International Journal of Entomology and Nematology

Vol. 5(1), pp. 121-127, March, 2019. © www.premierpublishers.org. ISSN: 2326-7262

Research Article
Naturally Occurring Insecticides from the Marine Sponge
Xestospongia testudinaria to Control the Whitefly Bemicia
tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
Mohamed El Hosieny Mostafa1,*, Alaa Fathy2, Hajar Saeed Alorfi3, Amr Negm4, Mamdouh Abdel-
Mogib5
1PlantProtection Research Institute, Agriculture Research Center, 12618, Egypt
2,4,5Departmentof Chemistry, Faculty of Science, Mansoura University, Mansoura, 35516, Egypt
3Department of Chemistry, Faculty of Science, King Abdulaziz University, PO. Box 80203, Jeddah 21589, Saudi Arabia

The insecticidal potential of the marine sponge Xestospongia testudinaria extracts against the
whitefly Bemicia tabaci and the aphid Aphis gossypii under laboratory conditions were
examined after 24 and 72 hrs. of treatments. The bioassay-guided isolation afforded nine
structurally diverse marine compounds: xestosterol (1), 18-bromooctadeca-(9E,17E)-dien-7,15-
diynoic acid (2), 18,18-dibromo-(9E)-octadeca-9,17-dien-5,7-diynoic acid (3), 16-
bromo(7E,11E,l5E)hexadeca-7,11,l5-trien-5,13-diynoic acid (4), methyl 18-bromooctadeca-
(9E,17E)-dien-7,15-diynoate (5), 2-methylmaleimide-5-oxime (6), maleimide-5-oxime (7), 2'-
deoxythymidine (8) and 2'-deoxyuridine (9). The chemical structures were determined on the
basis of detailed spectroscopic analyses (ESI-MS and 1H, 13C-NMR). Maleimide-5-oxime (7) was
the most potent one against 3rd instar nymphs of B. tabaci under laboratory conditions after 24
and 72 hrs of treatments, with LC50 values of 175.26 and 7.62 ppm, respectively. While against
Aphis gossypii, 18-bromooctadeca-(9E,17E)-dien-7,15-diynoic acid (2) recorded the highest LC50
values of 1.54 and 0.001 ppm after 24 and 72 hrs of treatments against Aphis gossypii,
respectively. Structure-activity relationship revealed the effectiveness of maleimides and
brominated polyacetylenic lipids.

Keywords: Marine extracts, Insecticidal activity, Sucking pests

INTRODUCTION

Among cotton sucking pests, whiteflies Bemisia tabaci and
aphid Aphis gossypii are of major importance. These
sucking pests are responsible for indirect and direct yield
losses by sucking the sap from leaves and also by
transmitting different viruses. Both species secrete
honeydew on leaves resulting in sooty mold growth, which
can reduce the quality and marketability of harvested
products. Honeydew falling on open bolls make the lint
sticky which creates problems during ginning (Torres et al.
2003; Zidan, 2012).
Consequently, there is a great interest in developing new
sustainable and eco-friendly insecticides to manage pest
populations with less toxicological risks through using
natural products (Mostafa et al. 2017). The marine
environment is an exceptional rich reservoir of biologically
active metabolites, which may be safer alternatives in
contrast to synthetic pesticides as they are known to have
minimal environmental impact and danger to consumers
(Peng et al. 2003; Abou-Elela et al. 2009; Joseph et al.
2010).

The high pesticides toxicity and their residues in soil, water

resources and crops as a result of their non-biodegradable
properties create hazard impacts on human health,
environment and beneficial non-targeted organisms
(Joseph et al. 2010; Dawidar et al. 2014; Mostafa et al.
2017).
*Corresponding Author: Mohamed El Hosieny Mostafa,
Plant Protection Research Institute, Agriculture Research
Center, 12618, Egypt. Email: melhosieny80@gmail.com

