Arabian Journal for Science and Engineering

https://doi.org/10.1007/s13369-021-05592-w

RESEARCH ARTICLE-BIOLOGICAL SCIENCES

Alkaloids Rich Extracts from Brown Algae Against Multidrug‑Resistant

Bacteria by Distinctive Mode of Action
Rabia Alghazeer1 · Salah Azwai2 · Aboubaker M. Garbaj3 · Amr Amr4 · Sana Elghmasi5 · Maammar Sidati6 ·
Ervia Yudiati7 · Mahasn G. Kubbat8 · Areej A. Eskandrani9 · Ghalia Shamlan10 · Wafa S. Alansari11

Received: 6 October 2020 / Accepted: 21 March 2021

© King Fahd University of Petroleum & Minerals 2021

Abstract
Algal alkaloids are widely used for their pharmacological properties as antimicrobial agents. This study determined the
antibacterial activities of algal alkaloid-rich extracts against isolates of multidrug-resistant Staphylococcus aureus and entero-
haemorrhagic Escherichia coli (EHEC) O157, as well as the probable mode of action underlying their antibacterial effect. The
total alkaloids were extracted from two Libyan brown algae, namely Sargassum hornschuchii and Cystoseira compressa and
tested against six different isolates from the bacteria mentioned above using the agar-well diffusion method, and their mode
of action on isolates was evaluated by several bacterial physiological indicators, including intracellular potassium ion efflux
and nucleotide leakage. Also, the extracts’ hemolytic activity was assessed as an indicator of their cytotoxicity on red blood
cells. Although not to the same extent, both alkaloid extracts presented antibacterial activities against all tested isolates with
no evidence of bacterial regrowth. The alkaloid extract from S. hornschuchii exerted the best effect on bacteria growth with
minimum inhibitory concentration values ranging between 125 and 500 mg/mL. The results showed that the alkaloid extracts
significantly induced a distinct release of nucleotide and potassium ions out of the cell membrane, indicating that they cause
a change in the fluidity or permeability or both of the cell membrane. Moreover, the results revealed that there were very low
cytotoxic effects. Therefore, algal alkaloids may contribute to the development of potential antibacterial agents in the future.

Keywords Alkaloids · Antibacterial activity · Cystoseira compressa · Escherichia coli · Sargassum hornschuchii ·
Staphylococcus aureus

7
* Rabia Alghazeer Department of Marine Science, Faculty of Fisheries
R.alghazeer@uot.edu.ly and Marine Science, Diponegoro University, Semarang,
Indonesia
1
Biochemistry Division, Chemistry Department, Faculty 8
General of Advanced Laboratory of Chemical Analysis,
of Sciences, University of Tripoli, Tripoli 50676, Libya
Libyan Authority to Search Science and Technology, Tripoli,
2
Department of Microbiology and Parasitology, Faculty Libya
of Veterinary Medicine, University of Tripoli, Tripoli, Libya 9
Chemistry Department, Faculty of Science, Taibah
3
Department of Food Hygiene and Control, Faculty University, Medina 30002, Saudi Arabia
of Veterinary Medicine, University of Tripoli, Tripoli, Libya 10
Department of Food Science and Nutrition, College
4
Department of Chemistry, The Libyan Academy, Tripoli, of Food and Agriculture Sciences, King Saud University,
Libya Riyadh 11362, Saudi Arabia
5 11
Department of Biochemistry, Faculty of Medicine, University Biochemistry Department, Faculty of Science, University
of Tripoli, Tripoli, Libya of Jeddah, Jeddah 21577, Saudi Arabia
6
Biotechnology Lab, Marin Biology Research Center,
Tajura-East of Tripoli, Tripoli, Libya

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 Arabian Journal for Science and Engineering

