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ISSN:2682-4043
DOI:10.36462/H.BioSci.202506

Research Article
Open Access

1 Department of Zoology, Faculty of Sciences,

Menoufia University.

* To whom correspondence should
be addressed: Hodaahmed83@sci-
ence.menofia.edu.eg

Editor: Muhammad M. Adeel, Arthritis Clinical
Immunology program Oklahoma Medical Research
Foundation, Oklahoma City, United States.

Reviewer(s):
Khrokalo Liudmyla, Physical Chemistry
Department National Technical University of
Ukraine "Igor Sikorsky Kyiv Polytechnic Institute"
Kyiv 03056 Ukraine.

Grace Magalhaes-Ghiotto, Department of
Biotechnology, Genetics and Cell Biology,
Biological Sciences Center, State University of
Maringá, Maringá, Paraná 87020-900, Brazil.

Received: May 11, 2025

Accepted: August 2, 2025

Published: August 12, 2025

Citation: AbdelAzeem HH, Osman GY,
Sheir SK. Apoptotic and histopathological
impacts of Moringa oleifera seed oil on the
land snail Cornu aspersum (O. F. Müller,
1774). 2025 Aug. 12;8:bs202506

Copyright: © 2025 AbdelAzeem HH et al.. This is
an open access article distributed under the terms
of the Creative Commons Attribution License,
which permits unrestricted use, distribution, and
reproduction in any medium, provided the original
author and source are credited.
Data Availability Statement: All relevant data are
within the paper and supplementary materials.
Funding: The authors have no support or funding to
report.
Competing interests: The authors declare that they
have no competing interests.

Apoptotic and histopathological impacts of Moringa oleifera seed oil on
the land snail Cornu aspersum (O. F. Müller, 1774)

Hoda H. AbdelAzeem*1
>< �, Gamalat Y. Osman1

><�, Sherin K. Sheir1
><�

Abstract

Cornu aspersum (O. F. Müller, 1774) is among the most harmful pests for many crops.
Using natural molluscicides is essential to reduce the adverse effects of chemical ones
on the biosystem. The toxicity of Moringa oleifera seed oil was determined, and LC50

and LC90 were 20.06% and 29.5%, respectively, after 72 hr of exposure. Moringa oil
diminished the survival rate of Cornu aspersum (O. F. Müller, 1774) by 50% compared
to the control (98.6%). Moreover, Moringa oil significantly increased the apoptosis and
necrosis of digestive and ovotestis gland cells (P ≤ 0.02). By the end of the experiment,
the proportion of apoptotic cells rose dramatically to 62.6% and 50.3% in the digestive
and ovotestis glands, respectively, compared to the control (10.8% and 12.3%). In
addition, the percentage of necrotic cells significantly increased to 21.5% and 24.7%
(P ≤ 0.05), while the control values were 5.6% and 4.5%, respectively, at the 8%
concentration. Regarding the digestive gland, Moringa oil caused vacuolation, nuclear
pyknosis, and haemocyte infiltration. Deficiency of mature ova and spermatozoa,
fibrosis, degeneration, and necrosis were recorded in the ovotestis. Moringa oil has
proved its effectiveness as a natural molluscicide.

Keywords: Cornu aspersum (O. F. Müller, 1774), Moringa oleifera Seed Oil, survival rate,

Apoptosis, Histology.

Introduction
Mollusca are characterized as the second-largest phylum in the animal kingdom due to their great

reproductive capacity-especially slugs and snails-which enables them to spread quickly throughout

agricultural fields, making population control extremely challenging [1; 2]. The damage caused

by these snails is attributed to their rasping feeding habits and the deposition of waste products

such as slime and feces. Crop losses of 50–90% in soybean and corn have been reported in India

[3; 4]. Land snails are notorious pests that cause significant damage to various crops and vegetation

throughout Egypt [5; 6; 7]. These Land snails (gastropods) can damage multiple plant species, leading

to major economic losses in agricultural fields, gardens, orchards, and greenhouses [8; 9; 10; 11].

Several authors have recorded different snail species and investigated their harmful effects: Cornu

aspersum (O. F. Müller, 1774), Achatina fulica, Helix vestalis, and Theba pisana and Monacha

sp. and Oxychilus sp. [12; 13; 14; 15]. These pests have caused serious damage to vegetables,

banana crops, tomatoes, ornamental plants, mulberries, grapevines, and germinated seeds [16]. Land

mollusks also harm potatoes, grains, lettuce, cabbage, carrots, maize, clover, and other horticultural

and field crops. They feed on roots, seedlings, seeds, and tubers of nearly every fruit, vegetable, oil

plant, and ornamental species in gardens, greenhouses, and fields. Damage includes killing seedlings

(leading to poor stands) and destroying the leaves of young plants. Molluscs’ detrimental effects,

however, differ depending on the crop. For example, some consume freshly planted wheat seeds,

scrape strips of leaves from maize and many small grain plants, and make craters in the cotyledons

and ragged holes in the leaves of soybean crops [17; 1; 18].

Pest control for snails requires a continuous and integrated approach. Physical methods such

as hand collection and device innovation, the use of simple chemicals like sodium chloride, and

the application of registered molluscicides such as methomyl are commonly employed, often in

combination with biological control methods [18; 19; 20]. However, the extensive use of pesticides

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https://creativecommons.org/licenses/by/4.0/
mailto:Hodaahmed83@science.menofia.edu.eg
https://orcid.org/0000-0002-7599-8077
mailto: gamalat.osman@science.menofia.edu.eg 
https://orcid.org/0000-0001-9646-558X
mailto:Shyreen.shaair@science.menofia.edu.eg
https://orcid.org/0000-0002-0036-1514
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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

by agricultural workers in many countries has raised seri-
ous health concerns due to the toxicity of these chemicals [21].
Moreover, they often affect non-target organisms and disrupt
the ecosystem [22]. Given the side effects associated with each
method, there is an urgent need for safe and cost-effective alterna-
tives. As a result, ongoing efforts are focused on discovering and
evaluating promising, effective, and eco-friendly molluscicides
against pest snail species. Natural plant derivatives, also known
as botanical pesticides, have gained attention for their environ-
mental safety. Oil extracts from these plants are excellent natural
products-they are biodegradable into non-toxic compounds, easy
to access and dispose of, and economically affordable [23].

