Zoonoses and Public Health

ORIGINAL ARTICLE

American Trypanosomiasis Infection in Fattening Pigs from

the South-East of Mexico
M. Jiménez-Coello1, K. Y. Acosta-Viana1, E. Guzman-Marin1 and A. Ortega-Pacheco2
1
Laboratorio de Biologia Celular, CA Biomedicina de Enfermedades Infecciosas y Parasitarias, C.I.R. ‘Dr. Hideyo Noguchi’, Universidad Autonoma
de Yucatan, Yucatan, Mexico
2
Departamento de Salud Animal y Medicina Preventiva. CA Salud Animal, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autonoma
de Yucatan, Yucatan, Mexico

Impacts
• Pigs are important reservoirs of Trypanosoma cruzi in commercial pigs
destined to human consumption.
• Infected pigs may be a source of human infection when uncooked meat is
consumed or manipulated during acute infections.
• Vector control should be implemented around all commercial pig farms in
the State to prevent exposition.

Keywords:
Trypanosoma cruzi; seroprevalence; pigs;
serology; zoonosis
Correspondence:
A. Ortega-Pacheco. Departamento de
Medicina y Cirugia, CA Salud Animal.
Facultad de Medicina Veterinaria y Zootecnia,
Universidad Autonoma de Yucatan, Km. 15.5
Carr. Merida-Xmatkuil AP 4-116 Merida,
Yucatan, Mexico. Tel.: +52 (999) 9423200,
ext 16; Fax: +52(999) 423205;
E-mail: opacheco@uady.mx
The work was carried out in Universidad
Autonoma de Yucatan, Yucatan, Mexico.
Received for publication August 2, 2011
doi: 10.1111/zph.12016
Summary
American Trypanosomiasis (AT) is an infectious parasitic disease produced by
the protozoa Trypanosoma cruzi (T. cruzi). Infection is acquired by vectorial
via but can also be transmitted congenitally, by ingestion of an infected host,
by transfusion with contaminated blood or transplant of organs from an
infected donor. Currently, AT is widely distributed from the South of the Uni-
ted States to South America. In Mexico, the presence of the parasite has been
reported throughout the country where several reservoirs such as dogs, opos-
sums, rats and cats have been identified. Yucatan is in the south-east of Mexico
where AT is endemic and has been reported since 1940s. There is little infor-
mation about the role of pigs as reservoirs of T. cruzi. The frequency of specific
antibodies against T. cruzi was determined in fattening pigs from Yucatan,
Mexico. After sampling in the 3 main areas of pig production in the state, IgG
ELISA and Western blot were performed to identify seropositive cases. Associa-
tion of farm size, farm area and production system with infected pigs was eval-
uated. From 273 sampled pigs, 5.4% (n = 15) positive cases were found. No
association with evaluated factors and infected pigs was found. Pigs are also
reservoirs of T. cruzi in the studied area. These findings are considered impor-
tant to improve vectorial control in the area in order to avoid the parasite
infection in animal populations destined for human consumption and avoid
further transmission to humans.

is through a haematophagous insect vector well estab-

Introduction
American trypanosomiasis or Chagas disease is a parasitic
disease caused by the flagellate protozoan Trypanosoma
cruzi and is found from the southern United States to
Argentina. This parasite can infect humans and more
than 150 wild and domestic mammals. The transmission
lished in all Latin-American countries, but can also be
transmitted less frequently by the transplacental route,
orally, and by blood transfusions or organ transplants.
Until now, there is no effective drug treatment or
immune to prevent or control infection, which gives it
special importance and seeks to prevent transmission in