1 Introduction 2 Materials and Methods

Marine algae are a promising source of novel natural prod- 2.1 Algal Species
ucts, such as glycosides, alkaloids, saponins, flavonoids,
steroid, and tannins, and their related metabolites appear Cystoseira compressa and Sargassum hornschuchii were
to be of significant interest in the food and pharmaceutical collected from the western coast of Libya (SA 01, N32
industries [1]. Alkaloids are novel algal biochemically active 53.764 E13 20.990; SA 02, N32 53.756 E13 21.064; SA
compounds attracting significant drug research interest due 03, N32 53.792 E13 21.070; SA 04, N32 53.804 E13
to their complex and diverse structure and biological activ- 21.028; SA 05, N32 53.777 E13 20.983) between June and
ity. Marine alkaloids are obtained from marine organisms, August 2017. Samples were identified at the Department
such as green–blue algae and macroalgae [2]. Several alka- of Marine Plankton and Algae, Marine Biology Research
loids are found in several kinds of algae, which have been Centre, Tripoli, Libya. Algae samples were cleaned by
published [3]. Specific algal alkaloids include 2-phenyleth- removing the epiphytes and necrotic parts. Algal sam-
ylamine, halogenated alkaloids, and indole derivatives with ples were rinsed with sterile water and shade dried for
different substitutions and functionalities [4–6]. 7–14 days, ground thoroughly to powder, and stored
Algal alkaloid extracts presented interesting bioactivities at 4 °C in opaque bottles until use. The study obtained
against a wide range of microorganisms [7, 8], in addition to favorable opinion from the Libyan National Committee
demonstrating antioxidant [9, 10], anticancer [11, 12], anti- for Biosafety and Bioethics (20/HS/66).
inflammatory [13], neuromodulatory, and neurotransmitory
functions [5, 14].
2.2 Alkaloids Extraction
The antibacterial potential of both natural and synthetic
alkaloids has been widely investigated. Marine extracts
Alkaloids-rich fraction of algae was isolated using the
show significant antibacterial activity that partially relates to
acid–base extraction method [22]. Powdered algae mate-
their rich source of alkaloids. For instance, the total alkaloid
rials (50 g) were extracted several times with methanol
extract of certain red and brown algae demonstrated inhibi-
(300 ml) until algae material gave a negative result for
tory activity against eight human pathogens [7]. Moreover,
alkaloids (Mayer’s test). The mixture was filtered using
pyrrole-imidazole alkaloids isolated from marine sponges
Whatman No. 1 filter paper, and the obtained methanolic
showed antibiotic resistance against the Gram-negative
extract was evaporated under reduced pressure. The crude
Acinetobacter baumannii and Gram-positive methicillin-
alkaloid mixture was then separated from neutral and
resistant Staphylococcus aureus [15].
acidic materials and water-soluble ingredients by extrac-
Several mechanisms have been suggested for the antimi-
tion with aqueous acetic acid (5%), followed by dichlo-
crobial action of isolated phytochemicals against pathogenic
romethane extraction (150 ml). The aqueous solution was
bacteria, but uncertainties continue to exist, indicating the
basified with ­Na2CO3 (10%) to pH 10 and then subjected
need for extensive in vitro investigations using sensitive
to further dichloromethane extraction. The dichlorometh-
enteric pathogenic microorganisms. Phytochemicals can kill
ane layer rich in alkaloids was separated and evaporated.
or inhibit bacterial growth in many ways, such as inducing
Dragendorff reagent was used to confirm the presence of
alteration in the membrane permeability, interacting with
alkaloids in the residue. The alkaloid-rich fraction was
metabolic processes, and modulating signal transduction
dissolved in 0.01% ethanol and stored at − 20 °C for sub-
or gene expression pathways [16, 17]. Alkaloids isolated
sequent analysis (Fig. 1).
from natural sources, such as indolizidine, isoquinoline, qui-
nolone, agelasine, and polyamine classes, have antibacterial
activity and can inhibit efflux pump [18, 19], protein synthe- 2.3 Bacterial Isolates
sis, DNA transcription, and prevent toxin production [20].
Our previous research examined the pharmaceutical S. aureus 122 (S1), S. aureus 128 (S2), S. aureus 287 (S3),
importance of bioactive molecules from Libyan brown mac- EHEC O157 49 (E1), EHEC O157 52 (E2), and EHEC
roalgae [7, 21]. Here, we determined the antibacterial activ- O157 55 (E3) were isolated from dairy and meat prod-
ity and cytotoxic effect of alkaloid-containing extracts on six ucts [23, 24]. The isolates of S. aureus were methicillin-
bacterial isolates from two multidrug-resistant S. aureus and resistant (MRSA) [24]. The isolates were identified by
enterohaemorrhagic Escherichia coli (EHEC) O157. conventional microbiological techniques and confirmed by
partial sequencing of 16S rDNA by the method described
by Azwai et al. [25].