Naturally Occurring Insecticides from the Marine Sponge Xestospongia testudinaria to Control the Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
Mostafa et al. 122
The barrel sponge Xestospongia testudinaria
(Haplosclerida, Petrosiidae) contains diverse interesting
secondary metabolites such as brominated polyacetylenic
lipids (Jiang et al. 2011; Liu et al. 2011; El-Gamal et al.
2016), alkaloids, sterols, terpenoids and xestoquinones
(Jiang et al. 2011). Considering these components, the
sponge was reported to possess remarkable biological
activities, including antifungal, antibacterial, Na+/K+
ATPase inhibition and HIV-1 integrase inhibition (Liang et
al. 2014), cytoxic activity (El-Gamal et al. 2016), anti-
inflammatory, antioxidant and immunomodulatory effects
(El-Shitany et al. 2015).
The current study aimed to assess the potential of marine
natural products extracted from the sponge Xestospongia
testudinaria as naturally occurring insecticidal agents and
to identify the bioactive ingredient of these extracts.
MATERIALS AND METHODS
Instruments
NMR spectra were recorded on either Bruker AMX 400 or
JEOL 500 MHz, 13C-NMR spectra were recorded at JEOL
125 MHz. Chemical shifts are given in δ (ppm) relative to
TMS as internal slandered. ESIMS spectra were
performed on UPLC MS-MS "H2O" 3100 "USA" with
Bruker microOTOF and TQ detector. GC/MS analysis was
conducted on a Varian GC interfaced to Finnegan SSQ
7000 for MS identification of the GC components, Mass
Selective Detector (SMD) with ICIS V2.0 data system.
Chemicals
Normal column chromatography was performed on silica
gel Merck grain size 0.2-0.063 mm; thin layer
chromatography and preparative TLC were carried out on
silica gel Merck GF 254 precoated plates on aluminum
sheets 20×20 cm. Pet. ether (60 -80°C), methylene
chloride, ethyl acetate and methanol solvents were
purchased from Adwic company, Egypt.
Sponge sample
The sponge Xestospongia testudinaria was collected from
deep water (fifteen m depth) of Sharm Obhur (21°29′31″N
39°11′24″E), Jeddah, Saudi Arabia, and was characterized
from Faculty of Maritime Studies, King Abdulaziz
University. A Voucher sample (JAD 02013) has been
charged at the Chemistry Department, Faculty of Science,
King Abdulaziz University.
Extraction and isolation
The freeze-dried sponge (180 g) was extracted using
solvent mixture CH2Cl2/MeOH (1:1 v/v, 2 x 6L) at room
temperature. A viscous reddish oil extract (15 g) was
obtained. A part of the extract (8 g) was subjected to
Naturally Occurring Insecticides from the Marine Sponge Xestospongia testudinaria to Control the Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