166 ª 2012 Blackwell Verlag GmbH • Zoonoses Public Health 59 (Suppl. 2) (2012) 166–169
M. Jiménez-Coello et al.
humans. Domestic and wild animals are important reser-
voir to maintain the circulation of the protozoa in the
environment (i.e. opossums, raccoons, dogs, cats, etc.).
Domestic and wild pigs (Sus scrofa) as other mammals
are susceptible to be infected with T. cruzi and become
reservoirs of the protozoa, but no information about its
role has been reported.
The aim of this study was to determine the prevalence
of IgG-specific antibodies against T. cruzi in commercial
fattening pig farms in an endemic area of Mexico.
Materials and Methods
Study area
The study was conducted in the state of Yucatan, located in
south-eastern Mexico (1930¢ and 2135¢ north latitude
and 9024¢ west longitude). The climate is tropical sub-
humid with heavy summer rains. The maximum monthly
temperature varies between 35 and 40C, with an average
temperature of 26.6C. The relative humidity varies
between 65% and 100% taking the mean value over 78%.
The annual rainfall is from 415 to 1290 mm (INEGI, 2002).
Size of the sample
A two-stage cross-sectional study was performed during
September to December 2009. Two-hundred and seventy-
three pigs from 39 farms, between 18 and 20 weeks of
age, were randomly selected. A proportional number of
farms from the three main pig production regions of the
State (West, Centre and South) were included. The sam-
ple size to estimate prevalence was determined by the for-
mula: n ¼ Dðz 2 pð1  pÞÞ=d2 . Considering a confidence
level of 95% (z = 1.96), absolute precision of 5% (d), a
design effect (D) of 2 and an estimated prevalence of
10% (p). The estimated prevalence was obtained from a
pilot study that included 45 animals. The number of ani-
mals sampled at each farm (b = 7) was calculated based
on the formula: b =  (ce/cd) (1 ) re/re), where ce cost
of a sample into two clusters (10), cd sample cost of two
units of interest in a same cluster (1), and the correlation
intraconglomerates (re) was (0.04) (Segura-Correa and
Honhold, 2000). The number of farms (m = n/b) was cal-
culated by dividing the sample size (n = 273) between the
number of animals sampled in each establishment
(b = 7).
Sample collection
Individual blood samples were collected using vacutainer
tubes without EDTA to obtain serum. Blood samples
were centrifuged at 2500 g for 10 min and were stored at
)20C until further evaluation.
ª 2012 Blackwell Verlag GmbH • Zoonoses Public Health 59 (Suppl. 2) (2012) 166–169
American Trypanosomiasis in Pigs from Mexico
Indirect enzyme-linked immunosorbent assay and
western blot assays
For the specific detection of IgG antibodies against
T. cruzi, a commercial indirect enzyme-linked immuno-
sorbent assay was used (WienerLab ChagaTest). The
technique used was adapted to that described by Jiménez-
Coello et al. (2010) using anti-IgG pig antibody labelled
with horseradish peroxidase (HRP; Santa Cruz Inc., Santa
Cruz, CA, USA) on 96-well plate coated with recombi-
nant proteins of T. cruzi. Serum samples were diluted to
a ratio of 1 : 100 in phosphate-buffered saline (PBS; pH
7.2), and the secondary goat anti-IgG pig antibody HRP
labelled was used at a dilution of 1 : 5000.
Sera from previously evaluated dogs and BALB/c artifi-
cially infected mice showing high anti-IgG antibodies
titres by ELISA (1 : 1024) and PCR positive against
T. cruzi were used as positive controls, and sera pool from
10 healthy pigs previously tested by triplicate with ELISA
IgM, IgG and PCR were used as negative controls. On the
basis of the ELISA, subjects were diagnosed as either
positive or negative for specific IgG antibodies to T. cruzi.
The optical density (OD) was measured in a spectropho-
tometer at 450 nm (Multiskan Multisoft Primary EIA)
and was used to compute the per cent positivity (PP)
using the following formula:
mean OD value of sample or negative control
 100:
mean OD value of positive control
Per cent positivity of 15% or above was considered as
positive.
For confirmation of the diagnosis ELISA seropositive
cases, the western blot (WB) technique was performed
(Ramos-Ligonio et al., 2006; Jiménez-Coello et al., 2008),
in which Tulahuen strain epimastigotes of T. cruzi were
used as antigen. Samples were considered positives to WB
based on an established criterion. A serum sample was
considered positive when it recognized at least five anti-
genic bands from a group of 10 with the highest frequency
of recognition; the result was considered indeterminate