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Arabian Journal for Science and Engineering

Fig. 1  Morphology of Cysto-

seira compressa and Sargassum
hornschuchii

Cystoseira compressa Sargassum hornschuchii

2.4 Inoculum Preparation subcultured once more in MHB and further incubated at

37 °C for 18–24 h. Negative cultures were also prepared.
Isolates were inoculated in 25-ml Erlenmeyer flask con-
taining Mueller-Hinton Broth (MHB, Hi-Media) and incu-
2.8 Determination of Percentage Inhibition
bated at 37 °C in a shaker (120 rpm) for 24 h, followed
of Diameter Growth
by adjusting the culture to a turbidity equivalent to 0.5
McFarland standard (1.5 × ­1 0 8 CFU/mL) using normal
The relative percentage inhibition of alkaloid samples with
saline.
respect to amoxicillin and clavulanic acid (positive control)
was obtained using the following formula:
2.5 Antimicrobial Assay
Percentage of relative inhibition of the extract
Antimicrobial activity assay of algal alkaloids was per-
[ ]
= (At−Ab)∕(Ac−Ab) × 100,
formed by the agar-well diffusion method [26]. Individual
extracts (1000 μg/mL) were loaded onto separate 8-mm where At is the diameter of the test sample, Ab is the diam-
diameter wells (200µL) in triplicates and incubated at 37 °C eter of the blank sample, and Ac is the diameter of the posi-
for 24 h. The diameter of inhibition zone (DIZ) for each tive control.
extract was measured. Streptomycin (10 µg) as reference
control and solvent were also assayed.
2.9 Measurement of the Released Potassium Ion
2.6 Calculating Minimum Inhibitory Concentration
The method by Hao et al. [29] was used to assess the cyto-
plasmic membrane permeabilizing properties of the alka-
The minimum inhibitory concentration (MIC) was deter-
loids-containing extracts against bacterial cell membranes
mined by the agar dilution method adopted by Daud
by measuring the amount of potassium ion efflux. For dif-
et al. [27]. Fourfold serial dilutions of the original extract
ferent time durations, isolated cells (2 × ­106 CFU/mL) were
(1000 μg/mL) were prepared in Mueller-Hinton agar to
cultured with extracts (2MIC) and solvent (negative con-
obtain concentration from 500 to 50 μg/mL. The wells were
trol) at 37 °C. Following incubation, cells were centrifuged,
inoculated with bacteria and incubated at 37 °C for 24 h. The
and clear supernatants were analyzed for potassium using a
MIC was determined visually in agar as the lowest concen-
flame photometer (BMW Technologies, UK).
tration of the extracts that totally inhibits bacterial growth
as compared with the control.
2.10 Measurement of Nucleotide Leakage
2.7 Calculating Minimum Bactericidal
Concentration The impact of alkaloids-containing extracts on bacterial cell
membrane integrity can be investigated by measuring the
The MBC of the plant extracts was assessed according to leakages of intracellular components such as nucleotides
Spencer and Spencer [28] with some modifications. Extracts from the damaged cell membrane. Nucleotide leakage from
with MIC, 2MIC, 3MIC, and 4MIC were subcultured into isolated bacterial cells was assessed according to the method
the Mueller-Hinton Broth (MHB, Hi-Media) tubes. The by Tang et al. [30]. The bacterial suspension at the logarith-
tubes were inoculated with bacteria and incubated at 37 °C mic growth phase (1 × 106 CFU/mL) was incubated with and
for 24 h. After the incubation, the lowest concentration with- without the alkaloid-containing extracts at 2MIC for differ-
out any visible growth was taken as the MBC. To confirm ent periods. Cells were then centrifuged, and the obtained
the MBC results, 1 mL of the experimental suspensions was supernatants were diluted. The absorbance of supernatants