normal silica gel column chromatography, eluted with
gradient increasing polarities from petroleum ether to ethyl
acetate then methanol. The effluent was incorporated to
five main fractions (A-E), based on their TLC pattern.
Fraction A was obtained with pet. ether/ethyl acetate (1:1)
purified on silica gel PTLC, developed by pet. ether/ethyl
acetate (17:1) to afford five compounds, (1) (42 mg, Rf
=0.69), (2) (37 mg, Rf =0.38), a binary mixture of (3), (4)
(28 mg, Rf =0.26) and (5) (24 mg, Rf =0.92). Fraction C
was obtained by 100% ethyl acetate purified on PTLC
silica gel using CH2Cl2/MeOH (9:1), to yield two
compounds (6) (31 mg, Rf =0.47) and (7) (35 mg, Rf
=0.39). Fraction D was obtained by ethyl acetate/MeOH
(9:1) separated using the same previous solvent system in
fraction C to yield two compounds (8) (36 mg, Rf =0.27)
and (9) (27 mg, Rf =0.20).
Xestosterol (1)
White powder. 1H NMR (CDCl3): δH 0.68 (3H, s, H-18),
1.00 (3H, s, H-19), 0.94 (3H, d, J=6.6 Hz, H-21), 1.82 (1H,
m, H-25), 0.80 (6H, t, J=7.3 Hz, H-29, H-30), 3.52 (1H, m,
H-3), 5.35 (1H, brs, H-6), 2.27 (1H, m, H-4a), 2.23 (1H, m,
H-4b), 4.68 (1H, brs, H-28a), 4.76 (1H, brs, H-28b). EI-MS;
m/z (rel. int.) 426 (5%) [M+], 411 (8%) [M-CH3]+, 314
(100%) [C22H34O]+., 281 (43%) [C21H29]+, 271 (28%)
[C19H27O]+, 229 (33%) [C17H24]+, 173 (13%) [C13H17]+, 55
(33%) [C4H7]+.
18-Bromooctadeca-(9E,17E)-diene-7,15-diynoic acid (2)
White powder. ESI-MS (m/z), [M-H]- 349 (C18H23O279Br),
351 (C18H23O281Br). 1H NMR (CDCl3): δH 6.57 (1H, d, J=14
Hz, H-18), 6.16 (1H, dt, J=14, 2.2 Hz, H-17), 6.01 (1H, m,
H-10), 5.45 (1H, brd, J=15.7 Hz, H-9), 2.46 (2H, t, J=7.3
Hz, H-2), 2.30 (2H, m, H-6), 2.25 (2H, m, H-14), 2.09 (H,
q, J=6.9 Hz, H-11a), 2.27 (H, q, J=6.9 Hz, H-11b), 1.67
(2H, m, H-3). 13C NMR: 167.0 (C=O), 110.2 (C-9 ), 142.6
(C-10), 88.6 (C-7), 79.2 (C-8 ), 117.9 (C-17), 117.2 (C-18
), 92.7 (C-15), 77.4 (C-16), 34.6 (C-2), 24.6 (C-3), 28.2 (C-
4 ), 28.3 (C-5), 19.3 (C-6), 32.4 (C-11), 28.0 (C-12), 27.7
(C-13), 19.2 (C-14).
18,18-dibromo-(9E)-octadeca-9,17-diene-5,7-diynoic
acid (3)
White solid, ESI-MS (m/z), [M-H]- 427 (C18H22O279Br2). 1H
NMR (CD3OD): δH 6.48 (1H, t, J=7.4 Hz, H-17), 6.24 (dt,
J=15.8, 7.1 Hz, H-10), 5.48 (brd, J=15.8 Hz, H-9), 2.26 (t,
2H, J=7.3 Hz, H-2), 2.30 (2H, td, J=7.1, 1.5 Hz, H-16), 1.78
(2H, quin, J=7.3 Hz, H-3), 2.30 (2H, td, J=7.3, 1.5 Hz, H-
4), 2.11 (2H, m, H-11), 1.43 (4H, m, H-12, 15).
16-bromo (7E,11E,l5E)hexadeca-7,11,l5-triene-5,13-
diynoic acid (4)
White solid, ESI-MS (m/z), [M-H]- 318.97 (C16H17O279Br).
1H NMR (CD OD): δ 6.79 (1H, d, J=14 Hz, H-16), 6.37
3 H
(1H, dd, J=14, 2.3 Hz, H-15), 6.14 (1H, dt, J=16, 6.8 Hz,