Table 1. Physiochemical properties of Moringa seed oil

Physical properties

Refractive index 1.471 ±0.00

Specific gravity 0.903 ±0.2

Acid value (mg/g) 0.58 ±0.21

Peroxide value (Meq/kg) 2.37 ±0.06

Saponification value (mg KOH/g) 160.32 ±0.4

Iodine value (g/100g) 65.27 ±0.21

Chemical properties (Fatty acids composition)

Name Concentration(%)

Palimitic acid C 18:0 6.61 ±0.5

Stearic acid C18:0 5.30 ±0.5

Oleic acid C 18:1 66.12 ±0.04

Linolinic acid C 18:3 1.03 ±0.4

Palmitoleic acid C 16:1 1.78 ±0.02

Vaccinic acid C16:1 5.86 ±1

Linoleic acid C18:2 1.47 ±0.1

Arachidic acid C 20:0 3.46 ±0.04

Gadolic acidic C20:1 n9 2.75 ±0.1

Behenic acid 22:0 6.36 ±0.03

Total unsaturated fatty acids 79.01 ±1.2

Total saturated fatty acids 21.73 ±1

(n=3 replicates, data are mean ±SD)

One of the most well-known and widely distributed species
is Moringa oleifera [24]. The seed oil extract contains fatty acids
similar to olive oil, with linoleic acid serving as a substitute [25].
It is rich in protein, and its leaves contain high levels of min-
erals such as iron, vitamins, and calcium, making it useful for
treating malnutrition [26]. Different parts of the tree including
the root, leaf, fruit, and seedalong with their extracts or oils,
possess various medicinal properties and have been used in both
non-food products and traditional medicine [27]. It possesses
antioxidant, anti-inflammatory, antianemic, and antidiabetic prop-
erties, supports the immune system, and combats neurological,
reproductive, cardiovascular, and bone illnesses. [28; 29]. Fur-

thermore, the bioactive constituents of botanical molluscicides,
such as flavonoids, saponins, and tannins found in Moringa spp.,
have been studied for their effects on snails [30; 31; 32]. Thus,
the aim of this study is to evaluate the molluscicidal efficacy of
Moringa oleifera seed oil against Cornu aspersum (O. F. Müller,
1774).

Materials and Methods
Experimental materials

Moringa oleifera seed oil was purchased from the Moringa
Unit at the National Research Center, Dokki, Egypt. The anal-
ysis of the oil, including its components and physicochemical
properties, was conducted at the same unit (Table 1). Methomyl
(Copter 90% SP), a carbamate compound (S-methyl N-[(methyl-
carbamoyl) oxy]thioacetimidate), with the molecular formula
C5H10N2O2S, was used as a standard pesticide. It was obtained
from Egyptchem International for Agrochemicals (Cairo, Egypt).
The required concentrations of Moringa oil were prepared by
dissolving the oil in a 1% Tween 80 solution (v/v) [33; 34]. The
chemical structure of Tween 80 is 2-[2-[3,4-bis(2-hydroxyethoxy)
oxolan-2-yl]-2-(2-hydroxyethoxy)ethoxy]ethyl octadec-9-enoate.

Determination of LC50 and LC90

To determine the LC50 and LC90 of Moringa oleifera seed
oil, acclimatized snails (mean weight: 4.4 ± 0.6 g) were divided
into three replicates, with 10 snails in each group. Each group
of 10 individuals was sprayed with 10 ml of its specific concen-
tration. A range of concentrations (1%, 3%, 6%, 9%, 15%, 21%,
27%, and 30%) of Moringa oil was freshly prepared using a 1%
Tween 80 solution (v/v) [35]. Snail mortality was observed and
recorded daily for up to 72 hours. After counting and removing
the dead individuals, the lethal concentrations (LC50 and LC90)
were calculated using Probit analysis in the SPSS statistical soft-
ware package (IBM Corp., Armonk, NY, USA). Two sub-lethal
concentrations, 4% and 8%, were selected as LC10 and LC20,
respectively.

Collection of snails and experimental design
Cornu aspersum (O. F. Müller, 1774) individuals were col-

lected from garden plants and decorative trees, then transported
in plastic boxes to the laboratory. The snails were maintained at
a temperature of 23 ± 2◦C, under a 12 h photoperiod and relative
humidity ranging from 75% to 85%. Each box contained a layer
of soil at the bottom and was covered with muslin cloth. The
snails were fed fresh lettuce leaves throughout the acclimatization
and experimental periods. Snails were randomly divided into
six groups, with three replicates per group, and each replicate
consisting of 15 snails. The experimental groups were as follows:
(a) Control group (unexposed). (b) Two groups exposed to 4%
and 8% Moringa oleifera seed oil. (c) Two groups exposed to
4% and 8% methomyl. (d) One group exposed to 1% Tween 80
solution. Snails were sprayed daily with 1 ml of the respective
treatment solution per snail (15 ml per replicate) over the course
of three weeks. Survival rate was recorded as the percentage

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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

Table 2. The effect of M. oleifera seed oil on the survival rate of Cornu aspersum (O. F. Müller, 1774).

Exposure period Survival rate

Control Methomyl Moringa Tween 80

4 8 4 8 1

Zero time 100 ±0 100 ±0 100 ±0 100 ±0 100 ±0 100 ±0

1st week 100 ±0 67.6 ±0.5 64.3 ±0.5 82 ±0.4 75.3 ±0.3 100 ±0

2nd week 100 ±0 54.3 ±1.5 47.6 ±1.1 72 ±0.5 63.3 ±1 98.9 ±0.5

3rd week 98.6 ±0.5 34.3 ±1.5a 30 ±1b 63.3 ±1 50 ±1ab 97.6 ±0.5

n = 3 replicates; data were reported as mean ±SD; a denotes a significant difference between the exposed and control groups and b vs

methomyl-exposed group when P ≤ 0.05.

of snails remaining alive at each observation point during the
exposure period. At the end of the experiment, the digestive
and ovotestis glands were dissected for apoptosis and necrosis
assessment using cell cycle analysis and AnnexinV/PI staining,
in addition to histological investigations.

Cell cycle analysis
Digestive and ovotestis glands (0.08 g) were homogenized in

1500 µl phosphate-buffered saline (cold) (PBS) and centrifuged at
1,000 rpm for 1 min at 4°C. The supernatant was collected for the
assay. 200 µl of cell suspension in citrate buffer was added, along
with propidium iodide (PI). There were 10,000 assessed nuclei
on average for each specimen, and 120 nuclei were scanned every
second. Accuri™ C6 flow cytometer analysis (Becton Dickinson,
United States) was used to assess apoptotic cells utilizing sub-G1

peak labeling with PI [36; 37].

