251
Neotrop. Helminthol., 8(2), 2014
2014 Asociación Peruana de Helmintología e Invertebrados Afines (APHIA)
ISSN: 2218-6425 impreso / ISSN: 1995-1043 on line
ORIGINAL ARTICLE / ARTÍCULO ORIGINAL
PARASITIC NEMATODE INFECTION IN THE SOUTH AMERICAN RED-FOOTED TORTOISE
GEOCHELONE CARBONARIA FROM UPATA, BOLIVAR, VENEZUELA
PARASITISMO POR NEMATODES EN LA TORTUGA SURAMERICANA GEOCHELONE
CARBONARIA DE UPATA, BOLÍVAR, VENEZUELA
1 1 1 1 1
Arlett Pérez Mata , Aixa Aguirre , Jessica Quijada Piñango , Jesmil Lecuna Olivares , Angélica Bethencourt Camacho ,
2 3
Nataly Bautista &Otto Núñez .
Abstract
Keywords: albendazole - nematodes - tortoise - Venezuela.
Suggested citation Pérez Mata, A, Aguirre, A, Quijada Piñango, J, Jesmil Lecuna Olivares, J, Bethencourt Camacho, A, Bautista,
N & Núñez, O. 2014. Parasitic nematode infection in the South American Red-footed tortoise Geochelone carbonaria from
Upata, Bolivar, Venezuela. Neotropical Helminthology, vol. 8, n°2, jul-dec, pp. 251-257.
1Veterinary Parasitology Department, Faculty of Veterinary Sciences (FCV), Central University of Venezuela (UCV),
Campus Maracay, Maracay 2101, Aragua, Venezuela
2 3
Undergraduated student. FCV-UCV Veterinary Surgeon, private practice.
E-mail: arlettperez@gmail.com; Tel:00-58-243-8710125.
Geochelone carbonaria is one of the two species of tortoises that have been described in
Venezuela. We report herein a severe parasitic infection with strongylid and pinworm nematodes
in 15 red-footed tortoises, G. carbonaria from Upata, State of Bolivar, Venezuela. Animals were
obtained from the owner of a cattle ranch in the savannah and allocated in a restricted area. Two
months later, animals became ill, displaying anorexia, lethargy and diarrhea. Faeces were fluid,
mucoid and haemorragic. Five animals were found dead. At necropsy, caecum and colon were
enlarged due to the presence of hundreds of small nematode parasites in their lumen; mucosal
haemorragy and fibrin tracts in the liver were also observed. Faecal samples and part of the
intestinal content were analyzed in the laboratory. Faecal examination showed a severe strongylid
and pinworm infection. Intestinal content was abundantly infected by pinworm nematodes
identified, at genus level, as Atractis (Order Ascaridida: Cosmocercoidea), and Thelandros
(Order Oxyurida: Pharyngodonidae). This represents the first report of both genera in G.
-1
carbonaria in Venezuela. Animals were orally treated with Albendazole (10 mg·kg ), once a day
for three days. Nematodes were expelled and health condition of the animals improved
dramatically. Stress is a common cause of outbreak of parasitism in recently captured reptiles.
Pérez Matta et al.
Parasitic nematode in Geochelone
252
Resumen
Palabras clave: albendazole- nematodos - tortugas - Venezuela.
Geochelone carbonaria es una de las dos especies de Tortugas terrestres que han sido descritas en
Venezuela. Aquí reportamos un caso de infección parasitaria severa con nemátodos estrongilidos
y oxyuridos en 15 tortugas de la localidad de Upata, Estado Bolívar, Venezuela. Los animales
fueron capturados en la sabana por el propietario de una finca ganadera en la zona y encerrados en
un área restringida con cerca. Dos meses más tarde, los animales se mostraron enfermos,
presentando anorexia, letargia y diarrea. Las heces eran fluidas, mucoides y hemorrágicas. Cinco
animales fueron encontrados muertos. A la necropsia se observó un aumento de tamaño del colon
y ciego con presencia de cientos de pequeños nematodos en la luz de estos órganos. Muestras
fecales y parte del contenido intestinal fueron enviados y analizados en el laboratorio. El examen
coprológico demostró infección parasitaria severa con gran cantidad de huevos strongylidos y
oxyuridos. El contenido intestinal presentó abundante cantidad de nematodos, los cuales fueron
identificados a nivel de género como Atractis (Orden Ascaridida: Cosmocercoidea), and
Thelandros (Orden Oxyurida: Pharyngodonidae). Este representa el primer reporte de ambos
géneros para G. carbonaria en Venezuela. Los animales enfermos fueron tratados con Albendazol
-1
vía oral (10 mg·kg ), una vez al día por tres días. Grandes cantidades de nematodos fueron
expulsados y la condición de los animales mejoró drásticamente. El estrés es una causa común de
brotes de enfermedad parasitaria en reptiles recientemente capturados.