H-11), 5.95 (1H, dt, J=15.9, 6.8 Hz, H-8), 5.61 (1H, dd,
J=16, 2.2 Hz, H-12), 5.49 (1H, brd, J=15.9 Hz, H-7), 1.80
(2H, quin, J=7.3 Hz, H-3), 2.35 (2H, t, J=7.3 Hz, H-4), 2.19
(4H, m, H-9, 10).
Methyl-18-bromooctadeca-(9E,17E)-diene-7,15-
diynoate (5)
White powder. 1H-NMR (CDCl3): δH 6.50 (1H, d, J=14 Hz,
H-18), 6.10 (1H, d, J=14 Hz, H-17), 5.95 (1H, dt, J=15.6,
6.6 Hz, H-10), 5.38 (1H, brd, J=15.6 Hz, H-9), 2.26 (2H, m,
H-6), 2.26 (2H, m, H-14), 1.63 (2H, m, H-3), 3.59 (3H, s,
OCH3).
2-Methylmaleimide-5-oxime (6)
Colourless crystals .1H NMR (CD3OD): δH 7.20 (1H, d,
J=1Hz, H-4), 1.80 (d, J=1 Hz, CH3).
Maleimide-5-oxime (7)
Colourless crystals. 1H NMR (CD3OD): δH 5.59 (1H, d, J=7
Hz, H-3), 7.39 (1H, d, J=7 Hz, H-4). 13C NMR (CD3OD):
167.4 (C=O), 101.7 (C-3), 143.6 (C-4), 153.5 (C-5).
2'-deoxythymidine (8)
Colourless needles, 1H NMR (CD3OD): δH 1.85 (3H, d, J=
0.9 Hz, CH3), 7.79 (1H,d, J =1Hz, H-4), 6.25 (1H, t, J=7
Hz, H-7), 2.20 (1H, m, H-8a), 2.18 (1H, m, H-8b), 4.37
(1H, quin, J=3.4Hz, H-9), 3.87 (1H,q, J=3.4 Hz, H-10), 3.76
(1H, dd, J=12, 3.4 Hz, H-11a), 3.69 (1H, dd, J=12, 3.4 Hz,
H-11b).
2'-deoxyuridine (9)
Colourless needles, 1H NMR (CD3OD): δH 5.67 (1H, d,
J=8.1 Hz, H-3), 7.96 (1H, d, J= 8.1 Hz, H-4), 6.25 (1H ,t,
J= 6.8 Hz, H-7), 2.20 (1H, m, H-8a), 2.18 (1H, m, H-8b),
4.36 (1H, quin, J=3.4 Hz, H-9), 3.90 (1H, q, J=3.4 Hz, H-
10), 3.75 (1H, dd, J=12.1, 3.4 Hz, H-11a), 3.69 (1H, dd,
J=12.1, 3.4 Hz, H-11b), 13C NMR (CD3OD): 166.2 (C-2),
102.6 (C-3), 142.5 (C-4), 86.6 (C-7), 41.3 (C-8), 72.3 (C-
9), 88.9 (C-10), 62.8(C-11).
Insect collection and rearing
The culture of B. tabaci was maintained on unsprayed
potted plants of cotton variety, var. Giza 86 caged in a
separate muslin cloth cages (1.5×1.5 ×1.5 m) in the
greenhouse of Faculty of Agriculture farm, Mansoura
University, Egypt. For this purpose, whitefly adults were
collected from the field using an aspirator and were
released on the caged cotton plants in the greenhouse.
These cages were preserved in a greenhouse at 25-35°C,
55-75% R.H. and natural light.
A. gossypii were collected from naturally infested cotton
plants which were unsprayed before with any pesticides in
greenhouse of the farm of Faculty of Agriculture,
Mansoura University, Egypt.
Naturally Occurring Insecticides from the Marine Sponge Xestospongia testudinaria to Control the Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
Int. J. Entomol. Nematol. 123
Bioassays
Bioassays were conducted on 3rd instar nymphs of B.
tabaci using spray method technique as described by
Cruz-Estrada et al. (2013). Cotton leaves with nymphs
were detached and rounded sections (~2 cm in diameter)
that had at least 30 nymph individuals were marked. All
fractions and isolated compounds were formulated as an
emulsion in water containing 0.1% triton X-100. A series of
five diluted concentrations of each naturally components
were prepared and sprayed immediately after preparation
using a plastic hand atomizer. Control treatment was
sprayed with 0.1% triton X-100 in water. The marked
leaves of each treatment and control were deposited
individually after spraying process on a bed of 2% agar in
9 cm-diameter Petri dishes to keep moisturized. Each
treatment was replicated three times. Petri dishes
containing leaves were incubated under laboratory
conditions at 24 ± 3 ºC, 75 ± 8 % R.H. and 12:12 hrs L:D.
Mortality evaluations were carried out after 24 and 72 hrs
of extracts application.
One hundred individuals of A. gossypii were counted on
an infested leaf and then placed in a Petri-dish (15 cm in
diameter). Each infested leaf with the chosen number was
treated using spraying method as described before in B.
tabaci bioassay. Each treatment was replicated three
times. Mortality was evaluated after 24 and 72 hrs of
treatment.
Statistical analysis
The average of mortality percentages were corrected
using Abbott's formula (Abbott, 1925) and statistically
analyzed according to Finney (1971) to determine LC50,
LC90 and slope values. Toxicity index was calculated using
Sun's equation for different fractions or isolated
compounds by comparing these materials with the most
effective one (Sun, 1950).
RESULTS AND DISCUSSION
Our interest of identifying new leads that could serve as
prototype for eco-friendly natural insecticides has
motivated us to examine the insecticidal activity of the
marine sponge X. testudinaria natural components. Strong
insecticidal properties of marine sponge, X. testudinaria
were obtained after 24 and 72 hrs of exposure with LC50
728.53 and 31.15 ppm for B. tabaci, and 3.41 and 0.019
for A. gossypii, respectively. Bioassay-guided fractionation
has led to separate nine marine natural products, identified
and examined for their insecticidal activity against 3rd instar
nymphs of B. tabaci and A. gossypii.
Column and thin layer chromatographic separation
techniques led to isolate nine compounds (Fig.1)
belonging to different classes. Five compounds belonging
to sterols and brominated polyacetylenic lipids were
Mostafa et al. 124