• G0/G1 peak: Normal diploid cells.

• S phase (Synthesis): Cells synthesizing DNA.

• G2/M peak (Gap 2 /Mitosis): Cells ready to divide.

• Sub-G1: Apoptotic cells with fragmented DNA.

Assay of PI/annexin-V dual staining
Discrimination of the apoptotic profile was determined using

the Apoptosis Detection Kit I (Cat. No. 556547BD, Pharmin-
gen™, Bioscience, New Zealand) according to the manufacturer’s
instructions. The principle of this procedure relies on the inter-
action between the membrane phospholipid phosphatidylserine
of apoptotic cells and Annexin V, a phospholipid-binding pro-
tein that requires calcium. To differentiate between viable and
non-viable cells, propidium iodide (PI) was employed as a con-
ventional flow cytometric viability probe. Non-viable or damaged
cells with permeable membranes allow PI to enter, while viable
cells with intact membranes exclude PI. The BD Accuri™ C6
flow cytometer was used to analyze the cells immediately. Four
distinct cell populations were identified: (1) Early apoptotic cells
(bound to Annexin V only), (2) Late apoptotic/necrotic cells
(bound to both Annexin V and PI), (3) Necrotic cells (stained
with PI only), (4) Viable cells (unstained). The percentage of

fluorescent cells in each quadrant was calculated after the fluores-
cence distribution was presented in a two-color dot plot analysis
[38].

Histological investigation
After three weeks, the digestive and ovotestis glands were

dissected and fixed in Bouin’s fluid. After 24 hr of fixation, dehy-
dration was performed through a series of alcohols and cleared in
xylene. Paraffin blocks were sectioned at 5 µm thickness on glass
slides for hematoxylin and eosin staining [39; 40]. A good wash
with tap water was performed after staining. A photo-automated
camera (Optika, Italy) was used to capture images of the histo-
logical sections, which were then analyzed for histopathological
syndromes.

Statistical analysis
The Statistical Package for the Social Sciences (SPSS; IBM

Corp., Armonk, NY, USA) was used to analyze the data, which
were presented as mean ± standard deviation. One-way ANOVA
was performed, followed by the Least Significant Difference
(LSD) post-hoc test to determine the significance between the
control and different concentrations of the tested materials. A
significance level of P < 0.05 was considered statistically signifi-
cant.

Results
Toxical effect of Moringa oleifera seed oil

The values (%) of LC50 and LC90 were 20.06 (95% confi-
dence limit for log = 1.3) and 29.5 (95% confidence limit for log
= 1.4), respectively, after 72 hr of exposure. The slopes of LC50

and LC90 were 2.5 and 3, respectively. The sublethal concen-
trations used were 4% Moringa oil (40 mL/L of 1% Tween 80)
and 8% Moringa oil (80 mL/L of 1% Tween 80) (v/v) solutions.
Additionally, 4% methomyl (4 g / 100 mL) and 8% methomyl
(4 g / 100 mL) (w/v) solutions were used.

The survival rate of Cornu aspersum (O. F. Müller, 1774) under the
effect of Moringa oil

The survival rate of Cornu aspersum decreased gradually
according to the concentration and exposure duration. The reduc-
tion was more significant in methomyl-exposed snails, followed

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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

Table 3. Percentage of cells in different cell cycle phases after treatment

Concentration Exposure Period %Sub G1 % Cells/Cell Cycle G0/G1 S G2/M

Control 1st week 12.8 ±0.1 91.9 ±0.1 51.03 ±1 4.4 ±0.1 24.5 ±0.8

2nd week 11.8 ±0.2 95.8 ±0.3 52 ±1 4.1 ±0.3 24.8 ±1

3rd week 11.7 ±0.4 96 ±0.4 52.1 ±1.2 4 ±0.2 24.3 ±0.5

4% Moringa oil 1st week 14.9 ±0.6 97.2 ±2.9 70.1 ±3.3 2.1 ±0.2 10.4 ±1.9

2nd week 15.02 ±0.1ab 83.2 ±1.4 40.1 ±0.7 15.03 ±0.05 15.5 ±0.7

3rd week 12.4 ±1.2 90.9 ±2.9 60.1 ±2.07 4.1 ±0 15.3 ±1.4

8% Moringa oil 1st week 18.2 ±0.3ab 99.2 ±1.3 78.3 ±1.4 1.8 ±0.2 0.9 ±0.1

2nd week 10.4 ±0.05 91.03 ±1.1 57.1 ±0.6 3.7 ±0.05 20.6 ±0.8

3rd week 13.3 ±0.9 90.2 ±2.03 59.5 ±2.4 4.03 ±0.1 14.5 ±3.2

4% Methomyl 1st week 21.3 ±0.3ab 96.3 ±1.1 64.3 ±1.06 1.3 ±0.05 9.93 ±0.1

2nd week 14.1 ±0.5b 92.1 ±0.8 51.4 ±1.1 6.5 ±0.05 21.1 ±1.6

3rd week 14.7 ±0.6 93.8 ±1.05 52.06 ±2.2 1.8 ±0.05 26.1 ±2.8

8% Methomyl 1st week 22.1 ±0.1ab 96.9 ±0.6 63.6 ±1.5 1.6 ±0.05 10.1 ±1.7

2nd week 12.8 ±0.6 90.7 ±3.03 41.9 ±2.4 13.4 ±0.3 23.6 ±3.9

3rd week 14.1 ±0.5b 92.6 ±1.5 45.4 ±1.2 5.2 ±0.1 28.9 ±2.4

1% Tween 1st week 12.8 ±2.7 94.03 ±1.7 76.1 ±1.2 2.23 ±0.2 3.5 ±0.1

2nd week 9.2 ±0.7 87.8 ±4.9 65.6 ±4.2 13.3 ±0.1 0.8 ±0.1

3rd week 14.02 ±1 86.3 ±4.4 54.3 ±3.9 3.7 ±0.4 16.1 ±0.2

Data are stated as mean ± SD, n = 3 replicates; significant differences are represented as a when compared with the control and b when

compared with methomyl at P ≤ 0.05.