when the sample recognized of 1–4 antigenic bands and
was negative when the serum sample showed no reactivity
(Teixeira et al., 1994; Jiménez-Coello et al., 2008).
Statistical analysis
Univariate analysis (X2) was used to determine the associ-
ation of possible risk factors with the presence of specific
antibodies to Chagas’ disease. Risk factors considered
were farm size (£400 or >400), production system
(full cycle or fattening farms) and farm region (West,
Central or Sotuh). Odds ratio (OR) and 95% confidence
167
American Trypanosomiasis in Pigs from Mexico
Table 1. Risk factors evaluated to determine association with seropositivity against Trypanosoma cruzi in fattening pigs from Yucatan, Mexico
Risk Factor Category Positives Prevalence (%)
Farm Size £400 5 5.05
>400 10 5.43
Production Full cycle 9 4.3
System Fattening 6 7.2
Farm region West 5 5.4
Center 4 4.2
South 6 6.9
intervals were also calculated using the EpiInfo program
(Dean et al., 1994). The level of significance was set at
P < 0.05.
Results
From 273 sampled pigs, there were found a 5.4%
(n = 15) positive cases. All positive cases were from dif-
ferent farms. There was no association between seroposi-
tivity and any of the evaluated risk factors (P > 0.05;
Table 1).
Discussion
American trypanosomiasis is a widespread infection in
the Americas in a large number of domestic mammals
that serve as reservoirs that maintain the infection
because they are in direct contact with vectors and
human settlements. There are no formal epidemiological
studies that demonstrate the role of the domestic pig as a
reservoir of T. cruzi. However, natural and experimental
infections have been described (Salazar-Schettino et al.,
1997; Solaymani-Mohammadi and Petri, 2006). These
reports point to the pig as a potential reservoir host
for infections of T. cruzi in humans, which have an
important role in the maintenance of domestic and
peridomestic cycle of infection.
The prevalence of infection with T. cruzi found in this
study is considered low. However, this is the first formal
epidemiological report showing the presence of this pro-
tozoan distribution in a major pig production area from
Mexico. Previously, the presence of natural infection in
pigs in Mexico has been described (Salazar-Schettino et
al., 1997). The presence of pigs naturally infected with
T. cruzi has been reported in other Latin American coun-
tries like Brazil (Costa et al., 1998) and Paraguay (Fujita
et al., 1994).
The infection of pigs with T. cruzi in the study area is
probably due to exposure to the vector Triatoma dimidi-
ata, which has been described as the main vector trans-
mitting T. cruzi in south-eastern Mexico where the
disease is considered endemic (Guzman-Tapia et al.,
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M. Jiménez-Coello et al.
OR IC (95%) P-value
0.9 0.32–2.67 0.89
0.58 0.21–1.64 0.32
1 0.67
0.78 0.21–2.8
1.2 0.40–4.00
2005), unlike the vector transmitter in domestic pigs in
Brazil (Panstrongylus geniculatus and Triatoma brazilienzis;
Valente et al., 1998).
Apparently, this protozoal infection in pigs does not
have considerable economic importance because it does
not affect the production parameters (i.e. daily weight
gain). The pathogenesis of the disease in pigs has not
been demonstrated, but as in other mammals, clinical
signs during the acute phase may result in fatal conse-
quences. On the other hand, because of the short lifetime
of fattening pigs, the indeterminate or chronic phase of
the disease is never reached. However, the parasite strains
circulating in the region show heart and muscle tissue
tropism of infected hosts (amastigote intracellular stage)
(Barrera et al., 2001). Although the presence of infection
in pigs is low, the consumption of undercooked pork
meat should be avoided, and precautions in handling raw
meat are recommended because during the acute stage of
infection, the parasite is circulating in the bloodstream
(trypomastigote stage in the blood).
It is concluded that the domestic pig in the area of
study is an important reservoir of T. cruzi because it is in
an endemic area where the vector is widely distributed
and recommended vector control measures in pork pro-
duction farms to reduce the risk of transmission for this
zoonotic disease.
Acknowledgements
We gratefully acknowledge to D. Saez-Peniche and M.E.
Rosado-Barrera for their technical assistance.
Conflicts of interest
The authors have not declared any potential conflicts.
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