Sequencing of the rDNA ITS–1 proved that the lymnaeid snail species Galba truncatula is present in Argentina and that it belongs to the haplotype HC, the same as that responsible for the fascioliasis transmission in the human hyperendemic area with the highest human prevalences and intensities known, the Northern Bolivian Altiplano.La secuenciación del ITS–1 del ADNr demostró que la especie de gasterópodo lymnaeido Galba truncatula se encuentra en Argentina y que pertenece al haplotipo HC, el mismo responsable de la transmisión de la fascioliasis en el área de hiperendemia humana con las mayores prevalencias e intensidades de fascioliasis conocidas, el Altiplano Norte Boliviano

Artigas, P.

Bargues, M. D.

Gómez, H. G.

Mas-Coma, Santiago

Mera y Sierra, R. L.

Animal Biodiversity and Conservation

Revistes Catalanes amb Accés Obert

191Animal Biodiversity and Conservation 29.2 (2006)
© 2006 Museu de Ciències NaturalsISSN: 1578–665X
Bargues, M. D., Mera y Sierra, R. L., Gómez, H. G., Artigas P. & Mas–Coma, S., 2006. Ribosomal DNA ITS–1
sequencing of Galba truncatula (Gastropoda, Lymnaeidae) and its potential impact on fascioliasis transmission
in Mendoza, Argentina. Animal Biodiversity and Conservation, 29.2: 191–194.
Abstract
Ribosomal DNA ITS–1 sequencing of Galba truncatula (Gastropoda, Lymnaeidae) and its potential impact
on fascioliasis transmission in Mendoza, Argentina.— Sequencing of the rDNA ITS–1 proved that the
lymnaeid snail species Galba truncatula is present in Argentina and that it belongs to the haplotype HC, the
same as that responsible for the fascioliasis transmission in the human hyperendemic area with the highest
human prevalences and intensities known, the Northern Bolivian Altiplano.
Key words: Galba truncatula, Lymnaeid vectors, Human and animal fascioliasis, Transmission, Mendoza,
Argentina.
Resumen
Secuenciación del ITS–1 del ADN ribosomal de Galba truncatula (Gastropoda, Lymnaeidae) y su impacto
potencial en la transmisión de la fascioliasis en Mendoza, Argentina.— La secuenciación del ITS–1 del ADNr
demostró que la especie de gasterópodo lymnaeido Galba truncatula se encuentra en Argentina y que
pertenece al haplotipo HC, el mismo responsable de la transmisión de la fascioliasis en el área de
hiperendemia humana con las mayores prevalencias e intensidades de fascioliasis conocidas, el Altiplano
Norte Boliviano.
Palabras clave: Galba truncatula, Vectores Lymnaeidae, Fascioliasis humana y animal, Transmisión,
Mendoza, Argentina.
(Received: 15 XI 06; Final acceptance: 15 I 07)
M. D. Bargues, R. L. Mera y Sierra, H. G. Gómez, P. Artigas & S. Mas–Coma, Depto. de Parasitología, Fac.
de Farmacia, Univ. de Valencia, Av. Vicente Andrés Estellés s/n, 46100 Burjassot (Valencia), España
(Spain).– R. L. Mera y Sierra, Fac. de Ciencias Médicas, Univ. Nacional de Cuyo, Av. Libertador 80 Centro
Universitario, P. O. Box 33, 5500 Mendoza, Argentina (Argentina).
Corresponding author: M. D. Bargues. E–mail: M.D.Bargues@uv.es
Ribosomal DNA ITS–1 sequencing
of Galba truncatula
(Gastropoda, Lymnaeidae)
and its potential impact on
fascioliasis transmission
in Mendoza, Argentina
M. D. Bargues, R. L. Mera y Sierra, H. G. Gómez,
P. Artigas & S. Mas–Coma
192 Bargues et al.
Introduction
Fascioliasis is a well–known parasitic disease
transmitted by freshwater snail species of the
family Lymnaeidae (Gastropoda) (Mas–Coma et
al., 2005). Fasciola hepatica is believed to be of
European origin, with the lymnaeid Galba
truncatula as the original vector species. G.
truncatula is a species which reproduces by selfing
and spread to other continents, most probably
together with the commercial export of livestock
(Mas–Coma et al., 2001).
When carrying out animal fascioliasis studies
in the province of Mendoza, Argentina, Mera y
Sierra (2001) collected lymnaeids which he clas-
sified as belonging to the species G. truncatula,
by comparing with morphological descriptions
(Oviedo et al., 1995; Samadi et al., 2000).
This was the first time that G. truncatula was
cited in Argentina. Studies performed during sub-
sequent years showed that morphology is not
sufficient to differentiate lymnaeid species be-
longing to the so–called Galba–Fossaria group
(Bargues et al., 2001; Durand et al., 2002).
Problems in morphological differentiation of
lymnaeid species have already been detected in
Mendoza (Mera y Sierra et al., 2005, 2006;
Artigas et al., 2005).
The present paper aimed to verify the species
classification of the lymnaeids collected by Mera
y Sierra (2001) in Mendoza, by analysis of the
complete sequence of the first internal transcribed
spacer (ITS–1) of the nuclear ribosomal DNA
(rDNA). This molecular marker has recently proved
its usefulness in Lymnaeidae (Bargues & Mas–
Coma, 2005; Bargues et al., 2006).
Materials and methods
The rDNA ITS–1 sequence was obtained from each
of 5 lymnaeid specimens collected in the locality of
El Salto, province of Mendoza, Argentina. DNA
extraction procedure steps were performed accord-
