Natural Product Research

Formerly Natural Product Letters

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Effect of extracts from several sponges of Yucatan

Coast on Giardia lamblia and preliminary chemical
investigation of the bioactive extract of Haliclona
(Reinera) tubifera

Juan Chalé-Dzul, Lorena V. León-Deniz, Sara Medina-Gómez, Rosa E. Moo-

Puc, Gumersindo Mirón-López, Patricia Gómez-López & Gonzalo J. Mena-
Rejón

To cite this article: Juan Chalé-Dzul, Lorena V. León-Deniz, Sara Medina-Gómez, Rosa E. Moo-
Puc, Gumersindo Mirón-López, Patricia Gómez-López & Gonzalo J. Mena-Rejón (2023) Effect of
extracts from several sponges of Yucatan Coast on Giardia lamblia and preliminary chemical
investigation of the bioactive extract of Haliclona (Reinera) tubifera, Natural Product Research,
37:23, 4023-4027, DOI: 10.1080/14786419.2022.2161540

To link to this article: https://doi.org/10.1080/14786419.2022.2161540

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Published online: 28 Dec 2022.

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NATURAL PRODUCT RESEARCH
2023, VOL. 37, NO. 23, 4023–4027
https://doi.org/10.1080/14786419.2022.2161540

SHORT COMMUNICATION

Effect of extracts from several sponges of Yucatan Coast

on Giardia lamblia and preliminary chemical investigation
of the bioactive extract of Haliclona (Reinera) tubifera
Juan Chal

n-Denizb, Sara Medina-Go
e-Dzula, Lorena V. Leo
mezc,
a
Rosa E. Moo-Puc , Gumersindo Miro
n-Lo

pez , Patricia Go
c

mez-Lo
pezd and
Gonzalo J. Mena-Rejo
nc

a
Unidad de Investigacio
n M
edica Yucat
an, Unidad M
edica de Alta Especialidad, Centro M
edico
“Ignacio Garc
ıa T
ellez”, IMSS, M
erida, Yucat
an, Mexico; bDepto. de Biolog
ıa Marina, Facultad de
Medicina de Veterinaria y Zootecnia, Universidad Aut
onoma de Yucat
an, Yucat
an, CP, Mexico;
c
Facultad de Qu
ımica, Universidad Aut
onoma de Yucat
an, M
erida, Yucat
an, Mexico; dInstituto de
Ciencias del Mar y Limnolog
ıa, Unidad Acad
emica de Ecolog
ıa y Biodiversidad Acu
atica, Universidad
Nacional Aut
onoma de M
exico, Circuito Exterior S/N Col., Coyoac
an, Mexico

ABSTRACT ARTICLE HISTORY

In this study, twenty-four organic extracts from six marine sponge Received 29 July 2022
species, collected at shallows of Yucatan, Mexico, were evaluated Accepted 14 December 2022
against Giardia lamblia trophozoites and Vero cells. The dichloro-
methane and hexane extracts of Haliclona tubifera exhibited the KEYWORDS
highest antigiardiasic activity (IC50 ¼ 1.00 and 2.11 mg/mL, antigiardiasic activity;
Giardia lamblia; Halichondria
respectively), as well as high selectivity (SI ¼ 41.8 and > 47.4, tubifera; marine sponges;
respectively), while ethyl acetate and methanol extracts of selectivity;
Cinachyrella alloclada, and methanol extract of Suberites auran- Yucatan peninsula
tiaca showed moderate activity. Contrastingly, the extracts of
Halichondria magniculosa and Oceanapia nodosa were considered
non actives. Consequently, the dichloromethane extract of H. tubi-
fera were subject to an exploratory chemical study, isolating chol-
esterol, two benzaldehyde derivatives, three benzoic acid
derivatives, cytosine, and thymine.

CONTACT Gonzalo J. Mena-Rej
on mrejon@correo.uady.mx; Lorena V. Le
on-Deniz lorena.leon@correo.uady.mx

Supplemental data for this article can be accessed online at https://doi.org/10.1080/14786419.2022.2161540
ß 2022 Informa UK Limited, trading as Taylor & Francis Group
4024 J. CHALÉ-DZUL ET AL.