INTRODUCTION tortoises, as G. carbonaria can be infected by
several species of nematodes, but their
pathogenicity in wild animals is controversial,
even though parasitism is a real concern in
captive animals (Mader, 1996; Messonier,
1996).
Oxyurids, ascarids and strongylid nematodes are
the most common nematodes in tortoises and
wild specimens usually can carry these parasites,
especially pinworms (oxyurids). In Spain
sixteen oxyurid species were found in Testudo
graeca Linnaeus 1758, including Tachygonetria
and Pharyngodon genus (Chávarri et al., 2012).
Atractis marquezi was described from
Geochelone nigrita Quoy & Gaimard, 1824 by
Bursay & Flanngan (2002), and Rideout et al.
(1987) reported mortality in G. carbonaria and
G. pardalis from an US zoo, due to Oxyurid
nematodes (Proatractis genus). In Venezuela,
the nematode species Chapiniella larensis
(different from Chapiniella variabilis Chapin,
1924), and two nematodes of Kathlanidae:
Labiduris zschokkei Linstow, 1899 and
Labiduris gulosa (Rudolphi, 1819) Schneider,
1866 were described by Díaz-Ungría & Gallardo
Geochelone (syn. Chelonoidis) carbonaria
Spix, 1824 (Testudines: Testudinidae) is one of
the two species of tortoises that have been
reported in Venezuela, where is commonly
named “Morrocoy”. This tortoise inhabits a
wide range of habitats in Venezuelan territory,
with preference to the lowlands, open savannahs
and gallery forest (Fernández, 1991). There are
no reports of its presence in the Andes. The other
species, G. denticulata, has a more restricted
distribution, and its presence has been reported
only in Amazonas and Bolivar States, preferring
tropical forest habitats. Some of the zoological
characteristic features of G. carbonaria include:
Carapace relatively flat top, typically black,
interrupted only by a small yellow area around
the areola of each scute. The limbs are black with
bright red tips on many of the scales, contrasting
sharply with the yellow head markings
(Pritchard & Trebbau, 1985). G. carbonaria is a
very popular pet and its meat has also been used
in several regions of Venezuela to cook local
dishes. According with the CBC (2007), G.
carbonaria is an endangered species. Terrestrial
253
performs necropsies of the dead animals.
Necropsy showed hundreds of tiny parasites into
the intestinal content of caecum and colon,
mucoid diarroea, haemorragy of the mucosal
surface and the liver of one of the animals was
slightly pale.
Fecal samples from dead and alive animals and
part of the intestinal content was sent to the
Helminthology Laboratory, at the Faculty of
Veterinary Sciences, Central University of
Venezuela (FCV-UCV, Maracay, Aragua,
located 900 Km to the west of the farm). Faecal
samples were examined using direct smears and
fecal flotation techniques with a sugar solution.
Intestinal content was diluted 50:50 with 70%
alcohol and examined under a stereomicroscope
(Karl Zeiss Stemi 2000®). All nematodes
recovered were first stored at 70% alcohol and
then examined without clarification and/or
thereafter clarified with lactophenol and
observed under binocular microscope (Nikon
Eclipse E200®). Microphotographs of
specimens were made with a digital camera
(Canon Cibershot®) attached to the microscope
(Nikon Eclipse E200®) using 10x, 25x and 40x
objectives. Parasites were identified using
proper keys (Yamaguti, 1961; Chabaud, 1978).
After identification nematodes were deposited
in the parasitological collection of CIPV-MARA
(1967) and Díaz-Ungría (1973) in Testudo (sic)
denticulata.