isolated from fraction A, purified and identified using
different spectral analysis.
Compound (1) was isolated as a white powder, with a
molecular formula C30H50O obtained from EI-MS spectrum
at m/z 426 [M]+. Examination of 1H NMR spectrum as well
as the MS fragmentation pattern suggested that
compound (1) is Xestosterol that was reported previously
by El-Gamal et al. (2016) from the same sponge.
Purification of fraction D resulted in separation of two
compounds (8) and (9), as colorless needles. Careful
examination of 1H and 13C NMR spectra gave a pyrimidine
base moiety in accordance with the nucleosides 2'-
deoxythymidine (8) and 2'-deoxyuridine (9) (Kamori et al.
1980)., which reported for the first time from X.
testudinaria.

X. testudinaria is rich in brominated polyacetylenic

compounds which possess a wide range of biological
activities (Liu et al. 2011). 1H and 13C NMR spectra of
compound (2) contained signals of two units of trans
disubstituted ethylene, adjacent to an acetylenic bond, in
between four adjacent methylene groups, beside, a
pentamethylene group accommodated between a
carboxyl and an acetylenic groups. The NMR data
together with [M-H]- ions at m/z 349 and 351, in
approximately equal abundance, in the negative ESI-MS
(corresponding to the molecular formula C18H23BrO2) allow
us to confirm that compound (2) is 18-bromooctadeca-
(9E,17E)-diene-7,15-diynoic acid (2). Spectral analyses of
compound (2) were identically matched with that
previously reported from X. testudinaria by Pham et al.
(1999) and Bourguet-Kondracki et al. (1992).

1H NMR spectrum of ester (5) contained signals identical

to those of the acid (2) except one singlet signal appeared
at δH 3.59 (3H, s, OCH3), confirming that compound (5) is
the methyl ester of (2) so, (5) is methyl 18-bromooctadeca-
(9E,17E)-diene-7,15-diynoate, which was identified from
the same sponge previously by Qu and Tucker (1991).