Figure 1. Representative flow cytometry histogram showing Moringa
oil’s effect on cell cycle distribution of Cornu aspersum (O. F. Müller,
1774) after the third week of exposure. The digestive gland is repre-
sented by panels (a), (b), and (c): (a) control; (b) 8% Moringa oil; (c)
8% methomyl. The ovotestis gland is represented by panels (d), (e), and
(f): (d) control; (e) 8% Moringa oil; (f) 8% methomyl.

by those exposed to M. oleifera seed oil. A substantial decrease
(P = 0.01) was observed in the third week of exposure, with
survival rates of 50% for M. oleifera oil and 30% for methomyl,
compared to 98.6% in the control. Methomyl exposure signifi-
cantly reduced the survival rate compared to M. oleifera seed oil
(Table 2).

Cell cycle distribution after Moringa oil exposure
The proportion of cells in the G0/G1, S, and G2/M phases

of the cell cycle was determined in both digestive and ovotestis
glands after 1, 2, and 3 weeks. Exposure to 4% and 8% M.
oleifera seed oil resulted in a significant increase in fragmented
cells (sub-G1) compared to the control (P ≤ 0.003) and methomyl
(P ≤ 0.01). Notably, the number of G0/G1 phase cells increased
to 78.3 ± 1.4% in the 8% Moringa oil group, compared to
51.03 ± 1% in the control and 63.6 ± 1.5% in the methomyl
group. Consequently, the proportions of S and G2/M phase cells
decreased at the same concentration (Table 3, Figure 1).

In the digestive gland, 8% Moringa oil significantly increased
sub-G1 phase cells compared to the control (P ≤ 0.001) and

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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

Table 4. Percentage of cells in different cell cycle phases after treatment

Concentration Exposure Period %Sub G1 % Cells/Cell Cycle G0/G1 S G2/M

Control 1st week 9.3 ±1 96.5 ±1.3 52.2 ±0.1 3.1 ±0.2 11.4 ±1

2nd week 10.4 ±0.3a 95.4 ±0.8 52.2 ±0.1 3.5 ±0.3 10.9 ±1

3rd week 10.4 ±0.5 95.3 ±0.4 52.5 ±0.2 3 ±0.2 10.8

4% Moringa oil 1st week 15.4 ±0.4 93.6 ±0.5 74.1 ±1.1 10.5 ±0.1 1.1 ±0.1

2nd week 17.06 ±0.5a 88.5 ±3.2 74.1 ±2.4 10.5 ±0.2 0.9 ±0.1

3rd week 15.2 ±0.05 93.6 ±0.2 75.1 ±0.5 10.3 ±0.05 1.2 ±0.3

8% Moringa oil 1st week 16.2 ±0.04 92 ±0 67.1 ±0.2 9.1 ±0.05 2.9 ±0.2

2nd week 30.2 ±0.1a 82.2 ±0.5 67.1 ±0.4 14.1 ±0.1 3 ±0.1

3rd week 15.7 ±0.7 96.1 ±1.4 67.4 ±1.5 13.1 ±0.2 2.8 ±2.4

4% Methomyl 1st week 18.1 ±0.1a 91.5 ±0.5 63.6 ±0.7 17.9 ±0.05 14 ±0.8

2nd week 12.9 ±0.1 86.2 ±0.2 63.7 ±0.5 17.6 ±0.1 14.1 ±0.9

3rd week 15.1 ±0.1 90.4 ±0.8 63.6 ±0.2 12 ±0.05 14.2 ±1.05

8% Methomyl 1st week 18.07 ±0.8a 91.3 ±1.4 68.6 ±3 12 ±0.05 13.4 ±1.8

2nd week 14.6 ±0.05 93.3 ±2.8 68.9 ±2.7 12.1 ±0.05 13.3 ±0.1

3rd week 17.9 ±0.4a 94.7 ±1.4 68.8 ±2.3 11.9 ±0.1 13.4 ±0.7

1% Tween 1st week 11.2 ±0.1 96.3 ±0.4 56 ±0.2 25.3 ±0.3 0.6 ±0.05

2nd week 11.5 ±0.2 96.6 ±0.7 59 ±0.2 25.7 ±0.7 0.7 ±0.1

3rd week 14.6 ±0.2 87.3 ±0.8 52.5 ±0.5 5.3 ±0.1 16.5 ±0.8

Data are expressed as mean ± SD, n = 3 replicates; significant differences are represented by a vs. control when P ≤ 0.05.

methomyl (P ≤ 0.005). The G0/G1 phase cell counts increased
to 75.1 ± 0.5% (4% Moringa) and 67.4 ± 1.5% (8% Moringa)
versus 52.5 ± 0.2% in the control. Furthermore, G2/M phase per-
centages decreased with both Moringa oil concentrations (Table
4, Figure 1).

Detection of apoptosis by annexin-V/PI
Using flow cytometry, apoptotic cells were identified by incu-

bating them with FITC-labeled annexin V and PI. To differentiate
between necrotic and apoptotic cells, PI was utilized. The ex-
posure of snails to Moringa oil caused a significant increase in
apoptotic cells (early and late apoptosis) in digestive gland cells
at both concentrations (P ≤ 0.001) and in the two concentrations
of methomyl (P ≤ 0.01) compared with the control group. At the
end of the experiment, the percentages of apoptotic cells were
50.3% and 63.9% at 8% Moringa oil and methomyl, respectively,
while that of the control was 12.3%. Depending on the material,
the increase in apoptotic cells in both concentrations of Moringa
oil was significant compared with methomyl (P ≤ 0.001).

In ovotestis gland cells, a significant increase in apoptotic
cells was observed at both concentrations of Moringa oil (P ≤

0.003) and at the two concentrations of methomyl (P ≤ 0.02).
At the third week of exposure, the percentages of apoptotic cells
were 62.6% and 50.8% at 8% Moringa oil and methomyl, respec-
tively, while the control was 10.8%. Depending on the time of
exposure, the increase in apoptotic cells in the third week was
significant when compared with that in the first week (P = 0.01).
Cells that were stained with PI were identified as necrotic, and
they significantly increased in the digestive gland cells after ex-
posure to both Moringa oil concentrations (P ≤ 0.005) and
methomyl (P ≤ 0.02) compared with the control. The percent-
ages were 21.5% and 14.9% at 8% Moringa oil and methomyl,
respectively, while that of the control was 5.6%. In the ovotestis
gland, a significant increase was noticed at both Moringa oil
concentrations and methomyl (P ≤ 0.03) compared with the con-
trol. Regarding exposure time, the increase in the third week was
24.7% and 31.1% (P = 0.02) at the 8% concentration of Moringa
oil and methomyl, respectively, compared with their values in
the first week (5.5% and 3.7%), when the control was 4.5%. As
a result, the number of viable cells was significantly decreased
in Moringa oil and methomyl (70.03% and 60.2%, respectively)
compared with the control (84.6%) in the third week of exposure

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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

Figure 2. Fluorocytograms of one representative experiment from three
separate trials following a three-week exposure to M. oleifera seed oil.