The aim of this report was to describe a severe
parasitic infection with pinworms (Oxyurids),
ascarids (Atractidae) and strongylid nematodes
in 15 wild G. carbonaria specimens from Upata,
Bolivar, Venezuela, and to establish the first
report for Atractis, and Thelandros genus in
Venezuelan G. carbonaria.
Fifteen adult specimens of G. carbonaria (nine
females and six males) with carapace length
ranging between 23 and 34 cm (media 26.91 cm,
S: 3.25 cm) were captured for local people in the
savannah near a cattle ranch (ordered by the
owner of that ranch) located in Upata, Bolivar,
Venezuela and allocated in a little yard confined
with a fence. Faeces and food wastes were not
frequently removed from that allocation. Two
months later, animals became ill, showing
initially anorexia and lethargy, and later mucoid
diarrhea. No treatment was applied and five
tortoises (three males, two females) were found
dead three weeks later. A local veterinarian was
called and took samples from faeces and
MATERIAL AND METHODS
Neotrop. Helminthol., 8(2), 2014
Figura 1. Atractis sp., Head. 250X.
254
Figura 2. Thelandros sp. 125X. Figura 3. Thelandros sp. detail of Tail, 400X.
composed of a cylindrical corpus, anteriorly
differentiated distinct pharyngeal part, a
subespherical or elongated isthmus and a valved
bulb (Figure 1). Male with a coiled tail,
constriction after anus. forming a large conical
process without caudal alae. Female with vulva
apart from anus, pointed tail. Ten Oxyurid
nematodes were identified as genus Thelandros.
This identification was based on: the presence of
the genital cone as well as the quitinized
structure V-shaped at the posterior end (Figures
2, 3).
While parasitological procedures were
performed, animals were treated orally by the
local veterinarian with a bovine formulation of
-1
albendazole (Valbazen®, 10 mg·Kg once a day
for three days). All animals successfully
recovered and start to feed after treatment, and
diarrhea stopped. Hundreds of nematodes were
expulsed after treatment.
FCV-UCV under nº CIPVMARA-N8900100-
2013 (Atractis), and CIPVMARA-N9000101-
2013 (Thelandros), Sauricolinae specimen were
too damaged to be conserved.
Two strongylid nematodes (only male)
recovered from faecal samples were identified at
subfamiliy level as Sauricolinae (Nematoda:
Strongylidae). This identification was based on:
the presence of cuticular striae widely spaced,
slender spicules, and the host. Nevertheless,
specimens were in bad condition and it was not
possible to stablish neither genus nor species.
Another group of nematodes were identified at
genus level as Atractis (Order Ascaridida,
Superfamily Cosmocercoidea) and presenting
the following characteristic features: esophagus
Pérez Matta et al.
Parasitic nematode in Geochelone
RESULTS
255
reptiles refers mainly to Oxfendazole and
Fenbendazole (Frye, 1973, Gianetto et al., 2007)
and in a lesser extent, Albendazole as Valbazen®
at the dosage of 50mg/kg PO, once (Mader,
1996), we reported herein a good efficacy for the
bovine presentation of Albendazole at the lower
dosage of 10mg/kg repeated for three days to treat
this kind of multiple helminthic infection in
tortoise. Because the use of ivermectin is not
recommended in turtles (Teare & Bush, 1983), it
is important to remark the successful treatment of
the animals with Albendazole at this lower dose.
Authors found several limitations on the
development of this work. First of all, this is a
clinical case produced 800 km away from
university. Authors were not implied in capture
and storage of animals. The Veterinarian (Otto
Núñez, coauthor) who served as contact with
parasitology laboratory did not know the
conditions under which animals were captured.
He worked in extreme conditions far away from
any pharmacy or drugstore, and made the
necropsies on the field under very rough
conditions.
Treatment of the animals with albendazole did not
represent an injury for the tortoise and was an
emergency treatment which ends successfully.
Regarding with the identification of nematodes
we found, taxonomical keys for identification of
tortoise oxyurids at species level were not
available. So, we could only stablish the genera of
specimens. Nevertheless is remarkable that
before the present work there had not been in our
country any report of these genera in Venezuelan
tortoises exception made for those species
previously mentioned by Díaz-Ungría &
Gallardo (1968) and Díaz-Ungría (1973). So,
even the establishment of genera is especially
important for Venezuelan records. With all
limitations this study represented a unique
opportunity to study the helminth fauna of this
endangered species of tortoise in Venezuela. As a
final conclusions we can say that Venezuelan
tortoise G. carbonaria can be naturally
parasitized for Thelandros sp and Atractis sp, and
Albendazole could be a good choise for treat this
helminth infection on this host. Additional
studies, including the use of molecular tools are
needed to identify nematode founds at species
level.