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 Arabian Journal for Science and Engineering

was determined at 260 nm using an ultraviolet–visible spec- derivatives as well as halogenated alkaloids [37]. However,
trophotometer (PerkinElmer, LAMBDA 25, USA). most of the studied alkaloids from marine sources was iso-
lated and their antibacterial properties were investigated and
2.11 Hemolytic Activity were from marine bacteria and sponges [38, 39] and few
marine alkaloids of algal origin were investigated [40]. In
The hemolytic activity (cytotoxicity) of the alkaloid-con- the current study, antibacterial activity of the algal alka-
taining extracts was established using human red blood loid-containing extracts against the six isolates of the two
cells (RBCs) from healthy, non-smoking volunteers [31]. multidrug-resistant microbes EHEC O157 and S. aureus is
RBCs were centrifuged at 3000 rpm for 10 min, washed, and summarized in Table 1. The data are presented as the diam-
re-suspended in saline. Algal extracts at concentrations of eter of clear zone (mm) produced by the alkaloid-contain-
6.25, 12.5, 25, 50, and 100 mg/mL were mixed with 100 mL ing extract of C. compressa and S. hornschuchii. All tested
RBCs to obtain a 200 mL 4% erythrocyte concentration and extracts were positively active, as was demonstrated using
were incubated at 37 °C for 1 h. Cultures were centrifuged, Gram-negative and Gram-positive isolates, the results were
and the absorbance of the supernatants (released hemo- in agreement with previous finding wherein the antibacterial
globin) was measured at 414 nm. Zero and 100% hemolysis activity of algal alkaloids is well documented [5, 7].
was determined in normal saline and 0.1% Triton X-100, The positive effect of the two extracts on Gram-negative
respectively, using an ultraviolet–visible spectrophotometer bacteria may be due to their ability to affect electrostatic
(PerkinElmer, LAMBDA 25, USA). The extract’s hemolytic interactions between lipopolysaccharides (LPS) and pro-
effect was calculated using the following formula, and the teins, which lead to a disturbance in the integrity of the outer
result was expressed as percentage hemolysis. membrane of Gram-negative bacteria [41].

Abs 414 nm of sample solution−Abs 414 nm of saline

% Hemolysis = ×100 =
Abs 414 nm of 0.1% Triton X − 100−Abs 414 nm of saline

2.12 Statistical Analysis The alkaloid-containing extract of S. hornschuchii

demonstrated its best results against S. aureus isolates
Data were expressed as means ± standard deviations (SD) (P < 0.001), whereas those of C. compressa was the most
of triplicate determinations. All statistical analyses were active against the EHEC O157 isolates. The maximum activ-
performed using SPSS v.16 (Statistical Program for Social ity was noted against S. aureus 287, with zones of inhibition
Sciences, SPSS Corporation, Chicago, IL). Statistical dif- of 41.5 mm for C. compressa alkaloid-containing extract
ferences between extract activities were determined using and 41 mm for S. hornschuchii alkaloid extract (Table 1).
one-way analysis of variance. Differences were considered These observations are in line with earlier findings by Tüney
statistically significant at P < 0.05.

Table 1  Antimicrobial effects and relative percentage inhibition of

3 Results and Discussion alkaloid-containing extracts against food-borne Gram-positive and
Gram-negative bacteria (Staphylococcus aureus (three isolates) and
3.1 In Vitro Antimicrobial Activity Escherichia coli O157 EHEC (three isolates)