1. Introduction
Human giardiasis is an infectious disease caused by the flagellate protozoan Giardia
lamblia, affecting close to 1 billion persons each year, of which around 200 million
develop symptoms (Riches et al. 2020).
Nowadays, it is possible to find this parasitosis disease throughout either developed
or developing countries, but its prevalence is 10-fold higher in the last ones.
Unfortunately, the inhabitants of underprivileged socioeconomic areas of those coun-
tries bear most of the burden (Al-Jawabreh et al. 2019).
Non-bloody foul-smelling diarrhoea, burping, bloating, epigastric pain, cramps, nau-
sea, vomiting, and weight loss are the typical symptoms of an infection by G. lamblia.
Most acute giardiasis cases last 4–6 weeks, but others can become chronic (Escobedo
et al. 2010). The chronic stage of giardiasis leads to malabsorption syndrome causing
a malnutrition state and delaying the physical growth and cognitive-intellectual devel-
opment of small children (Halliez and Buret 2013).
Currently, the first-line treatment of human giardiasis relies on 5-nitroimidazoles, as
well as on benzimidazoles or nitazoxanide. Nevertheless, in not a few cases, it is neces-
sary to use secondary agents such as furazolidone, chloroquine, quinacrine, or paro-
momycin (Riches et al. 2020).
Regrettably, all antigiardial drugs have important limitations. One of these limita-
tions is the variety of adverse effects these drugs provoke in patients. Some of
these effects can be transient, like gastrointestinal discomfort, headache, vertigo,
irritability or nausea. In contrast, others, such as leukopenia, hepatitis,
pancreatitis, neurotoxicity, and teratogenic and mutagenic effects, are severe (Riches
et al. 2020).
Increasing drug resistance is another limitation of antigiardial drugs. Indeed, this is
a critical issue for anti-Giardia treatment because, during the last decade, the rates of
treatment failure associated with parasite resistance increased significantly (Mørch and
Hanevik 2020).
This scenario makes it necessary to keep a continuous search for sources of new
effective and safe antigiardial drugs. In this context, it is well-known that natural
 NATURAL PRODUCT RESEARCH 4025

products are a promising source of compounds with potential application in pharmacy

and medicine.
Consequently, extracts from different parts of many terrestrial plants have been
screened for their activity against Giardia trophozoites leading to the isolation of a
substantial number of compounds with antigiardial activity (Calzada and Bautista
2020). In contrast, very few marine organisms have been investigated regarding their
content in antigiardial compounds.
The coastal and marine territory of the Yucatan Peninsula, in southeastern Mexico
and surrounded by the Gulf of Mexico and the Caribbean Sea, presents high habitat
diversity, where 50 sponge species grow. Shallows and coastal lagoons of Yucatan,
a state located at the north portion of Yucat
an peninsula, harbour nine of those 50
species (Torruco-Go
mez and Gonz
alez-Sol
ıs 2010). The potential of these nine spe-
cies as a source of compounds with antigiardial activity, as in most marine sponge
species, remains unexplored. Therefore, as a part of our bioprospecting studies of
marine invertebrates of the Yucatan state, Mexico, the organic extracts from
Cinachyrella alloclada, Clathria (Clathria) foliacea, Halichondria magniconulosa,
Haliclona (Reinera) tubifera, Suberites aurantiaca, and Oceanapia nodosa were
screened for their in vitro activity against Giardia lamblia trophozoites. Furthermore,
the dichloromethane extract from H. tubifera, was subject to a preliminary chem-
ical study.

