibi o y ac i i y
being shown agains o he o ganisms. The lowes inhibi ion was obse ed agains Mic ococcus lu eus. The inhibi ion
zone anged om 6.33 o 17.67 mm.
Du ing his s udy, we ound ha nei he he ex ac no he ac ions o he wo plan s exhibi ed any an ibac e ial
ac i i y agains E. coli. This s ain was esis an o all ex ac s. S udies ha e been con inued on he wo G am-posi i e
bac e ial s ains. The minimum inhibi o y concen a ions (MIC), and minimum bac e icidal concen a ions (MBC), and
hen he an ibio ic powe (AP) we e de e mined o he ex ac s and ac ions o each plan . Resul s we e desc ibed in
he ollowing able.
Table 3 MICs (mg/mL), MBCs(mg/mL) and an ibio ic po ency (AP) o F. i osa
Ex ac s and ac ions
S. au eus ATCC 29213
E. Foecalis ATCC25922
ID
MIC
MBC
AP
ID
MIC
MBC
AP
F M
8±0
2,5±0
10
4
7±0
5±0
7,5
1,5
F 1
15±0
10±0
10
1
13,5±1,5
10±0
10
1
F 2
7±0
2,5±0
7,5
3
7±0
2,5±0
10
4
F 3
NA
NA
NA
ND
NA
NA
NA
ND
F 4
NA
NA
NA
ND
NA
NA
NA
ND
ID : inhibi o y diame e s (mm) ; NA : no ac i e ; ND no de e mined
I should be no ed ha he wo bac e ia used o he an imic obial es a e highly sensi i e o ex ac s and ac ions
de i ed om Flueggea i osa lea powde , wi h MICs anging om 2.5 o 10 mg/mL and MBCs om 7.5 o 10 mg/mL.
In pa icula , he an ibio ic po ency (o powe ) (AP ≤ 4) a ies be ween he me hanol ex ac (F M) and he hexane
(F 1) and dichlo ome hane (F 2) ac ions, which display AP alues : 1, 1.5, 3, and 4. S. au eus and E. aecalis showed
high sensi i i y o he hexane pa i ion, wi h an inhibi o y diame e o 15 mm and 13.5 mm, espec i ely. The me hanol
ex ac and dichlo ome hane ac ion exhibi ed he g ea es an ibio ic po ency (1.5, 3 and 4). The wo s ains we e
esis an o he e hyl ace a e ac ion (F 3) and aqueous esidue (F 4). These indings suppo hose in he li e a u e in
which S. i osa's mos ac i e ex ac was he chlo o o m one, showing ac i i y agains 13 es o ganisms wi h MIC alues
anging om 15.6 μg/mL o o e 1000 μg/mL. The pe oleum spi i , chlo o o m and e hanol ex ac s o S. i osa
combined wi h s anda d an ibio ics showed e ec s agains an ibio ic- esis an s ains o S. au eus. Mo eo e , p e ious
epo s ha e documen ed he an ibac e ial p ope ies o secu in and i oallosecu nin, which a e alkaloids de i ed om
he lea es o F. i osa [48, 49]. In 2020, Ana ado e al. in hei s udy showed ha S. au eus was highly sensi i e o he
e hyl ace a e ex ac o Secu inega i osa bu esis an o he n-hexane ex ac [50]. O he esea ch has demons a ed
ha a me hanol ex ac o S. i osa oo ba k (6.25 o 25 mg/kg body mass, by in ape i oneal injec ion) conside ably
supp essed (P < 0.05) ace ic acid-induced s omach con ac ions and educed o malin-induced neu ogenic pain (phase
2). I also signi ican ly slowed he eac ion ime (P < 0.01) o mice o he mal pain induced by a hea ing pla e. A he
doses es ed, he ex ac educed paw swelling by 12%, 52%, and 52% by he hi d hou [51]. Recen ly, an e hanolic
ex ac de i ed om he ae ial pa s o F. i osa demons a ed an isickle cell ac i i y by no malising he shape o
abno mal ci cula ing e y h ocy es, a p ope y known as an i alcemic o an isickling ac i i y [52]. P e iously, he same
ype o ac i i y was demons a ed wi h an aqueous me hanolic lea ex ac , which inhibi ed sodium me abisulphi e-
induced sickling o HbSS ed blood cells in a concen a ion-dependen manne [53].