(Figure S1, Figure S2, Figure 2).

Histopathological signs after exposure to Moringa oil
Histological sections of the normal (unexposed) digestive

gland of Cornu aspersum (O. F. Müller, 1774) revealed that it is
mainly composed of digestive tubules, each lined with columnar
epithelial cells of various types, including digestive and excretory
cells. The digestive cells are typically characterized by numerous
green and yellow cytoplasmic granules. These tubules enclose
a distinct central lumen and are separated by interlobular con-
nective tissue containing hemolymphatic sinuses rich in actively
circulating hemocytes. Externally, the tubules are encased by a
distinct circular muscle layer.

Exposure to 8% M. oleifera oil caused pronounced pathologi-
cal alterations in the digestive gland, including marked vacuoliza-
tion and pyknosis of nuclei. Hemocytic infiltration, a clear sign
of inflammation, was also observed. Severe epithelial damage, in-
cluding extensive vacuolization, degeneration, and necrosis, was
evident following exposure to both 4% and 8% concentrations of
methomyl. Additionally, Moringa oil induced progressive degen-
eration of muscle fibers (Figure 3). The ovotestis of unexposed
snails consists of numerous small follicles (acini) lined with ger-
minal epithelium, which is clearly differentiated into primary and
secondary spermatogonia/oogonia and mature spermatozoa/ova.
After three weeks of exposure, the high concentration (8%) of
Moringa oil caused notable alterations in the histological archi-
tecture of the ovotestis. Histopathological signs included degen-
eration of ova and spermatozoa, inhibition of ova maturation,
appearance of dense fibrous tissue, and necrosis. Methomyl (8%)
exposure led to a substantial reduction or complete absence of
mature ova and degeneration of spermatozoa (Figure 4).

Discussions

Figure 3. Light photomicrographs of sections through the digestive
gland of Cornu aspersum (O. F. Müller, 1774) after three weeks of
exposure. (a & b) Control, (c, d, e, & i) exposed snails to 4% and
8% M. oleifera seed oil, and (f, g, h, & j) exposed snails to 4% and
8% methomyl. Dt, digestive tubule; Dc, digestive cell; Se, secretory
cell; Ct, connective tissue; L, lumen; Py, pyknosis; V, vacuolization;
Dg, degeneration; Uf, undigested food; IG, increasing the granules
in digestive cells; N, necrosis; Hi, hemocytic infiltration; Mf, muscle
fibers.

Effect of Moringa oleifera seed oil on the survival rate of Cornu
aspersum (O. F. Müller, 1774)

The present study recorded the influential role of Moringa
oil in reducing the survival rate of Cornu aspersum snails. As
primary consumers, land snails serve as models for rapid physio-
logical responses to dietary habits and breeding conditions. Pre-
vious research [41; 42] confirmed the sensitivity of Mollusca to
minimal dietary changes due to their rapid metabolism. Moringa
oil is composed of unsaturated and saturated fatty acids. Expo-
sure of snails to this oil may alter their fatty acid profile, which is
sufficient to change the physiological state of gastropods [43]. A
previous study also indicated that fluctuations in polyunsaturated
fatty acids can serve as a valuable ecotoxicological test in snails
[44]. Benzylamine extracted from M. oleifera has demonstrated
molluscicidal potency by reducing survival and reproductive rates
in Biomphalaria alexandrina snails [45]. Plant extracts contain-
ing essential oils, flavonoids, terpenes, saponins, and tannins
have been reported as effective agents in snail control [46; 47].
The negative impact of such extracts on the land snail Monacha
obstructa has also been documented [48].

Apoptotic effect of M. oleifera seed oil on Cornu aspersum
The findings indicated the apoptotic potential of M. oleifera

oil on both digestive and ovotestis gland tissues. This can be
explained by the fact that mollusks possess a unique composi-
tion of fatty acids. Given this, previous studies have shown that
unsaturated and saturated fatty acids exert different effects on
steatosis and cell death, influencing apoptosis in a variety of ex-
perimental systems and being identified as the most harmful lipid
types [49; 50; 51]. The presence of polyunsaturated fatty acids
in cellular membranes influenced membrane permeability. Previ-
ous research showed that apoptosis was caused by palmitic acid

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AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

Figure 4. Light photomicrographs of sections through the ovotestis
gland of Cornu aspersum (O. F. Müller, 1774) after three weeks of
exposure. (a) Control; (b, c, & f) exposed snails to 8% M. oleifera
oil; (d & e) exposed snails to 8% methomyl. f, follicle; fw, follicular
wall; ov, ovum; sp, spermatozoa; dv, developing oocyte; ss, stages
of spermatogonia; d, degeneration; ft, fibrous tissue; n, necrosis; vo,
vitellogenic oocyte. Reduction/lack of mature ova and spermatozoa
(black arrows).

through caspase-dependent Beclin 1 cleavage. The researchers
concluded that essential fatty acids induce apoptosis of tumor
cells by overexpressing cytochrome P450 [52]. Mitochondria
play a major role in apoptosis by releasing cytochrome c and
caspase 3, which activate apoptotic enzymes [53]. Fatty acids
that stimulate lipotoxicity were also reported to play a crucial role
in pathogenesis [54]. Moreover, the toxicity of metals has been
linked to disturbances in lipid membranes [55]. The observed
changes in fatty acid levels represent metabolic adjustments initi-
ated by external stimuli [56]. A recent study explored the ability
of Moringa oleifera leaf extract to promote apoptosis in adipocyte
cells (3T3-L1 cells) by increasing caspase 3 activity and regu-
lating the expression of BAX and BCL2 genes [57]. Moringa
seed extract has been shown to induce apoptosis and cell cycle
arrest in cancer cells, with fatty acids like caprylic acid, oleic
acid, and stearic acid contributing to apoptosis in lung, leukemia,
and ovarian cancer cells [58]. Autophagy and hepatic steatosis
were reported in the mouse liver after high-fat diets [59]. An-
other study demonstrated the harmful effects of Moringa extract
(containing saponins, flavones, and flavonols) on Biomphalaria
glabrata embryos, adults, and Schistosoma mansoni adult worms
[60].