Test organism Cystoseira compressa Sargassum hornschuchii

There has been increased attention paid to marine macroal- DIZ (mm) RPI (%) DIZ (mm) RPI (%)
gae as a reliable source of bioactive compounds produc-
tion widely used as antibacterial [32], antifungal [33], and S1 33.5 ± .50a 70 25.5 ± 0.5b 123.33
antiviral activities [34]. Alkaloids are providing a variety of S2 34.5 ± 0.5a 121.87 35.5 ± 0.5a 130
drugs used in pharmaceutical and food applications among S3 41.5 ± 0.5a 141.17 41.0 ± 0.01a 196.42
compounds of marine origin [35, 36]. E1 27 ± 1.0a 23.68 23.5 ± 0.5b 42.10
Most of the secondary metabolites of red, green and E2 34.5 ± 0.5a 153.57 35.5 ± 0.5a 146.42
brown algae with antibacterial activity have been identified E3 35.5 ± 0.5a 123.33 33.5 ± 0.5a 136.66
such as phlorotannins, fatty acids, peptides, terpenes, polya- DIZ diameter of inhibition zone, RPI relative percentage of inhibi-
cetylenes, sterols, indole alkaloids. Function groups in those tion. Data are presented as the mean ± SD, n = 3; E1: EHEC O157 49;
compounds play an indispensable role in their antibacterial E2: EHEC O157 52; E3: EHEC O157 55. S1: Staphylococcus aureus
activity. Algae of marine origin possess 44 types of alka- 122; S2: S. aureus 128; and S3: S. aureus 287. Different letters in
the same raw denote statistical significance between the two extracts
loids including indole, naphthyridine, and phenylethylamine (P < 0.05)

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et al. [42] and Aziz et al. [43], who found that algal extracts
predominantly inhibited Gram-positive bacteria.
The antibacterial results of the tested extracts were
compared with the positive control (streptomycin, 10 µg)
to evaluate their relative percentage inhibition (Table 1).
The alkaloid-containing extracts of C. compressa and S.
hornschuchii exhibited their maximum relative percentage
inhibition on S. aureus 287 (196.42% and 141.17%, respec-
tively), S. aureus 128 (130% and 121.87%, respectively),
and S. aureus 122 (123.33% and 70%, respectively). Both
tested samples demonstrated its maximum percentage rela-
tive inhibition against EHEC O157 52 (E2) (146.42% for C.
compressa and 153.57% for S. hornschuchii), whereas mini-
mum inhibition was noted against EHEC O157 49 (42.10%
for C. compressa and 23.68% for S. hornschuchii) (Table 1).
Higher relative percentage inhibition of some tested alka-
loids containing extracts concerning reference antibiotic may
suggest that those extracts are a good candidate for bacterial
strains resistant to conventional antibiotics [44].
An earlier study involving alkaloid extracts of certain
brown algae demonstrated zone of inhibition diameter values
lesser than those obtained in this study [21]. However, these
values cannot be compared because the alkaloid extracts of Fig. 2  Minimum inhibitory concentration (MIC) demonstrated by
C. compressa and S. hornschuchii are different from those alkaloid-containing extracts against a Staphylococcus aureus (S1, S2,
and S3) and b Escherichia coli EHEC 0157 strains (E1, E2, and E3).
previously tested. The recorded zone of inhibition diam-
Data are presented as the mean ± SD, n = 3. *P < 0.05 is considered
eter results could be affected by the antimicrobial agent’s statistically significant as compared to C. compressa extract
growth and diffusion rates, the initial inoculum density of
the microorganism, or the number of alkaloids in different
algal species. Table 2  Minimal inhibitory concentration (MIC) and minimal bac-
tericidal (MBC) ratio for alkaloid-containing extracts against tested
bacterial strains
3.1.1 The MIC and MBC
Cystoseira com- Sargassum
Here, antibacterial activities were considered significant pressa horns-
chuchii
when MIC was ≤ 150 μg/mL, moderate when MIC was
200–500 μg/mL, and inactive when MIC was 1000 μg/mL. MBC/MIC
Accordingly, the antibacterial activity of S. hornschuchii S. aureus isolates
alkaloid-containing extract had a significant inhibitory effect S1 2 3
on S. aureus, whereas the alkaloid-containing extracts of S2 1 2
both algae demonstrated moderate activity against all EHEC S3 2 2
O157 isolates. The least MIC values were found against S. E. coli O157 EHEC isolates
aureus isolates for both extracts, with MICs ranging from E1 2 3
125 to 250 µg/mL, compared with Gram-negative bacteria, E2 2 2
with MICs ranging from 250 to 500 µg/mL (Fig. 2). The E3 1 1
alkaloid-containing extract of S. hornschuchii demonstrated
E1: EHEC O157 49; E2: EHEC O157 52; E3: EHEC O157 55. S1: S.
lower MIC values against E1 and E2 (MIC, 250 µg/mL) than aureus 122; S2: S. aureus 128; and S3: S. aureus 287
the alkaloid-containing extract of C. compressa.
Antibacterial drug discovery research is interested in
agents with greater bacteriostatic than bactericidal effect as against all the bacterial strains (Table 2). The antibacterial
they can be utilized for short-term therapies [45]. Bacte- activity was considered bactericidal when the MBC-to-MIC
ricidal agents whose concentrations were not in excess of ratio was ≤ 1 or ≤ 2 and considered bacteriostatic if the ratio
four times their MIC were found to contribute to ≥ 99.9% was 3 or 4. Alkaloid extracts of C. compressa exhibited bac-
reduction in bacterial inoculum [46]. Alkaloid extracts from tericidal effect against all tested strains, whereas those of
the tested algae demonstrated an MBC-to-MIC ratio of ≤ 2 S. hornschuchii exhibited bactericidal effect against EHEC