The obtained binary mixture of (3) and (4) was examined

by negative ESI-MS that gave [M-H]- ions at m/z 426.99,
428.87 and 430.82 in ratio (1:2:1) corresponding to the
molecular formula (C18H22Br2O2) of (3), and [M-H]- ions at
m/z 318.97 and 321 in ratio (1: 1) corresponding to the
molecular formula (C16H17Br O2) of (4). The assignments
of the 1H NMR signals revealed the presence of isolated
terminal trisubstituted ethylene and endiyne systems
corresponding to the molecular formula of (3) while
compound (4) was characterized by additional ethylenic
system beside the trimethylene group between a carboxyl
and an acetylenic groups. This (3) and (4) were identified
as 18,18-dibromo-(9E)-octadeca-9,17-dien-5,7-diynoic
acid, previously published by Akiyama et al. (2013), and
16-bromo(7E,11E,l5E)hexadeca-7,11,l5-trien-5,13-
diynoic acid, published before by El-Gamal et al. (2016),
respectively.
Compounds (6) and (7), as colorless crystals, were
isolated from fraction C. 1H and 13C NMR spectra indicate
the maleimide structure. So the maleimide oximes (6) and
(7) were identified to be 2-methyl maleimide-5-oxime (6)
and maleimide-5-oxime (7), published previously by El-
Gamal et al. (2016).
Fig. 1: Isolated compounds from Xestospongia
testudinaria
Insecticidal activity of Xestospongia testudinaria
fractions and isolated compounds to 3rd instar
nymphs of whiteflies Bemisia tabaci
Natural products of marine origin are actually being
studied as alternative eco-friendly insecticides to manage
crop pests (Peng et al. 2003; El Sayed et al. 1997).
Among the different X. testudinaria fractions which
evaluated against 3rd instar nymphs of B. tabaci under
laboratory conditions, Fraction C was the most potent at
LC50 and LC90 levels (Table 1) followed by fractions A, D,
Crude extract, B and finally E, respectively after 24 hrs of
treatment. While after 72 hrs of treatment Crude extract
exhibited the highest degree of efficiency followed by
fractions D, C, E, A then the lowest active fraction B,
respectively (Table 1).

Naturally Occurring Insecticides from the Marine Sponge Xestospongia testudinaria to Control the Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
 Int. J. Entomol. Nematol. 125

The insecticidal activity of any natural extracts may be
attributed to chemical composition of their secondary
metabolites content. The most effective fractions A, C and
D were investigated chemically by using chromatographic
and spectral analyses to search for promising active
ingredients could serve as natural insecticides. Nine
isolated and purified compounds were examined against
3rd instar nymphs of B. tabaci under laboratory conditions
after 24 and 72 hrs of treatments. Results obtained in table
(2) revealed that compound (7) which was isolated from
fraction C (the most effective fraction) was the most potent
on the basis of toxicity index followed by compounds (6),
(3,4), (2), (5), (9), (8) and (1) respectively after 24 hrs of
treatment. The potency arrangement after 72 hrs of
treatment were compound (7) followed by compounds (6),
(3, 4), (2), (5), (1), (9) and (8), respectively (Table 2).

Table (1): Toxicity of Xestospongia testudinaria fractions against 3rd instar nymphs of Bemisia tabaci after 24 and
72hrs of treatment
24 hrs 72 hrs
LC50 (ppm) LC90 (ppm) LC50 (ppm) LC90 (ppm)
Fractions Toxicity Toxicity
and confidence and confidence limits Slope ± SE X2 and confidence and confidence Slope ± SE X2
index a index a
limits at 95% at 95% limits at 95% limits at 95%
Crude 728.53 5028.05 31.15 209.93
1.528±0.304 1.09 27.77 1.547±0.453 2.20 100.0
Extract 497.91 1405.37 2226.16 29990.13 3.90 58.73 138.66 488.64
369.86 4213.14 65.29 920.39
Fraction A 1.213±0.263 1.63 54.70 1.115±0.281 0.82 47.71
247.05 637.74 1731.93 34621.91 18.33 112.51 488.34 4590.91
787.22 10510.07 123.15 812.26
Fraction B 1.139±0.276 1.21 25.70 1.564±0.289 0.73 25.29
479.27 2257.93 3163.47 275164.07 75.76 172.10 508.47 1974.87
202.32 2410.38 48.32 275.60
Fraction C 1.191±0.357 1.04 100.0 1.695 ±0.387 1.31 64.46
122.80 378.19 864.11 104131.49 17.11 76.88 191.40 534.31
387.57 2755.42 47.28 328.67
Fraction D 1.505±0.387 0.37 52.2 1.522±0.396 0.31 65.88
261.11 893.33 1091.11 41243.56 12.55 78.59 215.22 867.23
818.35 31055.12 50.93 230.91
Fraction E 0.812±0.258 0.31 24.72 1.952±0.490 0.82 61.15
424.68 6246.68 4772.47 80.872 E+6 19.18 76.63 161.54 490.97
a Toxicity index = LC50 of the most effective fraction/ LC50 of the tested fractions × 100

Table (2): Toxicity of Xestospongia testudinaria isolated compounds against 3rd instar nymphs of Bemisia tabaci
after 24 and 72hrs of treatment
compounds