Several species of oxyurid, and cosmocercoid
nematodes (formerly included in Oxyurida)
have been described from turtles, most of them
described in T. graeca and other species of
Testudo (Buckley, 1970; Johnson, 1973; Bursay
& Flannagan, 2002; Traversa et al., 2005;
Chávarry et al., 2012).There are two genus for
oxyurid nematodes found in Podocnemys (a
freshwater turtle genus) from Colombia
(Gibbons et al., 1995). None of the nematodes
we found in this report fulfill the characteristics
of the description of nematodes formerly
described in G. denticulata from Venezuela by
Díaz-Ungría & Gallardo (1968) and Díaz-
Ungría (1973). Unfortunately, the two
strongylid nematodes that were recovered from
faeces were not well preserved during
transportation and therefore, it was impossible
to establish the genus. Nevertheless, it is
important to highlight that this is the first report
of clinical illness in G. carbonaria due to
oxyurid nematode infection and the first record
of Thelandros and Atractis genera, infecting G.
carbonaria in our country. The pathogenicity of
these helminths in tortoises is unclear, and most
of the authors suggest that animals might carry
great amount of helminths without showing any
clinical sign (Frye, 1973; Holt et al., 1979;
Mader, 1996). In contrast, there is a report of
mortality due to a heavy infection with Atractis
sp nematodes (Rideout et al., 1987). We believe
that although tortoises can carry a high burden of
nematodes without displaying clinical signs
when living in nature, the situation might change
in captivity, and factors such as: changes in
feeding habits, crowding effect, wrong
allocation and stress of captivity; probably
might exert an important impact on the
composition and abundance of helminth fauna in
tortoises, as have been discussed by other
authors (Rideout et al., 1987; Rataj et al., 2011;
Chávarry et al., 2012).
Although no anthelmintic have been formulated
for tortoises and studies on their efficacy or
toxicity are limited (Claussen & Forstner, 1981;
Teare & Bush, 1983; Giannetto et al., 2007), and
most reports of benzimidazole drugs used in
DISCUSSION
Neotrop. Helminthol., 8(2), 2014
256
Authors are fully grateful to Héctor Zerpa
(Physiology Department, FCV-UCV), for
revision of English manuscript and Gisela García
(Histology Dept, FCV-UCV) for her gently aid
with microphotographies. Conflict of interest:
Authors declare we have no interest conflict for
this paper.
ACKNOWLEDGEMENTS
REFERENCIAS BIBLIOGRÁFICAS
Buckley, J. 1970. Two remarkable nematodes
from a South American tortoise.
Transactions of Royal Society of Tropical
Medicine and Hygiene, vol. 64, pp. 25.
Bursay, C & Flannagan, J. 2002. Atractis
marquezi n.sp (Nematoda: Atractidae)
and a revision of Atractis Dujardin, 1845,
sensu Baker, 1987. The Journal of
Parasitology, vol. 88, pp. 320-324.
Chávarri, M, Berriatua, E, Giménez, A, Gracia,
E, Martínez-Carrasco, C, Ortiz, J & De
Ybáñez, R. 2012. Differences in helmintth
infections between captive and wild
spurthighed tortoises Testudo graeca in
southern Spain: a potencial risk of
reintroduccion of this species. Veterinary
Parasitology, vol. 187, pp. 491-497.
Claussen, J & Forstner, M. 1981. Helminths of
tortoises and deworming trials with
d r u g s . B e r l M u n c h T i e r a r z t l
Wochenscher, vol. 94, pp. 411-414.
Chabaud, AC. 1978 Identification of Genera of
the superfamilies Cosmocercoidea,
S e u r a t o i d e a , H e t e r a k o i d e a y
Subuluroidea. Anderson, R, Chabaud, A
& Willmott, S. (Ed.) 1978. CIH keys to the
nematodes of vertebrates, (nº6). CAB Int.,
Royal Bucks, England. 37p.