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O157 52, EHEC O157 55, S. aureus 287, and S. aureus 128 inhibiting of the bacterial membrane [52]. In this study, the
(Table 2). These findings are in line with earlier results, total nucleotide leakage and potassium ions released from
which support the bactericidal activity of alkaloids [47–49], bacterial cells were determined to study the mechanism of
although the activity can be species-dependent [44, 50]. alkaloid extracts’ antibacterial activity. As shown in Fig. 3,
distinct nucleotide leakage was observed in extracts-treated
3.1.2 Mode of Action cells compared with untreated cells. These findings imply
leaching of DNA and RNA from the cells, indicating that
Each antimicrobial active compounds has its one mode of alkaloids increase cell membrane permeability either by
action against specific microorganism targeting different site effecting positive charges on anionic lipids of the membrane
antimicrobial site. Antibacterial compounds may inhibit cell or by changing the net charge on the system leading to mem-
wall synthesis and block peptidoglycan essential for cell sur- brane destabilization [53, 54]. The amount of potassium ion
vive [51]. Besides, they can prevent transpeptidation as well efflux from the isolated cells at different incubation times
as damage cytoplasmic membrane that eventually lead to is presented in Fig. 4. The level of released ions increased

Fig. 3  Effect of minimum inhibitory concentration (MIC) of alkaloid- 49 (E1), EHEC O157 52 (E2); EHEC O157 55 (E3); S. aureus 122
containing extracts of Sargassum hornschuchii and Cystoseira com- (S1); S. aureus 128 (S2), and S. aureus 287 (S3). Each point repre-
pressa on the amount of total nucleotide released from EHEC O157 sents the mean values of three experiments (n = 3)

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Fig. 4  Effect of the number of alkaloid-containing extracts on the aureus 128 (S2), and S. aureus 287 (S3). Each point represents the
amount of potassium ions (K+) released from EHEC O157 49 (E1), mean values of three experiments (n = 3)
EHEC O157 52 (E2); EHEC O157 55 (E3); S. aureus 122 (S1); S.

in a time-dependent manner. In all samples, the amount of inhibition [55, 56]. Studies have demonstrated that the alka-
potassium ion efflux from extracts-treated cells increased loid cryptolepine can cause morphological changes and cell
significantly at 12 h of incubation compared with untreated lysis of S. aureus [57, 58]. Few studies have reported that
cells. The maximum alteration in the amount of released alkaloids can intercalate with DNA, leading to DNA synthe-
potassium was observed in E3 and S1 cells treated with the sis inhibition [59–61].
alkaloid-containing extracts of S. hornschuchii (Fig. 4).
Although the alkaloids’ mechanism of action remains 3.2 Anti‑hemolytic Activity
unclear, some studies suggested that the mechanism
is dependent on the type of alkaloids and the bacterial Natural active compounds are one of the most important
strain. The bioactivity of alkaloids is related to their abil- sources for drug discovery and development, and it is
ity to donate or accept protons and form hydrogen bonds important to investigate their toxicity. The hemolytic activ-
with enzymes, proteins, and receptors, resulting in growth ity of molecules is an indicator of their cytotoxicity as the