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
38
The ollowing esul s we e ob ained om he me hanol ex ac and ac ions o he N. lae is plan ( able 4).
Table 4 MICs (mg/mL), MBCs(mg/mL) and an ibio ic po ency (AP) o N. lae is
Ex ac s and ac ions
S. au eus ATCC 29213
E. Foecalis ATCC25922
ID
MIC
MCB
AP
ID
MIC
MCB
AP
NlM
NA
NA
NA
NA
NA
NA
NA
ND
Nl1
NA
NA
NA
NA
10±0
NA
NA
ND
Nl2
13±0
2,5±0
10
4
NA
NA
NA
ND
Nl3
NA
NA
NA
NA
NA
NA
NA
ND
Nl4
NA
NA
NA
NA
NA
NA
NA
ND
ID : inhibi o y diame e s ; NA : no ac i e ; ND no de e mined
O e all, all ac ions excep he dichlo ome hane ac ion emained inac i e agains he bac e ia used. The E. aecalis
s ain showed highly esis an o all ex ac s and ac ions. Only Nl2 inhibi ed bac e ial ac i i y on S. au eus (MIC = 2.5;
MBC = 10 mg/mL). This ex ac exhibi ed a bac e icidal e ec wi h an ibio ic po ency equi alen o 4. Ou wo k yielded
di e en esul s o hose ob ained by Suleiman e al. in 2024 in he e hyl ace a e lea ex ac o N. lae is. They
demons a ed ha he ex ac om he ae ial pa s exhibi ed an ibac e ial ac i i y agains he pa hogenic s ains K.
pneumoniae, P. ae uginosa, S. au eus, E. coli and S. yphi a a ying concen a ions depending on he bac e ial species
[54]. O he esea che concluded ha mic obes could no g ow a a ying doses o plan ex ac s and ha no signi ican
di e ences (P > 0.05) we e obse ed in he inhibi ion zones as he ex ac concen a ions inc eased [55]. In con as ,
Okeke e al. ound ha highe concen a ions esul ed in signi ican g ow h inhibi ion. The esul s sugges ha he b oad
spec um o an ibac e ial ac i i y exhibi ed by F. i osa can be a ibu ed o he p esence o di e se ac i e seconda y
me aboli es in he ex ac and i s a ious pa i ions o ac ions [56]. The inhibi o y e ec could be a esul o annins
and la onoids, which ha e been employed as an imic obials o a long ime. On he N. lae is side, whe e he bac e ia
showed s ong esis ance o he ex ac s and ac ions, pe haps he sol en s used did no su icien ly ex ac he
me aboli es esponsible o he ac i i ies. The nex s ep in he wo k will be o ca y ou a phy ochemical analysis o each
ex ac and ac ion o each plan . This could help us o be e unde s and and explain he di e en esul s ob ained.