ing to methods outlined previously (Bargues et al.,
2001). Total DNA was isolated according to the
phenol–chloroform extraction and ethanol precipi-
tation method, the ITS–1 fragment was amplified
by the Polymerase Chain Reaction (PCR),
sequencing performed by the dideoxy chain–termi-
nation method, and sequences were aligned using
CLUSTAL–W version 1.8 (Bargues et al., 2001;
Mas–Coma et al., 2001). Homologies were per-
formed using BLAST (http://www.ncbi.nlm.nih.gov/
BLAST). The following ITS–1 sequences present in
GenBank were used for comparisons: Galba
truncatula haplotype A (AJ243018), haplotype B
(AJ296270), and haplotype C (AJ272052) (Mas–
Coma et al., 2001).
Results
The five ITS–1 sequences from the 5 lymnaeid
specimens analyzed presented the same length of
504 base pairs (bp) and a scarcely biased GC
content of 57.5%. The nucleotide sequence was in
all cases the same and it is shown in table 1. The
Argentinian lymnaeids presented an ITS–1 sequence
showing a 99.6% similitude with the G. truncatula
haplotype A (HA) from Europe. The nucleotide differ-
ences detected were only two mutations: the transi-
tion G / A in position 74 and the transversion T / G in
position 75. Concerning G. truncatula haplotype B
Table 1. Complete nucleotide sequence of the first internal transcribed spacer ITS–1 of the nuclear
ribosomal DNA of lymnaeids from the population of El Salto, Mendoza, Argentina.
Tabla 1. Secuencia nucleotídica completa del primer espaciador transcrito interno ITS–1 del ADN
ribosomal nuclear de los lymnaeidos de la población de El Salto, Mendoza, Argentina.
  1 ATCATTAACG AGCAGCCAAC CGAGCGTTGA CTACTTTGTT GTCTCAGTCA GTCAGTCAGT
 61 CAGGCCCCGC GGCGTGCACG CATGAAGCGC TGTCGCGGGG CTGTGTCCGC TTCGTCTTTC
121 GGGGTACCTA TTACTGTCCT CGATGCGACC CACGGTGACG GCTTAGAGCC CGTGTGCTCG
181 CCGGGTCGCG ACGGTTCAAA GAGTGGCCGG CTTGGCTCAG CTCGAGAGTC AGCCGGCGAC
241 CGCCCCGCCG TCGCAAAAAA ACAGGAGGTT AGTCCGGGGT ACCTATGCCC TGCCCGCGCT
301 CGCTCTCGCG CCGGCAAGGC GGTAGCTCCA GCTCGCTATT TGGCCGCGAG GTTCAAAGAG
361 ACGACCGTGC CTTAACTTGC TCTCTCCGTG GGCAACGGTC GCCGCCCCGG GCCTCCTAAA
421 ATTTCCTTTA ATAAAACGAA ATTATTTTTT AAAAATGTGT GTCGGCTCGA TCGTGGCACA
481 CGAAAAACAA ACAAAAGTC TTAT
Animal Biodiversity and Conservation 29.2 (2006) 193
(HB) from Morocco, the similitude was of 99.4%
and a total of three mutations were detected: two
transitions G / A and T / C in positions 74 and 132,
respectively, and one transversion T / G in position
75 (table 2). When comparing the ITS–1 sequence
of the Mendoza lymnaeid specimens with that of G.
truncatula haplotype C (HC) from the Northern Bo-
livian Altiplano, no nucleotide differences appeared
(table 2).
Discussion
The present work demonstrates that the lymnaeids
of Mendoza belong to G. truncatula haplotype HC.
The presence of G. truncatula HC in the Andean
area of Argentina represents a great potential risk
of fascioliasis. The haplotype HC of this lymnaeid
is the same as that responsible for transmission of
the disease in the endemic area with the highest
prevalences and intensities known in humans, the
Northern Bolivian Altiplano (Mas–Coma et al., 1999).
Although of a somewhat lower level, prevalence
and intensity situations found in other Andean ar-
eas of Peru are similar (Mas–Coma et al., 2005).
The province of Mendoza is also located in the
Andean area and although the altitudes are not as
high as those of hyperendemic zones in Bolivia and
Peru, temperatures are similar because of the south-
ern latitude (Fuentes et al., 1999). This suggests a
high disease transmission capacity in Mendoza.
The wide ecological features of G. truncatula do
allow it to come close to human settings (Mas–
Coma et al., 1999). This explains why this lymnaeid
is in the background of many human infections.
Moreover, G. truncatula is markedly linked to areas
where livestock is present, enabling its passive
transportation by domestic animals from one place
to another. The disease–spreading risk in Mendoza
is evident.
Acknowledgements
Studies funded by Projects No. BOS2002–01978
and No. SAF2006–09278 of the Ministry of Educa-
tion and Science, Madrid, and Red de Investigación
de Centros de Enfermedades Tropicales–RICET
(Projects No. C03/04, No. ISCIII2005–PI050574 and
RD06/0021/0017, Programa de Redes Temáticas de
Investigación Cooperativa), FIS, Ministry of Health,
Madrid. Studies in Argentina funded by Universidad
Nacional de Cuyo. Technical support provided by the
Servicio Central de Secuenciación para la
Investigación Experimental (SCSIE) of the
Universidad de Valencia (Dr. A. Martínez).
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Secuenciación del ITS-1 del ADN ribosomal de Galba truncatula (Gastropoda, Lymnaeidae) y su impacto potencial en la transmisión de la fascioliasis en Mendoza, Argentina

Secuenciación del ITS-1 del ADN ribosomal de Galba truncatula (Gastropoda, Lymnaeidae) y su impacto potencial en la transmisión de la fascioliasis en Mendoza, Argentina

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