2. Results and discussion

Twenty-four extracts belonging to six sponge species collected in Yucatan coastal shal-
lows (Supplementary material, Table S1) were screened in vitro against Giardia lamblia
trophozoites (IC50) and mammalian normal cells (CC50) (Supplementary material).
According to a previous report in this study, an extract was considered to have anti-
giardial activity if it showed an IC50  10 mg/mL (Cantillo-Ciau et al. 2010). Also, it was
considered that the antiprotozoal activity exhibited by an extract was not due to
in vitro cytotoxicity if its selectivity index (SI ¼ CC50/IC50)  10 (Vonthron-S
en
echeau
et al. 2003). The results of the bioassays are summarized in Supplementary material,
Table S2.
Seven extracts showed IC50’s ranging from 1.0 to 9.74 mg/mL; consequently, they
were considered to have antigiardial activity. The most active extracts were the
dichloromethane and hexane extracts from H. tubifera, followed by the hexane extract
of C. foliae, exhibiting IC50‘s ¼ 1.00, 2.11 and 3.44 mg/mL, respectively. Surprisingly,
both extracts of H. tubifera were highly selective toward G. lamblia; indeed, the hexane
extract resulted in the most selective one with a SI > 47.4 while the dichloromethanic
extract showed a SI ¼ 41.8 being the second one.
Importantly, the H. tubifera dichloromethane extract meets both used criteria to
define an antiprotozoal compound as a hit (Pink et al. 2005), which are a remarkable
antiprotozoal activity (IC50  1 lg/mL) as well as an excellent selectivity index (SI 
10). Furthermore, the IC50 of this extract was only 4.2-fold higher than that shown by
positive control, metronidazole, the current drug used in the treatment of giardiasis.
4026 J. CHALÉ-DZUL ET AL.

In contrast to the low-polarity extracts, the ethyl acetate extract was three-fold less
active against G. lamblia, besides exerting significant effects (Suffness and Pezzuto
1991) on normal mammalian cells (CC50 ¼ 25.6 lg/mL). Conversely, the methanolic
extract did not affect protozoan and Vero cells.
The ethyl acetate and methanol extracts of C. alloclada, and methanol extract of S.
aurantiaca showed moderate antigiardial activity (CI50’s 6.42–9.74 mg/mL) and, curi-
ously, their cytotoxic was low; while the extracts of H. magniconulosa and O. nodosa
were non active (Supplementary Table S1).
Polyacetylenes have been the only class of compounds with antigiardiasic proper-
ties isolated from marine sponges (Quin ~oa and Crews 1988; Sala et al. 2019).
Interestingly, Haliclona genus has been recognized to be a source of compounds
derived from the polyketide pathway, including polyacetylenes (Zhou et al. 2015) and
sphingoids (Biegelmeyer et al. 2015).
Based on those facts, the dichloromethane extract of H. tubifera was selected for
exploratory chemical analysis. Then, this extract (1.5 g) was eluted on a column of sil-
ica gel with gradient mixtures of hexanes/CH2Cl2 and CH2Cl2/EtOAc to afford, after
TLC analysis, six main fractions (A-F). After successive chromatographic separations
were obtained cholesterol (1) from fraction B, 4-hydroxybenzaldehyde (2) and 3-
hydroxy-4-methoxybenzaldehyde (3) from fraction C, benzoic (4), 2-phenylacetic (5),
and 4-hydroxybenzoic (6) acids from fraction E, thymine (7) from fraction F, and cyto-
sine (8) from fraction G. The structures of isolated compounds were confirmed by 1H
and 13C NMR spectroscopy.
Regrettably, it was not possible the isolation of polyacetylenes; nevertheless, the
finding of aromatic compounds suggests that H. tubifera is a potential source of com-
pounds derived from the polyketide pathway.

3. Experimental
See Supplementary material.

4. Conclusions
To the best of our knowledge, for the first time, extracts from marine sponge species
growing in shallows of Yucatan, a state belonging to Yucatan Peninsula, Mexico, were
screened for their in vitro activity against G. lamblia trophozoites. The dichloromethane
extract of H. tubifera meets the criteria for an antiprotozoal compound to be consid-
ered a hit. The presence of benzaldehyde and benzoic acid derivatives in H. tubifera
dichloromethane extract strongly suggests that this species is a potential source of
polyketides. Finally, a further major study of this extract, and the hexane extracts of
the same species and of Clathria foliacea, should be performed to isolate the com-
pounds responsible for their antigiardial activity.

Acknowledgements

mez is grateful to CONACYT for the master fellowship No. 615041.
S. Medina-Go
 NATURAL PRODUCT RESEARCH 4027

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
This work was funded by grant FQUI-2016-008 provided by Facultad de Qu
ımica, Universidad
Auto
noma de Yucat
an.

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