Díaz-Ungria, C & Gallardo, MF. 1968.
Nematodes de reptiles Venezolanos, con
descripción de varias especies nuevas.
Boletín de la Sociedad Venezolana de
Ciencias Naturales, vol. 27, pp. 550-570.
Díaz-Ungría, C. 1973. Helmintos endoparásitos
de Venezuela. Ed. Centro Experimental de
Estudios Superiores, Escuela de Ciencias
Veterinarias, 126 p.
Fernández, M. 1991. Apuntes y observaciones
s o b re m o r ro co y e s ve n e z o l a n o s .
Manuscrito no publicado consultado en
zoológico de Caricuao, Caracas, Distrito
capital, Venezuela.
Frye, F. 1973. Husbandry, Medicine and Surgery
of captive reptiles. V M Publish., USA.
298 p.
Catálogo de la Biodiversidad colombiana
(CBC). 2007. Geochelone carbonaria
S p i x , 1 8 2 4 . I n :
http://www.siac.net.co/sib/catalogoespec
ies/especie.do?idBuscar=457&method=d
isplayAAT Chamorro-Rengifo, J &
Cubillos-Rodríguez, PA. Sistema de
Información sobre Biodiversidad de
Colombia SiB F. Actualizaci�n:
25062007. Consulta: enero 2013.
Giannetto, S, Brianti, E, Poglayen, G, Sorgi, C,
Capelli, G, Pennisi, M & Coci, G. 2007.
Efficacy of oxfendazole and fenbendazole
against tortoise (Testudo hermanni).
Parasitology Research, vol. 100, pp.
1069-1073.
Gibbons, L, Khalil, L & MarinKelle, C. 1995. A
new nematode genus Podocnematractis
for Atractis ortleppi Thapar, 1925.
(Cosmocercoidea: Atractidae) and the
description of another new species, P.
colombiaensis from turtles Podocnemis
s p p . i n C o l o m b i a . S y st e ma t ic
Parasitology, vol. 30, pp.47-56.
Holt, P, Cooper, J & Needham, R. 1979. Disease
s of tortoises: a survey of seventy cases. J
Small Anim Pract, vol. 20, pp. 269-86.
Johnson, S. 1973. Some oxyurid nematodes of
the genera Mehdiella and Thaparia from
the tortoise Testudo hermanni. Folia
Parasitologica (Praha), vol.20, pp.141-
148.
Mader, D. 1996. Reptile Medicine / Surgery.
W.B. Saunders Co., Phil, USA, 512 p.
Messonier, S. 1996. Common reptile diseases
and treatment. Blackwell Science Ed.
USA. 178 p.
Pritchard, R & Trebbau, P. 1985. Geochelone
(Chelonoidis) carbonaria. In: Turtles of
Venezuela. Ohio Society for the Study of
Pérez Matta et al.
Parasitic nematode in Geochelone
257
Neotrop. Helminthol., 8(2), 2014
Amphibians and Reptiles. pp. 207-229.
Rataj AV, Lindtner-Knific R, Vlahović K, Mavri
U & Dovč A. 2011. Parasites in pet
reptiles. Acta Veterinaria Scandinavica,
vol. 30, pp. 33.
Rideout, B, Montall, J, Phillips, L & Gardner, C.
1987. Mortality of captive tortoises due to
viviparous nematodes of the genus
Proatractis (Family Atractidae). Journal
of WildLife Disease, vol. 23, pp.103-108.
Teare, J & Bush, M. 1983. Toxicity and efficacy
of ivermectin in chelonians. Journal of the
A m e r i c a n Ve t er i n a r y M e d i c a l
Association, vol. 183, pp. 1195-1197.
Traversa, D, Capelli, G, Iorio, R, Bouamer, S,
Cameli, A & Giangaspero, A. 2005.
E p i d e m i o l o g y a n d b i o l o g y o f
nematodofauna affecting Testudo
hermanni, Testudo graeca and Testudo
marginata in Italy. Parasitolology
Research, vol. 98, pp. 14-20.
Yamaguty, S. 1961. Sistema Helminthum, Vol
III. The nematodes of vertebrates.
Interscience Publishers, INC., NY, USA.
pp. 679.
Received November 13, 2013.
Accepted June 6, 2014.