24 hrs 72 hrs
Fraction

Isolated

LC50 (ppm) LC90 (ppm) LC50 (ppm) LC90 (ppm)

2 Toxicity Toxicity
and confidence and confidence Slope ± SE X and confidence and confidence Slope ± SE X2
index index*
limits at 95% limits at 95% limits at 95% limits at 95%
1184.86 12876.55 133.33 1143.09
(1) 1.237±0.389 0.31 14.79 1.373±0.388 0.17 5.72
679.22 7596.47 3290.75 3467.9E+4 40.48 211.22 639.35 6562.62
285.73 2625.25 17.51 301.45
Fraction

(2) 1.331±0.367 0.84 61.34 1.037±0.306 1.07 43.57

190.12 603.03 985.37 61000.36 1.18 40.43 159.84 1549.76
A

219.50 7418.96 14.57 146.61

(3, 4) 1.283±0.267 3.71 79.85 1.278±0.402 0.16 52.35
115.70 1016.79 463.93 9758.7E+3 0.90 30.51 90.85 518.31
312.39 11729.78 21.61 170.64
(5) 0.814±0.255 0.05 56.10 1.428±0.414 1.12 35.29
170.24 1572.29 2018.83 1474.6E+4 3.91 37.77 106.33 619.94
Fraction C

196.86 6618.10 9.52 93.59

(6) 0.840±0.258 0.06 89.03 1.291±0.413 0.20 80.09
104.72 1208.98 1123.53 6839.2 E+3 0.91 17.96 54.65 557.75
175.26 4674.79 7.62 56.07
(7) 0.899±0.259 0.10 100.0 1.479±0.470 0.12 100.00
98.43 739.82 963.66 1148.4 E+3 0.63 14.76 35.73 177.29
939.85 6709.41 455.98 16577.55
Fraction

(8) 1.501±0.468 0.05 18.65 0.821±0.252 0.12 1.67

501.37 9059.49 1806.62 1337.8 E+3 255.35 1515.41 3273.84 912.9 E+3
D

812.99 10598.94 267.40 3881.77

(9) 1.149±0.278 1.15 21.56 1.103±0.356 0.38 2.85
493.81 2359.20 3198.99 272463.37 115.13 444.33 1409.79 267.5 E+3

Insecticidal activity of Xestospongia testudinaria Table (4) showed the susceptibility of A. gossypii to the
fractions and isolated compounds to Aphis gossypii nine isolated compounds after 24 and 72 hrs of treatment.
Data revealed that compound (2) which was isolated from
The results showed that the most toxic fraction was A fraction A (most active fraction) exhibited a high degree of
followed by C, B, Crude extract, D and finally E, efficiency at LC50 level against A. gossypii after 24 hrs of
respectively at LC50 level after 24 hrs of initial application. treatment, followed by compounds (9), (3, 4), (5), (8), (7),
Taking the toxicity index into consideration after 72 hrs of (6) and the least one (1), respectively. Comparing the
treatment Crude extract was the most toxic against A. toxicity indexes of the isolated compounds after 72 hrs of
gossypii followed by fractions A, C, B, E and D, exposure, it was found that compound (2) was the most
respectively (Table 3). effective followed by compounds (3, 4), (9), (5), (8), (7),
(1) and the least one (6), respectively.

Naturally Occurring Insecticides from the Marine Sponge Xestospongia testudinaria to Control the Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis gossypii (Glover)
Mostafa et al. 126