This s udy also in es iga ed he oxici y o he ex ac s and ac ions ha eac ed wi h bac e ia ( able 5). To achie e his,
A emia salina L. la ae we e used o e alua e he oxici y o each ex ac . We used Camp o hecin as a posi i e con ol,
a well-jus i ied choice. Camp o hecin and i s de i a i es a e used in chemo he apy because hey a e cy o oxic o
mammalian cells. I a ge s opoisome ase I, a highly conse ed enzyme, all o whose esidues in con ac wi h he d ug
in a c ys alline s uc u e o opo ecan/human opoisome ase I [57, 58] a e conse ed in insec s. Se e al s udies ha e
been conduc ed on his molecule, wi h a ying esul s [59-61]. I is inc easingly used in p elimina y oxici y s udies o
plan ex ac s and ac ions, as well as isola ed o syn hesized compounds. The le hal hal -concen a ions (LC50) o each
solu ion we e de e mined (see he able below). This es is a p elimina y in i o oxici y me hod ha is no en i ely
compa able o a es on human cells in cul u e. Al hough he e is a co ela ion be ween he wo ypes o cells, Ca ballo
e al. sugges ed pe o ming bo h es s be o e d awing conclusions, which would gi e a eliabili y o 75% compa ed o
50% o he es on sh imp la ae alone [62].
Table 5 Toxici y o ex ac s and pa i ion o plan s
F. i osa
N. lae is
Ex ac s and ac ions
LC50 (mg/mL)
Toxici y
Ex ac s and ac ions
LC50 (mg/mL)
Toxici y
F M
4.00
no oxic
NlM
3.01
no oxic
F 1
1.11
no oxic
Nl1
2.99
no oxic
F 2
1.02
no oxic
Nl2
0.30
no oxic
Con ol
LC50 = 0.13 mg/mL
non oxic
In ou s udy, and acco ding o he oxici y scale used, camp o hecin has an LC50 alue o 0.13 mg/mL, which is also
highe han he limi o 0.1 mg/mL [42] ha indica es whe he a subs ance unde s udy is oxic o no . I was ound ha
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(03), 031-043
39
all ou ex ac s and ac ions had an LC50 > 0.30 mg/mL. This esul is consis en wi h o he s ha ha e been ca ied ou
on o he pa s o he plan . In 2022, an acu e oxici y s udy showed ha he F. i osa oo ex ac s es ed did no cause
any mo ali y in mice up o 5000 mg/kg body weigh [63]. This p elimina y oxici y es is pe o med o gain an
unde s anding o he po en ial ha m ul e ec s o using hese ex ac s. I also p o es ha he bac e icidal e ec s (PA) o
hese ex ac s on he s ains a e no caused by he sol en s, which we e ho oughly e apo a ed using app op ia e
me hods. Howe e , gi en he co ela ion be ween cy o oxici y in sh imp la ae and in 9PS and 9KB cells (human
nasopha yngeal ca cinoma) on he one hand [64] and in A-549 lung ca cinoma cells and HT-29 colon cells on he o he
[62], i can be concluded, pending u he in es iga ion, ha he es ed ex ac s do no exhibi cy o oxic ac i i y and can
he e o e be used wi hou isking oxici y in he sho and medium e m.
4. Conclusion
This s udy demons a ed ha , o he me hanolic lea ex ac s and hei ac ions es ed, Flueggea i osa exhibi ed
signi ican ly highe an ibac e ial ac i i y agains G am-posi i e and G am-nega i e bac e ial pa hogens han
Newbouldia lae is. The supe io e icacy o F. i osa, pa icula ly e iden in se e al o i s ac ions, sugges s he p esence
o po en bioac i e compounds wi h b oad-spec um po en ial. In con as , N. lae is showed li le o no ac i i y agains
he bac e ial s ains es ed. None o he plan ex ac s o ac ions exhibi ed la al oxici y. These esul s con i m he
adi ional use o plan s by he gene al public o ea ing in ec ious diseases. Ou wo k also co obo a es he indings
o o he pha macological s udies, which suppo i s alue as a sou ce o new an ibac e ial agen s.
Compliance wi h e hical s anda ds
Acknowledgmen s
Au ho s hank Doc o Hyacin he AGNIMONHAN, P o esso s Lamine BABA-MOUSSA, Haziz SINA, Joachim GBENOU,
Eléono e YAYI, Fe nand GBAGUIDI and e e yone who con ibu ed o his s udy.
Disclosu e o con lic o in e es
No con lic o in e es in ou manusc ip
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