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Table 3  Hemolytic activity of alkaloid-containing extracts against 3.3 Conclusion

human erythrocytes

Sample/control Concentration Hemolytic activity (%) The antibacterial activity of the crude alkaloid-containing
extracts of C. compressa and S. hornschuchii indicates
Negative control* 8.5% 0.00 ± 0.002
that the extracts possess a strong antibacterial property to
Positive control** 0.1% 100.0 ± 0.084
prevent the growth of Gram-positive and Gram-negative
Sargassum hornschuchii 6.25 mg/mL 0.43 ± 0.14
pathogens effectively. The extracts induced a greater zone
12.5 mg/mL 0.67 ± 0.06
of inhibition against isolates compared with the positive
25 mg/mL 1.46 ± 0.15
controls. Extracts-treated cells demonstrated an increase
50 mg/mL 3.30 ± 0.13
in the number of potassium ions and nucleotides released
100 mg/mL 5.64 ± 0.22
compared to untreated cells. However, the tested alkaloid-
Cystoseira compressa 6.25 mg/mL 0.66 ± 0.11
containing extracts must be studied in animal models to
12.5 mg/mL 1.38 ± 0.21
determine their efficacy in vivo. The results also showed
25 mg/mL 2.65 ± 0.04
that algal alkaloid-containing extracts possess a consider-
50 mg/mL 4.00 ± 0.20
ably low hemolytic activity. These marine alkaloids may
100 mg/mL 8.75 ± 0.17
contribute to the development of potential antibacterial
*
NaCl, **Triton X-100; values are presented as mean ± SD (n = 3); S: agents in the future as summaries in mini-reviews by
Sargassum, C: Cystoseira. The extract’s hemolytic activity was evalu- (Bajpai, 2016) [52]. However, there is great scope for fur-
ated by measuring hemoglobin release of 4% suspensions of fresh ther studies in the role of alkaloids extracted from Libyan
human erythrocytes
brown algae pharmaceutically, which can be involved in
manufacturers of herbal products targeted many human
sample may interact with biological entities at the cellular degenerative diseases.
level [31]. It has been found that the hemolytic activity
of each extract is related to their chemical composition Authors’ contributions RA conceived, designed, and organized the
and may result from extracts lytic proprieties or mem- study. SA, AMG, AA, SE, MS, EY, and MGK contributed to the con-
duct of the study. SA, AMG, AA, SE, AAE, GS, and WSA performed
brane instability induced by the extracts. In this study, the the experiments, RA, AAE, GS, and WSA analyzed the data. RA, SA,
two tested alkaloid extracts’ cytotoxicity was studied by AMG, AA, SE, MS, EY, and MGK drafted the manuscript and cri-
hemolysis assay using RBCs from healthy, non-smoking tiqued the output for intellectual content. All authors discussed the
volunteers. The results presented in Table 3 demonstrate results and commented on the manuscript.
that the tested extracts’ anti-hemolytic activity against
Funding This research did not receive any specific grant from funding
human erythrocytes was dose-dependent. agencies in the public, commercial, or non-profit sectors.
Compared with Triton X-100, which was used as the
positive control, the alkaloid-containing extracts showed Declarations
no hemolytic cytotoxicity as they obtained a little eryth-
rocyte disruption impact at low or high doses during Conflict of interest The authors declare that they have no conflict of
prolonged administration. The percentage of hemolysis interest.
increased with an increase in the alkaloid concentration
[9]. The percentage of hemolysis observed with the C.
compressa extract was more significant than that observed
with the S. hornschuchii extract. For instance, at 100 mg/ References
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