Table (3): Toxicity of Xestospongia testudinaria fractions against Aphis gossypii after 24 and 72hrs of treatment
24 hrs 72 hrs
LC50 (ppm) LC90 (ppm) LC50 (ppm) LC90 (ppm)
Fractions Toxicity Toxicity
and confidence and confidence Slope ± SE X2 and confidence and confidence Slope ± SE X2
index index
limits at 95% limits at 95% limits at 95% limits at 95%
Crude 3.41 203.20 0.019 4.19
0.722±0.161 2.07 61.69 0.549±0.162 0.95 100.00
Extract 0.83 7.23 73.97 1966.61 0.0001 0.110 1.24 63.34
2.10 336.78 0.033 23.88
Fraction A 0.581±0.157 1.86 100.00 0.449±0.142 0.69 57.58
0.18 5.61 90.64 14992.79 0.0001 0.198 5.02 4103.33
2.62 108.18 0.101 4.88
Fraction B 0.793±0.165 1.12 80.25 0.760±0.149 0.73 19.00
0.67 5.48 46.41 592.27 0.016 0.267 2.09 18.91
2.33 441.26 0.068 71.41
Fraction C 0.563±0.156 1.93 90.02 0.424±0.135 1.08 27.94
0.19 6.25 107.90 32069.00 0.0002 0.348 11.42 60981.81
9.26 101.75 0.999 62.94
Fraction D 1.231±0.355 0.28 22.70 0.712±0.174 4.13 1.90
1.87 16.23 55.33 633.11 0.098 2.779 23.75 528.57
19.65 2987.03 0.224 45.45
Fraction E 0.587±0.159 0.39 10.69 0.556±0.173 1.23 8.48
7.80 52.89 451.44 1081.47 E+3 0.0004 1.21 15.80 737.94

Table (4): Toxicity of Xestospongia testudinaria isolated compounds against Aphis gossypii after 24 and 72hrs
of treatment
compounds

24 hrs 72 hrs
Fraction

Isolated

LC50 (ppm) LC90 (ppm) LC50 (ppm) LC90 (ppm)

Toxicity Toxicity
and confidence and confidence Slope ± SE X2 and confidence and confidence Slope ± SE X2
index index
limits at 95% limits at 95% limits at 95% limits at 95%
9.83 2353.32 1.20 514.21
(1) 0.539±0.157 3.27 15.70 0.487±0.157 0.69 0.08
1.87 27.06 342.96 1733.03E+3 0.011 4.74 98.87 2781.17E+2
Fraction A

1.54 582.05 0.001 13.13

(2) 0.498±0.155 2.47 100.00 0.311±0.150 0.38 100.00
0.03 5.02 115.02 2507.04E+2 0.000033 0.031 0.43 400.99
3.93 431.37 0.38 140.48
(3, 4) 0.628±0.187 2.61 39.27 0.500±0.112 1.08 0.26
0.64 9.28 91.61 96808.25 0.048 1.15 32..79 3773.21
4.04 342.01 0.59 224.24
(5) 0.665±0.160 0.82 38.20 0.497±0.116 0.92 0.17
0.90 9.42 96.09 8122.56 0.06 1.97 47.36 8162.98
6.95 563.36 2.42 316.82
Fraction D Fraction C

(6) 0.671±0.158 2.70 22.22 0.605±0.187 4.99 0.04

2.15 14.35 156.45 14098.24 0.20 6.17 70.17 81269.60
19484.14
5.38 1.03 63.82
(7) 55.37 1471.66 0.360±0.115 3.71 28.72 0.716±0.168 1.70 0.10
0.70 31.34 0.11 2.78 26.80 392.52
E+7
4.1221 1340.9252 0.69 155.24
(8) 0.510±0.155 1.45 37.46 0.545±0.162 0.52 0.15
0.2867 12.1713 213.795 1015.97E+3 0.010 2.59 45.78 6872.91
1.74 202.44 0.57 25.45
(9) 0.620±0.159 0.12 88.09 0.776±0.181 1.18 0.18
0.17 4.61 64.41 4066.04 0.045 1.68 11.70 95.98

Structure-activity relationship of the isolated nine Akiyama T, Takada K, Oikawa T, Matsuura N, Okada YIS,
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Accepted 15 March 2019
Citation: Mostafa ME, Fathy A, Alorfi HS, Negm A, Abdel-
Mogib M (2019). Naturally Occurring Insecticides from the
Marine Sponge Xestospongia testudinaria to Control the
Whitefly Bemicia tabaci (Genn.) and the Aphid Aphis
gossypii (Glover). International Journal of Entomology and
Nematology, 5(1): 121-127.
Copyright: © 2019 Mostafa et al. This is an open-access
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