Histopathological effect of M. oleifera seed oil on Cornu aspersum
Exposure to seed oil induced histopathological changes in

both digestive and ovotestis glands. This observation aligns with
previous findings [61], where damage such as fragmentation and
vacuolization was reported in the digestive and excretory cells of
B. alexandrina and B. truncatus snails following treatment with
Egyptian wild plant extracts. Exposure to aqueous seed extract

of M. oleifera caused severe damage in digestive cells, including
tip loss, degeneration, and increased numbers of secretory cells
[62]. Additional histopathological features included vacuolation
of digestive and secretory cells. Degeneration and rupture of
sperm and ova in B. truncatus were observed following expo-
sure to cerium oxide nanoparticles synthesized using Moringa
seeds [63]. Chlorophyllin from deep-frozen M. oleifera leaves
caused deformation of secretory cells and rupture of connective
tissue between tubules, leading to degeneration of digestive cells
in B. truncatus. When combined with magnesium or copper,
this compound also caused histological malformations due to
photosensitization [64; 65]. In Cornu aspersum, degeneration of
digestive tubules and damage to the basement membrane and hep-
atopancreas were reported following exposure to thiamethoxam
(200 mg/L) [66].

Conclusions
Moringa oleifera seed oil has significant molluscicidal po-

tency, exhibiting both apoptotic and histopathological effects on
the ovotestis and digestive glands of Cornu aspersum (O. F. Müller,
1774). This study investigated the effects of Moringa oil as a
whole against the snail Cornu aspersum. Further studies will aim
to isolate the main active constituents of the oil and assess their
effects on non-target organisms to ensure greater eco-friendliness
compared with conventional chemical molluscicides.

List of Abbreviations
• PBS: Phosphate-buffered saline

• PI: Propidium iodide

• COI: Cytochrome oxidase subunit I

• ND1: NADH dehydrogenase subunit 1

• DNA: Deoxyribonucleic acid

Supplementary
Figure S1: Exposure to M. oleifera seed oil increased cell

apoptosis and necrosis of the digestive gland of Cornu aspersum
(O. F. Müller, 1774) after exposure for3 weeks: (a) first week,
(b) second week, and (c) third week. Values expressed as mean
±SD. c The significant difference in necrosis between control
and exposed snails when P ≤ 0.02. The significant difference in
apoptosis was indicated as follows: a,b, between Moringa exposed
groups vs. control and methomyl exposed groups when P ≤ 0.05,
ANOVA.

Figure S2: Exposure to M. oleifera seed oil increased cell
apoptosis and necrosis of the ovotestis gland of Cornu aspersum
(O. F. Müller, 1774) after exposure 3 weeks: (a) first week; (b)
second week; and (c) third week. Values expressed as mean
±SD. c The significant difference in necrosis between control
and exposed snails when P ≤ 0.03. The significant difference
in apoptosis was indicated as follows: a,b between the Moringa
exposed group vs. control and methomyl exposed groups when
P ≤ 0.05, ANOVA.

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Reference
1. Zala M, Sipai S, Bharpoda T, Patel B. Molluscan pests and

their management: A review. AGRES-An International e
Journal. 2018;7(2):126-32.

2. Routray S, Dey D. Snails and slugs as crop pests. Rashtriya
Krishi. 2016;11(1):40-1.

3. Carlsson NOL, Brönmark C, Hansson LA. Invading her-
bivory: The golden apple snail alters ecosystem functioning
in Asian wetlands. Ecology. 2004;85:1575-80.

4. Askary TH, Khan AA, Waliullah MIS, Banday SA, Iqbal
U, Mir MM. Slug pest management through nematodes
in agricultural and horticultural crops. In: Boeri F, Chung
JA, editors. Nematodes. Nova Science Publisher; 2012. p.
197-212.

5. Shahawy WA. Field trials on land gastropods infesting some
ornamental plants at Kafr El-Sheikh Governorate. Journal of
Plant Protection and Pathology. 2019;10(1):7-11.

6. Mahmoud MMA, Omar MMA, Kurany HS. Ecological
studies on some terrestrial snails and slugs at Sohag gov-
ernorate, Egypt. Archives of Agriculture Sciences Journal.
2021;4(1):195-204.

7. Abd El-Wahed SIM, Ibrahim HAM. Molluscicidal assess-
ment of certain toxicants: Impact on biochemical alterations
and electrophoretic protein patterns in Massylaea vermicu-
lata (O. F. Müller, 1774) snails. Environmental Toxicology
and Pharmacology. 2025;113:104619.

8. South A. Terrestrial slugs. Biology, Ecology, Control. Lon-
don: Chapman & Hall Ltd.; 1992.

9. Kumar PA. Reviewon molluscs as an agricultural pest and
their control. International Journal of Agricultural and Food
Science. 2020;4(4):383-9.

10. Mobarak SA. Anti-fertility effect of three inorganic salts
against land snail Massylaea vermiculata (O.F. Müller 1774)
and their field efficiency. Journal of Basic and Applied
Zoology. 2021;82:1-8.

11. Sanderson G, Sirgel W. Helicidae as pests in Australian
and South African Grapevines. In: Barker GM, editor. Mol-
luscs as Crop Pests. London, UK: CABI Publishing, CAB
International; 2002. p. 255-70.

12. Jav Aregowda JA. Incidence of the giant African snail,
Achatina fulica (Bowdich), on horticulture crops. Pest Man-
agement and Economic Zoology. 2004;12(2):221-2.

13. Arafa AAI. Studies on terrestrial molluscs in some Delta
Governorate. Egypt: Al-Azhar University; 2006.

14. Lokma MHE. Studies on some terrestrial gastropods injuri-
ous to field crops at Sharkia Governorate. Zagazig Univer-
sity; 2007.

15. Tandingan De Ley I, Schurkman J, Wilen C, Dillman AR.
Mortality of the invasive white garden snail Theba pisana
exposed to three US isolates of Phasmarhabditis spp. PLoS
One. 2020;15(1):e0228244.

16. Mohammed MS, Al-Zarrouk MR, Ellwate NB. Assessment
of the damage caused by Eobania vermiculata and its con-
trol by metaldehyde on Beta vulgaris subsp. Cicla. Libyan
Journal of Plant Protection. 2025;15:29-41.

17. Ibrahim HA, Fadl MG, Hassan IA, Gad ME, El Mogy SA,
Khalifa MM, et al. Potential molluscicidal and antimicro-
bial activities of rare earth elements against the land snail
Theba pisana and certain microorganisms. Scientific Reports.
2025;15(1):20281.

18. Sallam A, El-Wakeil N. Biological and ecological studies on
land snails and their control. In: Integrated Pest Management
and Pest Control-Current and Future Tactics. InTech; 2012.
http://dx.doi.org/10.5772/29701.

19. Das A, Dolai SA. Successful “Mobile slug and snail con-
trol device” by physical method: an innovative idea and its
application. International Journal of Agricultural Research.
2015;5(6):25-30.

20. Howlett SA. Terrestrial slug problems: classical biological
control and beyond. CABI Reviews. 2021:1-10.

21. World Health Organization. Public Health Impact of Pes-
ticides Used in Agriculture. World Health Organization;
1990.

22. Serrão JE, Plata-Rueda A, Martínez LC, Zanuncio JC. Side-
effects of pesticides on non-target insects in agriculture: a
mini-review. Scientific Nature. 2022;109(17).

23. Lahlou M. Methods to study the phytochemistry and bioac-
tivity of essential oils. Phytotherapy Research: International
Journal Devoted to Pharmacological and Toxicological Eval-
uation of Natural Product Derivatives. 2004;18(6):435-48.

24. Morton JF. The horseradish tree, Moringa pterygosperma
(Moringaceae)a boon to arid lands. Economic Botany.
1991;45:318-33.

25. Madukwe EU, Ugwuoke AL, Ezeugwu JO. Effectiveness of
dry Moringa oleifera leaf powder in treatment of anaemia.
International Journal of Medicine and Medical Sciences.
2013;5(5):226-8.

26. Fahey JW. Moringa oleifera: a review of the medical evi-
dence for its nutritional, therapeutic, and prophylactic prop-
erties. Part 1. TFL Journal. 2005;1(5):1-15.

Highlights in BioScience Page 8 of 10 August 2025|Volume 8

http://dx.doi.org/10.5772/29701
http://bioscience.highlightsin.org/


AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

27. Ogbunugafor HA, Eneh FU, Ozumba AN, Igwo-Ezikpe MN,
Okpuzor J, Igwilo IO, et al. Physico-chemical and antioxi-
dant properties of Moringa oleifera seed oil. Pakistan Journal
of Nutrition. 2011;10(5):409-14.

28. Mahmood KT, Mugal T, Haq IU. Moringa oleifera: a
natural giftA review. Journal of Pharmaceutical Sciences.
2010;2(11):775-81.

29. Morales-Nava R, Gutiérrez-Uribe JA. Anti-Inflammatory
Properties of Moringa oleifera. In: Biological and Pharma-
cological Properties of the Genus Moringa. CRC Press; 2021.
p. 157-72.

30. Rocha-Filho CAA, Albuquerque LPA, Silva LRS, Silva PCB,
Coelho LC, Navarro DM, et al. Assessment of toxicity of
Moringa oleifera flower extract to Biomphalaria glabrata,
Schistosoma mansoni, and Artemia salina. Chemosphere.
2015;132:188-92.

31. Abiona JA, Abioja MO, Fabinu OO, Ehimiyein AO, Ladokun
AO, Olapeju YA, et al. Effect of Moringa oleifera on live
weight and reproductive tract dimension of giant African
land snail (Archachatina marginata). A Z (Animal Research
International). 2018;21(2):81-90.

32. Ibrahim M, Ghoname AI, Mansour SM, El-Dafrawy MS.
Effect of some medicinal plant extracts as molluscici-
dal and apoptotic agents on Biomphalaria alexandrina
snails. Egyptian Journal of Aquatic Biology and Fisheries.
2020;24(2):291-300.

33. Klopell FC, Lemos M, Sousa JPB, Comunello E, Maestro
EL, Bastos JK, et al. Nerolidol, an antiulcer constituent from
the essential oil of Baccharis dracunculifolia DC (Aster-
aceae). Natural Product Research. 2007;62:537-42.

34. Abdel-Rahman AHE. Usage of some botanical oils to
control the land snail Monacha sp (Gastropoda: Helici-
dae). Egyptian Journal of Plant Protection Research Institute.
2020;3(4):1241-54.

35. Parvate YA, Thayil L. Toxic Effect of Clove Oil on the
Survival and Histology of Various Tissues of Pestiferous
Land Snail Achatina fulica. Journal of Experimental Biology
and Agricultural Sciences. 2017;5(4):492-505.

36. Cohen JJ, Al-Rubeai M. Apoptosis-targeted therapies: the
next big thing in biotechnology. Trends in Biotechnology.
1995;13:281-3.

37. Reichard A, Asosingh K. Best practices for preparing a
single cell suspension from solid tissues for flow cytometry.
Cytometry Part A. 2018;95(2):219-26.

38. Badr BM, Moustafa NA, Eldien HMS, Mohamed AO,
Ibrahim HM, ElElaimy IA, et al. Increased levels of type 1

interferon in a type 1 diabetic mouse model induce the elim-
ination of B cells. Cellular Physiology and Biochemistry.
2015;35(1):137-47.

39. Romeis B. Mikroskopische Technik. 17th ed. Munich:
Urban und Schwarzenberg; 1989.

40. Suvarna KS, Layton CJ, Bancroft D. Bancrofts Theory
and Practice of Histological Techniques. Elsevier Health
Sciences; 2018.

41. Pirini M, Manuzzi MP, Pagliarani A, Trombetti F, Borgatti
AR, Ventrella V. Changes in fatty acid composition of
Mytilus galloprovincialis fed on microalgal and wheat germ
diets. Comparative Biochemistry and Physiology Part B.
2007;147:616-26.

42. Radwan MA, El-Gendy S, Gad AF. Oxidative stress biomark-
ers in the digestive gland of Theba pisana exposed to heavy
metals. Archives of Environmental Contamination and Toxi-
cology. 2010;58:828-35.

43. Kowalczyk-Pecka D, Pecka S, Kowalczuk-Vasilev E.
Changes in fatty acid metabolism induced by zinc in snails
Helix pomatia. Ecotoxicology and Environmental Safety.
2017;138:223-30.

44. Zhu N, Dai X, Lin DS, Connor WE. The lipids of
slugs and snail: evolution, diet and biosynthesis. Lipids.
1994;29(12):869-75.

45. Ibrahim AM, Youssef AA, Youssef AA, Sami M, Nasr SM.
Biological, biochemical and genotoxicological alterations of
Benzylamine on Biomphalaria alexandrina and its Schisto-
soma mansoni larvicidal potential. Pesticide Biochemistry
and Physiology. 2024;201:105855.

46. Teixeira T, Rosa JS, Rainha N, Baptista J, Rodrigues A.
Assessment of molluscicidal activity of essential oils from
five Azorean plants against Radix peregra (Müller, 1774).
Chemosphere. 2012;87:1-6.

47. Albuquerque LP, Pontual EV, Santana GMS, Silva LRS,
Aguiar JS, Coelho LCBB, et al. Toxic effects of Micro-
gramma vacciniifolia lectin on Artemia salina, human cells,
and Biomphalaria glabrata. Acta Tropica. 2014;138:23-7.

48. Ibrahim HAM, El-Mesalamy AF, Baghdadi SAE, Elhanbaly
R. Histopathological effects of methomyl and crude extracts
of Jatropha curcas against Monacha obstructa. Biotechnol-
ogy in Agriculture. 2022;9:65.

49. Mei S, Ni HM, Manley S, Bockus A, Kasse KM, Luyendyk
JP, et al. Differential roles of unsaturated and saturated fatty
acids on autophagy and apoptosis in hepatocytes. ASPET.
2011;339(2):487-98.

Highlights in BioScience Page 9 of 10 August 2025|Volume 8

http://bioscience.highlightsin.org/


AbdelAzeem HH et al., 2025 Apoptotic and histopathological impacts of Moringa oleifera seed oil

50. Listenberger LL, Han X, Lewis SE, Cases S, Farese RVJ,
Ory DS, et al. Triglyceride accumulation protects against
fatty acid-induced lipotoxicity. Proceedings of the National
Academy of Sciences USA. 2003;100:3077-82.

51. Ricchi M, Odoardi MR, Carulli L, Anzivino C, Ballestri S,
Pinetti A, et al. Differential effect of oleic and palmitic acid
on lipid accumulation and apoptosis in hepatocytes. Journal
of Gastroenterology and Hepatology. 2009;24:830-40.

52. Das UN. Essential fatty acids, lipid peroxidation and
apoptosis. Prostaglandins Leukot Essent Fatty Acids.
1999;61(3):157-63.

53. Fischer U, Jänicke R, Schulze-Osthoff K. Many cuts to ruin:
a comprehensive update of caspase substrates. Cell Death &
Differentiation. 2003;10:76.

54. Pirger Z, Rácz B, Kiss T. Dopamineinduced programmed
cell death in snail salivary gland cells. Cell Biology.
2009;101:105-16.

55. Nowakowska A, Rogalska J, Caputa M. Adaptability of
antioxidant defence system in Helix pomatia: effect of forced
aestivation. Journal of Molluscan Studies. 2016;82(1):205-7.

56. Balusamy SR, Perumalsamy H, Ranjan A, Park S, Ramani
SA. Dietary Moringa oleifera leaves induce fat cell apop-
tosis in 3T3-L1 adipocytes. Journal of Functional Foods.
2019;59:251-60.

57. Adebayo IA, Arsad H, Samian MR. The inhibitory role of
metabolites of Moringa oleifera seeds in cancer cells. In:
Phytomedicine: Pharmacologically Active Products from
Plants; 2021. p. 533-54.

58. Rocha-Filho CAA, Albuquerque LPA, Silva LRS, Silva PCB,
Coelho LC, Navarro DM, et al. Assessment of toxicity of
Moringa oleifera flower extract to Biomphalaria glabrata,
Schistosoma mansoni and Artemia salina. Chemosphere.
2015;132:188-92.

59. Mei S, Ni HM, Manley S, Bockus A, Kasse KM, Luyendyk
JP, et al. Differential Roles of Unsaturated and Saturated
Fatty Acids on Autophagy and Apoptosis in Hepatocytes.
ASPET. 2011;339(2):487-98.

60. Rocha-Filhoa CAA, Albuquerque LPA, Silva LRS, Silva
PCB, Coelho LC, Navarro DM, et al. Assessment of toxicity
of Moringa oleifera flower extract to Biomphalaria glabrata,
Schistosoma mansoni and Artemia salina. Chemosphere.
2015;132:188-92.

61. Yousef AAA, El-Kassas NE. Ultrastructure and histopatho-
logical effects of some plant extracts on digestive gland of
Biomphalaria alexandrina and Bulinus truncatus. JOBAZ.
2013;66:27-33.

62. Ibrahim AM, Abdalla AM. Impact of Moringa oleifera seed
aqueous extract on some biological, biochemical, and histo-
logical aspects of Biomphalaria alexandrina snails. Environ
Sci Pollut Res. 2017.

63. Abdel-Tawab H, Ibrahim AM, Hussein T, Mohamed F. Mech-
anism of action and toxicological evaluation of engineered
layered double hydroxide nanomaterials in Biomphalaria
alexandrina snails. Environ Sci Pollut Res Int. 2022;29.

64. Ibrahim AM, Bakry FA. Assessment of the molluscicidal
impact of extracted chlorophyllin on some biochemical pa-
rameters in the nervous tissue and histological changes in
Biomphalaria alexandrina and Lymnaea natalensis snails.
Invertebr Neurosci. 2019;19.

65. Dokmak HAAS, El-Emam MA, Mossalem HS, et al. Impact
of the photosensitizers copper and magnesium chlorophyllin
on biological and biochemical parameters of Bulinus trunca-
tus snail. Egypt J Aquat Biol Fish. 2021;25:525-40.

66. Hamlet SA, Bensoltane S, Djekoun M, Yassi F, Berrebbah
H. Histological changes and biochemical parameters in the
hepatopancreas of terrestrial gastropod Cornu aspersum as
biomarkers of neonicotinoid insecticide exposure. Afr J
Biotechnol. 2012;11(96):16277-83.

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	Abstract
	Introduction
	Materials and Methods
	Experimental materials
	Determination of LC50 and LC90
	Collection of snails and experimental design
	Cell cycle analysis
	Assay of PI/annexin-V dual staining
	Histological investigation
	Statistical analysis

	Results
	Toxical effect of Moringa oleifera seed oil
	The survival rate of Cornu aspersum (O. F. Müller, 1774) under the effect of Moringa oil
	Cell cycle distribution after Moringa oil exposure
	Detection of apoptosis by annexin-V/PI
	Histopathological signs after exposure to Moringa oil

	Discussions
	Effect of Moringa oleifera seed oil on the survival rate of Cornu aspersum (O. F. Müller, 1774)
	Apoptotic effect of M. oleifera seed oil on Cornu aspersum
	Histopathological effect of M. oleifera seed oil on Cornu aspersum

	Conclusions
	List of Abbreviations
	Supplementary

